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Sample records for culture supernatant fluids

  1. Effect of Physical and Chemical Changes on the Antimicrobial Activity of Culture Supernatant Fluid of Lactic Acid Bacteria

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    Leila Goudarzi (MSc

    2016-04-01

    Full Text Available Background and Objective: Lactic acid bacteria are Gram-positive, catalase-negative, nonsporulating, either rod- or coccus-shaped bacteria that have beneficial effects on their hosts by producing antimicrobial substances such as lactic acid, hydrogen peroxide, bacteriocins and biosurfactants. Bacteriocins are antimicrobial peptides that are produced by bacteria and can inhibit the growth of other bacteria. Methods: In this experimental study, bacteriocin production by Lactobacilli as known probiotic strains was evaluated in different physicochemical conditions. Antagonistic activity was evaluated using quantitative method of Microscale Optical Density Assay (MODA. After neutralization of acid and treatment with various enzymes, temperature, pH and NaCl conditions, the antimicrobial activity of culture supernatant fluid of Lactobacillus acidophilus and L. plantarum was investigated against pathogenic Proteus. Results: The culture supernatant fluid of Lactobacilli was sensitive to proteolytic enzymes with relatively good stability to temperature. The antimicrobial activity was also present due to production of bacteriocin under different NaCl conditions (1 to 4% NaCl and pH range of 5 to 8. Conclusion: It seems that the antimicrobial liquid of Lactobacillus strains contains bacteriocin, which shows antimicrobial effects against pathogenic strains of Proteus. To investigate further this effect, some complementary studies should be performed.

  2. Cell-free fetal DNA in amniotic fluid supernatant for prenatal diagnosis.

    Science.gov (United States)

    Soltani, M; Nemati, M; Maralani, M; Estiar, M A; Andalib, S; Fardiazar, Z; Sakhinia, E

    2016-04-30

    In widespread conviction, amniotic fluid is utilized for prenatal diagnosis. Amniotic fluid supernatant is usually discarded, notwithstanding being a good source of fetal DNA. The aim of the present study was to assess cell-free fetal DNA extracted from amniotic fluid supernatant for application in prenatal diagnosis such as gender determination and early diagnosis of β-thalassemia. Samples of amniotic fluid of 70 pregnant women were collected and went through routine tests along with tests for cell-free fetal DNA from amniotic fluid supernatant. The DNA in the amniotic fluid supernatant was extracted and analyzed for gender determination by PCR and Real-time PCR. ARMS-PCR was applied to test early diagnosis of IVS II-I mutation (common β-thalassemia mutation) and E7V mutation for sickle cell anemia using DNA extracted from the amniotic fluid supernatant. Using the cell-free fetal DNA extracted from the amniotic fluid supernatant, the sensitivity of PCR and Real-time PCR for gender detection was compared with the routine cytogenetic method. The fetus tested for sickle cell anemia and β-thalassemia was observed to be healthy but heterozygous for IVS II-I mutation. The findings indicated that cell-free fetal DNA from amniotic fluid supernatant can be a good source of fetal DNA and be used in early prenatal diagnosis since because of its fast and accurate application. Therefore, it would be suggested that the amniotic fluid supernatant's disposal is prevented because if the tests needs to be repeated, cell-free fetal DNA extracted from the amniotic fluid supernatant can be used as an alternative source for prenatal diagnosis.

  3. A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants

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    Eva Hiery

    2015-11-01

    Full Text Available Clavibacter michiganensis, subsp. michiganensis is a Gram-positive plant pathogen infecting tomato (Solanum lycopersicum. Despite a considerable economic importance due to significant losses of infected plants and fruits, knowledge about virulence factors of C. michiganensis subsp. michiganensis and host-pathogen interactions on a molecular level are rather limited. In the study presented here, the proteome of culture supernatants from C. michiganensis subsp. michiganensis NCPPB382 was analyzed. In total, 1872 proteins were identified in M9 and 1766 proteins in xylem mimicking medium. Filtration of supernatants before protein precipitation reduced these to 1276 proteins in M9 and 976 proteins in the xylem mimicking medium culture filtrate. The results obtained indicate that C. michiganensis subsp. michiganensis reacts to a sucrose- and glucose-depleted medium similar to the xylem sap by utilizing amino acids and host cell polymers as well as their degradation products, mainly peptides, amino acids and various C5 and C6 sugars. Interestingly, the bacterium expresses the previously described virulence factors Pat-1 and CelA not exclusively after host cell contact in planta but already in M9 minimal and xylem mimicking medium.

  4. A Proteomic Study of Clavibacter Michiganensis Subsp. Michiganensis Culture Supernatants.

    Science.gov (United States)

    Hiery, Eva; Poetsch, Ansgar; Moosbauer, Tanja; Amin, Bushra; Hofmann, Jörg; Burkovski, Andreas

    2015-11-12

    Clavibacter michiganensis, subsp. michiganensis is a Gram-positive plant pathogen infecting tomato (Solanum lycopersicum). Despite a considerable economic importance due to significant losses of infected plants and fruits, knowledge about virulence factors of C. michiganensis subsp. michiganensis and host-pathogen interactions on a molecular level are rather limited. In the study presented here, the proteome of culture supernatants from C. michiganensis subsp. michiganensis NCPPB382 was analyzed. In total, 1872 proteins were identified in M9 and 1766 proteins in xylem mimicking medium. Filtration of supernatants before protein precipitation reduced these to 1276 proteins in M9 and 976 proteins in the xylem mimicking medium culture filtrate. The results obtained indicate that C. michiganensis subsp. michiganensis reacts to a sucrose- and glucose-depleted medium similar to the xylem sap by utilizing amino acids and host cell polymers as well as their degradation products, mainly peptides, amino acids and various C5 and C6 sugars. Interestingly, the bacterium expresses the previously described virulence factors Pat-1 and CelA not exclusively after host cell contact in planta but already in M9 minimal and xylem mimicking medium.

  5. Actinobacillus pleuropneumoniae culture supernatants interfere with killing of Pasteurella multocida by swine pulmonary alveolar macrophages.

    OpenAIRE

    Chung, W. B.; Bäckström, L; McDonald, J.; Collins, M T

    1993-01-01

    The effect of Actinobacillus pleuropneumoniae culture supernatant on swine pulmonary alveolar macrophage (PAM) functions was studied. The A. pleuropneumoniae culture supernatant was toxic to PAMs when tested by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and lactate dehydrogenase (LDH) release assays. Biological activity of the supernatant was ascribed to cytotoxins. Both the LDH and MTT assays were used for measurement of crude A. pleuropneumoniae cytotoxin concentrati...

  6. Early stationary phase culture supernatant accelerates growth of sputum cultures collected after initiation of anti-tuberculosis treatment.

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    Kolwijck, E; Friedrich, S O; Karinja, M N; van Ingen, J; Warren, R M; Diacon, A H

    2014-07-01

    We investigated the effect of Mycobacterium tuberculosis culture supernatant added to sputum cultures collected during the first 8 weeks of anti-tuberculosis treatment. With ongoing treatment duration, time to culture positivity decreased significantly in supernatant-enriched cultures, possibly due to stimulation of dormant or slowly metabolizing M. tuberculosis cells.

  7. The culture of Chlorella vulgaris in a recycled supernatant: Effects on biomass production and medium quality

    KAUST Repository

    Hadj-Romdhane, F.

    2013-03-01

    Reusing supernatant of microalgae culture medium can have inhibitory or toxic effects on the biomass production because of the release of organic metabolites by cells in the culture medium during their growth. This work investigated the impact of Chlorella vulgaris medium recycling on culture productivity, cells quality and accumulation of excreted metabolites in the culture medium. No significant impact on the C. vulgaris growth was observed after 63days of recycling, the productivity remained stable at around 0.55kgm-3day-1. Organic matters accumulated in supernatant were identified as biopolymers (BP) poor in nitrogen and with a size above 40kDa (probably polysaccharides), and small organic molecules (SOM) richer in nitrogen with a molecular size ranging from 1 to 3kDa. The concentration of biopolymers in the supernatant increased till to a maximum and then decreased, possibly consumed by bacteria, whereas small organic compounds accumulated in the medium. © 2013 Elsevier Ltd.

  8. Bioactivities of Culture Supernatants from Retroviral Packaging Cells Carrying the Mouse Fas Ligand Gene

    Institute of Scientific and Technical Information of China (English)

    LIU Lingbo; ZOU Ping; GUO Rong; XIAO Juan; XU Zhiliang

    2001-01-01

    The bioactivities of culture supernatants from retroviral packaging cells carrying the mouse Fas ligand (mFasL) gene was investigated. FasLcDNA was cloned into PLXIN with an internal ribosome entry site to link two cistrons through gene recombination technology, PLXIN and the recombinant vector PLFIN were separately transfected into PA317 retrovirus packing cell line by lipofectamine 2000, and the resistant clones were selected with G418 selective medium. The integration of genome DNA was assayed by genomic DNA PCR. NIH3T3 cells were transduced by the culture supernatants from PA317 carrying the mFasLcDNA gene, and were selected with G418 selective medium, so as to select the PLFIN-PA317 clone capable of producing higher titer of supernatants. The levels of mFasL protein on NIH3T3 cells membrane were assayed by flow cytometry (FCM). The biological activity of mFasL on NIH3T3 cells membrane was investigated by the inducing apoptosis of Fas+ Yac-1 cells co-cultured with NIH3T3 cells expressing Fas ligand. To explore the direct mFasL cytotoxicity of culture supernatants from retroviral packaging cells carrying the mFasL gene, the culture supernatants from PLFIN-PA317 and PLXIN-PA317 were separately co-cultured with Yac-1cells in parallel. The recombinant PLFIN was successfully constructed. The highest titer of supernatants from twelve resistant clones was 8. 5 × 105 colony-forming-unit (CFU)/ml. The NIH3T3cells transfected by above supernatants had a higher level of mFasL (53.81±6.9 %), and significantly induced the apoptosis of Fas+ Yac-1 cells (56. 78±4.5 %), as both were cocultured for 5 h at1 : 1 ratio, whereas it is 7. 08±3.4 % in control group (P<0. 01). Supernatant from PLFINPA317 could also directly induce the apoptosis of Yac-1 within 5 h of incubation. Thus, the culture supernatants from PLFIN-PA317 possessed both infectivity and cytotoxicity of mFasL.

  9. Treatment of serum with supernatants from cultures of Candida albicans reduces its serum-dependent phagocytosis

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    Santos Aderbal Antonio dos

    2002-01-01

    Full Text Available Candida albicans is a potent activator of the complement system, and heat labile opsonins produced by activation of C3 (C3b and iC3b enhance phagocytosis of C. albicans mediated by complement receptors. In this study we treated mouse serum with supernatants from cultures of a protease producer strain of C. albicans and evaluated the ability of this serum to enhance phagocytosis of C. albicans. Cell-free supernatants from cultures of C. albicans were concentrated 5 fold and added to mouse serum for 30 min at 37ºC, before using this serum for opsonization of glutaraldehyde-fixed yeast cells. We observed that normal mouse serum increased about 3 fold the phagocytosis of C. albicans by mice peritoneal macrophages, whereas supernatant-treated serum did not increase phagocytosis. This effect of supernatants on serum was prevented by addition of pepstatin (5 µg/ ml; an inhibitor of C. albicans acid proteases to the medium. Serum treated with supernatants from cultures of a protease-deficient mutant of C. albicans also increased about 3 fold phagocytosis of the yeast. These results suggest that a protease produced by C. albicans causes proteolysis of serum opsonins, thereby reducing the phagocytosis of the yeast.

  10. EFFECT OF GLYCOSYLATION AT ASN302 OF PRO-UROKINASE ON ITS STABILITY IN CULTURE SUPERNATANT

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective To investigate the effect of glycosylation at Asn302 of pro-urokinase (pro-UK) on the stability in culture supernatant.Methods Nonglycosylated pro-UK was constructed by site-directed mutagenesis of Asn302 to Ala302. The proUK mutant and native pro-UK were transfected into dhfr-CHO cells, and serum-free culture supernatant was harvested and incubated at 4℃ and 37℃, respectively. The pro-UK activity in culture supernatant was measured by the optical density (OD) increase with time ( 12 hours) at 405 nm. Without thermolysin activation, the percentage of single chain pro-UK was measured.Results After 48 hours of incubation at 4℃, the activities of pro-UK mutant and native pro-UK decreased 3.7%and 2. 9% respectively, and at 37℃ decreased 37.9 % and 23.5 %, respectively. The total activity of native pro-UK was significantly higher than that of nonglycosylated mutant at 37℃. The single-chain percentage of native pro-UK was higher than that of nonglycosylated mutant at both 4℃ and 37℃.Conclusion Higher temperature increases the proteolysis of pro-UK. The glycosylation site on Asn302 is beneficial to pro-UK stability in culture supernatant.

  11. Dormant cells of Staphylococcus aureus are resuscitated by spent culture supernatant.

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    Ben Pascoe

    Full Text Available We describe the first in vitro model of dormancy in Staphylococcus aureus, showing that cells are generated which can be resuscitated by addition of spent medium supernatant taken from cultures of the same organism. Over 30 days, culturable counts in dormant cultures of S. aureus SH1000 fell from 10(6-10(7 cfu/ml to 600-fold increase in bacterial growth. Resuscitation was a specific effect, greatly reduced by boiling or addition of trypsin to the spent supernatant. Supernatant also effected a reduction in lag phase of dormant cultures. SEM demonstrated the presence of small coccoid cells in dormant cultures. The results are similar to those seen with resuscitation promoting factors (Rpfs in actinobacteria. This is the first time resuscitation has been demonstrated in Staphylococcus aureus, which is an important human pathogen. A better understanding of control and reactivation of dormant cells could lead to major improvements in managing staphylococcal infections; resuscitation could be an important step in restoring susceptibility to antibiotic treatment.

  12. Platelet-Rich Gel Supernatants Stimulate the Release of Anti-Inflammatory Proteins on Culture Media of Normal Equine Synovial Membrane Explants.

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    Ríos, Diana L; López, Catalina; Carmona, Jorge U

    2015-01-01

    The aims were as follows: (1) to evaluate the effects at 48 and 96 h of two concentrations (25 and 50%) of leukocyte and platelet-rich gel (L-PRG) and pure PRG (P-PRG) supernatants on the production/degradation in normal equine synovial membrane explants (SME) of platelet derived growth factor isoform BB, transforming growth factor beta-1, tumor necrosis factor alpha, interleukin (IL-) 4 (IL-4), IL-1 receptor antagonist (IL-1ra), and hyaluronan (HA) synthesis and (2) to correlate these molecules with their respective PRG supernatant treatments. SME from 6 horses were cultured for 96 h with L-PRG and P-PRG supernatants at 25 and 50% concentrations, respectively. SME culture media were changed each 48 h and used for determination by ELISA of the molecules, which were also determined in synovial fluid. 25% L-PRG supernatant produced a sustained release over time of IL-1ra and a gradual release of HA, whereas 50% L-PRG supernatant produced a sustained increase over time of IL-4 and HA. 50% P-PRG supernatant produced an increased and sustained production of IL-1ra and IL-4. The cellular composition and the articular concentration (volume) of a platelet-rich plasma preparation could affect the anti-inflammatory and anabolic joint responses in horses with osteoarthritis.

  13. Use of spray-dried zirconia microspheres in the separation of immunoglobulins from cell culture supernatant.

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    Subramanian, A; Carr, P W; McNeff, C V

    2000-08-18

    A method suitable for the isolation of monoclonal antibodies (MAbs) on novel zirconia microspheres (20-30 microm) is described. Zirconia microspheres were generated by spray drying colloidal zirconia. Spray-dried zirconia microspheres were further classified and characterized by X-ray diffraction, BET porosimetry and scanning electron microscopy. Spray-dried zirconia microspheres were modified with ethylenediamine-N,N'-tetra(methylenephosphonic) acid (EDTPA) to create a cation-exchange chromatographic support. The chromatographic behavior of a semi-preparative column packed with EDTPA-modified zirconia microspheres was evaluated and implications for scale-up are provided. EDTPA-modified zirconia microspheres were further used to purify MAbs from cell culture supernatant. Analysis by enzyme linked immunosorbent assay and gel electrophoresis demonstrate that MAbs can be recovered from a cell culture supernatant at high yield (92-98%) and high purity (>95%) in a single chromatographic step.

  14. An efficient process of generating bispecific antibodies via controlled Fab-arm exchange using culture supernatants.

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    Paul, Suparna; Connor, Judy; Nesspor, Tom; Haytko, Peter; Boakye, Ken; Chiu, Mark L; Jiang, Haiyan

    2016-05-01

    Bispecific antibody generation is actively pursued for therapeutic and research antibody development. Although there are multiple strategies for generating bispecific antibodies (bsAbs); the common challenge is to develop a scalable method to prepare bsAbs with high purity and yield. The controlled Fab-arm exchange (cFAE) method combines two parental monoclonal antibodies (mAbs), each with a matched point mutation, F405L and K409R in the respective CH3 domains. The conventional process employs two steps: the purification of two parental mAbs from culture supernatants followed by cFAE. Following a reduction/oxidation reaction, the bispecific mAb is formed with greater than 95% heterodimerization efficiency. In this study, cFAE was initiated in culture supernatants expressing the two parental mAbs, thereby eliminating the need to first purify the parental mAbs. The bsAbs formed in culture supernatant was then purified using a Protein A affinity chromatography. The BsAbs generated in this manner had efficiency comparable to the conventional method using purified parental mAbs. BsAbs prepared by two different routes showed indistinguishable characteristics by SDS capillary electrophoresis, analytical size exclusion, and cation exchange chromatography. This alternative method significantly shortened timelines and reduced resources required for bsAb generation, providing an improved process with potential benefits in large-scale bsAb preparation, as well as for HTP small-scale bsAb matrix selection.

  15. Culture - joint fluid

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    Joint fluid culture ... fungi, or viruses grow. This is called a culture. If these germs are detected, other tests may ... is no special preparation needed for the lab culture. How to prepare for the removal of joint ...

  16. Peritoneal fluid culture

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    Culture - peritoneal fluid ... sent to the laboratory for Gram stain and culture. The sample is checked to see if bacteria ... The peritoneal fluid culture may be negative, even if you have ... diagnosis of peritonitis is based on other factors, in addition ...

  17. Flow Cytometry Sorting to Separate Viable Giant Viruses from Amoeba Co-culture Supernatants

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    Khalil, Jacques Y. B.; Langlois, Thierry; Andreani, Julien; Sorraing, Jean-Marc; Raoult, Didier; Camoin, Laurence; La Scola, Bernard

    2017-01-01

    Flow cytometry has contributed to virology but has faced many drawbacks concerning detection limits, due to the small size of viral particles. Nonetheless, giant viruses changed many concepts in the world of viruses, as a result of their size and hence opened up the possibility of using flow cytometry to study them. Recently, we developed a high throughput isolation of viruses using flow cytometry and protozoa co-culture. Consequently, isolating a viral mixture in the same sample became more common. Nevertheless, when one virus multiplies faster than others in the mixture, it is impossible to obtain a pure culture of the minority population. Here, we describe a robust sorting system, which can separate viable giant virus mixtures from supernatants. We tested three flow cytometry sorters by sorting artificial mixtures. Purity control was assessed by electron microscopy and molecular biology. As proof of concept, we applied the sorting system to a co-culture supernatant taken from a sample containing a viral mixture that we couldn't separate using end point dilution. In addition to isolating the quick-growing Mimivirus, we sorted and re-cultured a new, slow-growing virus, which we named “Cedratvirus.” The sorting assay presented in this paper is a powerful and versatile tool for separating viral populations from amoeba co-cultures and adding value to the new field of flow virometry. PMID:28111619

  18. Bioleaching of metals from steel slag by Acidithiobacillus thiooxidans culture supernatant.

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    Hocheng, Hong; Su, Cheer; Jadhav, Umesh U

    2014-12-01

    The generation of 300–500 kg of slag per ton of the steel produced is a formidable amount of solid waste available for treatment. They usually contain considerable quantities of valuable metals. In this sense, they may become either important secondary resource if processed in eco-friendly manner for secured supply of contained metals or potential pollutants, if not treated properly. It is possible to recover metals from steel slag by applying bioleaching process. Electric arc furnace (EAF) slag sample was used for bioleaching of metals. In the present study, before bioleaching experiment water washing of an EAF slag was carried out. This reduced slag pH from 11.2 to 8.3. Culture supernatants of Acidithiobacillus thiooxidans (At. thiooxidans), Acidithiobacillus ferrooxidans (At. ferrooxidans), and Aspergillus niger (A. niger) were used for metal solubilization. At. thiooxidans culture supernatant containing 0.016 M sulfuric acid was found most effective for bioleaching of metals from an EAF slag. Maximum metal extraction was found for Mg (28%), while it was least for Mo (0.1%) in six days. Repeated bioleaching cycles increased metal recovery from 28% to 75%, from 14% to 60% and from 11% to 27%, for Mg, Zn and Cu respectively.

  19. Analysis of IL-2-like factor in lymphocyte culture supernatant of olive flounder, Paralichthys oliveaceus

    Institute of Scientific and Technical Information of China (English)

    WU Riqin; ZHANG Peijun; LI Jun; XU Yongli

    2005-01-01

    To study immune mechanism of fish lymphocyte we performed a proliferation assay and ELISA using monoclonal antibody against human IL-2. The result showed that an interleukin-2 (IL-2)-like factor was detected in the supernatant of plant haemoglutinin (PHA)-stimulated lymphocyte culture from peripheral blood,spleen and head kidney of olive flounder, Paralichthys olivaceus. The quantities of IL-2-1ike factor in the supematant from different lymphoid tissues were quite different. The IL-2 like factor in the supernatant from cultured head kidney lymphocytes was much higher than those of peripheral blood lymphocytes and spleen lymphocytes (P<0.01). The IL-2 activity was found in either mouse thymocyte proliferation assay or flounder head kidney lymphocyte proliferation assay and shown to have obvious enhancing effect on proliferation of the above two types of cell. The recombinant human IL-2 (rhIL-2) was able to stimulate flounder thymocyte proliferation and used to detect the IL-2 receptor (IL-2R) on the surface of flounder lymphocyte. The cross-reaction between the lymphocytes of flounder peripheral blood and CD25(IL-2R) was detected with flow cytometry and shown that the percentage of CD25-positive cell in peripheral blood was 7.74± 0.67%.

  20. Cerebrospinal fluid (CSF) culture

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    ... is a laboratory test to look for bacteria, fungi, and viruses in the fluid that moves in ... culture medium. Laboratory staff then observe if bacteria, fungi, or viruses grow in the dish. Growth means ...

  1. Comparison among three anion exchange chromatographic supports to capture erythropoietin from cell culture supernatant

    Institute of Scientific and Technical Information of China (English)

    Lourdes HERNNDEZ; Diobel STEWART; Lourdes ZUMALACRREGUI; Daniel AMARO

    2015-01-01

    Affinity and ion exchange conventional chromatography have been used to capture erythropoietin ( EPO)from mammalian cell culture supernatant. Currently,chromatographic adsorbent perfusion is available, however a limited number of applications have been found in the literature. In this work,three anion exchange chromatographic supports( gel,membrane and monolithic)were evaluated in the capture step of the recombi-nant erythropoietin purification process. The influences of load and flow rate on each support performance were analyzed. Also the purity of the EPO molecules was determined. A productivity analysis,as a decision tool for larger scale implementation,was done. As a conclusion,the evaluated supports are technically suitable to cap-ture EPO with adequate recovery and good purity. However,the monolithic column admits high operating velocity,showing the highest adsorption capacity and productivity.

  2. Quantitative analysis of particles, genomes and infectious particles in supernatants of haemorrhagic fever virus cell cultures

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    Hedlund Kjell-Olof

    2011-02-01

    Full Text Available Abstract Information on the replication of viral haemorrhagic fever viruses is not readily available and has never been analysed in a comparative approach. Here, we compared the cell culture growth characteristics of haemorrhagic fever viruses (HFV, of the Arenaviridae, Filoviridae, Bunyaviridae, and Flavivridae virus families by performing quantitative analysis of cell culture supernatants by (i electron microscopy for the quantification of virus particles, (ii quantitative real time PCR for the quantification of genomes, and (iii determination of focus forming units by coating fluorescent antibodies to infected cell monolayers for the quantification of virus infectivity. The comparative analysis revealed that filovirus and RVFV replication results in a surplus of genomes but varying degrees of packaging efficiency and infectious particles. More efficient replication and packaging was observed for Lassa virus, and Dengue virus resulting in a better yield of infectious particles while, YFV turned out to be most efficient with only 4 particles inducing one FFU. For Crimean-Congo haemorrhagic fever virus (CCHFV a surplus of empty shells was observed with only one in 24 particles equipped with a genome. The complete particles turned out to be extraordinarily infectious.

  3. Purification and characterization of a novel fibrinolytic enzyme from culture supernatant of Pleurotus ostreatus.

    Science.gov (United States)

    Liu, Xiao-Lan; Zheng, Xi-Qun; Qian, Peng-Zhi; Kopparapu, Narasimha-Kumar; Deng, Yong-Ping; Nonaka, Masanori; Harada, Naoki

    2014-02-28

    A fibrinolytic enzyme was produced by an edible mushroom of Pleurotus ostreatus using submerged culture fermentation. The enzyme was purified from the culture supernatant by applying a combination of freeze-thaw treatment, ammonium sulfate precipitation, hydrophobic interaction, and gel filtration chromatographies. The enzyme was purified by a 147-fold, with a yield of 7.54%. The molecular masses of the enzyme an determined by gel filtration and SDSPAGE were 13.6 and 18.2 kDa, respectively. The isoelectric point of the enzyme was 8.52. It hydrolyzed fibrinogen by cleaving the α and β chains of fibrinogen followed by the γ chains, and also activated plasminogen into plasmin. The enzyme was optimally active at 45°C and pH 7.4. The enzyme activity was completely inhibited by EDTA, whereas protease inhibitors of TPCK, SBTI, PMSF, aprotinin and pepstatin showed no inhibition on its activity. The partial amino acid sequences of the enzyme as determined by Q-TOF2 were ATFVGCSATR, GGTLIHESSHFTR, and YTTWFGTFVTSR. These sequences showed a high degree of homology with those of metallo-endopeptidases from P. ostreatus and Armillaria mellea. The purified enzyme can also be applied as a natural agent for oral fibrinolytic therapy or prevention of thrombosis.

  4. Two-dimensional gel electrophoresis for controlling and comparing culture supernatants of mammalian cell culture productions systems.

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    Wimmer, K; Harant, H; Reiter, M; Blüml, G; Gaida, T; Katinger, H

    1994-01-01

    A recombinant Chinese hamster ovary cell line, producing human erythropoietin, was cultivated in a continuous mode in a stirred tank reactor applying different dilution rates. In order to monitor the stability of this expression system, product and non-product proteins of the cell culture supernatant were analyzed by two-dimensional electrophoresis. The consistency of the isoforms of the recombinant product was determined by western blot combined with specific staining. The same cell line was propagated in a high cell density cultivation system based on macro-cell-aggregates. The patterns of secreted proteins of the cell line cultivated in the different systems were compared in order to detect modifications in protein expression of the product and of non product proteins relevant for cell culture supernatant. Hardly any alterations in two-dimensional pattern were detectable. The isoforms of erythropoietin, as well as the overall pattern of secreted proteins, detectable with the two-dimensional electrophoresis method were remarkably stable under different cultivation conditions.

  5. Isolation and Purification of an Early Pregnancy Factor–Like Molecule from Culture Supernatants Obtained from Lymphocytes of Pregnant Women

    OpenAIRE

    1998-01-01

    Purpose:Our purpose was to determine whether lymphocytes synthesize proteins during pregnancy, to observe whether one of the proteins synthesized has early pregnancy factor (EPF)–like activity and to isolate and purify this molecule from culture supernatants obtained from stimulated lymphocytes of pregnant women.

  6. Overcoming bottlenecks of enzymatic biofuel cell cathodes: crude fungal culture supernatant can help to extend lifetime and reduce cost.

    Science.gov (United States)

    Sané, Sabine; Jolivalt, Claude; Mittler, Gerhard; Nielsen, Peter J; Rubenwolf, Stefanie; Zengerle, Roland; Kerzenmacher, Sven

    2013-07-01

    Enzymatic biofuel cells (BFCs) show great potential for the direct conversion of biochemically stored energy from renewable biomass resources into electricity. However, enzyme purification is time-consuming and expensive. Furthermore, the long-term use of enzymatic BFCs is hindered by enzyme degradation, which limits their lifetime to only a few weeks. We show, for the first time, that crude culture supernatant from enzyme-secreting microorganisms (Trametes versicolor) can be used without further treatment to supply the enzyme laccase to the cathode of a mediatorless BFC. Polarization curves show that there is no significant difference in the cathode performance when using crude supernatant that contains laccase compared to purified laccase in culture medium or buffer solution. Furthermore, we demonstrate that the oxygen reduction activity of this enzymatic cathode can be sustained over a period of at least 120 days by periodic resupply of crude culture supernatant. This is more than five times longer than control cathodes without the resupply of culture supernatant. During the operation period of 120 days, no progressive loss of potential is observed, which suggests that significantly longer lifetimes than shown in this work may be possible. Our results demonstrate the possibility to establish simple, cost efficient, and mediatorless enzymatic BFC cathodes that do not require expensive enzyme purification procedures. Furthermore, they show the feasibility of an enzymatic BFC with an extended lifetime, in which self-replicating microorganisms provide the electrode with catalytically active enzymes in a continuous or periodic manner.

  7. Rapid Extracellular Biosynthesis of Silver Nanoparticles by Cunninghamella phaeospora Culture Supernatant

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    Ghareib, Mohamed; Tahon, Medhat Abu; Saif, Mona Mostafa; El-Sayed Abdallah, Wafaa

    2016-01-01

    The development of green approaches for the biosynthesis of silver nanoparticles (AgNPs) is of prime significance in the field of nanotechnology research. A fast and eco-friendly protocol for the biosynthesis of extracellular AgNPs using culture supernatant (CS) from the fungus Cunninghamella phaeospora was studied in this work. This CS was proved as a potential new source for the extracellular biosynthesis of AgNPs. The AgNPs were formed at 100 oC and pH 9 within four min of contact between CS and 1mM silver nitrate (AgNO3) solution. Nitrate reductase (NR) was confirmed to play a pivotal role in the biosynthesis of AgNPs. The enzyme expressed its highest activity at 80 oC and pH 9. At 100 oC the enzyme retained 70% of its original activity for one hour. The half-life (T1/2) of the enzyme activity was calculated to be 1.55 h confirming its thermostability. The produced AgNPs were characterized by UV-Vis spectroscopy, high resolution-transmission electron microscope (HR-TEM) and x-ray diffraction (XRD). These NPs showed an absorption peak at 415 nm in UV-Vis spectrum corresponding to the plasmon resonance of AgNPs. Transmission electron micrographs revealed the production of monodispersed spherical NPs with average particle size 14 nm. XRD spectrum of the NPs confirmed the formation of metallic crystalline silver. It was also suggested that the aromatic amino acids play a role in the biosynthesis process. The current research provided an insight on the green biosynthesis of AgNPs including some mechanistic aspects using a new mycogenic source.

  8. [Effects of culture supernatant of human amnion mesenchymal stem cells on biological characteristics of human fibroblasts].

    Science.gov (United States)

    Wu, Qi'er; Lyu, Lu; Xin, Haiming; Luo, Liang; Tong, Yalin; Mo, Yongliang; Yue, Yigang

    2016-06-01

    To investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts. (1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured. The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope. (2) Two batches of hAMSCs of the third passage were obtained, then the expression of vimentin of cells was observed with immunofluorescence method, and the expression of cell surface marker CD90, CD73, CD105, and CD45 was detected by flow cytometer. (3) hAMSCs-CS of the third passage at culture hour 72 were collected, and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay. (4) Human fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured. Human fibroblasts of the third passage were used in the following experiments. Cells were divided into blank control group and 10%, 30%, 50%, and 70% hAMSCs-CS groups according to the random number table (the same grouping method below), with 48 wells in each group. Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS), while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS. The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12, 24, 48, and 72, respectively, and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments. (5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4), with 4 wells in each group, at post

  9. Comparison of EBV DNA viral load in whole blood, plasma, B-cells and B-cell culture supernatant.

    Science.gov (United States)

    Ouedraogo, David Eric; Bollore, Karine; Viljoen, Johannes; Foulongne, Vincent; Reynes, Jacques; Cartron, Guillaume; Vendrell, Jean-Pierre; Van de Perre, Philippe; Tuaillon, Edouard

    2014-05-01

    Epstein-Barr virus (EBV) genome quantitation in whole blood is used widely for therapeutic monitoring of EBV-associated disorders in immunosuppressed individuals and in patients with EBV-associated lymphoma. However, the most appropriate biological material to be used for EBV DNA quantitation remains a subject of debate. This study compare the detection rate and levels of EBV DNA from whole blood, plasma, enriched B-cells, and B-cell short-term culture supernatant using quantitative real-time PCR. Samples were collected from 33 subjects with either HIV infection or B-cell lymphoma. Overall, EBV DNA was detected in 100% of enriched B-cell samples, in 82% of B-cell culture supernatants, in 57% of plasma, and 42% of whole blood samples. A significant correlation for EBV viral load was found between enriched B-cell and B-cell culture supernatant material (ρ = 0.92; P cells (ρ = -0.02; P = 0.89), whole blood and plasma (ρ = 0.24; P = 0.24), or enriched B-cells and plasma (ρ = 0.08; P = 0.77). Testing of enriched B-cells appeared to be the most sensitive method for detection of EBV DNA as well as for exploration of the cellular reservoir. Quantitation of EBV DNA in plasma and B-cell culture supernatant may be of interest to assess EBV reactivation dynamics and response to treatment as well as to decipher EBV host-pathogen interactions in various clinical scenarios.

  10. Improving the performance of a biofuel cell cathode with laccase-containing culture supernatant from Pycnoporus sanguineus.

    Science.gov (United States)

    Fokina, Oleksandra; Eipper, Jens; Winandy, Lex; Kerzenmacher, Sven; Fischer, Reinhard

    2015-01-01

    Laccases are multicopper oxidoreductases that can be used in biofuel cells to improve cathode performance by cathodic oxygen reduction. Here we present a laccase from the ligninolytic white-rot fungus Pycnoporus sanguineus that, in contrast to the Trametes versicolor laccase, can be produced in the absence of inducers in a standard culture medium. After 7days of cultivation the activity of this laccase in culture supernatant reached 2.5U/ml, which is high enough for direct application of the supernatant in biofuel cells. The highest current density of 115.0±3.5μA/cm(2) at 400mV vs. SCE was obtained at pH 5 with a buckypaper cathode with a laccase-containing culture supernatant. The enzyme also showed electrocatalytic activity at pH 6 and 7. These results not only present a new cost-efficient laccase for improving cathode performance, but also show that new laccases with different catalytic properties can be suitable for biofuel cells.

  11. THE EFFECT OF ANTISENSE OLIGONUCLEOTIDE ON THE INTERLEUKIN-5 IN THE SUPERNATANTS OF SPLEEN CELL CULTURES OF ASTHMATIC MICE

    Institute of Scientific and Technical Information of China (English)

    王美琴; 白春学; 钮善福; 方晓惠; 陈常庆; 陈波

    2001-01-01

    To explore the effect of antisense oligonucleotide on the production of IL-5 by mouse spleen T lymphocytes.Methods Based on the IL-5 cDNA sequence of mouse, a segment of antisense oligonucleotide was designed and synthesized. 5’-labeling of antisense oligonucleotide was signed by T4 PNK in order that the efficiency of stearylamine liposome in transfecting antisense oligonucleotide can be evaluated. Asthma model was duplicated with ovalbumin(OVA) absorbed to aluminum hydroxide. T lymphocytes of mice were separated by nylon fiber method, then T lymphocytes transfected with different concentration of antisense oligonucleotide with cation stearylamine liposme were incubated respectively in order to observe the effect of antisense oligonucleotide on Il-5 production by T lymphocytes. IL-5 levels in the supernatants of T lymphocyte cultures were determined by ELISA.Results Stearylamine liposome could markedly increase the efficiency of antisense oligonucleotide transfection. The transfection efficiency of antisense oligouncleotide increased approximately 12 times at a ratio of 1: 15m/m (antisense oligonucleotide to SA liposome). In healthy and asthma Balb/c mice, IL-5 was not detectable in the supernatants of T lymphocyte cultures without stimulated with OVA; however, IL-5 was increased markedly in the supernatants of T lymphocyte cultures stimulated with OVA. After transfection with different concentrations of antisense oligonucleotide, IL-5 levels in the supernatants of T lymphocyte cultures were significantly lower than those in control cultured without antisense oligonucleotide transfection. IL-5 levels decreased from 44.60±6.23 pg/ml to 30.70±7.362 pg/ml, 17.20±6.181 pg/ml and 8.16±2.34 pg/ml respectively. And IL-5 synthesis was inhibited by 31.17%, 61.43% and 81.7% respectively.Conclusion IL-5 synthesis could be obviously inhibited by antisense oligonucleotide and showed a markedly correlation between dose and effectiveness. It suggests the production

  12. Anion-exchange purification of recombinant factor IX from cell culture supernatant using different chromatography supports.

    Science.gov (United States)

    Ribeiro, Daniel A; Passos, Douglas F; Ferraz, Helen C; Castilho, Leda R

    2013-11-01

    Both recombinant and plasma-derived factor IX concentrates are used in replacement therapies for the treatment of haemophilia B. In the present work, the capture step for a recombinant FIX (rFIX) purification process was investigated. Different strong anion-exchange chromatography media (the resins Q Sepharose(®) FF and Fractogel(®) TMAE, the monolith CIM(®) QA and the membrane adsorber Sartobind(®) Q) were tested for their rFIX binding capacity under dynamic conditions. In these experiments, crude supernatant from CHO cells was used, thus in the presence of supernatant contaminants and mimicking process conditions. The highest dynamic binding capacity was obtained for the monolith, which was then further investigated. To study pseudoaffinity elution of functional rFIX with Ca(2+) ions, a design of experiments to evaluate the effects of pH, NaCl and CaCl2 on yield and purification factor was carried out. The effect of pH was not statistically significant, and a combination of no NaCl and 45mM CaCl2 yielded a good purification factor combined with a high yield of active rFIX. Under these conditions, activity yield of rFIX was higher than the mass yield, confirming selective elution of functional, γ-carboxylated rFIX. Scaling-up of this process 8 fold resulted in very similar process performance. Monitoring of the undesired activated FIX (FIXa) revealed that the FIXa/FIX ratio (1.94%) was higher in the eluate than in the loaded sample, but was still within an acceptable range. HCP and DNA clearances were high (1256 and 7182 fold, respectively), indicating that the proposed process is adequate for the intended rFIX capture step. Copyright © 2013 Elsevier B.V. All rights reserved.

  13. Antifungal performance of extracellular chitinases and culture supernatants of Streptomyces galilaeus CFFSUR-B12 against Mycosphaerella fijiensis Morelet.

    Science.gov (United States)

    Castillo, Benjamín Moreno; Dunn, Michael F; Navarro, Karina Guillén; Meléndez, Francisco Holguín; Ortiz, Magdalena Hernández; Guevara, Sergio Encarnación; Palacios, Graciela Huerta

    2016-03-01

    The tropical and mycoparasite strain Streptomyces galilaeus CFFSUR-B12 was evaluated as an antagonist of Mycosphaerella fijiensis Morelet, causal agent of the Black Sigatoka Disease (BSD) of banana. On zymograms of CFFSUR-B12 culture supernatants, we detected four chitinases of approximately 32 kDa (Chi32), 20 kDa (Chi20), and two with masses well over 170 kDa (ChiU) that showed little migration during denaturing electrophoresis at different concentrations of polyacrylamide. The thymol-sulphuric acid assay showed that the ChiU were glycosylated chitinases. Moreover, matrix assisted laser desorption ionization time-of-flight MS analysis revealed that the ChiU are the same protein and identical to a family 18 chitinase from Streptomyces sp. S4 (gi|498328075). Chi32 was similar to an extracellular protein from Streptomyces albus J1074 (gi|478687481) and Chi20 was non-significantly similar to chitinases from five different strains of Streptomyces (P > 0.05). Subsequently, Chi32 and Chi20 were partially purified by anion exchange and hydrophobic interaction chromatography and tested against M. fijiensis. Chitinases failed to inhibit ascospore germination, but inhibited up to 35 and 62% of germ tube elongation and mycelial growth, respectively. We found that crude culture supernatant and living cells of S. galilaeus CFFSUR-B12 were the most effective in inhibiting M. fijiensis and are potential biocontrol agents of BSD.

  14. Continuous precipitation of process related impurities from clarified cell culture supernatant using a novel coiled flow inversion reactor (CFIR).

    Science.gov (United States)

    Kateja, Nikhil; Agarwal, Harshit; Saraswat, Aditya; Bhat, Manish; Rathore, Anurag S

    2016-10-01

    Coiled Flow Inverter Reactor (CFIR) has recently been explored for facilitating continuous operation of several unit operations involved in downstream processing of biopharmaceuticals such as viral inactivation and protein refolding. The application of CFIR for continuous precipitation of clarified cell culture supernatant has been explored. The pH based precipitation is optimized in the batch mode and then in the continuous mode in CFIR using a design of experiments (DOE) study. Improved clearance of host cell DNA (52× vs. 39× in batch), improved clearance of host cell proteins (HCP) (7× vs. 6× in batch) and comparable recovery (90 vs. 91.5 % in batch) are observed along with six times higher productivity. To further demonstrate wider applicability of CFIR in performing continuous precipitation, two more case studies involving use of two different precipitation protocols (CaCl2 based and caprylic acid based) are also performed. In both cases, clearance of host cell DNA, HCP, and product recovery are found to be comparable or better in CFIR than in batch operations. Moreover, increase in productivity of 16 times (CaCl2 based) and eight times (caprylic acid based) is obtained for the two precipitation protocols, respectively. The data clearly demonstrate that CFIR can be seamlessly integrated into a continuous bioprocess train for performing continuous precipitation of clarified cell culture supernatant. To our knowledge this is the first report of such use. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Protein A is released into the Staphylococcus aureus culture supernatant with an unprocessed sorting signal.

    Science.gov (United States)

    O'Halloran, Dara P; Wynne, Kieran; Geoghegan, Joan A

    2015-04-01

    The immunoglobulin binding protein A (SpA) of Staphylococcus aureus is synthesized as a precursor with a C-terminal sorting signal. The sortase A enzyme mediates covalent attachment to peptidoglycan so that SpA is displayed on the surface of the bacterium. Protein A is also found in the extracellular medium, but the processes involved in its release are not fully understood. Here, we show that a portion of SpA is released into the supernatant with an intact sorting signal, indicating that it has not been processed by sortase A. Release of SpA was reduced when the native sorting signal of SpA was replaced with the corresponding region of another sortase-anchored protein (SdrE). Similarly, a reporter protein fused to the sorting signal of SpA was released to a greater extent than the same polypeptide fused to the SdrE sorting signal. Released SpA protected bacteria from killing in human blood, indicating that it contributes to immune evasion.

  16. Vip3A is responsible for the potency of Bacillus thuringiensis 9816C culture supernatant against Helicoverpa armigera and Spodoptera exigua.

    Science.gov (United States)

    Cai, Jun; Xiao, Liang; Yan, Bing; Bin, Guan; Chen, Yuehua; Ren, Gaixin

    2006-04-01

    Culture supernatant of Bacillus thuringiensis 9816C had high toxicity against Helicoverpa armigera and Spodoptera exigua. However, it lost insecticidal activities after being bathed in boiling water for 5 min. Acrystalliferous mutants of Bt9816C (Bt9816C-NP1 and Bt9816C-NP2) cured of its endogenous plasmids no longer possessed vip3A gene and toxicity. The 89 kD protein which existed in Bt9816C supernatant disappeared in the two mutants' supernatant; nevertheless, the two mutants still exhibited hemolytic and phospholipase C activity as Bt9816C did. The vip3A gene of Bt9816C, vip3Aa18, was cloned and expressed in Escherichia coli BL21. Bioassay demonstrated that the recombinant E. coli had high toxicity against S. exigua. Taken together, it suggested that Vip3A protein was responsible for the toxicity of Bt9816C culture supernatants.

  17. Pericardial fluid culture

    Science.gov (United States)

    ... the thin sac that surrounds the heart (the pericardium). A small amount of fluid is removed. You ... may be due to an infection of the pericardium. The specific organism causing the infection may be ...

  18. Biological synthesis of very small silver nanoparticles by culture supernatant of Klebsiella pneumonia: The effects of visible-light irradiation and the liquid mixing process

    Energy Technology Data Exchange (ETDEWEB)

    Mokhtari, Narges [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Daneshpajouh, Shahram; Seyedbagheri, Seyedali; Atashdehghan, Reza [Hydrometallurgy Research Unit, Research and Development Center, National Iranian Copper Industries Company, Sarcheshmeh, Rafsanjan (Iran, Islamic Republic of); Abdi, Khosro [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Sarkar, Saeed [Research Center for Science and Technology in Medicine, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Minaian, Sara [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of); Shahverdi, Hamid Reza [Department of Material Science, Faculty of Engineering, Tarbiat Modares University, Tehran (Iran, Islamic Republic of); Shahverdi, Ahmad Reza, E-mail: shahverd@sina.tums.ac.ir [Department of Pharmaceutical Biotechnology and Biotechnology Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran (Iran, Islamic Republic of)

    2009-06-03

    This study has investigated different visible-light irradiation's effect on the formation of silver nanoparticles from silver nitrate using the culture supernatant of Klebsiella pneumonia. Our study shows that visible-light emission can significantly prompt the synthesis of silver nanoparticles. Also, the study experimentally investigated the liquid mixing process effect on silver nanoparticle synthesis by visible-light irradiation. This study successfully synthesized uniformly dispersed silver nanoparticles with a uniform size and shape in the range of 1-6 nm with an average size of 3 nm. Furthermore, the study investigated the mechanism of the reduction of silver ions by culture supernatant of K. pneumonia, and used X-ray diffraction to characterize silver chloride as an intermediate compound. Silver chloride was prepared synthetically and used as a substrate for the synthesis of silver nanoparticles by culture supernatant of K. pneumonia. The silver nanoparticles have been prepared from silver chloride during this investigation for the first time.

  19. Modulation of the Culture Supernatant of Decidual Cells with Exogenous Cytokines on Killing Activity of Natural Killer Cells in Early Pregnancy

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Objective To investigate the important function of cytokines in early pregnancy and to provide basic and experimental evidence for understanding the mechanism of their action. Methods Add interferon-γ (IFN-γ) , interleukin- 2(IL- 2) , interleukin- 6(IL-6) and epidermal growth factor (EGF) to the confluent culturing decidual cells with three different concentrations and harvest the culture supernatant after 12, 24 and 48 h separately. Observe the effect of the supernatant on killing activity of NK cells with radioimmunological assay of 51Cr immersion. Results The culture supernatant of decidual cells can promote the killing activity of NK cells in various degrees, and the effect is independent of the type, concentration and acting time of cytokines. Conclusion In normal pregnancy, decidual cytokine network is in a dynamic equilibri um. Exogenous cytokines would be harm to normal pregnancy by interfering the equi librium state, but the exact mechanism needs further study.

  20. Modulation of the Culture Supernatant of Decidual Cells with Exogenous Cytokines on Killing Activity of Natural Killer Cells in Early Pregnancy

    Institute of Scientific and Technical Information of China (English)

    胡冬梅; 王丽莉; 何援利

    2000-01-01

    Objective To investigate the important function of cytohines in early pregnancy and to provide basic and experimental evidence for understanding the mechanism of their action.Methods Add interferon-γ (IFN-γ) ,interleuhin-2(IL-2) , interleuhin-6(IL-6) andepidermal growth factor(EGF) to the confluent culturing decidual cells with three different concentrations and harvest the culture supernatant after 12, 24 and 48 h separately. Observe the effect of the supernatant on killing activity of NK cells with radioimmunological assay of 51Cr immersion.Results The culture supernatant of decidual cells can promote the killing activity of NK cells in various degrees, and the effect is independent of the type, concentration and acting time of cytokines.Conclusion In normal pregnancy, decidual cytokine network is in a dynamic equilibri-um. Exogenous cytokines would be harm to normal pregnancy by interfering the equi-librium state, but the exact mechanism needs further study.

  1. Quantitative Determination of Lethal Toxin Proteins in Culture Supernatant of Human Live Anthrax Vaccine Bacillus anthracis A16R.

    Science.gov (United States)

    Zai, Xiaodong; Zhang, Jun; Liu, Ju; Liu, Jie; Li, Liangliang; Yin, Ying; Fu, Ling; Xu, Junjie; Chen, Wei

    2016-02-25

    Bacillus anthracis (B. anthracis) is the etiological agent of anthrax affecting both humans and animals. Anthrax toxin (AT) plays a major role in pathogenesis. It includes lethal toxin (LT) and edema toxin (ET), which are formed by the combination of protective antigen (PA) and lethal factor (LF) or edema factor (EF), respectively. The currently used human anthrax vaccine in China utilizes live-attenuated B. anthracis spores (A16R; pXO1+, pXO2-) that produce anthrax toxin but cannot produce the capsule. Anthrax toxins, especially LT, have key effects on both the immunogenicity and toxicity of human anthrax vaccines. Thus, determining quantities and biological activities of LT proteins expressed by the A16R strain is meaningful. Here, we explored LT expression patterns of the A16R strain in culture conditions using another vaccine strain Sterne as a control. We developed a sandwich ELISA and cytotoxicity-based method for quantitative detection of PA and LF. Expression and degradation of LT proteins were observed in culture supernatants over time. Additionally, LT proteins expressed by the A16R and Sterne strains were found to be monomeric and showed cytotoxic activity, which may be the main reason for side effects of live anthrax vaccines. Our work facilitates the characterization of anthrax vaccines components and establishment of a quality control standard for vaccine production which may ultimately help to ensure the efficacy and safety of the human anthrax vaccine A16R.

  2. Putative new heat-stable cytotoxic and enterotoxic factors in culture supernatant of Escherichia coli isolated from drinking water

    Directory of Open Access Journals (Sweden)

    DA Ribeiro

    2011-01-01

    Full Text Available Enteric infections caused by the ingestion of contaminated water, especially by Escherichia coli, are important to define the virulence properties of these bacteria. Due to frequent infantile diarrhea in the city of Ouro Preto, Minas Gerais state, Brazil, the phenotypic and genotypic diarrheagenic properties of E. coli isolated from drinking water were studied. The culture supernatants of 39 (40% among a total of 97 E. coli isolates from drinking water were positive by suckling mouse assay and induced cytotoxic effects on Vero cells. The enterotoxic and cytotoxic activities were present in the fraction with less than 10 kDa and were not lost when heated up to 60°C and 100°C for 30 minutes. PCR assays showed that among these 39 Vero cytotoxigenic E. coli, four (10.2% were positive for ST II (estB and two (5% positive for αHly (hlyA. Gene amplification of SLT (stx 1, stx 2, ST I (estA, LT (eltI, eltII, EAST1 (astA, EHly (enhly and plasmid-encoded enterotoxin (pet were not observed. This heat-stable cytotoxic enterotoxin of E. coli is probably a new putative diarrheagenic virulence factor, as a toxin presenting these characteristics has not yet been described.

  3. Human cerebrospinal fluid fatty acid levels differ between supernatant fluid and brain-derived nanoparticle fractions, and are altered in Alzheimer's disease.

    Directory of Open Access Journals (Sweden)

    Alfred N Fonteh

    Full Text Available Although saturated (SAFA, monounsaturated (MUFA, and polyunsaturated (PUFA fatty acids are important structural components of neuronal membranes and precursors of signaling molecules, knowledge of their metabolism in Alzheimer's disease (AD is limited. Based on recent discovery that lipids in cerebrospinal fluid (CSF are distributed in both brain-derived nanoparticles (NP and supernatant fluid (SF, we hypothesized that fatty acid (FA abundance and distribution into these compartments is altered in early AD pathology.We assayed the FA composition and abundance in CSF fractions from cognitively healthy (CH, mild cognitive impairment (MCI, and AD study participants using gas chromatography-mass spectrometry. In the SF fraction, concentration of docosahexaenoic acid [DHA, (C22:6n-3] was less in AD compared with CH, while alpha linolenic acid [α-LNA, (C18:3n-3] was lower in MCI compared with CH. In the NP fraction, levels of SAFAs (C15:0, C16:0 and a MUFA (C15:1 differentiated CH from MCI, while two MUFAs (C15:1, C19:1 and four PUFAs (C20:2n-6, C20:3n-3, C22:4n-6, C22:5n-3 were higher in AD compared with CH. Levels of even-chain free SAFA and total free FA levels were higher in AD, levels of odd-chain free SAFAs, MUFAs, n-3 PUFAs, and total PUFA, were lower in AD compared with CH. Free n-6 PUFA levels were similar in all three groups.FA metabolism is compartmentalized differently in NP versus SF fractions of CSF, and altered FA levels reflect the importance of abnormal metabolism and oxidative pathways in AD. Depleted DHA in CSF fractions in AD is consistent with the importance of n-3 PUFAs in cognitive function, and suggests that disturbed PUFA metabolism contributes to AD pathology. This study of FA levels in CSF fractions from different cognitive stages shows potential AD biomarkers, and provides further insight into cell membrane dysfunctions, including mechanisms leading to amyloid production.

  4. A novel toxin homologous to large clostridial cytotoxins found in culture supernatant of Clostridium perfringens type C.

    Science.gov (United States)

    Amimoto, Katsuhiko; Noro, Taichi; Oishi, Eiji; Shimizu, Mitsugu

    2007-04-01

    An unknown cytotoxin was identified in the culture supernatant of Clostridium perfringens type C. The cytotoxin, named TpeL, which was purified using mAb-based affinity chromatography, had a lethal activity of 62 minimum lethal dose (MLD) mg(-1) in mice and a cytotoxic activity of 6.2x10(5) cytotoxic units (CU) mg(-1) in Vero cells. The nucleotide sequence of TpeL was determined. The entire ORF had a length of 4953 bases, and the same nucleotide sequence was not recorded in the GenBank/EMBL/DDBJ databases. The molecular mass calculated from the deduced amino acid sequence was 191 kDa, and a signal peptide region was not found within the ORF. The deduced amino acid sequence exhibited 30-39 % homology to Clostridium difficile toxins A (TcdA) and B (TcdB), Clostridium sordellii lethal toxin (TcsL) and Clostridium novyi alpha-toxin (TcnA). The amino acid sequence of TpeL is shorter than these toxins, and the homologous region was located at the N-terminal site. Eighteen strains of C. perfringens types A, B and C were surveyed for the presence of the tpeL gene by PCR. The tpeL gene was detected in all type B (one strain) and C strains (five strains), but not in any type A strains (12 strains). TpeL was detected in culture filtrates of the five type C strains by dot-blot analysis, but not in the type B strain. It was concluded that TpeL is a novel toxin similar to the known large clostridial cytotoxins. Furthermore, the data indicated that TpeL is produced by many C. perfringens type C strains.

  5. [Effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells and ciprofloxacin on Staphylococcus aureus in vitro].

    Science.gov (United States)

    Zhou, B; Tu, H L; Ba, T; Wang, L F; Wang, S J; Nie, S Y

    2017-06-20

    Objective: To explore the effects of combined application of culture supernatant of human umbilical cord mesenchymal stem cells (hUCMSCs) and ciprofloxacin on Staphylococcus aureus (SA) in vitro. Methods: hUCMSCs were isolated from umbilical cord tissue of full-term healthy fetus after cesarean section and cultured. Cells in the third passage were used in the experiments after identification. SA strains isolated from wounds of burn patients in our burn wards were used in the experiments. Cells were divided into 0, 10, 100, and 1 000 ng/mL lipopolysaccharide (LPS) groups according to the random number table (the same dividing method below). Cells were cultured with culture medium of mesenchymal stem cells (MSCs) after being treated with medium containing the corresponding mass concentrations of LPS for 12 h. At post culture hour (PCH) 6, 12, and 24, 6 wells of culture supernatant of cells in each group were obtained to measure the content of LL-37 with enzyme-linked immunosorbent assay. Ninety blood agar plates were divided into ciprofloxacin control group (CC), ciprofloxacin+ supernatant group (CS), and ciprofloxacin+ supernatant+ LL-37 antibody group (CSL), with 30 blood agar plates in each group. Blood agar plates in group CC were coated with 1.5×10(8) colony forming unit (CFU)/mL bacteria solution prepared with normal saline. Blood agar plates in group CS were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline and culture supernatant of hUCMSCs (cultured by culture medium of MSCs, the same below) in double volume of normal saline. Blood agar plates in group CSL were coated with 1.5×10(8) CFU/mL bacteria solution prepared with normal saline, culture supernatant of hUCMSCs in double volume of normal saline, and 2.6 μL LL-37 antibody in the concentration of 2 μg/mL. At PCH 12, 24, and 48, 10 blood agar plates of each group were harvested to observe the distribution of SA colony on blood agar plate and to measure the diameter of

  6. Understanding ForteBio's Sensors for High-Throughput Kinetic and Epitope Screening for Purified Antibodies and Yeast Culture Supernatant.

    Science.gov (United States)

    Yu, Yao; Mitchell, Scott; Lynaugh, Heather; Brown, Michael; Nobrega, R Paul; Zhi, Xiaoyong; Sun, Tingwan; Caffry, Isabelle; Cao, Yuan; Yang, Rong; Burnina, Irina; Xu, Yingda; Estep, Patricia

    2016-01-01

    Real-time and label-free antibody screening systems are becoming more popular because of the increasing output of purified antibodies and antibody supernatant from many antibody discovery platforms. However, the properties of the biosensor can greatly affect the kinetic and epitope binning results generated by these label-free screening systems. ForteBio human-specific ProA, anti-human IgG quantitation (AHQ), anti-human Fc capture (AHC) sensors, and custom biotinylated-anti-human Fc capture (b-AHFc) sensors were evaluated in terms of loading ability, regeneration, kinetic characterization, and epitope binning with both purified IgG and IgG supernatant. AHC sensors proved unreliable for kinetic or binning assays at times, whereas AHQ sensors showed poor loading and regeneration abilities. ProA sensors worked well with both purified IgG and IgG supernatant. However, the interaction between ProA sensors and the Fab region of the IgG with VH3 germline limited the application of ProA sensors, especially in the epitope binning experiment. In an attempt to generate a biosensor type that would be compatible with a variety of germlines and sample types, we found that the custom b-AHFc sensors appeared to be robust working with both purified IgG and IgG supernatant, with little evidence of sensor-related artifacts.

  7. Characterization of monoclonal antibodies by a fast and easy liquid chromatography-mass spectrometry time-of-flight analysis on culture supernatant.

    Science.gov (United States)

    Henninot, Antoine; Terrier, Aurelie; Charton, Julie; Urbain, Rémi; Fontayne, Alexandre; Deprez, Benoit; Beghyn, Terence

    2015-12-15

    Rapid and efficient structural analysis is key to the development of new monoclonal antibodies. We have developed a fast and easy process to obtain mass spectrometry profiles of antibodies from culture supernatant. Treatment of the supernatant with IdeS generates three fragments of 25 kDa that can be analyzed by liquid chromatography-mass spectrometry time-of-flight (LC-MS TOF) in one run: LC, Fd, and Fc/2. This process gives rapid access to isoform and glycoform profiles. To specifically measure the fucosylation yield, we included a one-pot treatment with EndoS that removes the distal glycan heterogeneity. Our process was successfully compared with high-performance capillary electrophoresis with laser-induced fluorescence detection (HPCE-LIF), currently considered as the "gold standard" method. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. Cell-free culture supernatant of Bifidobacterium breve CNCM I-4035 decreases pro-inflammatory cytokines in human dendritic cells challenged with Salmonella typhi through TLR activation.

    Science.gov (United States)

    Bermudez-Brito, Miriam; Muñoz-Quezada, Sergio; Gomez-Llorente, Carolina; Matencio, Esther; Bernal, Maria J; Romero, Fernando; Gil, Angel

    2013-01-01

    Dendritic cells (DCs) constitute the first point of contact between gut commensals and our immune system. Despite growing evidence of the immunomodulatory effects of probiotics, the interactions between the cells of the intestinal immune system and bacteria remain largely unknown. Indeed,, the aim of this work was to determine whether the probiotic Bifidobacterium breve CNCM I-4035 and its cell-free culture supernatant (CFS) have immunomodulatory effects in human intestinal-like dendritic cells (DCs) and how they respond to the pathogenic bacterium Salmonella enterica serovar Typhi, and also to elucidate the molecular mechanisms involved in these interactions. Human DCs were directly challenged with B. breve/CFS, S. typhi or a combination of these stimuli for 4 h. The expression pattern of genes involved in Toll-like receptor (TLR) signaling pathway and cytokine secretion was analyzed. CFS decreased pro-inflammatory cytokines and chemokines in human intestinal DCs challenged with S. typhi. In contrast, the B. breve CNCM I-4035 probiotic strain was a potent inducer of the pro-inflammatory cytokines and chemokines tested, i.e., TNF-α, IL-8 and RANTES, as well as anti-inflammatory cytokines including IL-10. CFS restored TGF-β levels in the presence of Salmonella. Live B.breve and its supernatant enhanced innate immune responses by the activation of TLR signaling pathway. These treatments upregulated TLR9 gene transcription. In addition, CFS was a more potent inducer of TLR9 expression than the probiotic bacteria in the presence of S. typhi. Expression levels of CASP8 and IRAK4 were also increased by CFS, and both treatments induced TOLLIP gene expression. Our results indicate that the probiotic strain B. breve CNCM I-4035 affects the intestinal immune response, whereas its supernatant exerts anti-inflammatory effects mediated by DCs. This supernatant may protect immune system from highly infectious agents such as Salmonella typhi and can down-regulate pro

  9. Usefulness of whole blood, plasma, peritoneal fluid, and peritoneal fluid supernatant glucose concentrations obtained by a veterinary point-of-care glucometer to identify septic peritonitis in dogs with peritoneal effusion.

    Science.gov (United States)

    Koenig, Amie; Verlander, Lindsey Lane

    2015-11-01

    To evaluate the usefulness of a veterinary point-of-care glucometer for identification of septic peritonitis in dogs with peritoneal effusion (PE). Prospective clinical evaluation. 39 dogs with PE. Blood and peritoneal fluid convenience samples were collected concurrently in all dogs at the time of initial evaluation. A veterinary point-of-care glucometer was used to measure glucose concentration in heparinized whole blood, plasma, peritoneal fluid, and peritoneal fluid supernatant samples. Seventeen dogs had confirmed septic peritonitis, and 22 dogs had nonseptic PE. Sensitivity, specificity, positive and negative predictive values, and accuracy of identification of dogs with septic peritonitis were calculated for glucose concentration differences for whole blood versus peritoneal fluid (WB-PF), plasma versus peritoneal fluid (P-PF), and plasma versus peritoneal fluid supernatant (P-PFS). With a cutoff of > 20 mg/dL, the glucose concentration difference for WB-PF was an insensitive indicator of septic peritonitis (sensitivity, 41.2%; specificity, 100%). In comparison, the glucose concentration differences for P-PF and P-PFS had a higher sensitivity for septic peritonitis (88.2% and 82.4%, respectively) but a lower specificity (80% and 77.8%, respectively). With a glucose concentration difference cutoff of ≥ 38 mg/dL, specificity, positive predictive value, and accuracy of P-PF and P-PFS improved. Determination of the glucose concentration difference for WB-PF with the veterinary point-of-care glucometer was not useful in identifying all dogs with septic peritonitis. A glucose concentration difference of ≥ 38 mg/dL for P-PF or P-PFS, however, supported an accurate diagnosis of septic peritonitis in dogs with PE.

  10. Food preservative potential of gassericin A-containing concentrate prepared from a cheese whey culture supernatant from Lactobacillus gasseri LA39.

    Science.gov (United States)

    Nakamura, Kiyoshi; Arakawa, Kensuke; Kawai, Yasushi; Yasuta, Narimi; Chujo, Takahiro; Watanabe, Masamichi; Iioka, Hiroyuki; Tanioka, Masashi; Nishimura, Junko; Kitazawa, Haruki; Tsurumi, Koichi; Saito, Tadao

    2013-02-01

    Gassericin A (GA) is a circular bacteriocin produced by Lactobacillus gasseri LA39. In this study, GA-containing concentrate was prepared using a cross-flow membrane filtration device (30 kDa cut-off) from the culture supernatant of Lb. gasseri LA39 cultivated in a cheese whey-based food-grade medium. The bacteriocin activity titer in the concentrate was 16 times as high as that of the culture supernatant and was completely maintained through each incubation at 4°C for 3 months, 37°C for 2 months, 60°C for 5 h, and 100°C for 30 min. The GA-containing concentrate was used with glycine powder to make custard creams, and then four representative strains of custard cream spoilage bacteria (Bacillus cereus, Lactococcus lactis subsp. lactis, Achromobacter denitrificans and Pseudomonas fluorescens) were individually inoculated at c. 10(3) colony forming units/g in the custard creams. Throughout 30 days of incubation at 30°C, all of the inoculated bacteria were completely inhibited by the combination of 5% (w/w) of the GA-containing concentrate and 0.5% (w/w) glycine. This is the first highly practical application of GA to foods as a biopreservative, and the concentration method and the bacteriocin concentrate would contribute to biopreservation of several foods.

  11. High-efficient n-butanol production by co-culturing Clostridium acetobutylicum and Saccharomyces cerevisiae integrated with butyrate fermentative supernatant addition.

    Science.gov (United States)

    Luo, Hongzhen; Zeng, Qingwei; Han, Shuo; Wang, Zhaoyu; Dong, Qing; Bi, Yanhong; Zhao, Yuping

    2017-04-01

    Butanol is not only an important chemical intermediate and solvent in pharmaceutical and cosmetics industries, but also considered as an advanced biofuel. Although species of the natural host Clostridium have been engineered, butanol titers in the anaerobe seem to be limited by its intolerance to butanol less than 13 g/L. Here we aimed to develop a technology for enhancing butanol production by a co-culture system with butyrate fermentative supernatant addition. First, when adding 4.0 g/L butyrate into the acetone-butanol-ethanol (ABE) fermentation broth with single-shot at 24 h, the "acid crash" phenomenon occurred and the ABE fermentation performance deteriorated. Subsequently, we found that adding certain amino acids could effectively enhance butyrate re-assimilation, butanol tolerance and titer (from 11.1 to 14.8 g/L). Additionally, in order to decrease the raw material cost, butyrate fermentative supernatant produced by Clostridium tyrobutyricum was applied to butanol production in the Clostridium acetobutylicum/Saccharomyces cerevisiae co-culture system, instead of adding synthetic butyrate. Final butanol and total ABE concentrations reached higher levels of 16.3 and 24.8 g/L with increments of 46.8 and 37.8%, respectively. These results show that the proposed fermentation strategy has great potential for efficiently butanol production with an economic approach.

  12. Distinct galactofuranose antigens in the cell wall and culture supernatants as a means to differentiate Fusarium from Aspergillus species.

    Science.gov (United States)

    Wiedemann, Annegret; Kakoschke, Tamara Katharina; Speth, Cornelia; Rambach, Günter; Ensinger, Christian; Jensen, Henrik Elvang; Ebel, Frank

    2016-09-01

    Detection of carbohydrate antigens is an important means for diagnosis of invasive fungal infections. For diagnosis of systemic Aspergillus infections, galactomannan is commonly used, the core antigenic structure of which consists of chains of several galactofuranose moieties. In this study, we provide evidence that Fusarium produces at least two distinct galactofuranose antigens: Smaller amounts of galactomannan and larger quantities of a novel antigen recognized by the monoclonal antibody AB135-8. In A. fumigatus, only minor amounts of the AB135-8 antigen are found in supernatants and in the apical regions of hyphae. A galactofuranose-deficient A. fumigatus mutant lacks the AB135-8 antigen, which strongly suggests that galactofuranose is an essential constituent of this antigen. Using a combination of AB135-8 and a galactomannan-specific antibody, we were able to unambiguously differentiate A. fumigatus and Fusarium hyphae in immunohistology. Moreover, since Fusarium releases the AB135-8 antigen, it appears to be a promising target antigen for a serological detection of Fusarium infections. Copyright © 2016 Elsevier GmbH. All rights reserved.

  13. Culture supernatants of breast cancer cell line MDA-MB-231 treated with parthenolide inhibit the proliferation, migration, and lumen formation capacity of human umbilical vein endothelial cells

    Institute of Scientific and Technical Information of China (English)

    LI Cai-juan; GUO Su-fen; SHI Tie-mei

    2012-01-01

    Background Parthenolide has been tested for anti-tumor activities,such as anti-proliferation and pro-apoptosis in recent studies.However,little is known about its role in the process of tumor angiogenesis.This study aims to investigate the effects and potential mechanisms of parthenolide on the proliferation,migration and lumen formation capacity of human umbilical vein endothelial cells.Methods Different concentrations of parthenolide were applied to the human breast cancer cell line MDA-MB-231 cells.After 24-hour incubation,the culture supematants were harvested and used to treat human umbilical vein endothelial cells for 24 hours.Then an inverted fluorescence phase contrast microscope was used to evaluate the human umbilical vein endothelial cells.The secretion of vascular endothelial growth factor (VEGF),interleukin (IL)-8 and matrix metalloproteinases (MMP)-9 in the culture supernatant of the MDA-MB-231 cells was then measured with enzyme-linked immunosorbent assay (ELISA) assays.Results Suppression of proliferation,migration,and the lumen formation capacity of human umbilical vein endothelial cells was observed in the presence of the culture supernatants from the breast cancer cell line treated with different concentrations of parthenolide.Parthenolide decreased the levels of the angiogenic factors MMP-9,VEGF,and IL-8secreted by the MDA-MB-231 cells.Conclusions Parthenolide may suppress angiogenesis through decreasing angiogenic factors secreted by breast cancer cells to interfere with the proliferation,migration and lumen-like structure formation of endothelial cells,thereby inhibiting tumor growth.It is a promising potential anti-angiogenic drug.

  14. Performance of multiplex commercial kits to quantify cytokine and chemokine responses in culture supernatants from Plasmodium falciparum stimulations.

    Directory of Open Access Journals (Sweden)

    Gemma Moncunill

    Full Text Available BACKGROUND: Cytokines and chemokines are relevant biomarkers of pathology and immunity to infectious diseases such as malaria. Several commercially available kits based on quantitative suspension array technologies allow the profiling of multiple cytokines and chemokines in small volumes of sample. However, kits are being continuously improved and information on their performance is lacking. METHODOLOGY/PRINCIPAL FINDINGS: Different cytokine/chemokine kits, two flow cytometry-based (eBioscience® FlowCytomix™ and BD™ Cytometric Bead Array Human Enhanced Sensitivity and four Luminex®-based (Invitrogen™ Human Cytokine 25-Plex Panel, Invitrogen™ Human Cytokine Magnetic 30-Plex Panel, Bio-Rad® Bio-Plex Pro™ Human Cytokine Plex Assay and Millipore™ MILLIPLEX® MAP Plex Kit were compared. Samples tested were supernatants of peripheral blood mononuclear cells of malaria-exposed children stimulated with Plasmodium falciparum parasite lysates. Number of responses in range that could be detected was determined and reproducibility of duplicates was evaluated by the Bland-Altman test. Luminex® kits performed better than flow cytometry kits in number of responses in range and reproducibility. Luminex® kits were more reproducible when magnetic beads were used. However, within each methodology overall performance depended on the analyte tested in each kit. Within the Luminex® kits, the Invitrogen™ with polystyrene beads had the poorer performance, whereas Invitrogen™ with magnetic beads had the higher percentage of cytokines/chemokines with both readings in range (40%, followed by Bio-Rad® with magnetic beads (35%. Regarding reproducibility, the Millipore™ kit had the highest percentage (60% of cytokines/chemokines with acceptable limits of agreement (<30%, followed by the Invitrogen™ with magnetic beads (40% that had tighter limits of agreement. CONCLUSIONS/SIGNIFICANCE: Currently available kits for cytokine and chemokine

  15. Screening for epitope specificity directly on culture supernatants in the early phase of monoclonal antibody production by an ELISA with biotin-labeled antigen

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Jensen, Charlotte H; Gregersen, Annemette

    2004-01-01

    This report describes an assay for comparison of epitope specificity in groups of monoclonal antibodies against a given antigen. The only prerequisite is the biotin-labeled antigen. One of the monoclonal antibodies is captured onto a plastic surface via a rabbit anti-mouse Ig, and the other...... preincubated with biotinylated antigen. When the two antibodies react with the same epitope subsequent binding of the biotin-labeled antigen is abolished (inhibition). In the cases where no inhibition was observed, the two antibodies were considered to react with distinct, independent epitopes. The obvious...... advantages using this assay, are that it can be performed directly on culture supernatants in the early phase of monoclonal antibody production, and also works for antigens with repetitive epitopes. Moreover, the bonus effect, i.e., a signal in excess of the reference signal when sets of monoclonal...

  16. Purification of monoclonal antibodies, IgG1, from cell culture supernatant by use of metal chelate convective interaction media monolithic columns.

    Science.gov (United States)

    Rajak, Poonam; Vijayalakshmi, M A; Jayaprakash, N S

    2012-12-01

    Monoclonal antibodies (MAbs) have diverse applications in diagnostics and therapeutics. The recent advancement in hybridoma technology for large-scale production of MAbs in bioreactors demands rapid and efficient purification methods. Conventional affinity purification systems have drawbacks of low flow rates and denaturation of antibodies owing to harsh elution conditions. Here, we attempted purification of MAbs by use of a high-throughput metal-chelate methacrylate monolithic system. Monolithic macroporous convective interaction media-iminodiacetate (CIM-IDA) disks immobilized with four different metal ions (Cu²⁺, Ni²⁺, Zn²⁺ and Co²⁺) were used and evaluated for purification of anti-human serum albumin IgG1 mouse MAbs from cell culture supernatant after precipitation with 50% ammonium sulfate. Elution with 10 mM imidazole in the equilibration buffer (25 mM MMA = MOPS (Morpholino propane sulfonic acid) + MES (Morpholino ethane sulfonic acid) + Acetate + 0.5 M NaCl, pH 7.4) resulted in a purification of 25.7 ± 2.9-fold and 32.5 ± 2.6-fold in experiments done using Zn²⁺ and Co²⁺ metal ions, respectively. The highest recovery of 85.4 ± 1.0% was obtained with a CIM-IDA-Zn(II) column. SDS-PAGE, ELISA and immuno-blot showed that the antibodies recovered were pure, with high antigen-binding efficiency. Thus, metal chelate CIM monoliths could be a potential alternative to conventional systems for fast and efficient purification of MAbs from the complex cell culture supernatant.

  17. Comprehensive supernatant treatment

    Energy Technology Data Exchange (ETDEWEB)

    Egan, Z. [Oak Ridge National Lab., TN (United States)

    1996-10-01

    This task involves testing of sorbent materials for removing cesium, strontium, and technetium from the saline solutions in DOE storage tank supernatant at Oak Ridge and other sites. Staff at Oak Ridge National Laboratory (ORNL) are recovering and treating the liquid (supernatant) portions of Melton Valley Storage Tank (MVST) waste in a hot cell to separate and remove the radionuclides. Batch tests will be used to evaluate and select the most promising materials for supernatant treatment to reduce the amount of waste for final disposal. Small column tests will be made on selected sorbents to verify the batch data and to obtain additional data for process design. Efforts will be made to obtain samples of tank supernatant from Hanford for comparison.

  18. Lactobacillus rhamnosus and its cell-free culture supernatant differentially modulate inflammatory biomarkers in Escherichia coli-challenged human dendritic cells.

    Science.gov (United States)

    Bermudez-Brito, Miriam; Muñoz-Quezada, Sergio; Gomez-Llorente, Carolina; Romero, Fernando; Gil, Angel

    2014-05-28

    The intestinal immune system maintains a delicate balance between immunogenicity against invading pathogens and tolerance to the commensal microbiota and food antigens. Different strains of probiotics possess the ability to finely regulate the activation of dendritic cells (DC), polarising the subsequent activity of T-cells. Nevertheless, information about their underlying mechanisms of action is scarce. In the present study, we investigated the immunomodulatory effects of a potentially probiotic strain, Lactobacillus rhamnosus CNCM I-4036, and its cell-free culture supernatant (CFS) on human DC challenged with Escherichia coli. The results showed that the levels of pro-inflammatory cytokines such as IL-1β, IL-6, IL-8 and IL-12p70 were higher in the cells treated with live L. rhamnosus than in the cells treated with the CFS. In the presence of E. coli, the supernatant was more effective than the probiotic bacteria in reducing the secretion of pro-inflammatory cytokines. In addition, live L. rhamnosus potently induced the production of transforming growth factor (TGF)-β1 and TGF-β2, whereas the CFS increased the secretion of TGF-β1. However, in the presence of E. coli, both treatments restored the levels of TGF-β. The probiotic strain L. rhamnosus CNCM I-4036 and its CFS were able to activate the Toll-like receptor signalling pathway, enhancing innate immunity. The two treatments induced gene transcription of TLR-9. Live L. rhamnosus activated the expression of TLR-2 and TLR-4 genes, whereas the CFS increased the expression of TLR-1 and TLR-5 genes. In response to the stimulation with probiotic/CFS and E. coli, the expression of each gene tested was notably increased, with the exception of TNF-α and NFKBIA. In conclusion, the CFS exhibited an extraordinary ability to suppress the production of pro-inflammatory cytokines by DC, and may be used as an effective and safer alternative to live bacteria.

  19. Calcium carbonate mediates higher lignin peroxidase activity in the culture supernatant of Streptomyces Viridosporus T7A

    Directory of Open Access Journals (Sweden)

    J. M. B. MACEDO

    1999-06-01

    Full Text Available Lignin peroxidase (LiP production has been extensively studied due to the potential use of this enzyme in environmental pollution control. Important aspects of the production of the enzyme by S. viridosporus T7A which have been studied include the improvement of yield and enzyme stabilization. In experiments performed in agitated flasks containing culture media composed of yeast extract as the source of nitrogen, mineral salts and different carbon sources, the use of glucose resulted in the highest values for LiP activity (350 U/L, specific LiP activity (450 U/g and productivity (7 U/L/h. As the profile obtained with glucose-containing medium suggested enzyme instability, the effect of calcium carbonate was evaluated. The addition of CaCO3 in two different concentrations, 0.5% and 5.0%, resulted in higher values of maximum LiP activity, 600 and 900 U/L, respectively. The presence of this salt also anticipated enzyme activity peaks and allowed the detection of higher enzyme activities in the extracellular medium for longer periods of time. These results indicate a positive effect of calcium carbonate on LiP production, which is extremely relevant for industrial processes.

  20. Analysis of metal Bioleaching from thermal power plant fly ash by Aspergillus niger 34770 culture supernatant and reduction of phytotoxicity during the process.

    Science.gov (United States)

    Jadhav, Umesh U; Hocheng, Hong

    2015-01-01

    Aspergillus niger culture supernatant is used for bioleaching process. Before starting bioleaching process, fly ash was washed with distilled water. This removed 100 % sodium, 47 % (±0.45) boron, 38.07 % (±0.12) calcium, 29.89 % (±0.78) magnesium, and 11.8 % (±0.05) potassium. The pH was reduced from 10.5 to 8.5 after water washing. During bioleaching process, around 100 % metal removal was achieved in 4 h for all metals except chromium 93 % (±1.18), nickel 83 % (±0.32), arsenic 78 % (±0.52), and lead 70 % (±0.20). The process parameters including temperature, shaking speed, and solid/liquid ratio were optimized for bioleaching process. Experiments were conducted to evaluate effect of fly ash on growth of mung bean (Vigna radiata). At 20 g/100 ml fly ash concentration no germination of V. radiata seeds was observed. With an increasing concentration of untreated fly ash, a gradual decrease in root/shoot length was observed. After bioleaching process 78 % (±0.19) germination of V. radiata was observed with 20 g/100 ml fly ash. This study will help to develop an efficient process to remove the toxic metals from fly ash.

  1. Inhibition of miR122a by Lactobacillus rhamnosus GG culture supernatant increases intestinal occludin expression and protects mice from alcoholic liver disease.

    Science.gov (United States)

    Zhao, Haiyang; Zhao, Cuiqing; Dong, Yuanyuan; Zhang, Min; Wang, Yuhua; Li, Fengyuan; Li, Xiaokun; McClain, Craig; Yang, Shulin; Feng, Wenke

    2015-05-05

    Alcoholic liver disease (ALD) has a high morbidity and mortality. Chronic alcohol consumption causes disruption of intestinal microflora homeostasis, intestinal tight junction barrier dysfunction, increased endotoxemia, and eventually liver steatosis/steatohepatitis. Probiotic Lactobacillus rhamnosus GG (LGG) and the bacteria-free LGG culture supernatant (LGGs) have been shown to promote intestinal epithelial integrity and protect intestinal barrier function in ALD. However, little is known about how LGGs mechanistically works to increase intestinal tight junction proteins. Here we show that chronic ethanol exposure increased intestinal miR122a expression, which decreased occludin expression leading to increased intestinal permeability. Moreover, LGGs supplementation decreased ethanol-elevated miR122a level and attenuated ethanol-induced liver injury in mice. Similar to the effect of ethanol exposure, overexpression of miR122a in Caco-2 monolayers markedly decreased occludin protein levels. In contrast, inhibition of miR122a increased occludin expression. We conclude that LGGs supplementation functions in intestinal integrity by inhibition of miR122a, leading to occludin restoration in mice exposed to chronic ethanol.

  2. The Effect of Lactobacillus crispatus and Lactobacillus rhamnosusCulture Supernatants on Expression of Autophagy Genes and HPV E6 and E7 Oncogenes in The HeLa Cell Line.

    Science.gov (United States)

    Motevaseli, Elahe; Azam, Rosa; Akrami, Seyed Mohammad; Mazlomy, Mohammadali; Saffari, Mojtaba; Modarressi, Mohammad Hossein; Daneshvar, Maryam; Ghafouri-Fard, Soudeh

    2016-01-01

    The aim of this study was to clarify the mechanism by which lactobacilli exert their cytotoxic effects on cervical cancer cells. In addition, we aimed to evalu- ate the effect of lactobacilli on the expression of human papilloma virus (HPV) onco- genes. In this experimental study, using quantitative real-time polymer- ase chain reaction (PCR), we analyzed the expression of CASP3 and three autophagy genes [ATG14, BECN1 and alpha 2 catalytic subunit of AMPK (PRKAA2)] along with HPV18 E6 and E7 genes in HeLa cells before and after treatment with Lactobacillus crispatus and Lactobacillus rhamnosus culture supernatants. The expression of CASP3 and autophagy genes in HeLa cells was de- creased after treatment with lactobacilli culture supernatants. However, this de- crease was not significant for PRKAA2 when compared with controls. In addition, expression of HPV E6 was significantly decreased after treatment with lactobacilli culture supernatants. Lactobacilli culture supernatants can decrease expression of ATG14 and BECN1 as well as the HPV E6 oncogene. It has been demonstrated that the main changes occurring during cervical carcinogenesis in cell machinery can be reversed by suppression of HPV oncogenes. Therefore, downregulation of HPV E6 by lacto- bacilli may have therapeutic potential for cervical cancer. As the role of autophagy in cancer is complicated, further work is required to clarify the link between downregula- tion of autophagy genes and antiproliferative effects exerted by lactobacilli.

  3. Detoxification of azo dyes mediated by cell-free supernatant culture with manganese-dependent peroxidase activity: effect of Mn2+ concentration and H2O2 dose.

    Science.gov (United States)

    Contreras, Elsa; Urra, Johana; Vásquez, Carlos; Palma, Carolyn

    2012-01-01

    White-rot fungi (WRF) are capable of degrading complex organic compounds such as lignin, and the enzymes that enable these processes can be used for the detoxification of recalcitrant organopollutants. The aim of this study is to evaluate a system based on the use of an in vitro ligninolytic enzyme for the detoxification of recalcitrant dye pollutants. The dyes selected for investigation were the anionic and cationic commercial azo dyes, basic blue 41 (BB41), acid black 1 (AB1), and reactive black 5 (RB5). A supernatant, cell-free culture of WRF with manganese peroxidase activity was used to investigate its degradative capacity under various conditions, and concentrations of cofactors, H(2)O(2) and Mn(2+). The assays were carried out using a 2(2) experimental designs whose variables were concentration of Mn(2+) (33 and 1,000 μM) and semicontinuous dosage of the H(2)O(2) (0.02 and 0.10 μmol) added at a frequency of 0.2 min(-1). The response variables analyzed were the efficiency and the initial rate of the decolorization process. The dye concentrations considered ranged from 10 to 200 mg L(-1). AB1 and RB5 were decolorized over the entire interval of concentrations studied; reaching efficiencies between 15 and 95%. Decolorization of up to 100 mg L(-1), BB41 had less than 30% efficiency. The decay of the concentration of AB1 was interpreted by two-stage kinetics model, with the exception of the condition of 33 μM Mn(2+)-0.02 μmol of H(2)O(2) in which only one stage was observed. For all assays performed with 33 μM Mn(2+), the initial rate of the decolorization process was found to be dependent on the dosage of H(2)O(2). The results of this study can be applied to the development bioreactors for the degradation of recalcitrant pollutants from the textile industry and may be used as a model for expanding the use of extracellular enzyme supernatants in bioremediation. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

  4. Evaluation of cellobiose dehydrogenase and laccase containing culture fluids of Termitomyces sp. OE147 for degradation of Reactive blue 21

    Directory of Open Access Journals (Sweden)

    Rishabh Gangwar

    2016-12-01

    Full Text Available This study evaluates culture filtrate, rich in cellobiose dehydrogenase and laccases, of Termitomyces sp. OE 147, in decolouration and degradation of Reactive blue (RB 21. About 35% decolouration was achieved at low volumes of the culture supernatant without addition of external redox mediators. An optimized dye to culture fluid ratio (75 ppm: 0.1 ml at a pH of 4–5 resulted in removal of colour by 60%. The degradation products of RB21 were analysed by Electron Spray Ionization-Mass Spectrometry and several small molecules (of m/z 106–199 were detected. These were concluded to be o-Xylene, 2,3-Dihydro-1H-isoindole, Isoindole-1,3-dione, 2,Benzenesulfonyl-ethanol, (4-Hydroxy-phenyl-sulfamic acid, 2,3-Dihydro-1H-isoindole-5-sulfonic acid and proposed to result from joint action of cellobiose dehydrogenase, laccase, peroxidases and unidentified oxidoreductases present in the culture fluids. Based on the products formed and the known reactions of these enzymes, a degradation pathway was proposed for RB21. The culture fluid was also effective in decolouration (by about 50% and detoxification (by ∼25% of the combined effluent collected from a local mill indicating a treatment process that bypasses use of H2O2 and toxic mediators.

  5. Tetracycline and Glutathione Inhibit Matrix Metalloproteinase Activity: An In Vitro Study Using Culture Supernatants of L929 and Dalton Lymphoma Cell Lines

    Directory of Open Access Journals (Sweden)

    Gajanan Kendre

    2013-01-01

    Full Text Available Tetracycline and glutathione inhibited the protease activities of matrix metalloproteinase-2 and matrix metalloproteinase-9 expressed by mouse fibrosarcoma cells (L929 and Dalton lymphoma cells, respectively. The inhibitory activity of the tetracycline may be due to its ability to chelate metal ions such as calcium and zinc. Gelatin-zymography technique was used to demonstrate the inhibitory activity of both tetracycline and glutathione. The intensity of the bands corresponding to metalloproteinase activity in zymography gel was reduced in the presence of 50–100 μg/mL of tetracycline. The presence of 10–100 μg/mL of tetracycline in the medium increased the adherence of L929 cancer cells. These results clearly indicate the antimetastatic property of tetracycline. Reduced glutathione, a compound which is produced endogenously by the cells to maintain the redox status, was shown to inhibit the matrix metalloproteinase activity (in vitro. Therefore, it is assumed that decreased glutathione levels in synovial fluids or plasma might increase the activity of MMP. Reduced glutathione at 100 μg/mL inhibited the metalloproteinase activity in gelatin-zymographic gel. As both tetracycline and glutathione exhibited an inhibitory effect on matrix metalloproteinase activity, it was of great interest to check their clinical effects on various MMP associated pathological conditions such as cancer metastasis and arthritis. Here we report that tetracycline and reduced glutathione inhibited the activity of MMP2 completely and activity of MMP9 partly.

  6. Development of an indirect ELISA to detect Corynebacterium pseudotuberculosis specific antibodies in sheep employing T1 strain culture supernatant as antigen

    Directory of Open Access Journals (Sweden)

    Miriam F. Rebouças

    2013-11-01

    Full Text Available Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis (CLA, a chronic disease that affects goats and sheep, characterized by granuloma formation in subcutaneous and internal lymph nodes. CLA causes significant economic losses to commercial goat herds. In this study, we aimed to test secreted antigens secreted from T1 strain bacteria grown in brain heart infusion (BHI broth in an indirect ELISA system to determine the presence of specific immunoglobulins against C. pseudotuberculosis. We analyzed the BHI antigen electrophoretic profile and the recognition pattern by infected sheep sera samples. The ELISA results were compared with multiplex PCR assay and IFN-gamma production. The ELISA was able to discriminate between negative and positive animals, with a sensitivity of 89% and a specificity of 99%, using microbiological isolation as gold standard. When this assay was compared with multiplex PCR and specific IFN-gamma quantification, six discrepant results were found among thirty-two samples. We concluded that the ELISA using antigens secreted from C. pseudotuberculosis T1 strain growth in BHI broth culture can be used for the serodiagnosis of CLA in sheep.

  7. Cell culture supernatants for detection perforin ELISA

    African Journals Online (AJOL)

    Najwa

    2014-02-19

    Feb 19, 2014 ... suggested by Whitaker (1972), the resin was packaged gently in glass column, the .... crystals and incubated in shaker at room temperature for 30 min. ... a well used test for the medical statistics; P value <0.01 is considered.

  8. Effects of the supernatant from ankylosing spondylitis synovial cells culture on the osteogenic differentiation of ligament fibroblasts%强直性脊柱炎滑膜细胞培养上清液对韧带成纤维细胞成骨分化的影响

    Institute of Scientific and Technical Information of China (English)

    徐卫东; 梁志民; 袁恒锋

    2012-01-01

    Objective To investigate the effects of synovial cells culture fluid on the differentiation of ligament fibroblasts into osteoblasts. Methods Specimens of ligament and synovium were obtained from the hip joints with ankylosing spondylitis respectively. The primary cell culture of ligament fibroblasts and synovial cells were processed and then the supernatant of synovial cells in the first passage was collected and used to induce ligament cells. Osteogenic phenotype and alkaline phosphatase (ALP) activity were determined at the days of 0, 5,10,15 and 20 respectively. Calcium deposit was detected by Von Kossa staining at the 25th day. Results With time extending, the expression of ALP and osteocalcin increased significantly at day 10 and reached a peak at day 15. Calcium deposit could be detected at day 25. Conclusions The supernatant of synovial cells cuture plays a certain role on the differentiation of ligament fibroblasts into osteoblasts.%目的 探讨强直性脊柱炎(AS)滑膜细胞培养上清液对韧带成纤维细胞成骨分化的影响.方法 分别进行AS患者滑膜和韧带成纤维细胞原代培养,收集第1代的滑膜细胞培养液上清液,应用此上清液对韧带成纤维细胞进行诱导,分别于0、5、10、15、20 d检测碱性磷酸酶和骨钙素表达水平,并于25 d时进行钙结节染色.结果 在应用AS滑膜细胞培养上清液对韧带成纤维细胞进行诱导的第10天,碱性磷酸酶(ALP)和骨钙素(OC)表达水平明显升高,并在第15天达到高峰.第25天时进行钙结节Von Kossa染色,发现有钙结节形成.结论 AS滑膜细胞培养上清液对韧带成纤维细胞具有成骨诱导分化的能力.

  9. In vitro differentiation of MSCs into retina-like cells by the supernatant fluid of light-injured neurosensory retina%光损伤鼠视网膜片培养上清液诱导 MSCs 分化为视网膜样细胞的研究

    Institute of Scientific and Technical Information of China (English)

    白月; 徐国兴

    2014-01-01

    AIM: To explore the possibility of inducing rat mesenchymal stem cells ( MSCs) into retina-like cells by the supernatant fluid of light-injured neurosensory retina in vitro. METHODS: MSCs were isolated and attached to the wall of culture dishes by their specific adherent ability. Then the cells were characterized by flow cytometry.The neurosensory retina was isolated from retina of SD rat and it was tested by hematoxylin-eosin ( HE ) staining.The pathological changes of light-injured neurosensory retina was observed under transmission electron microscope. Three kinds of supernatant fluid of light -injured neurosensory retina of SD rats were prepared.The third passage of MSCs were cultured with these mixed medium for 7-8d, we used RT-PCR to see whether they could express rhodopsin, neuron-specific enolase (NSE), and glial fibrillary acidic protein ( GFAP ) , and positive cells were counted and analyzed. RESULTS: HE staining showed the retinal sheets included full-thickness neural retina.Neurosensory retina developed ultrastructural destructions by light injury.RT-PCR showed that the medium of mixed I expressed higher positive rate of rhodopsin (0.3915±0.00644), NSE (0.2019± 0.00682), GFAP (0.1972 ±0.00211) than the medium of mixed Ⅱ rhodopsin (0.0983 ±0.00319), NSE (0.1048 ± 0.00323), GFAP (0.1040±0.00254) and medium of mixedⅢrhodopsin(0.0044±0.00126), NSE (0.0498±0.00149), GFAP (0.0467±0.00333).The difference of intergroup has statistical significance. CONCLUTION:The supernatant fluid of light-injured neurosensory retina of SD rats can induce MSCs to differentiate into retina-like cells and provide new insights of stem cell therapy for retinopathy.%目的:应用大鼠视网膜片光损伤后的培养上清液,在体外诱导大鼠骨髓间充质干细胞( mesenchymal stem cells , MSCs)成为视网膜样细胞的可能性。  方法:贴壁筛选法分离、培养大鼠MSCs ,流式细胞仪对其细胞纯度鉴定。取材大鼠视网

  10. 恩度对肿瘤上清液诱导的人脐静脉内皮细胞增殖、凋亡的影响及其机制的初步研究%Effects and preliminary study on the mechanism of endostar on proliferation and apoptosis of the HUVECs induced by tumor supernatant fluid

    Institute of Scientific and Technical Information of China (English)

    高爽; 杨向红; 范一博; 曲秀娟; 刘云鹏

    2011-01-01

    目的:探讨恩度对肾癌ACHN细胞系上清液诱导的人脐静脉内皮细胞(HUVECs)增殖和凋亡的影响及其机制.方法:应用MTT法检测恩度对HUVECs增殖的抑制作用;应用Annexin-V/PI双染法流式细胞仪检测恩度对HUVECs凋亡的影响;Western blot检测恩度对Cbl-b,c-Cbl,磷酸化Akt(p-Akt)和磷酸化ERK(p-ERK)蛋白表达的影响.结果:恩度抑制肿瘤上清液诱导的HUVEC细胞增殖;促进HUVECs凋亡;恩度下调HUVEC中Cbl-b、c-Cbl、p-Akt及p-ERK的表达,且上述效应均呈浓度依赖性.结论:恩度对肿瘤上清液诱导的HUVECs的增殖具有抑制作用,对其凋亡具有促进作用.泛素连接酶、p-Akt及p-ERK通路可能是恩度作用的分子机制之一.%Objective: To explore the effects and the mechanism of endostar on proliferation and apoptosis of the human umbilical vein endothelial cells ( HUVECs ) induced by renal cell carcinoma ( ACHN cell lines ) supernatant fluid. Methods: To detect the effects of endostar on the proliferation of the HUVECs by MTT; detect the effects of endostar on the apoptosis of the HUVECs by Annexin V - FITC binding assay;detect the effects of endostar on the expression of Cbl - b, c - Cbl, p - Akt and p - ERK by western blot. Results: Endostar inhibited the proliferation of HUVECs induced by renal cell carcinoma supernatant fluid, promoteed the apoptosis of HUVECs reduced the expression of Cbl - b , c - Cbl, p - Akt and p - ERK in HUVECs, and the effects were depended on the concentration of endostar. Conclusion: Endostar inhibits the proliferation, promotes the apoptosis of HUVECs induced by renal cell carcinoma supernatant fluid, and ubiquitin ligases pathway, p - Akt pathway or p - ERK pathway may be the molecular mechanism of endostar.

  11. Detection of Neisseria meningitidis from Negative Blood Cultures and Cerebrospinal Fluid with the FilmArray Blood Culture Identification Panel

    OpenAIRE

    Pardo, Joe; Klinker, Kenneth P.; Borgert, Samuel J.; Butler, Brittany M.; Rand, Kenneth H.; Iovine, Nicole M.

    2014-01-01

    The FilmArray blood culture identification (BCID) panel is a rapid molecular diagnostic test approved for use with positive blood culture material. We describe a fatal case of meningococcemia with central nervous system (CNS) involvement detected using the BCID test with culture-negative blood and cerebrospinal fluid.

  12. Detection of Neisseria meningitidis from negative blood cultures and cerebrospinal fluid with the FilmArray blood culture identification panel.

    Science.gov (United States)

    Pardo, Joe; Klinker, Kenneth P; Borgert, Samuel J; Butler, Brittany M; Rand, Kenneth H; Iovine, Nicole M

    2014-06-01

    The FilmArray blood culture identification (BCID) panel is a rapid molecular diagnostic test approved for use with positive blood culture material. We describe a fatal case of meningococcemia with central nervous system (CNS) involvement detected using the BCID test with culture-negative blood and cerebrospinal fluid.

  13. Growth Inhibition of Cronobacter sakazakii in Experimentally Contaminated Powdered Infant Formula by Kefir Supernatant.

    Science.gov (United States)

    Kim, Dong-Hyeon; Chon, Jung-Whan; Kang, Il-Byeong; Kim, Hyunsook; Kim, Hong-Seok; Song, Kwang-Young; Seo, Kun-Ho

    2015-09-01

    Kefir is a type of fermented milk containing lactic and acetic acid bacteria and yeast. In this study, we evaluated the antimicrobial activity of kefir supernatant against Cronobacter sakazakii in powdered infant formula (PIF). In a spot-on-lawn test, the growth of 20 C. sakazakii strains, including 10 clinical and 10 food isolates, was completely inhibited in the presence of kefir supernatant. Significant differences in the diameters of inhibition zones were observed upon treatment with kefir compared with the results for Lactobacillus kefiri and Candida kefyr culture supernatants or solutions of lactic and acetic acid and ethyl alcohol in the agar well diffusion test (P < 0.05). The addition of 100 μl of kefir supernatant to 1 ml of nutrient broth completely inhibited the growth of C. sakazakii, as evaluated by spectrophotometry. The antimicrobial activity of kefir supernatant in experimentally contaminated PIF was also tested; we found no viable C. sakazakii cells remaining in PIF rehydrated with 30% kefir supernatant solution for 1 h, demonstrating that the antimicrobial activity of kefir supernatant against C. sakazakii could be applied in real food samples.

  14. Effects of LPS-stimulated monocyte culture supernatant on the osteogenic function of osteoblastic cells in vitro%脂多糖刺激单核巨噬细胞培养上清液对成骨细胞成骨特性的影响

    Institute of Scientific and Technical Information of China (English)

    王海燕; 邓辉; 徐春燕; 叶洁; 胡荣党

    2011-01-01

    目的 探讨建立牙周炎症的体外实验模型的可行性,为后续进行牙周炎症状态下应力对牙槽骨改建的研究奠定基础.方法 以牙龈卟啉单胞菌(Porphyromonas gingivalis,P.g)脂多糖(lipopolysaccharide,LPS)Pg-LPS刺激小鼠单核巨噬细胞(RAW264.7)的培养上清液作用于体外培养的小鼠成骨细胞(MC3T3-E1)24 h和48 h后,分别通过噻唑蓝(MTT)法和碱性磷酸酶(ALP)试剂盒检测MC3T3-E1的增殖活性和碱性磷酸酶活性,观察脂多糖刺激的小鼠单核巨噬细胞培养上清液对小鼠成骨细胞成骨功能的影响.结果 Pg-LPS刺激RAW264.7后其上清液含L-1β、IL-6 和TNF-α等炎症因子,且与LPS的浓度和刺激时间呈依赖性.此上清液对MC3T3-E1的增殖和ALP活性均有抑制作用,亦与上清液中以上炎症因子的浓度呈依赖性,与体内牙周炎症状态相似.结论 Pg-LPS体外刺激RAW264.7的培养上清液可以应用于建立体外牙周炎症模型,这将为后续研究牙周炎症状态下应力对牙槽骨改建的影响奠定初步的基础.%Objective To investigate the influence of Pg-LPS stimulated monocyte (RAW264.7) culture supernatant on the proliferation and alkaline phosphatase (ALP) activity of osteoblastic cells (MC3T3-E1) in vitro. Methods The culture supernatant of monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations(10%、20%、30%、40% and 50%) simultaneously for 24 h and 48 h in vitro.The cellular proliferation was assessed by MTT assay, Cellular ALP activity was examined using ALP measurement kit, and the results were statistically analyzed using SPSS 12.0 software package. Results LPS stimulated RAW264.7 to release inflammatory cytokines including IL-1β, IL-6 and TNF-α, whose secretion was in a time and dose dependent manner with the concentration of LPS. After the stimulation of different concentrations of inflammatory supernatant, the proliferation and ALP activity of MC3T3-El

  15. Content research on antibody titer and plasma protein of plasma and red blood cell suspension supernatant fluid%血浆与红细胞悬液上清液中抗体效价及血浆蛋白的含量研究

    Institute of Scientific and Technical Information of China (English)

    赵敏

    2012-01-01

      目的分析红细胞悬浮液与血浆对应的抗体效价及其蛋白含量.方法随机收集无偿献血者261名.针对各全血制备对应的血浆和红细胞悬浮液,检测各血上清的抗A、抗B效价及其对应的血浆蛋白含量并比较分析.结果 A型血浆抗B效价的效价为31.05±20.29,A型红细胞悬液抗B的抗体效价为5.86±3.60;B型血浆抗A的效价为21.69±18.34,B型红细胞悬液抗A的抗体效价为3.08±1.99;O型血浆抗A的效价为34.02±20.76,血浆抗B的抗体效价为31.98±19.32;O型红细胞悬液的抗体抗A效价为5.96±2.86,抗B抗体效价为5.32±2.45.A型血的血浆蛋白与红细胞悬液上清液存在显著差异,且血浆抗体的效价是红细胞上清效价均数的7倍;血浆蛋白则为红细胞悬液的5-8倍.结论与血浆比较,各血型红细胞悬浮液的抗体效价明显低,含有较少蛋白,可只用主侧配血.%  Objective To analyse the antibody titer and its protein content of red blood cell suspension and plasma corresponding. Methods Random collect voluntary blood donors 261. On the whole blood preparation corresponding plasma and erythrocytes suspension, detecting the blood supernatant of A resistance, resistance to B valence and the corresponding plasma protein content and comparative analysis. Results The results showed that the type A plasma resistance to B titer of valence was 31.05±20.29, type A red blood cell suspension resistance to B antibody titer was5.86±3.60; Type B plasma resistance to A titer was 21.69±18.34, type B cell suspension resistance to A antibody titer was 3.08±1.99; Type O blood plasma resistance to A titer was 34.02±20.76, plasma resistance to B antibody titer was 31.98±19.32; Type O red blood cell suspension antibody resistance to A titer was 5.96±2.86, resistance to B antibody titer was 5.32±2.45. Type A blood plasma protein and red blood cell suspension supernatant fluid exists significant difference, and plasma antibody

  16. Quantitative analysis of the toxicity of human amniotic fluid to cultured rat spinal cord.

    Science.gov (United States)

    Drewek, M J; Bruner, J P; Whetsell, W O; Tulipan, N

    1997-10-01

    It has been proposed that the myelodysplastic components of a myelomeningocele are secondarily damaged as the result of exposure to amniotic fluid, the so-called 'two-hit' hypothesis. The critical time at which this secondary insult might occur has not been clearly defined. The present study addresses this issue by quantitatively assessing the toxic effects of human amniotic fluid of various gestational ages upon organotypic cultures of rat spinal cord. Using an assay for lactate dehydrogenase efflux to evaluate toxicity in such spinal cord cultures, we found that the amniotic fluid became toxic at approximately 34 weeks' gestation. This toxic effect of amniotic fluid appears to emerge rather suddenly. Accordingly, it seems reasonable to suggest that prevention of exposure of vulnerable spinal cord tissue to this toxicity by surgical closure of a myelomeningocele defect prior to the emergence of toxicity in amniotic fluid may prevent injury to vulnerable myelodysplastic spinal cord tissue.

  17. Enrichment methodology to increase the positivity of cultures from body fluids

    Directory of Open Access Journals (Sweden)

    Alessandra Valle Daur

    Full Text Available Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially-severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1 the conventional culture method (agar plating and (2 the enrichment culture technique, using the Bact/Alert® blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents.

  18. Enrichment methodology to increase the positivity of cultures from body fluids

    Directory of Open Access Journals (Sweden)

    Alessandra Valle Daur

    2006-12-01

    Full Text Available Isolation and identification of etiological agents found in body fluids can be of critical importance for the recovery of patients suffering from potentially-severe infections, which are often followed by serious sequels. Eighty-two samples of different body fluids were analyzed using two different methods: (1 the conventional culture method (agar plating and (2 the enrichment culture technique, using the Bact/Alert® blood culture bottle. The number of positive cultures increased on average from 9.7% to 23.1% with the enrichment culture technique. Pseudomonas aeruginosa, Escherichia coli and Staphylococcus aureus were the most frequently isolated bacteria. The enrichment method could provide a more accurate means the identifying etiological agents.

  19. The Immunostimulation of F4ac ETEC-Culture Supernatant to Porcine Small Intestinal Epithelial Cells%F4ac型产肠毒素大肠杆菌菌液上清对仔猪小肠上皮细胞的免疫刺激作用

    Institute of Scientific and Technical Information of China (English)

    周传丽; 刘铮铸; 俞英; 张勤

    2014-01-01

    [Objective]Porcine enterotoxigenic Escherichia coli (ETEC) is a worldwide cause of bacteria induced diarrhoea in piglets. In the veterinary practices of pig production, serological identification of ETEC shows that F4ac is the most common serological type expressed in ETEC strains isolated from diarrheic piglets. So far, the mechanism by which ETEC produces diarrhoea in piglets has been clearly elucidated. However, the immunostimulation of ETEC-culture to host target cells has not been studied or described. In the present study, the immunostimulation of the supernatant of F4ac ETEC-culture to IPEC-J2 cells was studied.[Method]The culture of F4ac ETEC strain 200 was collected and centrifuged (4℃, 4 000 r/min for 15 min) after 12 hours in culture, and the supernatant was sterilized by passing it through a 0.22μm filter. The solution combined the sterilized supernatant with equivalent DMEM/F12 medium was used to challenge IPEC-J2 cells for 3 hours. The IPEC-J2 cells co-cultured with fresh LB medium and equivalent DMEM/F12 medium were as the control group. For both treatments, each experiment was repeated three times. Total RNAs of the stimulated and control IPEC-J2 cells were extracted using TRIZOL Reagent following the manufacturer’s instructions. According to the manufacturer’s instructions, complementary DNA (cDNA) was synthesized from RNA using the Prime Script® RT reagent Kit with gDNA Eraser (Perfect Real Time). The cDNA samples were then analyzed with real time RT-PCR using a LightCycler® 480 Real-Time PCR System. The real time RT-PCR reactions were performed in a final volume of 20μL with the Roche SYBR Green PCR Kit according to the manufacturer’s instructions. The pig house-keeping gene β-actin was used as the internal standards to correct the input of cDNA. Triplicate qRT-PCRs were performed on each cDNA and the average Ct was used for further analysis. The relative quantification values were calculated using the 2-ΔΔCt. Differential m

  20. Secretion of nerve growth factor, brain-derived neurotrophic factor, and glial cell-line derived neurotrophic factor in co-culture of four cell types in cerebrospinal fluid-containing medium

    Institute of Scientific and Technical Information of China (English)

    Sanjiang Feng; Minghua Zhuang; Rui Wu

    2012-01-01

    The present study co-cultured human embryonic olfactory ensheathing cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells in complete culture medium- containing cerebrospinal fluid. Enzyme linked immunosorbent assay was used to detect nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor secretion in the supernatant of co-cultured cells. Results showed that the number of all cell types reached a peak at 7–10 days, and the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor peaked at 9 days. Levels of secreted nerve growth factor were four-fold higher than brain-derived neurotrophic factor, which was three-fold higher than glial cell line-derived neurotrophic factor. Increasing concentrations of cerebrospinal fluid (10%, 20% and 30%) in the growth medium caused a decrease of neurotrophic factor secretion. Results indicated co-culture of human embryonic olfactory ensheathing cells, human Schwann cells, human amniotic epithelial cells and human vascular endothelial cells improved the expression of nerve growth factor, brain-derived neurotrophic factor, and glial cell line-derived neurotrophic factor. The reduction of cerebrospinal fluid extravasation at the transplant site after spinal cord injury is beneficial for the survival and secretion of neurotrophic factors from transplanted cells.

  1. Automated Static Culture System Cell Module Mixing Protocol and Computational Fluid Dynamics Analysis

    Science.gov (United States)

    Kleis, Stanley J.; Truong, Tuan; Goodwin, Thomas J,

    2004-01-01

    This report is a documentation of a fluid dynamic analysis of the proposed Automated Static Culture System (ASCS) cell module mixing protocol. The report consists of a review of some basic fluid dynamics principles appropriate for the mixing of a patch of high oxygen content media into the surrounding media which is initially depleted of oxygen, followed by a computational fluid dynamics (CFD) study of this process for the proposed protocol over a range of the governing parameters. The time histories of oxygen concentration distributions and mechanical shear levels generated are used to characterize the mixing process for different parameter values.

  2. Cultural factors influencing dietary and fluid restriction behaviour: perceptions of older Chinese patients with heart failure.

    Science.gov (United States)

    Rong, Xiaoshan; Peng, Youqing; Yu, Hai-Ping; Li, Dan

    2017-03-01

    To explore the cultural factors related to dietary and fluid restriction behaviours among older Chinese patients. Excess dietary sodium and fluid intake are risk factors contributing to the worsening and rehospitalisation for heart failure in older patients. Managing the complex fluid and diet requirements of heart failure patients is challenging and is made more complicated by cultural variations in self-management behaviours in response to a health threat. Qualitative study using semi-structured in interviews and framework analysis. The design of this study is qualitative descriptive. Semi-structured in-depth interviews were conducted with 15 heart failure patients. Data were analysed through content analysis. Seven cultural themes emerged from the qualitative data: the values placed on health and illness, customary way of life, preference for folk care and the Chinese healthcare system, and factors related to kinship and social ties, religion, economics and education. Dietary change and management in response to illness, including heart failure, is closely related to individuals' cultural background. Healthcare providers should have a good understanding of cultural aspects that can influence patients' conformity to medical recommendations. Heart failure patients need support that considers their cultural needs. Healthcare providers must have a good understanding of the experiences of people from diverse cultural backgrounds. © 2016 John Wiley & Sons Ltd.

  3. Larvicidal and Growth-Inhibitory Activity of Entomopathogenic Bacteria Culture Fluids Against Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    da Silva, João Luiz Rosa; Undurraga Schwalm, Fernanda; Eugênio Silva, Carlos; da Costa, Marisa; Heermann, Ralf; Santos da Silva, Onilda

    2017-01-06

    Dengue, Chikungunya, and Zika are important vector-borne diseases, and Aedes aegypti L. is their main transmitter. As the disease management is mainly based on mosquito control strategies, the search for alternative and cost-effective approaches is ongoing. The Gram-negative bacteria Xenorhabdus nematophila and Photorhabdus luminescens are symbiotically associated with entomopathogenic nematodes and are highly pathogenic for insect larvae. After we have recently confirmed the toxicity of these bacteria in Ae. aegypti larvae, we here evaluated the toxic activity of culture fluids on the development of this mosquito species. Larval susceptibility was assessed by exposing larvae to different concentrations of P. luminescens or X. nematophila culture fluids to confirm whether secondary metabolites might cause the mosquitos' death. Xenorhabdus nematophila culture fluid was more effective and stable during the mosquito pathogenicity bioassays compared to that of P. luminescens Larval mortality started a few hours after exposure of the insects to the fluids. Furthermore, the residual effect of larvicidal activity of X. nematophila fluid persisted at full efficiency for 4 d. Particularly, larval mortality was still higher than 50% for up to 8 d. Exposure of larvae to a sublethal dose of X. nematophila fluid delayed pupation as well as emergence of adult mosquitoes and caused cumulative larval mortality higher than 90% by day 14. Here, we describe for the first time the use of stable culture fluids and therefore secondary metabolites of P. luminescens and X. nematophila as a promising basis for the use as biopesticide for control of Ae. aegypti in the future.

  4. Trichoderma L-Lysine-α-Oxidase Producer Strain Culture Fluid Inhibits Impatiens Necrotic Spot Virus.

    Science.gov (United States)

    Smirnova, I P; Shneider, Yu A; Karimova, E V

    2016-01-01

    A method for PCR diagnosis of impatiens necrotic spot virus is developed. Concentrated culture fluid with active L-lysine-α-oxidase (0.54 U/ml) from Trichoderma harzianum Rifai fungus producer strain F-180 inhibits vitally hazardous impatiens necrotic spot phytovirus.

  5. Pregnancy-specific beta-1-glycoprotein (SP1) in cultured amniotic fluid cells.

    Science.gov (United States)

    Heikinheimo, M; Wahlström, T; Aula, P; Virtanen, I; Seppälä, M

    1980-12-01

    The synthesis of pregnancy-specific beta-1-glycoprotein (SP1) was studied in amniotic fluid cell cultures using RIA, immunoperoxidase, and immunofluorescence techniques. SP1 was found by RIA in all 11 sonicates and in 21 of 26 culture media. The SP1-immunoreactive material was immunologically similar to maternal serum SP1. Immunoperoxidase and indirect immunofluorescence staining were positive in large cells identified as epithelial amniotic cells by labeling with antikeratin antibodies. Fibroblast-like cells were occasionally found in cultures, but they did not contain demonstrable amounts of SP1. The physiological significance of the findings presented remains unclear.

  6. [Effect of burn blister fluid on human mesenchymal stem cell (MSC) in vitro and its phenotypic modulation in culture].

    Science.gov (United States)

    Liu, Li-Zhu; Rong, Xin-Zhou; Zhang, Tao; Yang, Rong-Hua; Wang, Zhen

    2009-06-01

    To study the effect of blister fluid obtained from burn patient on human MSCs in vitro and its phenotypic modulation in culture. Blister fluid from burn patients was collected at 12, 24, 48 post burn hour (PBH). The human MSCs were isolated, cultured, amplified and identified in vitro, then were divided into A (culture with 20% blister fluid collected at 12 PBH) , B (culture with 20% blister fluid collected at 24 PBH), C (culture with 20% blister fluid collected at 48 PBH), N (with ordinary culture medium) groups. The growth of MSCs and micro-organisms in blister fluid were observed. Positive expression rates of CD44 and CK7 were detected by flow cytometry after culture for 8 days. Bacterial and fungal growths were absent in 15 blister fluid samples. There was no obvious change in MSC morphology in each group. Compared with that of N group, the number of MSCs in A, B, C groups was decreased, especially in C group. CD44 positive expression rate in A, B, C groups was (83.0 +/- 3.1)%, (77.2 +/- 2.9)% and (65.1 +/- 2.3)%, respectively,which was obviously lower than that in N group [(89.5 +/- 3.2)%, P blister fluid can obviously inhibit the growth of human MSC cultured in vitro, and may promote modulation of its phenotype to certain extent.

  7. Antimicrobial-Loaded Bone Cement Does Not Negatively Influence Sonicate Fluid Culture Positivity for Diagnosis of Prosthetic Joint Infection.

    Science.gov (United States)

    Park, Kyung-Hwa; Greenwood-Quaintance, Kerryl E; Hanssen, Arlen D; Abdel, Matthew P; Patel, Robin

    2016-06-01

    We compared culture results to investigate the influence of antimicrobial-loaded cement on sonicate fluid culture positivity for the diagnosis of prosthetic joint infection. Fifty-four subjects were assessed. The sensitivities of sonicate fluid culture were 77.8% (14 of 18) in subjects with an antimicrobial-loaded cemented prosthesis and 58.3% (21 of 36) in subjects with an antimicrobial-free prosthesis.

  8. Diagnosis of spontaneous bacterial peritonitis: Role of tween 80 and triton X in ascitic fluid cultures

    Directory of Open Access Journals (Sweden)

    Iyer R

    2009-01-01

    Full Text Available A patient with alcoholic cirrhosis of the liver, portal hypertension with hepatic encephalopathy and spontaneous bacterial peritonitis (SBP was admitted in an obtunded condition. Attempts at delineating the aetiology of the SBP using conventional cultures as well as automated systems were not successful. The use of non-anionic surfactant agents such as Tween 80-incorporated blood agar and Triton X treatment of the specimens facilitated the growth of Klebsiella pneumoniae from the ascitic fluid, which otherwise would have been concluded to represent culture-negative neutrocytic ascites. Thus, the use of the aforementioned agents could be explored in elucidating the aetiology of body cavity infections when conventional methods fail.

  9. Coccidioidomycosis: adenosine deaminase levels, serologic parameters, culture results, and polymerase chain reaction testing in pleural fluid.

    Science.gov (United States)

    Thompson, George R; Sharma, Shobha; Bays, Derek J; Pruitt, Rachel; Engelthaler, David M; Bowers, Jolene; Driebe, Elizabeth M; Davis, Michael; Libke, Robert; Cohen, Stuart H; Pappagianis, Demosthenes

    2013-03-01

    In a patient with positive serum serology for coccidioidomycosis, the differential diagnosis of concurrent pleural effusions can be challenging. We, therefore, sought to clarify the performance characteristics of biochemical, serologic, and nucleic-acid-based testing in an attempt to avoid invasive procedures. The utility of adenosine deaminase (ADA), coccidioidal serology, and polymerase chain reaction (PCR) in the evaluation of pleuropulmonary coccidioidomycosis has not been previously reported. Forty consecutive patients evaluated for pleuropulmonary coccidioidomycosis were included. Demographic data, pleural fluid values, culture results, and clinical diagnoses were obtained from patient chart review. ADA testing was performed by ARUP Laboratories, coccidioidal serologic testing was performed by the University of California-Davis coccidioidomycosis serology laboratory, and PCR testing was performed by the Translational Genomics Research Institute using a previously published methodology. Fifteen patients were diagnosed with pleuropulmonary coccidioidomycosis by European Organization for the Research and Treatment of Cancer/Mycoses Study Group criteria. Pleural fluid ADA concentrations were coccidioidomycosis were not elevated in this study. The sensitivity and specificity of coccidioidal serologic testing in nonserum samples remained high, but the clinical usefulness of PCR testing in pleural fluid was disappointing and was comparable to pleural fluid culture.

  10. Detection of interleukin-10 and transforming growth factor-β1 in the culture supernatant of CD4+CD25+ T cells from patients with alopecia areata%斑秃患者外周血CD4+CD25+T细胞培养上清液白介素10和转化生长因子β1检测

    Institute of Scientific and Technical Information of China (English)

    马新华; 邵文俊; 金宛宛; 高宇

    2014-01-01

    Objective To evaluate the potential association of CD4+CD25+ T cells with alopecia areata.Methods Totally,this study enrolled 23 patients with progressive alopecia areata,25 patients with stable alopecia areata,and 25 healthy controls.Peripheral blood was isolated from these subjects followed by isolation of CD4+ CD25+ regulatory T cells,which were then cuhured with the presence of anti-CD3 and-CD28 monoclonal antibodies for four days.Subsequently,enzyme-linked immunosorbent assay was performed to measure the levels of interleukin (IL)-10 and transforming growth factor (TGF)-β1 in the culture supematant of these T cells.Results The levels of IL-10 and TGF-β1 were (31.68 ± 6.78) pg/ml and (32.29 ± 6.8) pg/ml respectively in the culture supernatant of CD4+CD25+ regulatory T cells from patients with progressive alopecia areata,significantly lower than those from the healthy controls ((57.34 ± 14.15) pg/ml and (57.43 ± 15.16) pg/ml,both P < 0.05) and patients with stable alopecia areata ((52.56 ± 13.02) pg/ml and (61.75 ± 14.10) pg/ml,both P < 0.05).However,no significant difference was observed in the supernatant levels of IL-10 or TGF-β1 between the healthy controls and patients with stable alopecia areata.Conclusions The secretion of IL-10 and TGF-β1 by CD4+CD25+ T cells is decreased in patients with progressive alopecia areata,which may contribute to the pathogenesis of alopecia areata.%目的 探讨CD4+CD25+T细胞与斑秃发病之间的关系.方法 收集了3组研究对象,其中健康对照组25例、稳定期斑秃患者25例、进展期斑秃患者23例.抽取所有对象外周血,提取CD4+CD25+T细胞,培养4d,收集培养上清液,ELISA法检测上清液IL-10和TGF-β1水平.结果 进展期斑秃患者外周血CD4+CD25+T细胞培养的IL-10和TGF-β1分别为(31.68±6.78) pg/ml和(32.29±6.80) pg/ml,明显低于健康对照组(57.34±14.15) pg/ml、(57.43±15.16) pg/ml和稳定期斑秃患者(52.56±13.02) pg/ml和(61.75±14.10) pg

  11. Modelling antagonic effect of lactic acid eacteria supernatants on some pathogenic bacteria

    OpenAIRE

    2009-01-01

    This work presents a statistical model of survival analysis for three pathogenic bacterial strains (Escherichia coli, Listeria monocytogenes and Staphylococcus aureus), when treated with neutralized and non-neutralized filtered supernatants broth from cultures of Lactobacillus acidhophilus, Lactobacillus rhamnosus and Lactobacillus sake. Survival analysis is a method employed to determine the period of time from an initial stage up to the occurrence of a particular event of interest, as death...

  12. Nuclear and mitochondrial DNA in blastocoele fluid and embryo culture medium: evidence and potential clinical use.

    Science.gov (United States)

    Hammond, Elizabeth R; Shelling, Andrew N; Cree, Lynsey M

    2016-08-01

    The ability to screen embryos for aneuploidy or inherited disorders in a minimally invasive manner may represent a major advancement for the future of embryo viability assessment. Recent studies have demonstrated that both blastocoele fluid and embryo culture medium contain genetic material, which can be isolated and subjected to downstream genetic analysis. The blastocoele fluid may represent an alternative source of nuclear DNA for aneuploidy testing, although the degree to which the isolated genetic material is solely representative of the developing embryo is currently unclear. In addition to nuclear DNA, mitochondrial DNA (mtDNA) can be detected in the embryo culture medium. Currently, the origin of this nuclear and mtDNA has not been fully evaluated and there are several potential sources of contamination that may contribute to the genetic material detected in the culture medium. There is however evidence that the mtDNA content of the culture medium is related to embryo fragmentation levels and its presence is predictive of blastulation, indicating that embryo development may influence the levels of genetic material detected. If the levels of genetic material are strongly related to aspects of embryo quality, then this may be a novel biomarker of embryo viability. If the genetic material does have an embryo origin, the mechanisms by which DNA may be released into the blastocoele fluid and embryo culture medium are unknown, although apoptosis may play a role. While the presence of this genetic material is an exciting discovery, the DNA in the blastocoele fluid and embryo culture medium appears to be of low yield and integrity, which makes it challenging to study. Further research aimed at assessing the methodologies used for both isolating and analysing this genetic material, as well as tracing its origin, are needed in order to evaluate its potential for clinical use. Should such methodologies prove to be routinely successful and the DNA recovered

  13. Analysis of fluid flow and wall shear stress patterns inside partially filled agitated culture well plates.

    Science.gov (United States)

    Salek, M Mehdi; Sattari, Pooria; Martinuzzi, Robert J

    2012-03-01

    The appearance of highly resistant bacterial biofilms in both community and hospitals environments is a major challenge in modern clinical medicine. The biofilm structural morphology, believed to be an important factor affecting the behavioral properties of these "super bugs", is strongly influenced by the local hydrodynamics over the microcolonies. Despite the common use of agitated well plates in the biology community, they have been used rather blindly without knowing the flow characteristics and influence of the rotational speed and fluid volume in these containers. The main purpose of this study is to characterize the flow in these high-throughput devices to link local hydrodynamics to observed behavior in cell cultures. In this work, the flow and wall shear stress distribution in six-well culture plates under planar orbital translation is simulated using Computational Fluid Dynamics (CFD). Free surface, flow pattern and wall shear stress for two shaker speeds (100 and 200 rpm) and two volumes of fluid (2 and 4 mL) were investigated. Measurements with a non-intrusive optical shear stress sensor and High Frame-rate Particle Imaging Velocimetry (HFPIV) are used to validate CFD predictions. An analytical model to predict the free surface shape is proposed. Results show a complex three-dimensional flow pattern, varying in both time and space. The distribution of wall shear stress in these culture plates has been related to the topology of flow. This understanding helps explain observed endothelial cell orientation and bacterial biofilm distributions observed in culture dishes. The results suggest that the mean surface stress field is insufficient to capture the underlying dynamics mitigating biological processes.

  14. Isolation of osteogenic progenitors from human amniotic fluid using a single step culture protocol

    Directory of Open Access Journals (Sweden)

    Salini Vincenzo

    2009-02-01

    Full Text Available Abstract Background Stem cells isolated from amniotic fluid are known to be able to differentiate into different cells types, being thus considered as a potential tool for cellular therapy of different human diseases. In the present study, we report a novel single step protocol for the osteoblastic differentiation of human amniotic fluid cells. Results The described protocol is able to provide osteoblastic cells producing nodules of calcium mineralization within 18 days from withdrawal of amniotic fluid samples. These cells display a complete expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2 within 30 days from withdrawal. In order to test the ability of these cells to proliferate on surfaces commonly used in oral osteointegrated implantology, we carried out cultures onto different test disks, namely smooth copper, machined titanium and Sandblasted and Acid Etching titanium (SLA titanium. Electron microscopy analysis evidenced the best cell growth on this latter surface. Conclusion The described protocol provides an efficient and time-saving tool for the production of osteogenic cells from amniotic fluid that in the future could be used in oral osteointegrated implantology.

  15. 乳酸菌发酵液中DL-3-苯乳酸高效液相色谱快速检测方法的建立%Development of HPLC Method to Assay DL-3-Phenyllactic Acid in Culture Supernatant from Lactic Acid Bacteria

    Institute of Scientific and Technical Information of China (English)

    张忠华; 李晓然; 李洪梅; 李海舟; 龚福明; 柳陈坚

    2013-01-01

    DL-3-Phenyllactic acid (PLA) is a novel organic acid produced by some lactic acid bacteria,which can inhibit the growth of a wide variety of food-borne pathogens.On the other hand,PLA is safe and does not affect the odor of food at low concentrations.However,its further application was limited by its high equipment cost and complicated process so that the screening and identifying the high-PLA-producing LAB strains is restricted.A rapid method is developed in this paper for the determination of DL-3-Phenyllactic acid in MRS culture supernatant from lactic acid bacteria by high performance liquid chromatography.The determination is carried out on Agilent Zorbax SB-C18 (4.6 mm × 150 mm,5 μm).0.5% H3PO4 (V/V) and 0.5%H3PO4-CH3CN solution (V/V) is used as mobile phase at a flow rate of 1mL/min.Detection wavelength is set at 210 nm.The calibration curve is linear when DL-3-phenyllactic acid concentration is in the range of 60 ~800 mg/L.The corresponding regression equation is y =14.723x + 0.132 5,r =0.999 9.The method is then used to recover MRS fermented supernatant at 50 ~ 150 mg/L.The effectiveness is proven by the results as the recoveries are all above 94.0% and the relative standard deviation of samples is 0.58%.%DL-3苯乳酸是近年来发现可以由部分乳酸菌分泌产生的一种新型有机酸,它能够有效抑制多种食源性致病菌,并且具有安全、无毒副作用、低浓度添加时不影响食品风味等优点.然而现有苯乳酸检测方法存在所需检测设备投入成本高、操作步骤繁琐等缺陷,从而严重限制高产苯乳酸菌株的筛选和利用.本研究建立乳酸菌发酵液中苯乳酸高效液相色谱法快速检测方法,采用Agilent Zorbax SB-C18(4.6 mm×150 mm,5μm)色谱柱;流动相A:0.5%磷酸水溶液(V/V),B:0.5%磷酸乙腈溶液(V/V).流速1 mL/min,检测波长210 nm.DL-3-苯乳酸浓度在60~800 mg/L之间线性回归良好(其方程为y=14.723x +0.132 5,r=0.9999).应用本

  16. Outcomes in culture positive and culture negative ascitic fluid infection in patients with viral cirrhosis: cohort study

    Directory of Open Access Journals (Sweden)

    Ali Ailia W

    2008-12-01

    Full Text Available Abstract Background Ascitic fluid infection (AFI in cirrhotic patients has a high morbidity and mortality. It has two variants namely, spontaneous bacterial peritonitis (SBP and culture negative neutrocytic ascites (CNNA. The aim of this study was to determine the outcome in cirrhotic patients with culture positive (SBP and culture negative neutrocytic ascites. Methods We analyzed 675 consecutive hepatitis B and/or C related cirrhosis patients with ascites admitted in our hospital from November 2005 to December 2007. Of these, 187 patients had AFI; clinical and laboratory parameters of these patients including causes of cirrhosis, Child Turcotte Pugh (CTP score were recorded. Results Out of 187 patients with AFI, 44 (23.5% had SBP while 143 (76.4% had CNNA. Hepatitis C virus (HCV infection was the most common cause of cirrhosis in 139 (74.3% patients. Patients with SBP had high CTP score as compared to CNNA (12.52 ± 1.45 vs. 11.44 ± 1.66; p 9/L as compared to CNNA (132 ± 91 × 109/L, p = 0.005. We found a high creatinine (mg/dl (1.95 ± 1.0 vs. 1.44 ± 0.85, (p = 0.003 and high prothrombin time (PT in seconds (24.8 ± 6.6 vs. 22.4 ± 7.2 (p = 0.04 in SBP as compared to CNNA. More patients with SBP (14/44; 31.8% had blood culture positivity as compare to CNNA (14/143; 9.8%, p = 0.002. Escherichia. Coli was the commonest organism in blood culture in 15/28 (53.5% patients. SBP group had a higher mortality (11/44; 25% as compared to CNNA (12/143; 8.4%, p = 0.003. On multiple logistic regression analysis, creatinine >1.1 mg/dl and positive blood culture were the independent predictors of mortality in patients with SBP. Conclusion Patients with SBP have a higher mortality than CNNA. Independent predictors of mortality in SBP are raised serum creatinine and a positive blood culture.

  17. Cell vacuolation induced by Haemophilus influenzae supernatants in HEp-2 cells

    Directory of Open Access Journals (Sweden)

    Maria del Rosario Espinoza-Mellado

    2013-12-01

    Full Text Available Haemophilus influenzae belongs to respiratory tract microbiota. We observed vacuoles formation in previous studies with H. influenzae culture supernatants, so in this work we characterised that cytotoxic effect. We observed an abundant production of acidic cytoplasmic vacuoles due to the presence of a “vacuolating factor” in H. influenzae supernatants which was characterised as thermolabile. Greatest vacuolating activity was observed when utilizing the fraction > 50 kDa. The presence of a large number of vacuoles in HEp-2 cells was verified by transmission electron microscopy and some vacuoles were identified with a double membrane and/or being surrounded by ribosomes. These results suggest similar behaviour to that of vacuolating effects described by autotransporter proteins an undescribed cytotoxic effect induced by H. influenzae .

  18. Using a co-culture microsystem for cell migration under fluid shear stress.

    Science.gov (United States)

    Yeh, Chia-Hsien; Tsai, Shen-Hsing; Wu, Li-Wha; Lin, Yu-Cheng

    2011-08-01

    We have successfully developed a microsystem to co-cultivate two types of cells with a minimum defined gap of 50 μm, and to quantitatively study the impact of fluid shear stress on the mutual influence of cell migration velocity and distance. We used the hydrostatic pressure to seed two different cells, endothelial cells (ECs) and smooth muscle cells (SMCs), on opposite sides of various gap sizes (500 μm, 200 μm, 100 μm, and 50 μm). After cultivating the cells for 12 h and peeling the co-culture microchip from the culture dish, we studied the impacts of gap size on the migration of either cell type in the absence or presence of fluid shear stress (7 dyne cm(-2) and 12 dyne cm(-2)) influence. We found that both gap size and shear stress have profound influence on cell migration. Smaller gap sizes (100 μm and 50 μm) significantly enhanced cell migration, suggesting a requirement of an effective concentration of released factor(s) by either cell type in the gap region. Flow-induced shear stress delayed the migration onset of either cell type in a dose-dependent manner regardless of the gap size. Moreover, shear stress-induced decrease of cell migration becomes evident when the gap size was 500 μm. We have developed a co-culture microsystem for two kinds of cells and overcome the conventional difficulties in observation and mixed culture, and it would have more application for bio-manipulation and tissue repair engineering.

  19. Surface Enhanced Raman Spectroscopy detection of p-coumaric acid from cell supernatant using gold-capped silicon nanopillar substrates

    DEFF Research Database (Denmark)

    Morelli, Lidia; Jendresen, Christian Bille; Burger, Robert;

    HCA spiked in culture medium, in the same concentration range (10-4 – 10-5 M) commonly found in cell supernatant. For supernatant analysis, triplicate cultures of FjTAL modified (P strains) and control (C strains) E.coli strains were carried out according to the methods described by[5] and shown in Fig.1....... Samples of cell supernatant were extracted from each culture at 0, 3, 24 and 48 h post seeding and their pHCA content was measured with HPLC[5]. For SERS analysis, aliquots of supernatant were diluted 10-fold with MilliQ water, and 1 μL droplets were dried on the SERS substrates. A MatLab analysis......A standard protocol for analysis of microbial factories requires the screening of several populations in order to find the best performing ones. This is done with standard analytical methods (e.g. HPLC) with an expensive and time-consuming process. Surface Enhanced Raman Spectroscopy (SERS...

  20. Detection of p-coumaric acid from cell supernatant using surfaceenhanced Raman spectroscopy

    DEFF Research Database (Denmark)

    Morelli, Lidia; Jendresen, Christian Bille; Zor, Kinga

    2016-01-01

    A standard protocol for analysis of microbial factories requires the screening of several populations in order to find the bestperforming ones. Standard analytical methods usually include high performance liquid chromatography (HPLC), thin layerchromatography (TLC) or spectrophotometry, which...... on detection ofp-coumaric acid (pHCA) in cell supernatant.SERS active substrates, based on leaning gold-capped silicon nanopillars, were used for detection. They were successfullyused to detect culture medium spiked with pHCA, and the effect of medium dilution was studied. For analysis of biologicalproduction...

  1. High prevalence of Kingella kingae in joint fluid from children with septic arthritis revealed by the BACTEC blood culture system.

    Science.gov (United States)

    Yagupsky, P; Dagan, R; Howard, C W; Einhorn, M; Kassis, I; Simu, A

    1992-05-01

    In an effort to improve detection of fastidious organisms, joint fluid aspirates of pediatric patients were inoculated into BACTEC 460 aerobic blood culture bottles, in addition to cultures on solid media. Culture records for the 1988 to 1991 period were reviewed to compare the performance of both methods for the recovery of pathogens. Overall, 216 children underwent a diagnostic joint tap, and 63 specimens grew significant organisms, including Kingella kingae in 14. While both methods were comparable for recovery of usual pathogens, with a single exception, K. kingae isolates were detected by the BACTEC system only. K. kingae appears to be a more common cause of septic arthritis in children than has been previously recognized. The BACTEC blood culture system enhances the recovery of K. kingae from joint fluid and improves bacteriologic diagnosis of pediatric septic arthritis.

  2. Particulate metal bioaccessibility in physiological fluids and cell culture media: Toxicological perspectives.

    Science.gov (United States)

    Leclercq, Bérénice; Alleman, Laurent Yves; Perdrix, Esperanza; Riffault, Véronique; Happillon, Mélanie; Strecker, Alain; Lo-Guidice, Jean-Marc; Garçon, Guillaume; Coddeville, Patrice

    2017-07-01

    According to the literature, tiny amounts of transition metals in airborne fine particles (PM2.5) may induce proinflammatory cell response through reactive oxygen species production. The solubility of particle-bound metals in physiological fluids, i.e. the metal bioaccessibility is driven by factors such as the solution chemical composition, the contact time with the particles, and the solid-to-liquid phase ratio (S/L). In this work, PM2.5-bound metal bioaccessibility was assessed in various physiological-like solutions including cell culture media in order to evidence the potential impact on normal human bronchial epithelial cells (NHBE) when studying the cytotoxicity and inflammatory responses of PM2.5 towards the target bronchial compartment. Different fluids (H2O, PBS, LHC-9 culture medium, Gamble and human respiratory mucus collected from COPD patients), various S/L conditions (from 1/6000 to 1/100,000) and exposure times (6, 24 and 72h) were tested on urban PM2.5 samples. In addition, metals' total, soluble and insoluble fractions from PM2.5 in LHC-9 were deposited on NHBE cells (BEAS-2B) to measure their cytotoxicity and inflammatory potential (i.e., G6PDH activity, secretion of IL-6 and IL-8). The bioaccessibility is solution-dependent. A higher salinity or organic content may increase or inhibit the bioaccessibiliy according to the element, as observed in the complex mucus matrix. Decreasing the S/L ratio also affect the bioaccessibility depending on the solution tested while the exposure time appears less critical. The LHC-9 culture medium appears to be a good physiological proxy as it induces metal bioaccessibilities close to the mucus values and is little affected by S/L ratios or exposure time. Only the insoluble fraction can be linked to the PM2.5-induced cytotoxicity. By contrast, both soluble and insoluble fractions can be related to the secretion of cytokines. The metal bioaccessibility in LHC-9 of the total, soluble, and insoluble fractions of the

  3. Establishment of Primary Cell Culture From Ascitic Fluid and Solid Tumor Obtained From Epithelial Ovarian Carcinoma Patients.

    Science.gov (United States)

    Kar, Rajarshi; Chawla, Diwesh; Gupta, Bindiya; Mehndiratta, Mohit; Wadhwa, Neelam; Agarwal, Rachna

    2017-08-16

    Ovarian cancer is the seventh leading cause of cancer death worldwide. This is mainly due to late diagnosis and high rate of relapse and resistance following chemotherapy. In the present study, we describe simple and cost-effective method to establish primary culture from ascitic fluid and solid tumor obtained from epithelial ovarian carcinoma patient, which may provide a better tool for in vitro testing of drug sensitivity and designing individualized treatment protocol. Complete Dulbecco modified Eagle medium (DMEM) was prepared by supplementing DMEM with 10% fetal bovine serum and antibiotics (ciprofloxacin and amphotericin B). Establishment of primary culture of ovarian cancer cells from ascites fluid and solid tumor was done by using complete DMEM media. Primary cultures of ovarian cancer cells were established from ascitic fluid and solid tumor tissue. Of the 7 ascitic fluid samples, we were able to establish 5 primary cultures of ovarian cancer cells. All the 7 samples were diagnosed as serous papillary adenocarcinoma. Some fibroblasts were also attached to culture flask on day 4; they were removed by exposing them to trypsin for a brief period. On day 7, grape-like clusters were visualized under inverted microscope. The cells became confluent on the 10th and 11th day and showed cobblestone appearance, which is a hallmark of ovarian cancer cells. Senescent irregularly shaped cells that have ceased dividing were seen after 8 to 10 passages. This study highlights the fact that establishing primary cultures from ascitic fluid or solid tumor tissue may help us to understand the molecular profile of the cancer cells, which allow us to select the best chemotherapeutic agent for ovarian cancer patients and thus take a step toward patient-tailored therapy so that patients are not exposed to drugs to which they are not likely to respond.

  4. Low Fluid Shear Culture of Staphylococcus Aureus Represses hfq Expression and Induces an Attachment-Independent Biofilm Phenotype

    Science.gov (United States)

    Ott, C. Mark; Castro, S. L.; Nickerson, C. A.; Nelman-Gonzalez, M.

    2011-01-01

    Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

  5. Legionella pneumophila Arthritis: use of medium specific for Mycobacteria for isolation of L. pneumophila in culture of articular fluid specimens.

    Science.gov (United States)

    Bemer, Pascale; Leautez, Sophie; Ninin, Emmanuelle; Jarraud, Sophie; Raffi, François; Drugeon, Henri

    2002-07-01

    We report the first case, to our knowledge, of acute purulent arthritis due to Legionella pneumophila in an immunosuppressed patient. L. pneumophila was isolated from samples of blood and articular fluid cultured with use of medium specific for mycobacteria (Bactec 13A medium).

  6. 人羊膜间充质干细胞培养上清液对人成纤维细胞生物学功能的影响%Effects of culture supernatant of human amnion mesenchymal stem cells on biological characteristics of human fibroblasts

    Institute of Scientific and Technical Information of China (English)

    吴骐而; 吕璐; 辛海明; 罗亮; 童亚林; 莫永亮; 岳毅刚

    2016-01-01

    Objective To investigate the effects of culture supernatant of human amnion mesenchymal stem cells (hAMSCs-CS) on biological characteristics of human fibroblasts.Methods (1) hAMSCs were isolated from deprecated human fresh amnion tissue of placenta and then sub-cultured.The morphology of hAMSCs on culture day 3 and hAMSCs of the third passage were observed with inverted phase contrast microscope.(2) Two batches of hAMSCs of the third passage were obtained,then the expression of vimentin of cells was observed with immunofluorescence method,and the expression of cell surface marker CD90,CD73,CD105,and CD45 was detected by flow cytometer.(3) hAMSCs-CS of the third passage at culture hour 72 were collected,and the content of insulin-like growth factor Ⅰ (IGF-Ⅰ),vascular endothelial growth factor (VEGF),epidermal growth factor (EGF),and basic fibroblast growth factor (bFGF) were detected by enzyme-linked immunosorbent assay.(4) Hunan fibroblasts were isolated from deprecated human fresh prepuce tissue of circumcision and then sub-cultured.Human fibroblasts of the third passage were used in the following experiments.Cells were divided into blank control group and 10%,30%,50%,and 70% hAMSCs-CS groups according to the random number table (the same grouping method below),with 48 wells in each group.Cells in blank control group were cultured with DMEM/F12 medium containing 2% fetal bovine serum (FBS),while cells in the latter 4 groups were cultured with DMEM/F12 medium containing corresponding volume fraction of hAMSCs-CS and 2% FBS.The proliferation activity of cells was detected by cell counting kit 8 and microplate reader at culture hour 12,24,48,and 72,respectively,and corresponding volume fraction of hAMSCs-CS which causing the best proliferation activity of human fibroblasts was used in the following experiments.(5) Human fibroblasts were divided into blank control group and 50% hAMSCs-CS group and treated as in (4),with 4 wells in each group,at post

  7. Genotoxic effects of cadmium chloride on human amniotic fluid cells cultured in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Fogu, G. [Sassari Univ., Sassari (Italy). Dept. of Physiological, Biochemical and Cellular sciences; Sassari Univ., Sassari (Italy). Centre of Clinical Genetics; Congiu, A. M.; Sini, M. C.; Ladu, R. [Sassari Univ., Sassari (Italy). Dept. of Physiological, Biochemical and Cellular sciences; Campus, P. M.; Sanna, R.; Soro, G. [Sassari Univ., Sassari (Italy). Centre of Clinical Genetics

    2000-12-01

    In this study it has been reported the results of cytogenetic tests, namely a search for chromosome aberrations (CA) and sister chromatid exchanges (SCEs), performed on human amniotic fluid cells cultured and treated with Cadmium chloride. The cells from primary cultures were exposed to CdCl{sub 2} at 1 {mu}M and 10 {mu}M for 24 h. At the higher dose, no metaphases were scored and at the lower dose (1 {mu}M) no effects were evident on cell proliferation, and no chromosome aberrations were found. In the subsequent experiments were used cells from subcultures exposed to 1 {mu}M and 5 {mu}M CdCl{sub 2}. At the 5 {mu}M dose was evident the induction of chromatid breaks, while the frequency of sister chromatid exchanges shows a small increase, not statistically significant at the dose of 1 {mu}M. In this study it was positively demonstrated that amniotic fluid cells grown in vitro are reliable for testing various mutagenic or teratogenic substances. With regard to cadmium treatment results, it is evident a clastogenic effect of cadmium chloride but not a significant induction of SCEs. [Italian] In questo studio abbiamo riportato i risultati dei tests citogenici di valutazione degli scambi fra cromatidi fratelli (SCEs) e di induzione di aberrazioni cromosomiche (CA) condotti su colture cellulari di liquido amniotico umano, trattate con cloruro di cadmio. Le cellule delle colture primarie venivano esposte al CdCl{sub 2} a concentrazioni di 1 {mu}M 10 {mu}M per 24 h. Alla dose piu' alta (10 {mu}M) non sono state osservate metafasi, mentre alla dose piu' bassa (1 {mu}M) non sono stati evidenziati ne' effetti sulla proliferazione cellulare ne' CA. Nei successivi esperimenti il CdC{sub 2} e' stato testato su subcolture cellulari alle dosi di 1 {mu}M e 5 {mu}M. Alla dose di 5 {mu}M sono state evidenziate rotture cromatidiche, mentre la frequenza di SCE alla dose di 1 {mu}M ha mostrato un piccolo incremento, statisticamente non significativo. In questo

  8. Use of the isolator 1.5 microbial tube for culture of synovial fluid from patients with septic arthritis.

    Science.gov (United States)

    Yagupsky, P; Press, J

    1997-09-01

    Synovial fluid specimens obtained from patients with arthritis were plated onto solid media (conventional cultures) or inoculated into an Isolator 1.5 microbial tube (Isolator cultures), and the yield and time to detection of organisms were compared. Overall, 144 specimens obtained from 137 patients were processed, and 31 (21.5%) cultures obtained from 29 patients were positive by at least one method. Staphylococcus aureus was isolated from 12 patients, Streptococcus pneumoniae and Kingella kingae were isolated from 4 patients each, group G streptococci were isolated from 3 patients, Staphylococcus epidermidis and members of the family Enterobacteriaceae were isolated from 2 patients each, and Streptococcus mitis and Peptostreptococcus prevotii were isolated from 1 patient each. Overall, the causative organism was detected in 31 of 31 (100.0%) Isolator cultures and 24 of 31 (77.4%) conventional cultures (P septic arthritis.

  9. Immunoproteomic analysis of antibody in lymphocyte supernatant in patients with typhoid fever in Bangladesh.

    Science.gov (United States)

    Charles, Richelle C; Liang, Li; Khanam, Farhana; Sayeed, M Abu; Hung, Chris; Leung, Daniel T; Baker, Stephen; Ludwig, Albrecht; Harris, Jason B; Larocque, Regina C; Calderwood, Stephen B; Qadri, Firdausi; Felgner, Philip L; Ryan, Edward T

    2014-03-01

    We have previously shown that an assay based on detection of anti-Salmonella enterica serotype Typhi antibodies in supernatant of lymphocytes harvested from patients presenting with typhoid fever (antibody in lymphocyte supernatant [ALS] assay) can identify 100% of patients with blood culture-confirmed typhoid fever in Bangladesh. In order to define immunodominant proteins within the S. Typhi membrane preparation used as antigen in these prior studies and to identify potential biomarkers unique to S. Typhi bacteremic patients, we probed microarrays containing 2,724 S. Typhi proteins with ALS collected at the time of clinical presentation from 10 Bangladeshis with acute typhoid fever. We identified 62 immunoreactive antigens when evaluating both the IgG and IgA responses. Immune responses to 10 of these antigens discriminated between individuals with acute typhoid infection and healthy control individuals from areas where typhoid infection is endemic, as well as Bangladeshi patients presenting with fever who were subsequently confirmed to have a nontyphoid illness. Using an ALS enzyme-linked immunosorbent assay (ELISA) format and purified antigen, we then confirmed that immune responses against the antigen with the highest immunoreactivity (hemolysin E [HlyE]) correctly identified individuals with acute typhoid or paratyphoid fever in Dhaka, Bangladesh. These observations suggest that purified antigens could be used with ALS and corresponding acute-phase activated B lymphocytes in diagnostic platforms to identify acutely infected patients, even in areas where enteric fever is endemic.

  10. Studies on Using Cattle and Sheep Hydatid Cyst Fluid Instead of the Fetal Calf Serum in Leishmania Culture

    Directory of Open Access Journals (Sweden)

    Hossein Rezvan

    2013-12-01

    Full Text Available Background: Leishmania is a single cell parasite causing leishmaniasis, which is a common disease between humans and animals. Due to the importance of in-vitro culture of the parasite in leishmania research, developing new methods for in-vitro cultivation of the parasite has always been a goal for leishmania researchers. The main objective of7T 5T7Tthis study was to use sheep and bovine hydatid cyst fluids as alternatives for fetal calf serum (FCS in leishmania in-vitro5T culture5T. Materials and Methods: 7TA total of 5T7T1 million leishmania promastigotes were added to 4 flasks as follow5T7T. A f5T7Tlask containing DMEM medium with 105T7T% 5T7Tfetal bovine serum5T7T, a f5T7Tlask containing DMEM and 10% sheep hydatid cyst fluid5T7T, a f5T7Tlask containing DMEM medium with 105T7T% 5T7Tbovine hydatid cyst fluid and a5T7T f5T7Tlask containing DMEM medium alone. After 2, 45T7T, 5T7T7, 95T7T, 11, 5T7T21 and 24 days, the number of parasites were counted and compared5T7T. Results: The result of this study showed that, DMEM medium enriched with 10% sheep hydatid cyst fluid in 168 hours and medium enriched with 10% bovine hydatid cyst fluid in 96 hours can act as a good alternative for fetal bovine serum in the culture Leishmania major. Conclusion: 5TThe results showed that sheep and bovine hydatid cyst fluid can be used as alternatives to FCS for dense cultivation of leishmania. The results also showed that5T, 5Tthe growth of promastigotes in medium enriched with bovine cyst fluid is more rapid than the medium enriched with sheep5T c5Tyst fluid5T in5T the beginning of cultivation.

  11. Induction of E-cadherin+ human amniotic fluid cell differentiation into oocyte-like cells via culture in medium supplemented with follicular fluid.

    Science.gov (United States)

    Liu, Te; Huang, Yongyi; Bu, Yanzhen; Zhao, Yanhui; Zou, Gang; Liu, Zhixue

    2014-07-01

    Pluripotent human amniotic fluid cells (HuAFCs) can differentiate into various types of somatic cell in vitro. However, their differentiation into oocyte-like cells has never been described to the best of our knowledge. In the present study, differentiation of E-cadherin+ and E-cadherin- HuAFC sub-populations into oocyte-like cells was induced via culture in medium containing bovine follicular fluid and β-mercaptoethanol. The E-cadherin+ HuAFCs expressed DAZL highly. Post-induction, cells with an oocyte-like phenotype were found among the E-cadherin+ HuAFCs, expressing markers specific to germ cells and oocytes (VASA, ZP3 and GDF9) and meiosis (DMC1 and SCP3). When specific small interfering RNA (siRNA) was used to suppress E-cadherin in the E-cadherin+ HuAFCs, the levels of DAZL expression were reduced. Post-induction, the morphology of the siRNA‑E‑cadherin HuAFCs was poorer and the expression levels of germ cell-specific markers were lower compared with those of the siRNA-mock HuAFCs. Therefore, E-cadherin+ HuAFCs could be more easily induced to differentiate into oocyte-like cells by bovine follicular fluid and β-mercaptoethanol. In addition, the E-cadherin+ HuAFCs exhibited potential characteristics of DAZL protein expression, and thus it was conjectured that bovine follicular fluid acts on DAZL protein and promotes E-cadherin+ HuAFC differentiation into oocyte-like cells.

  12. Mechanism of Elevated Vascular Endothelial Growth Factor Levels in Peritoneal Fluids from Patients with Endometriosis

    Institute of Scientific and Technical Information of China (English)

    刘义; 吕立群

    2004-01-01

    In order to investigate the mechanism of elevated vascular endothelial growth factor (VEGF) in peritoneal fluids from patients with endometriosis, macrophages were recovered from peritoneal fluids obtained at the time of diagnostic laparoscopy from infertile women with endometriosis (EMT group, n = 20) and without endometriosis (control group, n = 20). Macrophages were cultured in vitro. The VEGF levels of peritoneal fluid and the supernatant of macrophages culture were determined by enzyme linked immunoassay (ELISA). Meanwhile, the eutopic (n = 20) and ectopic endometrium (n=20) from endometriosis patients, and normal edometrium (n=20) from non-endometriosis patients were obtained for the analysis of VEGF expression by labeled Streptavidin Biotin (LSAB). It was found that VEGF levels in peritoneal fluid and macrophages culture supernatant were significantly higher in EMT group than in control group (P<0.01). In normal endometrium, VEGF showed a cyclic changes and similar in eutopic and ectopic endometrium from patients with endometriosis. There was no difference in the intensity of VEGF in endometrium between two groups within each menstrual phase. It is suggested that altered VEGF production by peritoneal macrophages and ectopic endometrium secretion may contribute to the elevated VEGF levels in the peritoneal fluid of patients with endometriosis.

  13. Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis.

    Science.gov (United States)

    Kashyap, R S; Ramteke, S S; Gaherwar, H M; Deshpande, P S; Purohit, H J; Taori, G M; Daginawala, H

    2010-01-01

    The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth) for the diagnosis of tuberculous meningitis (TBM) in cerebrospinal fluid (CSF). CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50) were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64%) out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI): 67-93%] and 36% specificity (95% CI: 57-98%) for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM). This coexistence may go undetected or potentially lead to erroneous reporting of results.

  14. Evaluation of BioFM liquid medium for culture of cerebrospinal fluid in tuberculous meningitis to identify Mycobacterium tuberculosis

    Directory of Open Access Journals (Sweden)

    Kashyap R

    2010-01-01

    Full Text Available The present study was designed to evaluate the sensitivity and specificity of liquid culture medium (BioFM broth for the diagnosis of tuberculous meningitis (TBM in cerebrospinal fluid (CSF. CSF samples from 200 patients (TBM group = 150 and non-TBM group = 50 were tested for culture of Mycobacterium tuberculosis in BioFM liquid culture medium. Out of 150 TBM cases, 120 were found to be culture positive, indicating a sensitivity of 80% in BioFM broth within 2-3 weeks of inoculation. Positive cultures were also observed for CSF from 32 (64% out of 50 non-TBM patients in BioFM liquid culture medium within 4 days of sample inoculation. Therefore, according to our study, BioFM broth system yielded 80% sensitivity [95% confidence interval (CI: 67-93%] and 36% specificity (95% CI: 57-98% for TBM diagnosis. Our results indicate that although BioFM broth allows the detection of positive cultures within a shorter time, it has a high potential for contamination or for the coexistence of M. tuberculosis and non-tuberculous meningitis (NTM. This coexistence may go undetected or potentially lead to erroneous reporting of results.

  15. Acaricidal activities of whole cell suspension, cell-free supernatant,and crude cell extract of Xenorhabdus stokiae against mushroom mite (Luciaphorus sp.)

    Institute of Scientific and Technical Information of China (English)

    Prapassom BUSSAMAN; Chirayu SA-UTH; Paweena RATTANAS ENA; Angsumarn CHANDRAPATYA

    2012-01-01

    Xenorhabdus bacterium has been used as a biological control agent against Luciaphorus sp.,a mushroom mite endemic in Thailand.To develop an effective formulation of Xenorhabdus stokiae,treatments using different parts of X.stokiae isolate PB09 culture,including whole cell suspension,cell-free supernatant,and crude cell extract,were performed.The results show that different parts ofX.stokiae isolate PB09 culture could induce variable effects on mite mortality and fecundity.Application with cell-free supernatant of X.stokiae culture resulted in both the highest mite mortality rate [(89.00+3.60)%] and the lowest mite fecundity [(41.33+23.69) eggs/gravid female].Whole cell suspension of X.stokiae isolate PB09 culture was found to be slightly less effective than its cell-free supernatant,suggesting that X.stokiae was more likely to release its metabolites with acaricidal activities to the surrounding culture media.Crude cell extract of X.stokiae was not effective against mites.Cell-free supernatant of X.stokiae isolate PB09 was the most effective biological control agent and it could be conveniently used in future formulations instead of live bacteria.

  16. Culture supernatants from V. cholerae O1 ElTor strains isolated from different geographic areas induce cell vacuolation and cytotoxicity Cepas de V. cholerae O1 biotipo ElTor aisladas de diferente origen geográfico inducen vacuolización celular y citotoxicidad

    Directory of Open Access Journals (Sweden)

    Jorge E Vidal

    2009-02-01

    Full Text Available OBJECTIVE: To investigate whether the HlyA-induced vacuolating effect is produced by V. cholerae O1 ElTor strains isolated from different geographic origins, including Mexico. MATERIAL AND METHODS: Supernatant-induced haemolysis, vacuolating activity and cytotoxicity in Vero cells were recorded. PCR, RFLP analysis and molecular cloning were performed. RESULTS: All ElTor strains analyzed induced cellular vacuolation. Ribotype 2 strains isolates from the U.S. gulf coast yielded the highest titer of vacuolating activity. Eight of nine strains were haemolytic, while all strains were PCR positive for the hlyA gene. We cloned the hlyA gene from two ElTor strains, a toxigenic (2514-88, ctxAB+ and a non-toxigenic Mexican strain (CM 91-3, ctxAB-. Supernatant from those recombinant E. coli strains induced haemolysis, cell vacuolation and cytotoxicity. RFLP-PCR analysis revealed similarities in the hlyA gene from all strains tested. CONCLUSION: The HlyA-induced vacuolating effect is a widespread phenotype of epidemic V. cholerae O1 ElTor strains.OBJETIVO: Analizar el efecto vacuolizante de cepas de V. cholerae O1 ElTor aisladas de diferente origen geográfico, incluyendo México. MATERIAL Y MÉTODOS: Se realizaron pruebas de hemolisis, vacuolización y citotoxicidad en células Vero, así como PCR, análisis por RFLP y clonación molecular. RESULTADOS: Todas las cepas indujeron el efecto vacuolizante. Las cepas del ribotipo 2, aisladas de las costas del Golfo en Estados Unidos, presentaron títulos altos de vacuolización. El gen hlyA fue amplificado en las nueve cepas mediante PCR, aunque sólo ocho fueron hemolíticas. Se clonó el gen hlyA de una cepa toxigénica (2514-88, ctxAB+ y de una cepa no toxigénica aislada en México (CM 91-3, ctxAB-. El sobrenadante de las clonas recombinantes indujo hemólisis, efecto vacuolizante y citotoxicidad. El RFLP mostró alta similitud del gen hlyA de las cepas estudiadas. CONCLUSIÓN: El efecto vacuolizante es un

  17. Clinical significance of histologic chorioamnionitis with a negative amniotic fluid culture in patients with preterm labor and premature membrane rupture

    Science.gov (United States)

    Park, Jeong Woo; Park, Kyo Hoon; Jung, Eun Young

    2017-01-01

    Objective To evaluate the effect of histological chorioamnionitis (HCA) with a negative amniotic fluid (AF) culture on adverse pregnancy and neonatal outcomes and inflammatory status in the AF compartment in women with preterm labor or preterm premature rupture of membranes (PPROM). Methods This is a retrospective cohort study of 153 women diagnosed as having a preterm labor or PPROM (20–34 weeks) who delivered singleton gestations within 48 hours of amniocentesis. AF obtained through amniocentesis was cultured, and interleukin (IL)-6, IL-8, and metalloproteinase-9 (MMP-9) levels were determined. The placentas were examined histologically. Results The prevalence of HCA with negative AF culture was 23.5% (36/153). The women with HCA but with a negative AF culture (group 2) and those with a positive AF culture (group 3) had a significantly lower mean gestational age at amniocentesis and delivery than those with a negative AF culture and without HCA (group 1). Women in group 3 had the highest levels of AF IL-6, IL-8, and MMP-9, followed by those in group 2, and those in group 1. Composite neonatal morbidity was significantly higher in groups 2 and 3 than in group 1, but this was no longer significant after adjusting for confounders caused mainly by the impact of gestational age. Discussion In the women who delivered preterm neonates, HCA with a negative AF culture was associated with increased risks of preterm birth, intense intra-amniotic inflammatory response, and prematurity-associated composite neonatal morbidity, and its risks are similar to the risk posed by positive AF culture. PMID:28278303

  18. The congruity between urine and cervical swab cultures in pregnant women with versus without amniotic fluid infection.

    Science.gov (United States)

    Mavrova-Risteska, L; Hrgovic, Z; Curzik, D; Kissler, S; Kaufmann, M; Gaetje, R

    2005-12-01

    We observed the congruity of bacteria found in urine and cervix of pregnant women with amniotic fluid infection (AFI) compared to healthy controls. Over three years, we prospectively analysed urine and cervical swabs cultures in 120 pregnant women in gestational week 16-20. The patient population was divided in two groups: group I patients had clinical symptoms of AFI. The rest of the patients were designated as healthy controls (group II). Congruity between findings in both groups was observed. In patients with bacterial growth, antibiotic treatment was initiated as recommended and once ended - culture probes were repeated. The rates of nonsignificant and significant bacteriuria were doubled in group I. Fifty-five percent (n = 33) of patients in group I had identical bacteria cultured both from the urine sample and cervical swab, in contrast to only 13.3 % in group II. Congruity was most pronounced for Klebsiella species and E. coli, the later being single most dominant isolate in regards to both cultures. After antimicrobial treatment, microbial eradication occurred in 15 patients (45.5 %). The observed incidences of abortions and preterm deliveries were significantly lower in patients with microbial eradication versus patients with microbial persistence. Patients with clinical symptoms of AFI have high risk for bacteriuria identical to bacterial culture from cervical swab. Antimicrobial treatment was effective only partially where indicated. Screening for eradication is recommended and consensus on the most appropriate therapy is needed.

  19. BIOTECHNOLOGICAL ASPECTS OF INTERFACIAL TENSIOMETRY AND RHEOMETRY OF XYLOTROPHIC BASIDIOMYCETES CULTURE FLUID

    Directory of Open Access Journals (Sweden)

    Chaika A. V.

    2013-12-01

    Full Text Available The tensio-rheometric characteristics of 63 strains belonging to 19 basidiomycetes species submerged culture filtrate were investigated by the axisymmetric pendent drop profile analysis. The method showed required high sensitivity with mycological material. It was found that the interfacial tensiometric and rheometric parameters depend significantly on culture species, hence it is proposed to use ones complex for systematic identification of cultures and as a selection criterion for biosurfactantsproducing strains of basidiomycetes. Correlations of tensio-rheometric characteristics both among themselves and with the culture growth and lipid peroxidation rates were found. This provides an integrated indicator of the submerged culture metabolic state. By the results of the study several strains of basidiomycetes — potential producers of biosurfactants with a high growth rate and intensity of lipid peroxidation were selected for biotechnological manufacture.

  20. Vasoactive intestinal polypeptide and peptide histidine methionine. Presence in human follicular fluid and effects on DNA synthesis and steroid secretion in cultured human granulosa/lutein cells

    DEFF Research Database (Denmark)

    Gräs, S; Ovesen, P; Andersen, A N;

    1994-01-01

    fluid and cells were obtained from patients undergoing in-vitro fertilization for tubal infertility. The concentrations of VIP and PHM in pre-ovulatory human follicular fluid were measured radioimmunochemically. Granulosa/lutein cells isolated from follicular fluid were cultured under serum......Vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM) originate from the same precursor molecule, prepro VIP. In the present study we examined the concentrations of VIP and PHM in human follicular fluid and their effects on cultured human granulosa/lutein cells. Follicular......-free conditions with VIP and PHM in varying concentrations (0.1, 10, 1000 nmol/l). [3H]Thymidine incorporation in the cells and oestradiol as well as progesterone concentrations in the culture medium were measured. The mean (+/- SEM) concentrations of VIP and PHM were 6.8 +/- 0.1 and 7.7 +/- 0.8 pmol...

  1. Comparison between traditional laboratory tests, permeability measurements and CT-based fluid flow modelling for cultural heritage applications.

    Science.gov (United States)

    De Boever, Wesley; Bultreys, Tom; Derluyn, Hannelore; Van Hoorebeke, Luc; Cnudde, Veerle

    2016-06-01

    In this paper, we examine the possibility to use on-site permeability measurements for cultural heritage applications as an alternative for traditional laboratory tests such as determination of the capillary absorption coefficient. These on-site measurements, performed with a portable air permeameter, were correlated with the pore network properties of eight sandstones and one granular limestone that are discussed in this paper. The network properties of the 9 materials tested in this study were obtained from micro-computed tomography (μCT) and compared to measurements and calculations of permeability and the capillary absorption rate of the stones under investigation, in order to find the correlation between pore network characteristics and fluid management characteristics of these sandstones. Results show a good correlation between capillary absorption, permeability and network properties, opening the possibility of using on-site permeability measurements as a standard method in cultural heritage applications.

  2. Changes of dynamical balance of free radicals induced by levodopa in rat glia-containing mesencephalic culture fluid

    Institute of Scientific and Technical Information of China (English)

    Wenxia Li; Shenggang Sun; Jingru Wang; Baoliang Sun; Yanbo Zhang

    2006-01-01

    BACKGROUND: Parkinson disease is neurodegenerative disorders, characterized by a progressive and selective degeneration of nigrostriatal dopaminergic pathway. Its main clinical symptoms include bradykinesia, rigidity, rest tremor and disturbances in balance. Levodopa (L-DOPA) is the "gold standard" for the symptomatic treat ment of Parkinson disease, but L-DOPA is toxic to dopaminergic neurons and the chronic administration of L-DOPA often causes the side effects of motor such as "on-off ", etc., and its mechanism still has not been completely clarified.OBJECTIVE: To observe the changes of the content of glutathione (GSH), activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) and content of malondialdehyde (MDA) in glia-containing mesencephalic culture fluid after L-DOPA of different concentrations were added.DESIGN: A comparative observation.SETTINGS: Department of Neurology, Affiliated Hospital of Taishan Medical University; Department of Neurology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology. MATERIALS: The experiment was carried out in the Basic Research Institute, Taishan Medical University from December 2003 to September 2004. Ten postnatal newborn SD rats (within 2 days) were selected, either male or female. Fetal bovine serum (DMEM/F12) was from Gibco Company; L-DOPA and glial fibrillary acidic protein (GFAP) from Sigma Company. Kits for determination of GSH, GSH-Px, SOD and MDA were purchased from Nanjing Jiancheng Bioengineering Research Institute.METHODS : Glia-containing mesencephalic culture fluid were placed in 24-well culture plate, and L-DOPA of 50, 100 and 500 μmol/L was added to each group, the blank control group was also set. The glia-containing mesencephalic culture fluid selected at 4, 24, 48 and 72 hours respectively to determine the GSH content with colorimetric quantitative technique, GSH-Px activity with colorimetric method, SOD activity with xanthine oxidase method and MDA

  3. The Utility of Blood Culture Fluid for the Molecular Diagnosis of Leptospira

    DEFF Research Database (Denmark)

    Dittrich, Sabine; Rudgard, William E.; Woods, Kate L.

    2016-01-01

    Leptospirosis is an important zoonosis worldwide, with infections occurring after exposure to contaminated water. Despite being a global problem, laboratory diagnosis remains difficult with culture results taking up to 3 months, serology being retrospective by nature, and polymerase chain reaction...

  4. Peritoneal fluid culture and antibiotic treatment in patients with perforated appendicitis in a Pacific Island

    Directory of Open Access Journals (Sweden)

    Alexia Boueil

    2015-10-01

    Conclusion: Although antibiotic use may be considered as an adjunct to surgical intervention of AP, the appropriate use of preoperative antibiotics and modifications according to culture results are essential to prevent infectious complications.

  5. Molecular and Culture-Based Bronchoalveolar Lavage Fluid Testing for the Diagnosis of Cytomegalovirus Pneumonitis.

    Science.gov (United States)

    Tan, Susanna K; Burgener, Elizabeth B; Waggoner, Jesse J; Gajurel, Kiran; Gonzalez, Sarah; Chen, Sharon F; Pinsky, Benjamin A

    2016-01-01

    Background.  Cytomegalovirus (CMV) is a major cause of morbidity and mortality in immunocompromised patients, with CMV pneumonitis among the most severe manifestations of infection. Although bronchoalveolar lavage (BAL) samples are frequently tested for CMV, the clinical utility of such testing remains uncertain. Methods.  Retrospective analysis of adult patients undergoing BAL testing via CMV polymerase chain reaction (PCR), shell vial culture, and conventional viral culture between August 2008 and May 2011 was performed. Cytomegalovirus diagnostic methods were compared with a comprehensive definition of CMV pneumonitis that takes into account signs and symptoms, underlying host immunodeficiency, radiographic findings, and laboratory results. Results.  Seven hundred five patients underwent 1077 bronchoscopy episodes with 1090 BAL specimens sent for CMV testing. Cytomegalovirus-positive patients were more likely to be hematopoietic cell transplant recipients (26% vs 8%, P definition, the sensitivity and specificity of PCR, shell vial culture, and conventional culture were 91.3% and 94.6%, 54.4% and 97.4%, and 28.3% and 96.5%, respectively. Compared with culture, PCR provided significantly higher sensitivity and negative predictive value (P ≤ .001), without significantly lower positive predictive value. Cytomegalovirus quantitation did not improve test performance, resulting in a receiver operating characteristic curve with an area under the curve of 0.53. Conclusions.  Cytomegalovirus PCR combined with a comprehensive clinical definition provides a pragmatic approach for the diagnosis of CMV pneumonitis.

  6. 羊水预处理在羊水细胞培养瓶法中应用效果研究%Application effect research with amniotic fluid pretreatment in amniotic fluid cell culture bottle

    Institute of Scientific and Technical Information of China (English)

    向文秀

    2013-01-01

    目的 研究羊水预处理在羊水细胞培养瓶法中的应用效果.方法 采用羊水预处理培养瓶法(实验组)和未经处理羊水细胞培养瓶法(对照组)对100例18-24周的孕中期孕妇和152例25-32周的孕晚期孕妇进行羊水细胞培养.结果 100例孕18-24周羊水细胞培养实验组培养成功率高于对照组(P<0.05),实验组羊水细胞培养时间比对照组短(P<0.001),实验组有效核分裂数比对照组多(P <0.001);152例孕25-32周羊水细胞培养实验组培养成功率高于对照组(P<0.01),实验组羊水细胞培养时间比对照组短(P <0.001),实验组有效核分裂数比对照组多(P <0.001).结论 羊水预处理羊水细胞培养瓶法羊水细胞培养成功率高,细胞培养时间短,有效核分裂数多,可进行羊水产前诊断孕妇孕周范围增宽.%Objective:Research with amniotic fluid pretreatment in amniotic fluid cell culture bottle of application effect.Methods:100 samples of amniotic fluid from pregnant women during 18-24th-week gestation and 152 samples of amniotic fluid from pregnant women during 25-32th-week gestation were cultured in fluid pretreatment amniotic fluid cell culture bottle and untreated amniotic fluid cell culture bottle.Results:In 100 samples of pregnant women during 18-24th-week gestation.Culture success rate of the experimental group than the control group (P < 0.05).Amniotic fluid cell culture time of the experimental group than the control group short (P < 0.001).Effective mitotic count of the experimental group than the control group (P < 0.001).In 152 samples of pregnant women during 25-32th-week gestation,Culture success rate of the experimental group than the control group (P <0.01),Amniotic fluid cell culture time of the experimental group than the control group short (P < 0.001).Effective mitotic count of the experimental group than the control group (P < 0.001).Conclusion:Amniotic fluid pretreatment in amniotic fluid cell

  7. Fluorescence Spectrum and Decay Measurement for Hsil VS Normal Cytology Differentiation in Liquid Pap Smear Supernatant

    Science.gov (United States)

    Vaitkuviene, A.; Gegzna, V.; Juodkazis, S.; Jursenas, S.; Miasojedovas, S.; Kurtinaitiene, R.; Rimiene, J.; Vaitkus, J.

    2009-06-01

    Cervical smear material contains endo and exocervical cells, mucus and inflammative, immune cells in cases of pathology. Just not destroyed keratinocytes lay on the glass for microscopy. Liquid cytology supernatant apart other diagnostics could be used for photodiagnostic. The spectroscopic parameters suitable for Normal and HSIL cytology groups supernatant differentiation are demonstrated. The dried liquid PAP supernatant fractions—sediment and liquid were investigated. Excitation and emission matrices (EEM), supernatant fluorescence decay measured under 280 nm diode short pulse excitation and fluorescence spectroscopy by excitation with 355 nm laser light were analyzed. The differences between Normal and HSIL groups were statistically proven in the certain spectral regions. Fluorescence decay peculiarities show spectral regions consisting of few fluorophores. Obtained results on fluorescence differences in Normal and HSIL groups' supernatant shows the potency of photodiagnosis application in cervical screening.

  8. Fluid Dynamic Modeling to Support the Development of Flow-based Hepatocyte Culture Systems for Metabolism Studies

    Directory of Open Access Journals (Sweden)

    Jenny M Pedersen

    2016-09-01

    Full Text Available Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics (CFD to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were 1 minimization of shear stress experienced by the cells to maximize viability, 2 rapid establishment of a uniform distribution of test compound in the chamber, and 3 delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices — RealBio® (RB and QuasiVivo® (QV — and a custom developed fluidized bed (FB bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results.Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able

  9. Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies

    Science.gov (United States)

    Pedersen, Jenny M.; Shim, Yoo-Sik; Hans, Vaibhav; Phillips, Martin B.; Macdonald, Jeffrey M.; Walker, Glenn; Andersen, Melvin E.; Clewell, Harvey J.; Yoon, Miyoung

    2016-01-01

    Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were (1) minimization of shear stress experienced by the cells to maximize viability, (2) rapid establishment of a uniform distribution of test compound in the chamber, and (3) delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices – RealBio® and QuasiVivo® (QV) – and a custom developed fluidized bed bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results. Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able to optimize the experimental

  10. Efficacy of nasopharyngeal culture in identification of pathogens in middle ear fluid in chronic otitis media with effusion

    Directory of Open Access Journals (Sweden)

    Eser O

    2009-01-01

    Full Text Available Purpose: Chronic otitis media with effusion (OME is the leading cause of hearing loss during childhood. In bacterial etiology of OME, the most frequent pathogens responsible are Haemophilus influenzae followed by Streptococcus pneumoniae and Moraxella catarrhalis . This study aimed at evaluating the accuracy of nasopharyngeal (NP specimens in the identification of pathogens in the middle ear fluid (MEF in patients with OME. Materials and Methods: In this cross sectional, case-control study, 95 MEFs and 53 NP secretion specimens were obtained from 53 children. As a control group, 102 NP specimens were taken from children having an operation other than an otological disease. Conventional culture methods and multiplex-PCR method have been used to determine the etiology of OME; NP carriage between cases and control groups were compared using conventional culture methods. Pearson Chi-Square and Fisher′s Exact tests were used in statistical analysis. Results : Bacteria were isolated by culture in 37.9% of MEF specimens, 14.7% of which belonged to the group H. influenzae , S. pneumoniae and M. catarrhalis. PCR was positive in 30.5% specimens targeting the same pathogens. There was a two-fold increase in carriage rate of S. pneumoniae and H. influenzae in patients than controls for each pathogen. Conclusion: PCR is a more reliable method to detect middle ear pathogens in MEF in comparison with the conventional culture methods. The NP colonization wasn′t found to be an indicator of the pathogen in MEF although middle ear pathogens colonize more in nasopharynx of diseased children.

  11. Microbial prevalence, diversity and abundance in amniotic fluid during preterm labor: a molecular and culture-based investigation.

    Directory of Open Access Journals (Sweden)

    Daniel B DiGiulio

    Full Text Available BACKGROUND: Preterm delivery causes substantial neonatal mortality and morbidity. Unrecognized intra-amniotic infections caused by cultivation-resistant microbes may play a role. Molecular methods can detect, characterize and quantify microbes independently of traditional culture techniques. However, molecular studies that define the diversity and abundance of microbes invading the amniotic cavity, and evaluate their clinical significance within a causal framework, are lacking. METHODS AND FINDINGS: In parallel with culture, we used broad-range end-point and real-time PCR assays to amplify, identify and quantify ribosomal DNA (rDNA of bacteria, fungi and archaea from amniotic fluid of 166 women in preterm labor with intact membranes. We sequenced up to 24 rRNA clones per positive specimen and assigned taxonomic designations to approximately the species level. Microbial prevalence, diversity and abundance were correlated with host inflammation and with gestational and neonatal outcomes. Study subjects who delivered at term served as controls. The combined use of molecular and culture methods revealed a greater prevalence (15% of subjects and diversity (18 taxa of microbes in amniotic fluid than did culture alone (9.6% of subjects; 11 taxa. The taxa detected only by PCR included a related group of fastidious bacteria, comprised of Sneathia sanguinegens, Leptotrichia amnionii and an unassigned, uncultivated, and previously-uncharacterized bacterium; one or more members of this group were detected in 25% of positive specimens. A positive PCR was associated with histologic chorioamnionitis (adjusted odds ratio [OR] 20; 95% CI, 2.4 to 172, and funisitis (adjusted OR 18; 95% CI, 3.1 to 99. The positive predictive value of PCR for preterm delivery was 100 percent. A temporal association between a positive PCR and delivery was supported by a shortened amniocentesis-to-delivery interval (adjusted hazard ratio 4.6; 95% CI, 2.2 to 9.5. A dose

  12. Beneficial effects of BV2 cell on proliferation and neuron-differentiating of mesenchymal stem cells in the circumstance of injured PC12 cell supernatant

    Institute of Scientific and Technical Information of China (English)

    Xiao-Guang LUO; Hong WANG; Jin ZHOU; Rong YAN; Zhe WU; Chao-Dong ZHANG; Qiu-Shuang WANG

    2006-01-01

    Objective The microglias is the representative of immune cells in the brain. It plays dual roles of both repairing and damaging in injured nervous system, and works as an inevitable component of the circumstance of injured neurons. This study was aiming at the effects of the microglias on the biological activities of mesenchymal stem cells (MSCs) inthe circumstance of injured neurons. Methods MSCs were obtained by primary culture. We adopted PC12 cells (PC12) and BV2 cells (BV2) to substitute for neurons and microglias, respectively. PC12 were injured by aged Aβ1-40 and the supernatant of the injured PC12 was used to set up the circumstance of injured neurons. Transwells were used for co-culture of BV2 and MSCs, which allowed the independent detection of cells after co-culture. Immunofluorescence was used to identify MSCs and neuron-differentiating cells with CD44 and neuron specific enolase (NSE) staining, respectively. MTT assay was adopted to measure the proliferation. Results In the circumstance of both BV2 presence and injured PC12 supernatant incubation, either the proliferation or the differentiation of MSCs reached the highest, which seemed to be contradictory, but we gave our explanations. With the BV2 co-culture, the proliferation of MSCs tend to be higher, but the neuron-differentiating MSCs were similar to those incubated without BV2 co-culture either in normal or injured in PC12 supernatant. With the incubation of injured PC12 supernatant, the neuron-differentiating cells were significantly higher than that of control (P < 0.05). Conclusion In the circumstance of injured neurons, microlgias tend to promote the MSCs proliferation. Although not helpful in neuron-differentiating, microglias did not exert any negative effect either.

  13. Anaerobic bioremediation of RDX by ovine whole rumen fluid and pure culture isolates.

    Science.gov (United States)

    Eaton, H L; Duringer, J M; Murty, L D; Craig, A M

    2013-04-01

    The ability of ruminal microbes to degrade the explosive compound hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in ovine whole rumen fluid (WRF) and as 24 bacterial isolates was examined under anaerobic conditions. Compound degradation was monitored by high-performance liquid chromatography analysis, followed by liquid chromatography-tandem mass spectrometry identification of metabolites. Organisms in WRF microcosms degraded 180 μM RDX within 4 h. Nitroso-intermediates hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX), hexahydro-1,3-dinitroso-5-nitro-1,3,5-triazine (DNX), and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX) were present as early as 0.25 h and were detected throughout the 24-h incubation period, representing one reductive pathway of ring cleavage. Following reduction to MNX, peaks consistent with m/z 193 and 174 were also produced, which were unstable and resulted in rapid ring cleavage to a common metabolite consistent with an m/z of 149. These represent two additional reductive pathways for RDX degradation in ovine WRF, which have not been previously reported. The 24 ruminal isolates degraded RDX with varying efficiencies (0-96 %) over 120 h. Of the most efficient degraders identified, Clostridium polysaccharolyticum and Desulfovibrio desulfuricans subsp. desulfuricans degraded RDX when medium was supplemented with both nitrogen and carbon, while Anaerovibrio lipolyticus, Prevotella ruminicola, and Streptococcus bovis IFO utilized RDX as a sole source of nitrogen. This study showed that organisms in whole rumen fluid, as well as several ruminal isolates, have the ability to degrade RDX in vitro and, for the first time, delineated the metabolic pathway for its biodegradation.

  14. Effect of Lactobacillus sp. isolates supernatant on Escherichia coli O157:H7 enhances the role of organic acids production as a factor for pathogen control

    Directory of Open Access Journals (Sweden)

    Larissa B. Poppi

    2015-04-01

    Full Text Available Many attempts have been made to establish the control of foodborne pathogens through Lactobacillus isolates and their metabolism products with success being obtained in several situations. The aim of this study was to investigate the antagonistic effect of eight Lactobacillus isolates, including L. casei subsp. pseudoplantarum, L. plantarum, L. reuteri and L. delbrueckii subsp. delbrueckii, on the pathogenic Escherichia colistrain O157:H7. The inhibitory effect of pure cultures and two pooled cultures supernatants of Lactobacillus on the growth of pathogenic bacteria was evaluated by the spot agar method and by monitoring turbidity. Antimicrobial activity was confirmed for L. reuteri and L. delbrueckii subsp. delbrueckii and for a pool of lactic acid bacteria. The neutralized supernatant of the pool exerted a higher antimicrobial activity than that of the individual strains. Furthermore, D-lactic acid and acetic acid were produced during growth of the Lactobacillus isolates studied.

  15. Comparison between traditional laboratory tests, permeability measurements and CT-based fluid flow modelling for cultural heritage applications

    Energy Technology Data Exchange (ETDEWEB)

    De Boever, Wesley, E-mail: Wesley.deboever@ugent.be [UGCT/PProGRess, Dept. of Geology, Ghent University, Krijgslaan 281, 9000 Ghent (Belgium); Bultreys, Tom; Derluyn, Hannelore [UGCT/PProGRess, Dept. of Geology, Ghent University, Krijgslaan 281, 9000 Ghent (Belgium); Van Hoorebeke, Luc [UGCT/Radiation Physics, Dept. of Physics & Astronomy, Ghent University, Proeftuinstraat 86, 9000 Ghent (Belgium); Cnudde, Veerle [UGCT/PProGRess, Dept. of Geology, Ghent University, Krijgslaan 281, 9000 Ghent (Belgium)

    2016-06-01

    In this paper, we examine the possibility to use on-site permeability measurements for cultural heritage applications as an alternative for traditional laboratory tests such as determination of the capillary absorption coefficient. These on-site measurements, performed with a portable air permeameter, were correlated with the pore network properties of eight sandstones and one granular limestone that are discussed in this paper. The network properties of the 9 materials tested in this study were obtained from micro-computed tomography (μCT) and compared to measurements and calculations of permeability and the capillary absorption rate of the stones under investigation, in order to find the correlation between pore network characteristics and fluid management characteristics of these sandstones. Results show a good correlation between capillary absorption, permeability and network properties, opening the possibility of using on-site permeability measurements as a standard method in cultural heritage applications. - Highlights: • Measurements of capillary absorption are compared to in-situ permeability. • We obtain pore size distribution and connectivity by using micro-CT. • These properties explain correlation between permeability and capillarity. • Correlation between both methods is good to excellent. • Permeability measurements could be a good alternative to capillarity measurement.

  16. How does the supernatant of Lactobacillus acidophilus affect the proliferation and differentiation activities of rat bone marrow-derived stromal cells?

    Science.gov (United States)

    Samadikuchaksaraei, A; Gholipourmalekabadi, M; Saberian, M; Abdollahpour Alitappeh, M; Shahidi Delshad, E

    2016-08-31

    Low proliferation rate and unwanted differentiation of bone marrow-derived stromal cells (rBMSCs) during the frequent passages have limited the use of such cells in clinical cell therapy. Recently, the researchers have focused on the effects of the components produced by some bacteria on proliferation of the stem cells. In this study, we discussed the possible effects of the Lactobacillus acidophilus supernatant on proliferation and differentiation of the rBMSCs. For this aim, the cells were isolated from rat bone marrow, characterized by culturing on tissue specific differentiation media and stained. The cells (passage two) were treated with different concentrations of the L. acidophilus supernatant (0, 0.1, 0.3, 0.9, 3, 9 and 30 &mgr;l/ml) for 14 days. The proliferation and differentiation capacity of the cells were then determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT assay) and tissue specific staining. The results showed a positive effect of the supernatant on the cell proliferation in 3 and 9 &mgr;l/ml concentrations, while did not affect the differentiation capacity of the rBMSCs. The current study strongly suggests the L. acidophilus supernatant as an alternative material that could be added to the media with aim of improvement in the proliferation rate of the rBMSCs without affecting their differentiation capacity.

  17. Positive peritoneal fluid fungal cultures in postoperative peritonitis after bariatric surgery.

    Science.gov (United States)

    Zappella, N; Desmard, M; Chochillon, C; Ribeiro-Parenti, L; Houze, S; Marmuse, J-P; Montravers, P

    2015-09-01

    Postoperative peritonitis (POP) is a common surgical complication after bariatric surgery (BS). We assessed the importance of positive fungal cultures in these cases of POP admitted to the intensive care unit. Clinical features and outcome were compared in 25 (41%) Candida-positive patients (6 (22%) fluconazole-resistant Candida glabrata) and 36 patients without Candida infection. Candida infections were more commonly isolated in late-onset peritonitis and were often associated with multidrug-resistant bacteria. Risk factors for intensive care unit mortality (19.6%) were diabetes and superobesity. Candida infections, including fluconazole-resistant strains, are common in POP after BS. These data encourage the empirical use of a broad-spectrum antifungal agent. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

  18. Comparison of Selective Culturing and Biochemical Techniques for Measuring Biological Activity in Geothermal Process Fluids

    Energy Technology Data Exchange (ETDEWEB)

    Pryfogle, Peter Albert

    2000-09-01

    For the past three years, scientists at the Idaho National Engineering and Environmental Laboratory have been conducting studies aimed at determining the presence and influence of bacteria found in geothermal plant cooling water systems. In particular, the efforts have been directed at understanding the conditions that lead to the growth and accumulation of biomass within these systems, reducing the operational and thermal efficiency. Initially, the methods selected were based upon the current practices used by the industry and included the collection of water quality parameters, the measurement of soluble carbon, and the use of selective medial for the determination of the number density of various types of organisms. This data has been collected on a seasonal basis at six different facilities located at the Geysers’ in Northern California. While this data is valuable in establishing biological growth trends in the facilities and providing an initial determination of upset or off-normal conditions, more detailed information about the biological activity is needed to determine what is triggering or sustaining the growth in these facilities in order to develop improved monitoring and treatment techniques. In recent years, new biochemical approaches, based upon the analyses of phospholipid fatty acids and DNA recovered from environmental samples, have been developed and commercialized. These techniques, in addition to allowing the determination of the quantity of biomass, also provide information on the community composition and the nutritional status of the organisms. During the past year, samples collected from the condenser effluents of four of the plants from The Geysers’ were analyzed using these methods and compared with the results obtained from selective culturing techniques. The purpose of this effort was to evaluate the cost-benefit of implementing these techniques for tracking microbial activity in the plant study, in place of the selective culturing

  19. Capillary size exclusion chromatography with picogram sensitivity for analysis of monoclonal antibodies purified from harvested cell culture fluid.

    Science.gov (United States)

    Rea, Jennifer C; Moreno, G Tony; Vampola, Lisa; Lou, Yun; van Haan, Bjorn; Tremintin, Guillaume; Simmons, Laura; Nava, Adrian; Wang, Yajun Jennifer; Farnan, Dell

    2012-01-06

    Size exclusion chromatography (SEC) is widely used in the characterization and quality control of therapeutic proteins to detect aggregates. Aggregation is a carefully monitored quality attribute from the earliest stages of clinical development owing to the possibility of eliciting an immunogenic response in the patient. During early stage molecule assessment for cell culture production, small-scale screening experiments are performed to permit rapid turn-around of results so as to not delay timelines. We report the development of a capillary SEC methodology for preliminary molecule assessment to support the evaluation of therapeutic candidates at an early stage of development. By making several key modifications to a commercially available liquid chromatography system, we demonstrate a platform process to perform capillary SEC with excellent precision, picogram sensitivity and good ruggedness. The limit of quantitation was determined to be approximately 15 pg; picogram sensitivity for SEC analysis of monoclonal antibodies had not been achieved prior to this work. In addition, we have developed a method to capture low levels of antibody (1 μg/mL) from harvested cell culture fluid (HCCF) for capillary SEC analysis. Up to 40% recovery efficiency was achieved using this micro-recovery method on 3 mL HCCF samples. Using early stage cell culture transient transfection samples, which typically have much lower titers than stable cell line samples, we demonstrate a consistent method for analyzing aggregates in low protein concentration HCCF samples for molecule assessment and early stage candidate screening. Copyright © 2011 Elsevier B.V. All rights reserved.

  20. Vasoactive intestinal polypeptide and peptide histidine methionine. Presence in human follicular fluid and effects on DNA synthesis and steroid secretion in cultured human granulosa/lutein cells

    DEFF Research Database (Denmark)

    Gräs, S; Ovesen, P; Andersen, A N;

    1994-01-01

    Vasoactive intestinal polypeptide (VIP) and peptide histidine methionine (PHM) originate from the same precursor molecule, prepro VIP. In the present study we examined the concentrations of VIP and PHM in human follicular fluid and their effects on cultured human granulosa/lutein cells. Follicular...

  1. Test procedures and instructions for Hanford tank waste supernatant cesium removal

    Energy Technology Data Exchange (ETDEWEB)

    Hendrickson, D.W., Westinghouse Hanford

    1996-05-31

    This document provides specific test procedures and instructions to implement the test plan for the preparation and conduct of a cesium removal test using Hanford Double-Shell Slurry Feed supernatant liquor from tank 251-AW-101 in a bench-scale column.Cesium sorbents to be tested include resorcinol-formaldehyde resin and crystalline silicotitanate. The test plan for which this provides instructions is WHC-SD-RE-TP-022, Hanford Tank Waste Supernatant Cesium Removal Test Plan.

  2. Test procedures and instructions for Hanford complexant concentrate supernatant cesium removal using CST

    Energy Technology Data Exchange (ETDEWEB)

    Hendrickson, D.W.

    1997-01-08

    This document provides specific test procedures and instructions to implement the test plan for the preparation and conduct of a cesium removal test, using Hanford Complexant Concentrate supernatant liquor from tank 241-AN-107, in a bench-scale column. The cesium sorbent to be tested is crystalline silicotitanate. The test plan for which this provides instructions is WHC-SD-RE-TP-023, Hanford Complexant Concentrate Supernatant Cesium Removal Test Plan.

  3. Global Profiling of Metabolite and Lipid Soluble Microbial Products in Anaerobic Wastewater Reactor Supernatant Using UPLC-MS(E).

    Science.gov (United States)

    Tipthara, Phornpimon; Kunacheva, Chinagarn; Soh, Yan Ni Annie; Wong, Stephen C C; Pin, Ng Sean; Stuckey, David C; Boehm, Bernhard O

    2017-02-03

    Identification of soluble microbial products (SMPs) released during bacterial metabolism in mixed cultures in bioreactors is essential to understanding fundamental mechanisms of their biological production. SMPs constitute one of the main foulants (together with colloids and bacterial flocs) in membrane bioreactors widely used to treat and ultimately recycle wastewater. More importantly, the composition and origin of potentially toxic, carcinogenic, or mutagenic SMPs in renewable/reused water supplies must be determined and controlled. Certain classes of SMPs have previously been studied by GC-MS, LC-MS, and MALDI-ToF MS; however, a more comprehensive LC-MS-based method for SMP identification is currently lacking. Here we develop a UPLC-MS approach to profile and identify metabolite SMPs in the supernatant of an anaerobic batch bioreactor. The small biomolecules were extracted into two fractions based on their polarity, and separate methods were then used for the polar and nonpolar metabolites in the aqueous and lipid fractions, respectively. SMPs that increased in the supernatant after feed addition were identified primarily as phospholipids, ceramides, with cardiolipins in the highest relative abundance, and these lipids have not been previously reported in wastewater effluent.

  4. The effect of moderately halophilic bacteria supernatant on proliferation and apoptosis of cancer cells and mesenchymal stem cells.

    Science.gov (United States)

    Sarvari, S; Seyedjafari, E; Amoozgar, M A; Bakhshandeh, B

    2015-06-10

    Many drug discoveries and developing of their applications has originated from microbial metabolites. The most efforts in development of new drugs are concerned with anti—cancer agents that cause better treatment results, less side effects, and more economical production. Several anti—tumor drugs have been recently extracted from natural microbial products. Among these various microbial diversity, Marin bacteria and Archaea have been considered as important and efficient organisms to serve as manufacturers of diverse bioactive compounds. Moderately halophilic microorganisms isolated from saline ponds and lakes of Iran show high capability for production of bioactive compounds like enzymes, dyes and anti—cancer agents. In this research, nine moderately halophilic bacteria isolates were screened to evaluate their anti—cancer agent productivity. After five days of culture on suitable mediums, supernatant samples were tested for in vitro anti—proliferative activity against Human Umbilical Vein Endothelial Cells (HUVEC) while same concentrations of supernatants were examined for evaluating of proliferative activity against Adipose—derived Mesenchymal stem cells (MSCs). Both assessments were carried out by MTT assay and double PI and DAPI staining. GASX17, GBWy6 and GBPX3 isolates just induced HUVEC cell deaths and exhibited anti—proliferative activity while R2S12 not only reduced HUVEC cell proliferation but also enhanced proliferation of MSCs. R2S12 , GASX17, GBWy6 and GBPX3 isolates were characterized biochemically and six hydrophilic components were detected. This research established new bioactive compounds that could be used as an effective treatment in chemotherapy.

  5. Comparison of peptide nucleic acid fluorescence in situ hybridization assays with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry for the identification of bacteria and yeasts from blood cultures and cerebrospinal fluid cultures.

    Science.gov (United States)

    Calderaro, A; Martinelli, M; Motta, F; Larini, S; Arcangeletti, M C; Medici, M C; Chezzi, C; De Conto, F

    2014-08-01

    Peptide nucleic acid fluorescence in situ hybridization (PNA FISH) is a molecular diagnostic tool for the rapid detection of pathogens directly from liquid media. The aim of this study was to prospectively evaluate PNA FISH assays in comparison with culture-based matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) identification, as a reference method, for both blood and cerebrospinal fluid (CSF) cultures, during a 1-year investigation. On the basis of the Gram stain microscopy results, four different PNA FISH commercially available assays were used ('Staphylococcus aureus/CNS', 'Enterococcus faecalis/OE', 'GNR Traffic Light' and 'Yeasts Traffic Light' PNA FISH assays, AdvanDx). The four PNA FISH assays were applied to 956 positive blood cultures (921 for bacteria and 35 for yeasts) and 11 CSF cultures. Among the 921 blood samples positive for bacteria, PNA FISH gave concordant results with MALDI-TOF MS in 908/921 (98.64%) samples, showing an agreement of 99.4% in the case of monomicrobial infections. As regards yeasts, the PNA FISH assay showed a 100% agreement with the result obtained by MALDI-TOF MS. When PNA FISH assays were tested on the 11 CSF cultures, the results agreed with the reference method in all cases (100%). PNA FISH assays provided species identification at least one work-day before the MALDI-TOF MS culture-based identification. PNA FISH assays showed an excellent efficacy in the prompt identification of main pathogens, yielding a significant reduction in reporting time and leading to more appropriate patient management and therapy in cases of sepsis and severe infections.

  6. Peritoneal Fluid Reduces Angiogenesis-Related MicroRNA Expression in Cell Cultures of Endometrial and Endometriotic Tissues from Women with Endometriosis

    Science.gov (United States)

    Braza-Boïls, Aitana; Gilabert-Estellés, Juan; Ramón, Luis A.; Gilabert, Juan; Marí-Alexandre, Josep; Chirivella, Melitina; España, Francisco; Estellés, Amparo

    2013-01-01

    Endometriosis, defined as the presence of endometrium outside the uterus, is one of the most frequent gynecological diseases. It has been suggested that modifications of both endometrial and peritoneal factors could be implicated in this disease. Endometriosis is a multifactorial disease in which angiogenesis and proteolysis are dysregulated. MicroRNAs (miRNAs) are small non-coding RNAs that regulate the protein expression and may be the main regulators of angiogenesis. Our hypothesis is that peritoneal fluid from women with endometriosis could modify the expression of several miRNAs that regulate angiogenesis and proteolysis in the endometriosis development. The objective of this study has been to evaluate the influence of endometriotic peritoneal fluid on the expression of six miRNAs related to angiogenesis, as well as several angiogenic and proteolytic factors in endometrial and endometriotic cell cultures from women with endometriosis compared with women without endometriosis. Methods Endometrial and endometriotic cells were cultured and treated with endometriotic and control peritoneal fluid pools. We have studied the expression of six miRNAs (miR-16, -17-5p, -20a, -125a, -221, and -222) by RT-PCR and protein and mRNA levels of vascular endothelial growth factor-A, thrombospondin-1, urokinase plasminogen activator and plasminogen activator inhibitor-1 by ELISA and qRT-PCR respectively. Results Control and endometriotic peritoneal fluid pools induced a significant reduction of all miRNAs levels in endometrial and endometriotic cell cultures. Moreover, both peritoneal fluids induced a significant increase in VEGF-A, uPA and PAI-1 protein levels in all cell cultures without significant increase in mRNA levels. Endometrial cell cultures from patients treated with endometriotic peritoneal fluid showed lower expression of miRNAs and higher expression of VEGF-A protein levels than cultures from controls. In conclusion, this “in vitro” study indicates that

  7. Purification of monoclonal antibodies from clarified cell culture fluid using Protein A capture continuous countercurrent tangential chromatography.

    Science.gov (United States)

    Dutta, Amit K; Tran, Travis; Napadensky, Boris; Teella, Achyuta; Brookhart, Gary; Ropp, Philip A; Zhang, Ada W; Tustian, Andrew D; Zydney, Andrew L; Shinkazh, Oleg

    2015-11-10

    Recent studies using simple model systems have demonstrated that continuous countercurrent tangential chromatography (CCTC) has the potential to overcome many of the limitations of conventional Protein A chromatography using packed columns. The objective of this work was to optimize and implement a CCTC system for monoclonal antibody purification from clarified Chinese Hamster Ovary (CHO) cell culture fluid using a commercial Protein A resin. Several improvements were introduced to the previous CCTC system including the use of retentate pumps to maintain stable resin concentrations in the flowing slurry, the elimination of a slurry holding tank to improve productivity, and the introduction of an "after binder" to the binding step to increase antibody recovery. A kinetic binding model was developed to estimate the required residence times in the multi-stage binding step to optimize yield and productivity. Data were obtained by purifying two commercial antibodies from two different manufactures, one with low titer (∼ 0.67 g/L) and one with high titer (∼ 6.9 g/L), demonstrating the versatility of the CCTC system. Host cell protein removal, antibody yields and purities were similar to those obtained with conventional column chromatography; however, the CCTC system showed much higher productivity. These results clearly demonstrate the capabilities of continuous countercurrent tangential chromatography for the commercial purification of monoclonal antibody products.

  8. [The Supernatant Obtained from Cultured Anip973 Cells Enhances the Biological Activities of HUVEC].

    Science.gov (United States)

    Zhang, Cuicui; Li, Yueya; Wang, Jing; Li, Kai

    2015-11-01

    背景与目的 肿瘤微血管内皮细胞与正常组织来源的微血管内皮细胞相比,具有对生长因子反应性高、粘附因子表达高等特点,造成肿瘤血管通透性高且新生旺盛,导致肿瘤的快速生长和转移。因此了解肿瘤微环境中内皮细胞发生、形态及功能上的异质性特点,对进行合理的、个性化的、以血管内皮细胞为靶点的抗血管新生治疗有一定的指导作用。本实验旨在研究高转移肺腺癌Anip973细胞培养上清液对人脐静脉内皮细胞(human umbilical vein endothelial cell, HUVEC)表面标记及其生物学行为的影响。方法 以含有不同浓度Anip973细胞上清液的培养基(RPMI-1640+10%胎牛血清)培养的HUVEC为实验组、单纯培养基培养的HUVEC为对照组,以CCK-8检测各组细胞增殖率、基质胶成管实验检测成管能力、Transwell检测迁移能力、流式细胞术检测细胞表面标记CD105、CD31以及凋亡标记Annexin V的表达,并分析细胞表面标记的表达与生物学行为之间的关系。结果 与对照组相比,经Anip973细胞上清液培养的HUVEC增殖更多、且在浓度为250 µL/mL作用24 h最明显(P=0.002);成管及迁移能力亦均较对照组增强,分别在浓度为125 µL/mL(P<0.001)、250 µL/mL(P=0.002)时达最强;细胞表面标记CD105表达阳性率升高、浓度为250 µL/mL(P=0.028)最明显;CD31表达阳性率呈浓度依赖性升高;凋亡细胞比例降低。相关性分析显示:CD105的表达阳性率与细胞增殖及迁移能力呈正相关关系、CD31的表达与生物学行为并无明显相关性。结论 Anip973细胞上清液能促进HUVEC细胞增殖、迁移、血管形成,CD105也随之变化、并可在一定程度上反应其生物学行为。.

  9. Characterization of tetanus toxin, neat and in culture supernatant, by electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van; Hulst, A.G.; Roberts, B.; Wils, E.R.J.

    2002-01-01

    A method was developed for the liquid chromatographic-mass spectrometric (LC-MS) identification of extremely neurotoxic toxins. The method combines sample treatment in a safety containment and analysis of detoxified material in a common laboratory facility. The method was applied to the

  10. Characterization of tetanus toxin, neat and in culture supernatant, by electrospray mass spectrometry

    NARCIS (Netherlands)

    Baar, B.L.M. van; Hulst, A.G.; Roberts, B.; Wils, E.R.J.

    2002-01-01

    A method was developed for the liquid chromatographic-mass spectrometric (LC-MS) identification of extremely neurotoxic toxins. The method combines sample treatment in a safety containment and analysis of detoxified material in a common laboratory facility. The method was applied to the characteriza

  11. Residual urinary extracellular vesicles in ultracentrifugation supernatants after hydrostatic filtration dialysis enrichment

    Science.gov (United States)

    Musante, Luca; Tataruch-Weinert, Dorota; Kerjaschki, Dontscho; Henry, Michael; Meleady, Paula; Holthofer, Harry

    2017-01-01

    ABSTRACT Urinary extracellular vesicles (UEVs) appear an ideal source of biomarkers for kidney and urogenital diseases. The majority of protocols designed for their isolation are based on differential centrifugation steps. However, little is still known of the type and amount of vesicles left in the supernatant. Here we used an isolation protocol for UEVs which uses hydrostatic filtration dialysis as first pre-enrichment step, followed by differential centrifugation. Transmission electron microscopy (TEM), mass spectrometry (MS), western blot, ELISA assays and tuneable resistive pulse sensing (TRPS) were used to characterise and quantify UEVs in the ultracentrifugation supernatant. TEM showed the presence of a variety of small size vesicles in the supernatant while protein identification by MS matched accurately with the protein list available in Vesiclepedia. Screening and relative quantification for specific vesicle markers showed that the supernatant was preferentially positive for CD9 and TSG101. ELISA tests for quantification of exosome revealed that 14%, was left in the supernatant with a particle diameter of 110 nm and concentration of 1.54 × 1010/ml. Here we show a comprehensive characterisation of exosomes and other small size urinary vesicles which the conventional differential centrifugation protocol may lose.

  12. Detection of Haemophilus influenzae type b antigens in body fluids, using specific antibody-coated staphylococci.

    Science.gov (United States)

    Suksanong, M; Dajani, A S

    1977-01-01

    Protein A-rich staphylococci coated with Haemophilus influenzae type b antiserum agglutinate specifically with homologous bacterial cells or with cell-free supernatant fluids of cultures of the organism. Antibody-coated staphylococci were used to detect soluble antigens in body fluids of patients infected with H. influenzae type b. Cerebrospinal fluid from 36 cases of meningitis caused by this orgainsm showed positive coagglutination tests in 86% of patients prior to initiation of therapy. Antigens could be detected in 46% of sterile cerebrospinal fluid specimens obtained from the same cases 1 to 10 days after therapy. Soluble antigens were also detectable in sera (58%) and urine specimens (67%) of patients with H. influenzae type b septicemia, when such specimens were tested within 10 days of onset of illness. No antigen could be detected in body fluids beyond 10 days. The coagglutination test was positive in 57% of all body fluids examined; contercurrent immunoelectrophoresis (CCIE) was positive in only 27%. All specimens positive by CCIE were also positive by coagglutination. No false-positive reactions were noted by either test in body fluids from controls. The coagglutination test is simple, specific, and more sensitive than the CCIE method and could be a valuable tool for detecting antigens in body fluids of patients with various infections.

  13. Evaluation of Hanford Tank Supernatant Availability for Technetium Management Project Studies in FY16

    Energy Technology Data Exchange (ETDEWEB)

    Rapko, Brian M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2015-09-30

    This report examines the need for actual Hanford tank waste solutions to support tasks in the Technetium Management Program in fiscal year (FY) 2016. One key need is to identify both samples where a majority of the soluble technetium is present as pertechnetate and samples where it is not. The total amount of tank supernatant needed from any given tank waste supernatant was determined by polling the tasks leaders for their technology testing needs in FY16 and then arbitrarily ascribing a 10% process loss associated with consolidation and the Cs-137 removal needed to reduce the dose to a level suitable for testing in radiological fumehoods. These polling results identified a need for approximately 2.1 to 3.6 kg of any particular targeted Hanford tank waste supernatant.

  14. Modelling antagonic effect of lactic acid eacteria supernatants on some pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Augustus Caeser Franke Portella

    2009-11-01

    Full Text Available This work presents a statistical model of survival analysis for three pathogenic bacterial strains (Escherichia coli, Listeria monocytogenes and Staphylococcus aureus, when treated with neutralized and non-neutralized filtered supernatants broth from cultures of Lactobacillus acidhophilus, Lactobacillus rhamnosus and Lactobacillus sake. Survival analysis is a method employed to determine the period of time from an initial stage up to the occurrence of a particular event of interest, as death or a particular culture growth failure. In order to evaluate the potential efficacy of the ahead mentioned lactic acid bacteria when used as bioprotective starters in foods, experimental data were statistically treated and expressed by simple representative curves. Following the methodology of Cox and Kaplan-Meier, it was possible to make the selection of the best bioprotective lactic starter, as a predictive tool for evaluation of shelf life and prevention of eventual risks in fresh sausages and other similar food products.Este trabalho apresenta um modelo estatístico de análise de sobrevivência para três bactérias patogénicas (Escherichia coli, Listeria monocytogenes e Staphylococcus aureus, quando tratados com sobrenadantes filtrados neutralizado e não neutralizado provenientes de culturas de Lactobacillus acidhophilus, Lactobacillus rhamnosus e Lactobacillus sake. A Análise de sobrevivência é um método utilizado para determinar o período de tempo a partir de uma fase inicial até a ocorrência de um determinado evento de interesse, como a morte ou a inibição de uma particular cultura, a fim de avaliar a eficácia potencial das referidas bactérias lácticas quando usadas como bioproteção em alimentos. Os dados experimentais foram tratados estatisticamente, seguindo a metodologia de Cox e Kaplan-Meier e foi possível fazer a seleção dos melhores fermentos láticos bioprotectivos, como uma ferramenta para avaliação preditiva, vida de

  15. Long acting β2-agonist and corticosteroid restore airway glandular cell function altered by bacterial supernatant

    Directory of Open Access Journals (Sweden)

    Nawrocki-Raby Béatrice

    2010-01-01

    Full Text Available Abstract Background Staphylococcus aureus releases virulence factors (VF that may impair the innate protective functions of airway cells. The aim of this study was to determine whether a long-acting β2 adrenergic receptor agonist (salmeterol hydroxynaphthoate, Sal combined with a corticosteroid (fluticasone propionate, FP was able to regulate ion content and cytokine expression by airway glandular cells after exposure to S. aureus supernatant. Methods A human airway glandular cell line was incubated with S. aureus supernatant for 1 h and then treated with the combination Sal/FP for 4 h. The expression of actin and CFTR proteins was analyzed by immunofluorescence. Videomicroscopy was used to evaluate chloride secretion and X-ray microanalysis to measure the intracellular ion and water content. The pro-inflammatory cytokine expression was assessed by RT-PCR and ELISA. Results When the cells were incubated with S. aureus supernatant and then with Sal/FP, the cellular localisation of CFTR was apical compared to the cytoplasmic localisation in cells incubated with S. aureus supernatant alone. The incubation of airway epithelial cells with S. aureus supernatant reduced by 66% the chloride efflux that was fully restored by Sal/FP treatment. We also observed that Sal/FP treatment induced the restoration of ion (Cl and S and water content within the intracellular secretory granules of airway glandular cells and reduced the bacterial supernatant-dependent increase of pro-inflammatory cytokines IL8 and TNFα. Conclusions Our results demonstrate that treatment with the combination of a corticosteroid and a long-acting β2 adrenergic receptor agonist after bacterial infection restores the airway glandular cell function. Abnormal mucus induced by defective ion transport during pulmonary infection could benefit from treatment with a combination of β2 adrenergic receptor agonist and glucocorticoid.

  16. Neuronal activation by mucosal biopsy supernatants from irritable bowel syndrome patients is linked to visceral sensitivity.

    Science.gov (United States)

    Buhner, Sabine; Braak, Breg; Li, Qin; Kugler, Eva Maria; Klooker, Tamira; Wouters, Mira; Donovan, Jemma; Vignali, Sheila; Mazzuoli-Weber, Gemma; Grundy, David; Boeckxstaens, Guy; Schemann, Michael

    2014-10-01

    Based on the discomfort/pain threshold during rectal distension, irritable bowel syndrome (IBS) patients may be subtyped as normo- or hypersensitive. We previously showed that mucosal biopsy supernatants from IBS patients activated enteric and visceral afferent neurons. We tested the hypothesis that visceral sensitivity is linked to the degree of neuronal activation. Normo- and hypersensitive IBS patients were distinguished by their discomfort/pain threshold to rectal balloon distension with a barostat. Using potentiometric and Ca(2+) dye imaging, we recorded the response of guinea-pig enteric submucous and mouse dorsal root ganglion (DRG) neurons, respectively, to mucosal biopsy supernatants from normosensitive (n = 12 tested in enteric neurons, n = 9 tested in DRG) and hypersensitive IBS patients (n = 9, tested in both types of neurons). In addition, we analysed the association between neuronal activation and individual discomfort/pain pressure thresholds. The IBS supernatants evoked Ca(2+) transients in DRG neurons and spike discharge in submucous neurons. Submucous and DRG neurons showed significantly stronger responses to supernatants from hypersensitive IBS patients as reflected by higher spike frequency or stronger [Ca(2+)]i transients in a larger proportion of neurons. The neuroindex as a product of spike frequency or [Ca(2+)]i transients and proportion of responding neurons correlated significantly with the individual discomfort/pain thresholds of the IBS patients. Supernatants from hypersensitive IBS patients caused stronger activation of enteric and DRG neurons. The level of activation correlated with the individual discomfort/pain threshold pressure values. These findings support our hypothesis that visceral sensitivity is linked to activation of peripheral neurons by biopsy supernatants.

  17. Boildown Study on Supernatant Liquid Retrieved from AW-106 in December 2012

    Energy Technology Data Exchange (ETDEWEB)

    Page, Jason S.

    2013-06-04

    This document reports the results of a boildown study using a composite created from supernatant liquid grab samples retrieved from tank 241-AW-106 in December of 2012. The composite was made using predetermined volumes of the grab samples which accounted for layering of the supernatant liquid in the tank. The finished composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (24 - 27 °C). The density of the test composite was measured in the hot cell immediately before the boildown study and was 1.266 g/mL at 27.1 °C.

  18. Highly Efficient In Vitro Production of Bovine Blastocyst in Cell-Free Sequential Synthetic Oviductal Fluid vs. TCM199 Vero Cell Co-Culture System

    Directory of Open Access Journals (Sweden)

    Sayyed Morteza Hosseini

    2008-01-01

    Full Text Available Background: The aim of this study was to establish a cell-free sequential culture system that cansupport high levels of in vitro embryo development and blastocyst formation from bovine zygotes.To this end, this investigation was carried out to evaluate the effects of glucose, serum and EDTAon bovine zygote in vitro development.Materials and Methods: Bovine presumptive zygotes were derived from oocytes matured, andfertilized in vitro and cultured in synthetic oviductal fluid sequential medium in a two-steps manner;SOF 1 for the first 3 days and SOF 2 for the second 5-6 days of in vitro embryo development. Inorder to evaluate the effect of different modifications of the basic medium on embryo development,glucose was added to the second phase (SOF A, serum was added to the first phase (SOF C andEDTA alone (SOF D or in combination with serum (SOF E was added into the first phase of invitro embryo culture. The results of each composition were compared with each other and with theresults of embryo development in TCM199 vero cell co-culture system.Results: Glucose addition to the second phase of embryo culture, improved the developmentalcompetency; however, the differences were not significant. Serum addition to the first phase ofembryo culture, significantly improved the developmental competency of embryos beyond thecleavage stage, compared to all the treatment and TCM199 co-culture groups. EDTA supplementationof culture medium, either alone or in combination with serum, significantly inhibits the embryodevelopment beyond the morula stage.Conclusion: The results indicated that culture of bovine presumptive zygotes in two steps cell-freeculture system, can support embryo development, and addition of serum throughout the culture andglucose to the second step significantly increased overall developmental competency compared toTCM199 co-culture system.

  19. The effect of NaCl substitution with KCl on proteinase activities of cell-free extract and cell-free supernatant at different pH levels and salt concentrations: Lactobacillus acidophilus and Lactobacillus casei.

    Science.gov (United States)

    Ayyash, M M; Sherkat, F; Shah, N P

    2013-01-01

    The aim of this study was to investigate the effect of substitution of NaCl with KCl at different pH levels and salt concentrations on proteinase activity of cell-free extract and cell-free supernatant of the probiotics Lactobacillus acidophilus and Lactobacillus casei. de Man, Rogosa, and Sharpe broth aliquots were mixed with 2 pure salts (NaCl and KCl) and 2 salt concentrations at 2 concentration levels (5 and 10%), inoculated with Lactobacillus acidophilus or Lactobacillus casei, and incubated aerobically at 37°C for 22 h. The cultures were then centrifuged at 4,000×g for 30 min, and the collected cell pellets were used to prepare cell-wall proteinases and the supernatants used as a source of supernatant (extracellular) proteinases. The proteolytic activity and protein content of both portions were determined. After incubation of both portions with 3 milk caseins (α-, β-, κ-casein), the supernatants were individually subjected to analysis of angiotensin-converting enzyme (ACE)-inhibitory activity and proteolytic activity using the o-phthalaldehyde method. Significant differences were observed in ACE-inhibitory and proteolytic activities between salt substitution treatments of cell-free extract and cell-free supernatant from both probiotic strains at the same salt concentration and pH level.

  20. Surface functionalization of tissue culture polystyrene plates with hydroxyapatite under body fluid conditions and its effect on differentiation behaviors of mesenchymal stem cells.

    Science.gov (United States)

    Iijima, Kazutoshi; Suzuki, Ryo; Iizuka, Ayako; Ueno-Yokohata, Hitomi; Kiyokawa, Nobutaka; Hashizume, Mineo

    2016-11-01

    The surfaces of polystyrene (PS) cell culture plates were functionalized with hydroxyapatite (HAp) under body fluid conditions utilizing protein adsorption layers and a pretreatment with an alternate soaking process (ASP) using solutions containing calcium and phosphate ions. Adsorption layers of human serum albumin (HSA) formed on the surface of each well of commercial 24-well PS plates by solution processes. CaCl2 and K2HPO4 solutions were alternately added to the wells, the plates were incubated to form the precursors, and this was followed by the addition of simulated body fluid (SBF) and a further incubation for 24h. These treatments resulted in the surfaces of the PS cell culture plates being completely covered with bone-like HAp. The coating of PS plates with HAp promoted the adhesion of mesenchymal stem cells (MSCs) and maintained cell growth that was as fast as that on tissue culture-treated PS (TCPS) plates. Osteogenic differentiation was greater, whereas adipogenic and chondrogenic differentiation was less in the culture on HAp-coated PS plates than in that on TCPS plates. The present method is useful for preparing HAp-coated PS plates at clean benches without the need for any expensive apparatus. HAp coated on PS plates by this method was a bone-like apatite with high bioactivity; therefore, the present HAp-coated PS plates are promising materials for assays of bone-related cells in the bone remodeling process.

  1. Fate of cyanobacteria in drinking water treatment plant lagoon supernatant and sludge

    Energy Technology Data Exchange (ETDEWEB)

    Pestana, Carlos J.; Reeve, Petra J.; Sawade, Emma [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Voldoire, Camille F. [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); École Européenne de Chimie, Polymères et Matériaux (ECPM), Strasbourg 67087 (France); Newton, Kelly; Praptiwi, Radisti [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Collingnon, Lea [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); École Européenne de Chimie, Polymères et Matériaux (ECPM), Strasbourg 67087 (France); Dreyfus, Jennifer [Allwater, Adelaide Services Alliance, Wakefield St, Adelaide, SA 5001 (Australia); Hobson, Peter [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia); Gaget, Virginie [University of Adelaide, Ecology and Environmental Sciences, School of Biological Sciences, Adelaide, SA 5005 (Australia); Newcombe, Gayle, E-mail: gayle.newcombe@sawater.com.au [Australian Water Quality Centre, South Australian Water Corporation, Adelaide, SA 5000 (Australia)

    2016-09-15

    In conventional water treatment processes, where the coagulation and flocculation steps are designed to remove particles from drinking water, cyanobacteria are also concentrated into the resultant sludge. As a consequence, cyanobacteria-laden sludge can act as a reservoir for metabolites such as taste and odour compounds and cyanotoxins. This can pose a significant risk to water quality where supernatant from the sludge treatment facility is returned to the inlet to the plant. In this study the complex processes that can take place in a sludge treatment lagoon were investigated. It was shown that cyanobacteria can proliferate in the conditions manifest in a sludge treatment lagoon, and that cyanobacteria can survive and produce metabolites for at least 10 days in sludge. The major processes of metabolite release and degradation are very dependent on the physical, chemical and biological environment in the sludge treatment facility and it was not possible to accurately model the net effect. For the first time evidence is provided to suggest that there is a greater risk associated with recycling sludge supernatant than can be estimated from the raw water quality, as metabolite concentrations increased by up to 500% over several days after coagulation, attributed to increased metabolite production and/or cell proliferation in the sludge. - Highlights: • Cyanobacteria in water treatment sludge significantly impact supernatant quality • Cyanobacteria can survive, and thrive, in sludge lagoon supernatant and in treatment sludge • Metabolite concentrations in cyanobacteria in sludge can increase up to 500% • The risk associated with supernatant recycling was assessed relative to available treatment barriers.

  2. Faecalibacterium prausnitzii supernatant improves intestinal barrier function in mice DSS colitis.

    Science.gov (United States)

    Carlsson, Anders H; Yakymenko, Olena; Olivier, Isabelle; Håkansson, Fathima; Postma, Emily; Keita, Asa V; Söderholm, Johan D

    2013-10-01

    OBJECTIVE. The intestinal microbiota plays a substantial role in the pathogenesis of inflammatory bowel disease (IBD). Faecalibacterium prausnitzii (FP) is underrepresented in IBD patients and have been suggested to have anti-inflammatory effects in mice. Increased intestinal permeability is common in IBD but the relationship between FP and intestinal barrier function has not been investigated. Our aim was to study treatment with FP supernatant on intestinal barrier function in a dextran sodium sulfate (DSS) colitis mice model. MATERIAL AND METHODS. C57BL/6 mice received 3% DSS in tap water ad libitum during five days to induce colitis. From day 3 the mice received a daily gavage with FP supernatant or broth during seven days. Ileum and colon were mounted in Ussing chambers for permeability studies with (51)Cr-EDTA and Escherichia coli K-12. Colon was saved for Western blot analyses of tight junction proteins. RESULTS. DSS-treated mice showed significant weight loss and colon shortening. Gavage with FP supernatant resulted in a quicker recovery after DSS treatment and less extensive colonic shortening. Ileal mucosa of DSS mice showed a significant increase in (51)Cr-EDTA-passage compared to controls. (51)Cr-EDTA passage was significantly decreased in mice receiving FP supernatant. No significant differences were observed in passage of E. coli K12. Western blots showed a trend to increased claudin-1 and claudin-2 expressions in DSS mice. CONCLUSIONS. Supernatant of FP enhances the intestinal barrier function by affecting paracellular permeability, and may thereby attenuate the severity of DSS-induced colitis in mice. These findings suggest a potential role of FP in the treatment of IBD.

  3. Development and optimization of quantitative PCR for the diagnosis of invasive aspergillosis with bronchoalveolar lavage fluid

    Directory of Open Access Journals (Sweden)

    Hackman Robert C

    2008-05-01

    Full Text Available Abstract Background The diagnosis of invasive pulmonary aspergillosis (IPA remains challenging. Culture and histopathological examination of bronchoalveolar lavage (BAL fluid are useful but have suboptimal sensitivity and in the case of culture may require several days for fungal growth to be evident. Detection of Aspergillus DNA in BAL fluid by quantitative PCR (qPCR offers the potential for earlier diagnosis and higher sensitivity. It is important to adopt quality control measures in PCR assays to address false positives and negatives which can hinder accurate evaluation of diagnostic performance. Methods BAL fluid from 94 episodes of pneumonia in 81 patients was analyzed. Thirteen episodes were categorized as proven or probable IPA using Mycoses Study Group criteria. The pellet and the supernatant fractions of the BAL were separately assayed. A successful extraction was confirmed with a human 18S rRNA gene qPCR. Inhibition in each qPCR was measured using an exogenous DNA based internal amplification control (IAC. The presence of DNA from pathogens in the Aspergillus genus was detected using qPCR targeting fungal 18S rRNA gene. Results Human 18S rRNA gene qPCR confirmed successful DNA extraction of all samples. IAC detected some degree of initial inhibition in 11 samples. When culture was used to diagnose IPA, the sensitivity and specificity were 84.5% and 100% respectively. Receiver-operating characteristic analysis of qPCR showed that a cutoff of 13 fg of Aspergillus genomic DNA generated a sensitivity, specificity, positive and negative predictive value of 77%, 88%, 50%, 96% respectively. BAL pellet and supernatant analyzed together resulted in sensitivity and specificity similar to BAL pellet alone. Some patients did not meet standard criteria for IPA, but had consistently high levels of Aspergillus DNA in BAL fluid by qPCR. Conclusion The Aspergillus qPCR assay detected Aspergillus DNA in 76.9% of subjects with proven or probable IPA when

  4. Survival of the North American strain of viral hemorrhagic septicemia virus (VHSV) in filtered seawater and seawater containing ovarian fluid, crude oil and serum-enriched culture medium

    Science.gov (United States)

    Kocan, R.M.; Hershberger, P.K.; Elder, N.E.

    2001-01-01

     The North American strain of viral hemorrhagic septicemia virus (NA-VHSV) could be recovered for up to 40 h in natural filtered seawater (27 ppt) with a 50% loss of infectivity after approximately 10 h at 15°C. Addition of 10 ppb North Slope crude oil to the seawater had no effect on virus survival. However, when various concentrations of teleost ovarian fluid were added to seawater, virus could be recovered after 72 h at 0.01% ovarian fluid and after 96 h at 1.0%. When cell culture medium supplemented with 10% fetal bovine serum was added to the seawater, 100% of the virus could be recovered for the first 15 d and 60% of the virus remained after 36 d. These findings quantify NA-VHSV infectivity in natural seawater and demonstrate that ovarian fluid, which occurs naturally during spawning events, significantly prolongs the survival and infectivity of the virus. The extended stabilization of virus in culture medium supplemented with serum allows for low titer field samples to be collected and transported in an unfrozen state without significant loss of virus titer.

  5. [Effects of Bushen Shuji Granule on IL-6 in the culture fluid sample of fibroblast cells from the synovial liquid of the ankylosing spondylitis patients].

    Science.gov (United States)

    Zhu, Jun-Ling; Yan, Xiao-Ping

    2012-02-01

    To explore the effects of Bushen Shuji Granule (BSG) on inhibiting the interleukin 6 (IL-6) level in the synovial fluid sample of fibroblast cells from the ankylosing spondylitis (AS) patients. Using serum pharmacologic method, the IL-6 level in the culture fluid sample of fibroblast cells was observed by ELISA method with different concentrations of medicated serum containing BSG. The IL-6 level at the mRNA level was detected using reverse transcriptase-polymerase chain reaction (RT-PCR). The vehicle serum and sulfasalazine (SSZ) serum were taken as controls. Results of ELISA showed the IL-6 level in the AS group was more than that in the vehicle serum group with obvious statistical difference. BSG could obviously inhibited the IL-6 level, showing statistical difference when compared with the vehicle serum group. Besides, obvious dose-dependent correlation existed between BSG and its inhibition on fibroblast proliferation. And the IL-6 level at the mRNA level in the AS group was higher than that in the vehicle serum group, showing statistical difference by semi-quantitative analysis. BSG could play its clinical role of anti-inflammation and anti-fibrosis through inhibiting the IL-6 level in the culture fluid sample of fibroblast cells.

  6. Comparison of excretory-secretory antigen and positive faecal supernatant antigen in the detection of Echinococcus granulosus infection in dogs by CIEP

    Directory of Open Access Journals (Sweden)

    P. R. Prathiush

    Full Text Available Coproantigen detection of Echinococcosis in dogs by counter immunoelectrophoresis was standardized. Adult Echinococcus granulosus worms were obtained from intestine of a necropsied positive dog. Excretory-secretory antigen was prepared by culturing adult worms in Medium 199 (pH 7.4. Faeces of positive dog were collected and fecal supernatant was prepared and used for coproantigen detection. CIEP was carried out using tris-borate buffer (pH 8.0 at a constant current of 8mA/slide for 60 minutes. CIEP detected infection with both the antigens. [Vet World 2009; 2(11.000: 421-422

  7. Thalidomide and pentoxifylline block the renal effects of supernatants of macrophages activated with Crotalus durissus cascavella venom

    Directory of Open Access Journals (Sweden)

    A.M.C. Martins

    2004-10-01

    Full Text Available Because thalidomide and pentoxifylline inhibit the synthesis and release of tumor necrosis factor-alpha (TNF-alpha, we determined the effect of these drugs on the renal damage induced by supernatants of macrophages activated with Crotalus durissus cascavella venom in order to identify the role of TNF-alpha in the process. Rat peritoneal macrophages were collected with RPMI medium and stimulated in vitro with C.d. cascavella venom (10 µg/ml in the absence and presence of thalidomide (15 µM or pentoxifylline (500 µM for 1 h and washed and kept in culture for 2 h. Supernatant (1 ml was tested on an isolated perfused rat kidney (N = 6 for each group. The first 30 min of each experiment were used as control. The supernatant was added to the perfusion system. All experiments lasted 120 min. The toxic effect of the preparation of venom-stimulated macrophages on renal parameters was determined. At 120 min, thalidomide (Thalid and pentoxifylline (Ptx inhibited (P < 0.05 the increase in perfusion pressure caused by the venom (control = 114.0 ± 1.3; venom = 137.1 ± 1.5; Thalid = 121.0 ± 2.5; Ptx = 121.4 ± 4.0 mmHg, renal vascular resistance (control = 4.5 ± 0.2; venom = 7.3 ± 0.6; Thalid = 4.5 ± 0.9; Ptx = 4.8 ± 0.6 mmHg/ml g-1 min-1, urinary flow (control = 0.23 ± 0.001; venom = 0.44 ± 0.01; Thalid = 0.22 ± 0.007; Ptx = 0.21 ± 0.009 ml g-1 min-1, glomerular filtration rate (control = 0.72 ± 0.06; venom = 1.91 ± 0.11; Thalid = 0.75 ± 0.04; Ptx = 0.77 ± 0.05 ml g-1 min-1 and the decrease in percent tubular sodium transport (control = 77.0 ± 0.9; venom = 73.9 ± 0.66; Thalid = 76.6 ± 1.1; Ptx = 81.8 ± 2.0%, percent tubular chloride transport (control = 77.1 ± 1.2; venom = 71.4 ± 1.1; Thalid = 77.6 ± 1.7; Ptx = 76.8 ± 1.2%, and percent tubular potassium transport (control = 72.7 ± 1.1; venom = 63.0 ± 1.1; Thalid = 72.6 ± 1.0; Ptx = 74.8 ± 1.0%, 30 min before and during the stimulation of macrophages with C.d. cascavella venom

  8. Development of a rapid lateral flow immunoassay test for detection of exosomes previously enriched from cell culture medium and body fluids

    Directory of Open Access Journals (Sweden)

    Myriam Oliveira-Rodríguez

    2016-08-01

    Full Text Available Exosomes are cell-secreted nanovesicles (40–200 nm that represent a rich source of novel biomarkers in the diagnosis and prognosis of certain diseases. Despite the increasingly recognized relevance of these vesicles as biomarkers, their detection has been limited due in part to current technical challenges in the rapid isolation and analysis of exosomes. The complexity of the development of analytical platforms relies on the heterogeneous composition of the exosome membrane. One of the most attractive tests is the inmunochromatographic strips, which allow rapid detection by unskilled operators. We have successfully developed a novel lateral flow immunoassay (LFIA for the detection of exosomes based on the use of tetraspanins as targets. We have applied this platform for the detection of exosomes purified from different sources: cell culture supernatants, human plasma and urine. As proof of concept, we explored the analytical potential of this LFIA platform to accurately quantify exosomes purified from a human metastatic melanoma cell line. The one-step assay can be completed in 15 min, with a limit of detection of 8.54×105 exosomes/µL when a blend of anti-CD9 and anti-CD81 were selected as capture antibodies and anti-CD63 labelled with gold nanoparticles as detection antibody. Based on our results, this platform could be well suited to be used as a rapid exosome quantification tool, with promising diagnostic applications, bearing in mind that the detection of exosomes from different sources may require adaptation of the analytical settings to their specific composition.

  9. Rapid identification and multiple susceptibility testing of pathogens from positive-culture sterile body fluids by a combined MALDI-TOF Mass Spectrometry and Vitek Susceptibility system

    Directory of Open Access Journals (Sweden)

    Ru Yue Tian

    2016-04-01

    Full Text Available Infections of the bloodstream, central nervous system, peritoneum, joints and other sterile areas are associated with high morbidity and sequelae risk. Timely initiation of effective antimicrobial therapy is crucial to improving patient prognosis. However, standard final identification and antimicrobial susceptibility tests (ASTs are reported 16–48 hours after a positive alert. For a rapid, effective and low-cost diagnosis, we combined matrix-assisted laser desorption/ionization time of flight mass spectrometry with a Vitek AST system, and performed rapid microbial identification (RMI and rapid multiple AST (RMAST on non-duplicated positive body fluid cultures collected from a hospital in Shanghai, China. Sterile body fluid positive culture and blood positive culture caused by Gram negative (GN or polymicrobial were applied to the MALDI–TOF measurement directly. When positive blood culture caused by Gram positive (GP bacteria or yeasts, they were resuspended in 1 ml brain heart infusion for 2h or 4h enrichment, respectively. Regardless of enrichment, the RMI (completed in 40 min per sample accurately identified GN and GP bacteria (98.9% and 87.2%, respectively, fungi (75.7% and anaerobes (94.7%. Dominant species in multiple cultures and bacteria that failed to grow on the routing plates were correctly identified in 81.2% and 100% of cases, respectively. The category agreements of RMAST results, determined in the presence of various antibiotics, were similarly to previous studies. The RMI and RMAST results not only reduce the turnaround time of the patient report by 18–36 hours, but also indicate whether a patient’s antibiotic treatment should be accelerated, ceased or de-escalated, and adjusted the essential drugs modification for an optimised therapy.

  10. Optmization of culture media for Bifidobacterium bifidum and Bifidobacterium crudilactis and study of the antimicrobial effect of culture supernatants

    OpenAIRE

    Bondue, Pauline; Delcenserie, Véronique

    2015-01-01

    Complex oligosaccharides from human milk (HMO) contribute to infant health. Bifidobacteriumbifidum mainly found in breast-fed infant microbiota has all the enzymatic machinery for degradation of HMO. On the other hand, whey is rich in complex bovin milk oligosaccharides (BMO) very similar to HMO, including 3’-sialyllactose (3’SL). They are very likely to be metabolised by B. bifidum too, but also by B. crudilactis, a bovine origin strain. Fermentation of HMO or BMO by bifidobacteria can resul...

  11. Identification of Fusarium oxysporum in the fluid of hydroponic culture using DNA analysis; DNA kaiseki ni yoru suiko saibai yoekichu no horenso ichobyokin

    Energy Technology Data Exchange (ETDEWEB)

    Oyama, K.; Kawabata, T. [Chugoku Electric Power Co. Inc., Hiroshima (Japan)

    1999-09-24

    In the case of circularly using a fluid of the hydroponic culture for crops such as vegetable, the destructive damage is caused when specified harmful germs multiply in the growing crop. therefore, it is necessary to find the germs in question out and deal with them as quickly as possible. In this paper, studies were made on a sure identification technology of F. oxysporum using DNA analysis and the measures to prevent crop damages caused by diseases. The results of the study are reported. (translated by NEDO)

  12. Identification and culture of neural stem cells isolated from adult rat subventricular zone following fluid percussion brain injury

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Objective To analyze proliferation and differentiation of glial fibrillary acid protein(GFAP)-and nestin-positive(GFAP+/nestin+)cells isolated from the subventricular zone following fluid percussion brain injury to determine whether GFAP+/nestin+ cells exhibit characteristics of neural stem cells.Methods Male Sprague-Dawley rats,aged 12 weeks and weighing 200-250 g,were randomly and evenly assigned to normal control group and model group.In the model group,a rat model of fluid percussion brain injury was es...

  13. Inhibition of Candida albicans biofilm formation and modulation of gene expression by probiotic cells and supernatant.

    Science.gov (United States)

    James, K M; MacDonald, K W; Chanyi, R M; Cadieux, P A; Burton, J P

    2016-04-01

    Oral candidiasis is a disease caused by opportunistic species of Candida that normally reside on human mucosal surfaces. The transition of Candida from budding yeast to filamentous hyphae allows for covalent attachment to oral epithelial cells, followed by biofilm formation, invasion and tissue damage. In this study, combinations of Lactobacillus plantarum SD5870, Lactobacillus helveticus CBS N116411 and Streptococcus salivarius DSM 14685 were assessed for their ability to inhibit the formation of and disrupt Candida albicans biofilms. Co-incubation with probiotic supernatants under hyphae-inducing conditions reduced C. albicans biofilm formation by >75 % in all treatment groups. Likewise, combinations of live probiotics reduced biofilm formation of C. albicans by >67 %. When live probiotics or their supernatants were overlaid on preformed C. albicans biofilms, biofilm size was reduced by >63 and >65 % respectively. Quantitative real-time PCR results indicated that the combined supernatants of SD5870 and CBS N116411 significantly reduced the expression of several C. albicans genes involved in the yeast-hyphae transition: ALS3 (adhesin/invasin) by 70 % (P biofilm formation) by >99 % (P formation of and removing preformed C. albicans biofilms. Our novel results point to the downregulation of several Candida genes critical to the yeast-hyphae transition, biofilm formation, tissue invasion and cellular damage.

  14. The Effect of Anaerobic and Aerobic Fish Sludge Supernatant on Hydroponic Lettuce

    Directory of Open Access Journals (Sweden)

    Simon Goddek

    2016-06-01

    Full Text Available The mobilization of nutrients from fish sludge (i.e., feces and uneaten feed plays a key role in optimizing the resource utilization and thus in improving the sustainability of aquaponic systems. While several studies have documented the aerobic and anaerobic digestion performance of aquaculture sludge, the impact of the digestate on plant growth has yet to be understood. The present study examines the impact of either an aerobic or an anaerobic digestion effluent on lettuce plant growth, by enriching a mixture of aquaculture and tap water with supernatants from both aerobic and anaerobic batch reactors. The lettuce plants grown in the hydroponic system supplied with supernatant from an anaerobic reactor had significantly better performance with respect to weight gain than both, those in the system where supernatant from the aerobic reactor was added, as well as the control system. It can be hypothesized that this effect was caused by the presence of NH4+ as well as dissolved organic matter, plant growth promoting rhizobacteria and fungi, and humic acid, which are predominantly present in anaerobic effluents. This study should therefore be of value to researchers and practitioners wishing to further develop sludge remineralization in aquaponic systems.

  15. Senescence-associated β-galactosidase staining in fish cell lines and primary cultures from several tissues and species, including rainbow trout coelomic fluid and milt.

    Science.gov (United States)

    Vo, Nguyen T K; Mikhaeil, Michael S; Lee, Lucy E J; Pham, Phuc H; Bols, Niels C

    2015-04-01

    Cell lines and primary cultures from several teleost tissues and species were stained for senescence-associated β-galactosidase (SA β-Gal), revealing four general outcomes. (1) For long-standing fish cell lines that can be considered immortal, little or no SA β-Gal staining was observed, regardless of the culture conditions. (2) For a new walleye cell line from the bulbus arteriosus (WEBA), most cells stained for SA β-Gal even after 40 passages. This suggested that high SA β-Gal activity was a unique property of WEBA, perhaps reflecting their endothelial character, rather than cellular senescence. (3) For cell lines developed from the walleye caudal fin and from somatic cells in rainbow trout coelomic fluid, no SA β-Gal staining was observed in the earliest cultures to over 70 passages later. This suggested that cells from these anatomical sites do not undergo senescence in vitro. (4) By contrast, for cell lines developed from the walleye brain and from somatic cells in rainbow trout milt, most cells in the early-stage cultures stained for SA β-Gal, but as these were developed into cell lines, SA β-Gal-negative cells became dominant. This suggested that if cellular senescence occurred in vitro, this happened early in these cultures and subsequently a few SA β-Gal-negative cells went onto to form the cell line. Overall, the presence of SA β-Gal-positive cells in cultures could be interpreted in several ways, whereas their absence predicted that in these cultures, cells would proliferate indefinitely.

  16. Role of culture of postoperative drainage fluid in the prediction of infection of the surgical site after major oncological operations of the head and neck.

    Science.gov (United States)

    Candau-Alvarez, A; Linares-Sicilia, M J; Dean-Ferrer, A; Pérez-Navero, J L

    2015-02-01

    Infection of the surgical site after major oncological operations of the head and neck increases mortality and morbidity. The aim of this prospective pilot study was to assess the efficacy of culturing the exudate from the drain after cervical neck dissection to see if it predicted such infection. We studied 40/112 patients with squamous cell cancer of the head and neck who were treated during the last two years and met our inclusion criteria. Six patients developed infections (15%). Reconstruction with pedicled rather than local or microvascular flaps, duration of operation of over 7 hours, the presence of a tracheostomy, and bilateral neck dissection were considered risk factors (p=0.01). Culture of drainage fluid on postoperative day 3 that grew no pathogens predicted that the site would not become infected, with a negative predictive value of 96%. Copyright © 2014 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

  17. Prenatal diagnosis of congenital toxoplasmosis: comparative value of fetal blood and amniotic fluid using serological techniques and cultures.

    Science.gov (United States)

    Fricker-Hidalgo, H; Pelloux, H; Muet, F; Racinet, C; Bost, M; Goullier-Fleuret, A; Ambroise-Thomas, P

    1997-09-01

    The prenatal diagnosis of congenital toxoplasmosis is mainly based on biological tests performed on fetal blood and amniotic fluid. We studied the performance of neonatal diagnosis procedures and the results of fetal blood and amniotic fluid analysis. Of 127 women who contracted toxoplasmosis and underwent prenatal diagnosis, the postnatal serological follow-up was long enough to definitively diagnose congenital toxoplasmosis in 19 cases and to exclude it in 27 cases. Prenatal diagnosis allowed the detection of 94.7 per cent (18/19) of the infected fetuses. The sensitivities of tests in amniotic fluid and fetal blood were equivalent, 88.2 per cent (15/17) and 87.5 per cent (14/16), respectively. In fetal blood, biological techniques were positive in 12/16 cases and in 2/16 cases, serological tests were the only positive sign. The specificities of tests in amniotic fluid and fetal blood were respectively 100 per cent (23/23) and 86.3 per cent (19/22) (three false-positive serological results). These results, added to the lower morbidity of amniocentesis compared with cordocentesis, might lead to cordocentesis being abandoned in the prenatal diagnosis of congenital toxoplasmosis.

  18. Low predictive value of positive transplant perfusion fluid cultures for diagnosing postoperative infections in kidney and kidney-pancreas transplantation.

    LENUS (Irish Health Repository)

    Cotter, Meaghan P

    2012-12-01

    Infection following transplantation is a cause of morbidity and mortality. Perfusion fluid (PF) used to preserve organs between recovery and transplantation represents a medium suitable for the growth of microbes. We evaluated the relevance of positive growth from PF sampled before the implantation of kidney or kidney-pancreas (KP) allografts.

  19. Perbandingan Angka Fertilitas dan Hambatan Perkembangan Embrio Mencit yang Dikultur dalam Medium M16 dan Human Tubal Fluid (THE COMPARISON OF MICE FERTILITY RATE AND EMBRYONIC DEVELOPMENT CELL BLOCK WHEN CULTURED IN M16 AND HUMAN TUBAL FLUID MEDIA

    Directory of Open Access Journals (Sweden)

    Widjiati .

    2013-07-01

    Full Text Available The aim of this research was to compare the fertility rate and embryonic development cell block ofmice when cultured in M16 and Human Tubal Fluid (HTF media, respectively. Two months old femaleBalbC mice were super ovulated using Pregnant Mare Serum Gonadotrophin (PMSG and Human ChorionicGonadotrophin (HCG prior to mating with vasectomies mice. At 17 hours post mating the mice wassacrificed for the collections of egg cells and spermatozoa. Egg cells were collected by tearing the fertilizationsac, while the sperm were collected from caudal epididymis. After the collection, both the egg cells andsperm were put in Petri dish containing M16 and HTF media and kept in 5% CO2 incubator at 370C for onehour prior to the in vitro fertilization (IVF was performed. In vitro fertilization was performed in 5% CO2 incubator at 370C and kept for 24 hours in M16 and in HTF culture media. The results showed thatfertilization rate was 98.09% and 99.57%; cell block embryonic development was 85.09% and 83.36%when cultured in M16 and HTF media, respectively. In conclusion, HTF media can be used for culturingmouse embryo.

  20. Enhanced osteogenesis of human alveolar bone-derived mesenchymal stem cells for tooth tissue engineering using fluid shear stress in a rocking culture method.

    Science.gov (United States)

    Lim, Ki-Taek; Kim, Jangho; Seonwoo, Hoon; Chang, Jung Uk; Choi, Hwajung; Hexiu, Jin; Cho, Woo Jae; Choung, Pill-Hoon; Chung, Jong Hoon

    2013-02-01

    This study instituted a simple approach to stimulate alveolar bone regeneration for tooth tissue engineering by controlling effects of low fluid dynamic shear stress (LFDSS) on growth and differentiation in vitro. Human alveolar bone-derived mesenchymal stem cells (hABMSCs) harvested from human mandibular alveolar bone were cultured with LFDSS to generate cultures containing bone-like formations. To distinguish between osteodifferentiation and bone-like formation, cells were cultured either with or without fluid shear stress. The calcium content and alkaline phosphatase (ALP) activity of hABMSCs were used as indicators of osteogenesis. Cell viability and proliferation after stimulating with LFDSS for 10-60 min/day were higher than with longer stimulations. Mineralized nodules formed when osteoblasts were cultured with an induction medium, a marker of osteogenic differentiation. ALP activity tended to increase after 10 and 60 min/day of stimulation. In addition, LFDSS conditions also increased gene expression of IBSP, RUNX2, COL-I, ALP, OCN, and OPN, as shown by reverse transcriptase-polymerase chain reaction. From the results of a proteomics array, LFDSS groups were intensely expressed with several factors (EGF, HGF, IGF, TGF, and PDGF). Furthermore, CD146 and Stro-1 expression increased in cells treated with 30 min/day and decreased in cells treated with 120 min/day, as determined by cell surface antigen analysis by fluorescence-activated cell-sorting analysis. These results strongly showed that LFDSS at the proper intensity and time enhanced the differentiation and maturation of hABMSCs. In conclusion, an appropriate level of LFDSS can potently and positively modulate proliferation and differentiation in hABMSCs.

  1. Levels of CD105(+) cells increase and cell proliferation decreases during S-phase arrest of amniotic fluid cells in long-term culture.

    Science.gov (United States)

    Wang, Ding; Chen, Rui; Zhong, Xuan; Fan, Yong; Lai, Weiqiang; Sun, Xiaofang

    2014-11-01

    The present study aimed to improve the characterization of amniotic fluid cells (AFCs) in order to optimize their use in chromosomal prenatal diagnosis and as seed or stem cells for tissue engineering. The AFCs used in the current study were obtained from three females in their second trimester of pregnancy. The cells were cultured independently and characterized by cell morphology, cell markers, cell cycle distribution and chromosome Giemsa banding in an early- and late-passage. The AFCs remained homogeneous in culture and expressed mesenchymal markers, but not endothelial markers along the culture process. In addition, compared with the early-passage cells, the late-passage cells exhibit an increase in CD105 expression, a decrease in cell division and a delay in the cell cycle, and a number of cells underwent cell cycle arrest. However, the cells retained a normal karyotype. Therefore, the current study characterized AFCs in a clinical culture and confirmed that AFCs are mesenchymal precursors. The results obtained may be useful for the application of AFCs in prenatal diagnosis.

  2. 4 Cases of Peritoneal Fluid Culture Fungal Growth Case Analysis%4例腹腔液培养真菌生长病例分析

    Institute of Scientific and Technical Information of China (English)

    王文琴; 韦小荣

    2014-01-01

    通过回顾分析4例我院外科报告消化道穿孔腹腔液培养有真菌生长的病历。总结腹腔液培养真菌生长的因素及相关预防措施。因而对加强临床医师的培训,提高其业务水平,院感科同时密切监测医院感染的发生,对于防止医院感染的发生有着重要的作用。%through the retrospective analysis of 4 cases of our hospital surgical report gastrointestinal perforation peritoneal fluid culture fungi growth records. Summary of peritoneal fluid culture factors of fungal growth and relevant preventive measures. To strengthen the clinician training, improve the business level, hospital, and close monitoring to prevent hospital infection, plays an important role in the occurrence of nosocomial infection.

  3. Platelet-released supernatant induces osteoblastic differentiation of human mesenchymal stem cells: potential role of BMP-2

    Directory of Open Access Journals (Sweden)

    M Alini

    2010-12-01

    Full Text Available Platelet-rich preparations have recently gained popularity in maxillofacial and dental surgery, but their beneficial effect is still under debate. Furthermore, very little is known about the effect of platelet preparations at the cellular level, and the underlying mechanisms. In this study, we tested the effect of platelet-released supernatant (PRS on human mesenchymal stem cell (MSC differentiation towards an osteoblastic phenotype in vitro. Cultures of MSC were supplemented with PRS and typical osteoblastic markers were assessed at up to 28 days post-confluence. PRS showed an osteoinductive effect on MSC, as shown by an increased expression of typical osteoblastic marker genes such as collagen Ialpha1, bone sialoprotein II, BMP-2 and MMP-13, as well as by increased 45Ca2+ incorporation. Our results suggest that the effect of PRS on human MSC could be at least partially mediated by BMP-2.Activated autologous PRS could therefore provide an alternative to agents like recombinant bone growth factors by increasing osteoblastic differentiation of bone precursor cells at bone repair sites, although further studies are needed to fully support our observations.

  4. Cell-free supernatants from probiotic Lactobacillus casei and Lactobacillus rhamnosus GG decrease colon cancer cell invasion in vitro.

    Science.gov (United States)

    Escamilla, Juanita; Lane, Michelle A; Maitin, Vatsala

    2012-08-01

    Probiotics have been shown to have a preventative role in colorectal carcinogenesis but research concerning their prophylactic potential in the later stages of colorectal cancer, specifically metastasis is limited. This study explored the potential of cell-free supernatants (CFS) from 2 probiotic Lactobacillus sp., Lactobacillus casei and Lactobacillus rhamnosus GG, to inhibit colon cancer cell invasion by influencing matrix metalloproteinase-9 (MMP-9) activity and levels of the tight junction protein zona occludens-1 (ZO-1) in cultured metastatic human colorectal carcinoma cells. HCT-116 cells were treated with CFS from L. casei, L. rhamnosus, or Bacteroides thetaiotaomicron (a gut commensal); or with uninoculated bacterial growth media. Treatment with CFS from both Lactobacillus sp. decreased colorectal cell invasion but treatment with CFS from B. thetaiotaomicron did not. CFS from both Lactobacillus sp. decreased MMP-9 and increased ZO-1 protein levels. L. rhamnosus CFS also lowered MMP-9 activity. To begin elucidating the secreted bacterial factor conveying these responses, Lactobacillus sp. CFS were fractionated into defined molecular weight ranges and cell invasion assessed. Fractionation revealed that the inhibitory activity was contained primarily in the >100 kDa and 50-100 kDa fractions, suggesting the inhibitory compound may be a macromolecule such as a protein, nucleic acid, or a polysaccharide.

  5. Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

    Science.gov (United States)

    He, Xiaolong; Zeng, Qing; Puthiyakunnon, Santhosh; Zeng, Zhijie; Yang, Weijun; Qiu, Jiawen; Du, Lei; Boddu, Swapna; Wu, Tongwei; Cai, Danxian; Huang, Sheng-He; Cao, Hong

    2017-01-01

    The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal integrity. In vivo experiments revealed that pre-treatment with LCS significantly decrease susceptibility of neonatal rats to oral E. coli K1 infection as reflected by reduced bacterial intestinal colonization, translocation, dissemination and systemic infections. Further, we found that LCS treated neonatal rats have higher intestinal expressions of Ki67, MUC2, ZO-1, IgA, mucin and lower barrier permeability than those in untreated rats. These results indicated that LCS could enhance neonatal resistance to systemic E. coli K1 infection via promoting maturation of neonatal intestinal defense. In conclusions, our findings suggested that LCS has a prophylactic effect against systemic E. coli K1 infection in neonates. Future studies aimed at identifying the specific active ingredients in LCS will be helpful in developing effective pharmacological strategies for preventing neonatal E. coli K1 infection. PMID:28262688

  6. Polymer films removed from solid surfaces by nanostructured fluids: microscopic mechanism and implications for the conservation of cultural heritage.

    Science.gov (United States)

    Raudino, Martina; Selvolini, Giulia; Montis, Costanza; Baglioni, Michele; Bonini, Massimo; Berti, Debora; Baglioni, Piero

    2015-03-25

    Complex fluids based on amphiphilic formulations are emerging, particularly in the field of conservation of works of art, as effective and safe liquid media for the removal of hydrophobic polymeric coatings. The comprehension of the cleaning mechanism is key to designing tailored fluids for this purpose. However, the interaction between nanostructured fluids and hydrophobic polymer films is still poorly understood. In this study, we show how the combination of confocal laser scanning microscopy (CLSM) and atomic force microscopy (AFM) provides interesting and complementary insight into this process. We focused on the interaction between an ethyl methacrylate/methyl acrylate 70:30 copolymer film deposited onto a glass surface and a water/nonionic surfactant/2-butanone (MEK) ternary system, with MEK being a good solvent and water being a nonsolvent for the polymer. Our results indicate a synergy between the organic solvent and the surfactant assemblies: MEK rapidly swells the outer layers of the polymer film allowing for the subsequent diffusion of solvent molecules, while the amphiphile decreases the interfacial energy between the polymeric coating and the liquid phase, favoring dewetting and dispersion of swollen polymer droplets in the aqueous phase. The chemical nature of the surfactant and the microstructure of the assemblies determine both the kinetics and the overall efficiency of polymer removal, as assessed by comparing the behavior of similar formulations containing an anionic surfactant (sodium dodecyl sulfate, SDS).

  7. Comparison of 16S rDNA-PCR Amplification and Culture of Cerebrospinal Fluid for Diagnosis of Bacterial Meningitis

    Directory of Open Access Journals (Sweden)

    Farshad Foroughi

    2010-12-01

    Full Text Available Objective:Early and accurate diagnosis of bacterial meningitis is of critical concern. Optimum and rapid laboratory facilities are not routinely available for detecting the etiologic agents of meningitis. The objective of this study was to compare polymerase chain reaction (PCR assay with culture for detection of bacteria in central nervous system (CNS samples from patients suspected to have meningitis. Methods: One-hundred CSF samples were obtained and divided into two parts. One part of samples was used for standard bacterial culture and gram staining. The remaining was used for DNA extraction. PCR assay was performed with universal primers for 16S rDNA gene of bacteria. Performance characteristics of the test were determined. Findings:The PCR method was able to detect bacteria in all 36 culture-positive and in 38 of 64 culture-negative cases showing sensitivity and specificity of 100% and 40.6% respectively. Positive predictive value was 48.6% and negative predictive value 100%, however, Kappa coefficient showed the correlation of the 2 methods to be at 0.33. Conclusion:There are advantages and disadvantages in performance characteristics of the conventional CSF culture and universal CSF 16S rDNA PCR. Therefore, it is recommended to use both methods in clinical practice, particularly in suspicious contaminated samples, with presumable presence of fastidious or slow growing bacteria because of antibiotic consumption.

  8. An In-Vitro Investigation of the Antibacterial Effects of the Methanol and Aqueous Extracts and the Supernatant of the Algae Chlorella vulgaris CCATM 210-1 on Multiantibiotic-Resistant Staphylococcus aureus Isolates Causing Urinary Tract Infections

    Directory of Open Access Journals (Sweden)

    Senobar Asadi

    2016-04-01

    Full Text Available Algae contain compounds, some of which have antibacterial properties. For example, the antibiotic Chlorellin can be mentioned, which is extracted from Chlorella species. The methanol and aqueous extracts, and the supernatant of the algae Chlorella vulgaris CCATM- 210-1 were used in this study. After culturing the Algae and preparing the supernatant and extracts, the antibacterial effects of the extracts and supernatant of this algae against multidrugresistant (MDR Staphylococcus aureus isolates causing urinary tract infections were determined using the microdilution and checkerboard titration methods. In the microdilution method, first, the aqueous extract of this algae at a concentration of 0.72 ± 0.22 mg/ml could inhibit the growth of the bacteria being tested. Then, the methanol extract and the supernatant of this algae at concentrations of 0.98 ± 0.68 and 1 ± 0.26 mg/ml, respectively, inhibited the growth of the bacteria being studied. The sum of the FIC of the methanol and aqueous extracts of the algae Chlorella vulgaris CCATM 210-1 in this study, showed the indifference of these two extracts towards each other.

  9. An In-Vitro Investigation of the Antibacterial Effects of the Acetone and Ethanol Extracts and the Supernatant of the Algae Chlorella vulgaris CCATM- 210-1 on Some of the Gram-Negative Bacterial Foodborne Pathogens

    Directory of Open Access Journals (Sweden)

    Yasaman Asadi

    2016-09-01

    Full Text Available Algae are rich sources of amino acids, terpenoids, florentines, alkanes, halogenated ketones, steroidal compounds, fatty acids, and phenols that can be used in the pharmaceutical industry. The acetone and ethanol extracts, and the supernatant of the algae Chlorella vulgaris CCATM- 210-1, were used in this study. After culturing the Algae, and preparing the supernatant and extracts, the antibacterial effects of the extracts and supernatant of this algae against some of the gram-negative bacterial foodborne pathogens were determined using the well plate and agar disk diffusion methods. According to the study conducted, the acetone and ethanol extracts and the supernatant of the algae Chlorella vulgaris CCATM- 210-1 showed acceptable antibacterial properties against some of the bacteria being studied, at concentrations of 1.25, 2.5, 5, and 10 mg/ml in the well plate and agar disk diffusion methods. The maximum inhibition zone diameters in the well plate and agar disk diffusion methods, were 36.8 and 35.4 mm, respectively, which were related to the bacterium Proteus mirabilis PTCC 1776 and were observed at a 10 mg/ml concentration of the acetone extract. By comparing the mean inhibition zone diameters among the acetone and ethanol extracts and a mixture of the two extracts using the well plate method, it can be concluded that the acetone and ethanol extracts of Chlorella vulgaris CCATM- 210-1 are indifferent to each other. This analysis was also true of the agar disk diffusion method.

  10. Rheological properties in supernatant of peach gum from almond(Prunus dulcis)

    Institute of Scientific and Technical Information of China (English)

    王森; 谢碧霞; 钟秋平; 杜红岩

    2008-01-01

    The rheological properties in the supernatant of peach gum from Prunnus dulcis were discussed in order to provide more scientific technical parameters and references for developing peach gum as a kind of medicinal gum.The rheological properties in the supernatant of peach gum were comparatively studied in different material ratios,temperatures,shaking times,pH values and salinities.The results show that,1) the mathematical model of shear rate with material ratio and shear stress is Y=0.069X12+0.035X2 -1.174,R2=0.942;2) the mathematical model of shear rate with temperature and shear stress is Y=4.936X12+0.023 2X2-1.688,R2=0.937;3) the mathematical model of shear rate with shaking time and shear stress is Y=0.005 192 X13-0.140 73X12+1.249 045X1+ 0.036 546 X2-3.644 29,R2=0.954 3;4) the effects of pH value on the rheological properties in the supernatant of peach gum are comparatively complicated with a varying range of 3-11 and the shear rate shows a change trend of saddle model;5) the mathematical model of shear rate with the concentration of NaCl and shear stress is Y=-0.037 44X1+0.012 93 X2,R2=0.998;6) the mathematical model of shear rate with the concentration of CaCl2 and shear stress is Y=0.025 789X1+0.016 19X2,R2 =0.999;and 7) the mathematical model of shear rate with the concentration of sorbic acid potassium and shear stress is Y=0.079 5X1+0.017 3X2,R2=0.998.The results show that the material ratio,temperature,shaking time,pH value significantly affect the rheological properties in the supernatant of peach gum,and the concentrations of NaCl and CaCl2 also significantly affect the rheological properties expect the concentration of sorbic acid potassium.

  11. Study of gelatin-agar intermolecular aggregates in the supernatant of its coacervate.

    Science.gov (United States)

    Singh, S Santinath; Bohidar, H B; Bandyopadhyay, S

    2007-05-15

    Intermolecular interaction leading to formation of aggregates between gelatin, a polyampholyte, and agar, a polysaccharide was studied in the supernatant of the complex coacervate formed by these biopolymers. Electrophoresis, laser light scattering and viscometry data were used to determine the interaction and the physical structure of these intermolecular soluble complexes by modeling these to be prolate ellipsoids of revolution (rod-like structures with well defined axial ratio and Perrin's factor). Solution ionic strength was found to reduce the axial ratio of these complexes implying the presence of screened polarization-induced electrostatic interaction between the two biopolymers.

  12. Maternal uniparental disomy of chromosome 2 in a baby with trisomy 2 mosaicism in amniotic fluid culture

    Energy Technology Data Exchange (ETDEWEB)

    Harrison, K.B. [Morristown Memorial Hospital, NJ (United States); Eisenger, K.; Brown, S. [Columbia Univ., NY (United States)] [and others

    1994-09-01

    We describe the first case of a baby with maternal uniparental disomy for chromosome 2. Growth failure, hypothyroidism and hyaline membrane disease were present at birth, and the first year of life was complicated by bronchopulmonary dysplasia. At 14 months, motor and intellectual development appear to be normal, but growth remains below the 10th percentile. The baby was investigated for uniparental disomy because trisomy 2 mosaicism had been detected in a second trimester amniocentesis. This is the first reported case in which amniotic fluid chromosome mosaicism has been associated with uniparental disomy. Implications for prenatal diagnosis are considered.

  13. Maternal uniparental disomy of chromosome 2 in a baby with trisomy 2 mosaicism in amniotic fluid culture

    Energy Technology Data Exchange (ETDEWEB)

    Harrison, K. [Morristown Memorial Hospital, NJ (United States); Eisenger, K.; Brown, S. [Columbia Univ., New York, NY (United States)] [and others

    1995-08-28

    We describe the first case of a baby with maternal uniparental disomy of chromosome 2. Growth failure, hypothyroidism, and hyaline membrane disease were present at birth, and the first year of life was complicated by bronchopulmonary dysplasia. At age 14 months, motor and intellectual development were normal, but growth remained below the 10th centile. The baby was investigated for uniparental disomy because trisomy 2 mosaicism had been detected in a second trimester amniocentesis. This is the first reported case in which amniotic fluid chromosome mosaicism has been associated with uniparental disomy. Implications for prenatal diagnosis are considered. 26 refs., 4 figs.

  14. Inactivation of human immunodeficiency virus type 1 in tissue culture fluid and in genital secretions by the spermicide benzalkonium chloride.

    Science.gov (United States)

    Wainberg, M A; Spira, B; Bleau, G; Thomas, R

    1990-01-01

    We have shown that the spermicidal agent benzalkonium chloride can exert a direct inhibitory effect on the viral reverse transcriptase activity of human immunodeficiency virus type 1 (HIV-1) when utilized at concentrations of 0.05% and higher. Exposure of HIV-1 to this disinfectant at concentrations of more than 0.05% was able to completely destroy viral infectivity, as assessed on susceptible target cells. We have further shown that HIV-1, which is present in both seminal and genital secretions, can be inactivated in such fluids by direct exposure to benzalkonium chloride. PMID:1688873

  15. Neurotoxicity of cerebro-spinal fluid from patients with Parkinson's disease on mesencephalic primary cultures as an in vitro model of dopaminergic neurons.

    Science.gov (United States)

    Kong, Ping; Zhang, Ben-Shu; Lei, Ping; Kong, Xiao-Dong; Zhang, Shi-Shuang; Li, Dai; Zhang, Yun

    2015-08-01

    Parkinson's disease is a degenerative disorder of the central nervous system. In spite of extensive research, neither the cause nor the mechanisms have been firmly established thus far. One assumption is that certain toxic substances may exist in the cerebro-spinal fluid (CSF) of Parkinson's disease patients. To confirm the neurotoxicity of CSF and study the potential correlation between neurotoxicity and the severity of Parkinson's disease, CSF was added to cultured cells. By observation of cell morphology, changes in the levels of lactate dehydrogenase, the ratio of tyrosine hydroxylase-positive cells, and the expression of tyrosine hydroxylase mRNA and protein, the differences between the two groups were shown. The created in vitro model of dopaminergic neurons using primary culture of mouse embryonic mesencephalic tissue is suitable for the study of neurotoxicity. The observations of the present study indicated that CSF from Parkinson's disease patients contains factors that can cause specific injury to cultured dopaminergic neurons. However, no obvious correlation was found between the neurotoxicity of CSF and the severity of Parkinson's disease.

  16. Treatment test of supernatant from sewage sludge by irradiation of high energy electron beams under supersaturation with oxygen

    Energy Technology Data Exchange (ETDEWEB)

    Hosono, Masakazu; Arai, Hidehiko (Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment); Aizawa, Masaki; Shimooka, Toshio; Yamamoto, Ichiro; Shimizu, Ken; Sugiyama, Masashi.

    1993-02-01

    Supernatant comes from dewaterization of sewage sludge, and contains biologically nondegradable organics. Therefore, it is hard to be treated by conventional activated sludge method. The development of a new technology is required to decrease the chemical oxygen demand (COD) effectively below 30 mg/l. Irradiation of high energy electron beams can convert nondegradable organics in water into substances which are biodegradable. However, sufficient dissolved oxygen in water is needed to induce oxidation effectively. In the present study, the treatment of supernatant was studied using an apparatus which can be irradiated by high intensity electron beams in flow system under supersaturation with oxygen by pressurization up to 3 atms. The dependence of oxygen concentration on the reduction in absorbance at 230 nm of azo dye (Acid Red 265) aqueous solution was examined, and it was clarified that sufficient oxygen was supplied in the solution up to about 14 kGy under 3 atms of oxygen. Radiation treatment of supernatant which came from the leather works was carried out using the above apparatus. However, as this supernatant contained high concentration of nitrite, the nitrite was removed by limited aeration activated sludge method. By this pretreatment, COD was reduced from 200 mg/l to 53 mg/l. Then, the biodegradability of supernatant irradiated under supersaturation with oxygen was examined. The final COD of the supernatant was reduced below 30 mg/l by the combined method of irradiation of 7 kGy and biological treatment. (author).

  17. Evolution of composition of dairy manure supernatant in a controlled dung pit.

    Science.gov (United States)

    Rico, C; García, H; Rico, J L; Fernández, J; Renedo, J

    2009-12-01

    Anaerobic conversion of dairy manure into biogas is an attractive way of managing this waste. It is well known that the hydrolysis of large molecules into small, directly biodegradable ones is the rate limiting step of the overall anaerobic process. The present work studies the development of the hydrolytic and acidogenic stages of dairy manure with different solid concentrations (40, 60 and 80 g VS/L) at ambient temperature (20 degrees C). The purpose was to determine the operational conditions that provide a liquid fraction with a high soluble chemical oxygen demand (COD) and a high volatile fatty acids (VFA) content in manure before the methanogenic stage starts up. At 20 degrees C, the evolution of the studied parameters showed that, in a controlled plug-flow dung pit, the hydrolytic and acidogenic stages progressed moderately in a continuous way during the 25 days that the experimentation lasted, whereas no methanization was observed. Supernatant COD and VFA concentrations increased 30% and 107%, respectively, for the 60 g VS/L samples. Manure was also operated at 35 degrees C with a similar increase in supernatant COD but a higher increase in VFA, 154%. For both operational temperatures, the predominant VFAs were, in this order, acetic, propionic and butyric acids. During the operation at 35 degrees C, the methanogenic stage started between days 20 and 25 for the samples with lower solids content, i.e. 40 and 60 g VS/L.

  18. cultural

    Directory of Open Access Journals (Sweden)

    Irene Kreutz

    2006-01-01

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

  19. Peroxynitrite decomposition catalyst prevents apoptotic cell death in a human astrocytoma cell line incubated with supernatants of HIV-infected macrophages

    Directory of Open Access Journals (Sweden)

    Perno Carlo

    2002-09-01

    Full Text Available Abstract Background Oxidative stress has shown to contribute in the mechanisms underlying apoptotic cell death occuring in AIDS-dementia complex. Here we investigated the role of peroxynitrite in apoptosis occurring in astroglial cells incubated with supernatants of HIV-infected human primary macrophages (M/M. Results Flow cytometric analysis (FACS of human cultured astrocytes shortly incubated with HIV-1-infected M/M supernatants showed apoptotic cell death, an effect accompanied by pronounced staining for nitrotyrosine (footprint of peroxynitrite and by abnormal formation of malondialdehyde (MDA. Pretreatment of astrocytes with the peroxynitrite decomposition catalyst FeTMPS antagonized HIV-related astrocytic apoptosis, MDA formation and nitrotyrosine staining. Conclusions Taken together, our results suggest that inibition of peroxynitrite leads to protection against peroxidative stress accompanying HIV-related apoptosis of astrocytes. Overall results support the role of peroxynitrite in HIV-related programmed death of astrocytes and suggest the use of peroxynitrite decomposition catalyst to counteract HIV-1-related neurological disorders.

  20. Quantitative culture of endotracheal aspirate and BAL fluid samples in the management of patients with ventilator-associated pneumonia: a randomized clinical trial

    Directory of Open Access Journals (Sweden)

    Ricardo de Amorim Corrêa

    2014-12-01

    Full Text Available OBJECTIVE: To compare 28-day mortality rates and clinical outcomes in ICU patients with ventilator-associated pneumonia according to the diagnostic strategy used. METHODS: This was a prospective randomized clinical trial. Of the 73 patients included in the study, 36 and 37 were randomized to undergo BAL or endotracheal aspiration (EA, respectively. Antibiotic therapy was based on guidelines and was adjusted according to the results of quantitative cultures. RESULTS: The 28-day mortality rate was similar in the BAL and EA groups (25.0% and 37.8%, respectively; p = 0.353. There were no differences between the groups regarding the duration of mechanical ventilation, antibiotic therapy, secondary complications, VAP recurrence, or length of ICU and hospital stay. Initial antibiotic therapy was deemed appropriate in 28 (77.8% and 30 (83.3% of the patients in the BAL and EA groups, respectively (p = 0.551. The 28-day mortality rate was not associated with the appropriateness of initial therapy in the BAL and EA groups (appropriate therapy: 35.7% vs. 43.3%; p = 0.553; and inappropriate therapy: 62.5% vs. 50.0%; p = 1.000. Previous use of antibiotics did not affect the culture yield in the EA or BAL group (p = 0.130 and p = 0.484, respectively. CONCLUSIONS: In the context of this study, the management of VAP patients, based on the results of quantitative endotracheal aspirate cultures, led to similar clinical outcomes to those obtained with the results of quantitative BAL fluid cultures.

  1. Comparative analisys in cultural test of biological fluids: routine Vs a new strumentation (Alifax HB&L Uroquattro

    Directory of Open Access Journals (Sweden)

    Mariapia Galullo

    2011-03-01

    Full Text Available The accuracy and rapidity of a microbiological diagnosis is crucial for the proper management of critical or neutropenic patients.To reduce the time of analysis, we compared the performances of an automated system with those of the conventional method (direct coltural analysis in agar medium BD and a enrichement for aerobic and anaerobic bacteria and fungi in BD bactec bottles. For this study, we evaluated the kind of specimens, the time of analysis and the positivity for several bacterial strains. Finally we compared the specificity and sensitivity of the automated system with those of the traditional coltures. A total of 50 specimens were analysed.All the specimens were from patiens hospitalized in the wards of Perugia’s hospital.We found that the results obtained with the Alìfax system differed from those of conventional/coltural method.We propose to utilize the Alìfax system for coltural analysis of urine where the cut off of signifìcativity is 50 cfu/mL and the infectious agent involved is often monomicrobic and aerobic.We will continue to use the arrichment of fluid in BD bottles which is very accurate.

  2. Compressibility of the fouling layer formed by membrane bioreactor sludge and supernatant

    DEFF Research Database (Denmark)

    Jørgensen, Mads Koustrup; Poorasgari, Eskandar; Christensen, Morten Lykkegaard

    the pressure range 0.08 - 0.13 bar. The fouling layer resistance increased while fouling layer compression was reversible. Conversely, the supernatant filtrations showed that the gel layer is compressible within the range 0.1 - 2 bar applied pressure. Calculated resistance of the gel layer indicated....... Compressibility of the gel layer was studied in a dead-end filtration system, whereas the compressibility of a fouling layer formed by MBR sludge was studied in a submerged system hollow sheet membrane by TMP stepping. It was shown that the fouling layer formed by the MBR sludge was highly compressible within...... that higher pressure causes higher change in resistance. The results of the gel compression study showed a lower compressibility of gel layers than the fouling layer formed by sludge flocs. At pressures between 0.1 to 0.5 bar there was no significant change in specific cake resistance with pressure compared...

  3. Pseudolinkage of the duplicate loci for supernatant aspartate aminotransferase in brook trout, Salvelinus fontinalis.

    Science.gov (United States)

    Wright, J E; May, B; Stoneking, M; Lee, G M

    1980-01-01

    Electrophoretic variation involving three alleles is described for the duplicated loci for supernatant aspartate aminotransferase (AAT-1,2), from muscle extracts of brook trout. Both loci exhibit largely disomic inheritance. Exceptional progeny types are proposed to be the result of a form of tetrasomic inheritance. Nonrandom segregation was found among the progeny of males doubly heterozygous for AAT markers; where so-called linkage phase was known, this nonrandom assortment was shown to be pseudolinkage (78.9 percent recombination). Analyses of joint segregation of triply heterozygous males for the AAT-(1,2) loci and for the single alpha glycerophosphate dehydrogenase locus (AGP-1) revealed true linkage of AGP-1 with one AAT locus (mean r = 11 percent), but pseudolinkage with the other AAT locus (r = 74 percent). Intraindividual variation for homoeologous multivalent pairing of two acrocentric with two metacentric chromosomes in males, but with bivalent pairing in females, is proposed to account for pseudolinkage and for the tetrasomically inherited types.

  4. Host markers in Quantiferon supernatants differentiate active TB from latent TB infection: preliminary report

    Directory of Open Access Journals (Sweden)

    Walzl Gerhard

    2009-05-01

    Full Text Available Abstract Background Interferon gamma release assays, including the QuantiFERON® TB Gold In Tube (QFT have been shown to be accurate in diagnosing Mycobacterium tuberculosis infection. These assays however, do not discriminate between latent TB infection (LTBI and active TB disease. Methods We recruited twenty-three pulmonary TB patients and 34 household contacts from Cape Town, South Africa and performed the QFT test. To investigate the ability of new host markers to differentiate between LTBI and active TB, levels of 29 biomarkers in QFT supernatants were evaluated using a Luminex multiplex cytokine assay. Results Eight out of 29 biomarkers distinguished active TB from LTBI in a pilot study. Baseline levels of epidermal growth factor (EGF soluble CD40 ligand (sCD40L, antigen stimulated levels of EGF, and the background corrected antigen stimulated levels of EGF and macrophage inflammatory protein (MIP-1β were the most informative single markers for differentiation between TB disease and LTBI, with AUCs of 0.88, 0.84, 0.87, 0.90 and 0.79 respectively. The combination of EGF and MIP-1β predicted 96% of active TB cases and 92% of LTBIs. Combinations between EGF, sCD40L, VEGF, TGF-α and IL-1α also showed potential to differentiate between TB infection states. EGF, VEGF, TGF-α and sCD40L levels were higher in TB patients. Conclusion These preliminary data suggest that active TB may be accurately differentiated from LTBI utilizing adaptations of the commercial QFT test that includes measurement of EGF, sCD40L, MIP-1β, VEGF, TGF-α or IL-1α in supernatants from QFT assays. This approach holds promise for development as a rapid diagnostic test for active TB.

  5. Senescence-Associated Molecular and Epigenetic Alterations in Mesenchymal Stem Cell Cultures from Amniotic Fluid of Normal and Fetus-Affected Pregnancy

    Directory of Open Access Journals (Sweden)

    Jūratė Savickienė

    2016-01-01

    Full Text Available Human amniotic-fluid-derived mesenchymal stem cells (AF-MSCs are interesting for their multilineage differentiation potential and wide range of therapeutic applications due to the ease of culture expansion. However, MSCs undergo replicative senescence. So far, the molecular mechanisms that underlie fetal diseases and cell senescence are still poorly understood. Here, we analyzed senescence-associated morphologic, molecular, and epigenetic characteristics during propagation of MSCs derived from AF of normal and fetus-affected pregnancy. AF-MSCs cultures from both cell sources displayed quite similar morphology and expression of specific cell surface (CD44, CD90, and CD105 and stemness (Oct4, Nanog, Sox2, and Rex1 markers but had interindividual variability in proliferation capability and time to reach senescence. Within passages 4 and 8, senescent cultures exhibited typical morphological features, senescence-associated β-galactosidase activity, increased levels of p16, and decreased levels of miR-17 and miR-21 but showed differential expression of p21, p53, and ATM dependently on the onset of cell senescence. These differences correlated with changes in the level of chromatin modifiers (DNMT1 and HDAC1 and polycomb group proteins (EZH2, SUZ12, and BMI1 paralleling with changes in the expression of repressive histone marks (H3K9me3 and H3K27me3 and stemness markers (Oct4, Nanog, Sox2, and Rex1. Therefore epigenetic factors are important for AF-MSCs senescence process that may be related with individuality of donor or a fetus malignancy status.

  6. Culture of mouse amniotic fluid-derived cells on irradiated STO feeders results in the generation of primitive endoderm cell lines capable of self-renewal in vitro.

    Science.gov (United States)

    Babic, Aleksandar M; Jang, Sunyoung; Nicolov, Eugenia; Voicu, Horatiu; Luckey, Chance J

    2013-01-01

    The cells present in amniotic fluid (AF) are currently used for prenatal diagnosis of fetal anomalies but are also a potential source of cells for cell therapy. To better characterize putative progenitor cell populations present in AF, we used culture conditions that support self-renewal to determine if these promoted the generation of stable cell lines from AF-derived cells (AFC). Cells isolated from E11.5 mouse were cultured on irradiated STO fibroblast feeder layers in human embryonic germ cell derivation conditions. The cultures grew multicellular epithelial colonies that could be repropagated from single cells. Reverse transcription semiquantitative polymerase chain reaction of established cell lines revealed that they belonged to the extraembryonic endoderm (ExEn) expressing high levels of Gata6, Gata4, Sox17, Foxa2 and Sox7 mRNA. Hierarchical clustering based on the whole transcriptome expression profile of the AFC lines (AFCL) shows significant correlation between transcription profiles of AFCL and blastocyst-derived XEN, an ExEn cell line. In vitro differentiation of AFCL results in the generation of cells expressing albumin and α-fetoprotein (AFP), while intramuscular injection of AFCL into immunodeficient mice produced AFP-positive tumors with primitive endodermal appearance. Hence, E11.5 mouse AF contains cells that efficiently produce XEN lines. These AF-derived XEN lines do not spontaneously differentiate into embryonic-type cells but are phenotypically stable and have the capacity for extensive expansion. The lack of requirement for reprogramming factors to turn AF-derived progenitor cells into stable cell lines capable of massive expansion together with the known ability of ExEn to contribute to embryonic tissue suggests that this cell type may be a candidate for banking for cell therapies. © 2013 S. Karger AG, Basel

  7. Senescence-Associated Molecular and Epigenetic Alterations in Mesenchymal Stem Cell Cultures from Amniotic Fluid of Normal and Fetus-Affected Pregnancy

    Science.gov (United States)

    Savickienė, Jūratė; Baronaitė, Sandra; Zentelytė, Aistė; Treigytė, Gražina

    2016-01-01

    Human amniotic-fluid-derived mesenchymal stem cells (AF-MSCs) are interesting for their multilineage differentiation potential and wide range of therapeutic applications due to the ease of culture expansion. However, MSCs undergo replicative senescence. So far, the molecular mechanisms that underlie fetal diseases and cell senescence are still poorly understood. Here, we analyzed senescence-associated morphologic, molecular, and epigenetic characteristics during propagation of MSCs derived from AF of normal and fetus-affected pregnancy. AF-MSCs cultures from both cell sources displayed quite similar morphology and expression of specific cell surface (CD44, CD90, and CD105) and stemness (Oct4, Nanog, Sox2, and Rex1) markers but had interindividual variability in proliferation capability and time to reach senescence. Within passages 4 and 8, senescent cultures exhibited typical morphological features, senescence-associated β-galactosidase activity, increased levels of p16, and decreased levels of miR-17 and miR-21 but showed differential expression of p21, p53, and ATM dependently on the onset of cell senescence. These differences correlated with changes in the level of chromatin modifiers (DNMT1 and HDAC1) and polycomb group proteins (EZH2, SUZ12, and BMI1) paralleling with changes in the expression of repressive histone marks (H3K9me3 and H3K27me3) and stemness markers (Oct4, Nanog, Sox2, and Rex1). Therefore epigenetic factors are important for AF-MSCs senescence process that may be related with individuality of donor or a fetus malignancy status. PMID:27803714

  8. Supernatant from a cloned helper T cell stimulates resting B cells to express transferrin and IL-2 receptors.

    Science.gov (United States)

    Diu, A; Leclercq, L; Dautry-Varsat, A; Theze, J

    1987-07-01

    We describe the properties of the supernatant from a murine cloned helper T cell (clone 52.3) which is able to polyclonally activate most resting B cells in the absence of any additional stimulus. We hypothesize that an activity which we call BCAF (B-cell-activating factor(s] exists in our supernatant which can activate resting B cells alone or in conjunction with other lymphokines. In the present report, we investigate changes in the surface antigen pattern induced on resting B cells by BCAF-containing supernatant. Analysis of the cells by flow cytometry shows that transferrin receptor and IL-2 receptor expression increase on a large fraction of B cells after 2 days of activation by the T-helper-cell clone supernatant. Monoclonal anti-transferrin receptor antibody inhibits cell division but does not affect blastogenesis, while IL-2 has no effect in our experimental system. Our present results confirm that BCAF-containing supernatants can act on most resting B cells and replace helper T cells in inducing B-cell activation and proliferation.

  9. Electrochemical study of Type 304 and 316L stainless steels in simulated body fluids and cell cultures.

    Science.gov (United States)

    Tang, Yee-Chin; Katsuma, Shoji; Fujimoto, Shinji; Hiromoto, Sachiko

    2006-11-01

    The electrochemical corrosion behaviour of Type 304 and 316L stainless steels was studied in Hanks' solution, Eagle's minimum essential medium (MEM), serum containing medium (MEM with 10% of fetal bovine serum) without cells, and serum containing medium with cells over a 1-week period. Polarization resistance measurements indicated that the stainless steels were resistant to Hanks' and MEM solutions. Type 304 was more susceptible to pitting corrosion than Type 316L in Hanks' and MEM solutions. The uniform corrosion resistance of stainless steels, determined by R(p), was lower in culturing medium than in Hanks' and MEM. The low corrosion resistance was due to surface passive film with less protective to reveal high anodic dissolution rate. When cells were present, the initial corrosion resistance was low, but gradually increased after 3 days, consistent with the trend of cell coverage. The presence of cells was found to suppress the cathodic reaction, that is, oxygen reduction, and increase the uniform corrosion resistance as a consequence. On the other hand, both Type 304 and 316L stainless steels became more susceptible to pitting corrosion when they were covered with cells.

  10. Evaluation of DNA extraction methods and their clinical application for direct detection of causative bacteria in continuous ambulatory peritoneal dialysis culture fluids from patients with peritonitis by using broad-range PCR.

    Science.gov (United States)

    Kim, Si Hyun; Jeong, Haeng Soon; Kim, Yeong Hoon; Song, Sae Am; Lee, Ja Young; Oh, Seung Hwan; Kim, Hye Ran; Lee, Jeong Nyeo; Kho, Weon-Gyu; Shin, Jeong Hwan

    2012-03-01

    The aims of this study were to compare several DNA extraction methods and 16S rDNA primers and to evaluate the clinical utility of broad-range PCR in continuous ambulatory peritoneal dialysis (CAPD) culture fluids. Six type strains were used as model organisms in dilutions from 10(8) to 10(0) colony-forming units (CFU)/mL for the evaluation of 5 DNA extraction methods and 5 PCR primer pairs. Broad-range PCR was applied to 100 CAPD culture fluids, and the results were compared with conventional culture results. There were some differences between the various DNA extraction methods and primer sets with regard to the detection limits. The InstaGene Matrix (Bio-Rad Laboratories, USA) and Exgene Clinic SV kits (GeneAll Biotechnology Co. Ltd, Korea) seem to have higher sensitivities than the others. The results of broad-range PCR were concordant with the results from culture in 97% of all cases (97/100). Two culture-positive cases that were broad-range PCR-negative were identified as Candida albicans, and 1 PCR-positive but culture-negative sample was identified as Bacillus circulans by sequencing. Two samples among 54 broad-range PCR-positive products could not be sequenced. There were differences in the analytical sensitivity of various DNA extraction methods and primers for broad-range PCR. The broad-range PCR assay can be used to detect bacterial pathogens in CAPD culture fluid as a supplement to culture methods.

  11. Boildown Study on Supernatant Liquid Retrieved from AW-106 in December 2012

    Energy Technology Data Exchange (ETDEWEB)

    Page, Jason S. [Washington River Protection Solutions, LLC, Richland, WA (United States)

    2013-06-04

    This document reports the results of a boil down study using a composite created from supernatant liquid grab samples retrieved from tank 241-AW-I06 in December of 2012. The composite was made using predetermined volumes of the grab samples which accounted for layering of the supernatant liquid in the tank. The finished composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (24 - 27°C). The density of the test composite was measured in the hot cell immediately before the boildown study and was 1.266 g/mL at 27.1 °C. The boiling temperature of the composite was measured at three different pressures (40, 60, and 80 Torr) throughout the volume reduction, and the results show steadily increasing boiling temperatures with increasing volume reduction and no significant discontinuities. Moderate foaming was observed at the onset of the boildown. The foaming disappeared during the first reduction step, and minimal foaming was observed throughout the rest of the study. The bulk densities at 18.0 °C (D{sub Bulk}{sup 18 °C}) and quantities of settled and centrifuged solids were measured on samples of the boildown concentrates. Estimated values of the bulk densities at the 60-Torr boiling temperatures (D{sub Bulk}{sup 60 Torr}) were also calculated. Solids were first observed at boildown temperatures when the % VWR reached 39.3%. The quantity of solids in the composite quickly increased after this initial formation; the amount of centrifuged solids increased by 22% as the %WVR increased from 39.3 to 44.1 %. A small amount of solids did appear in the samples collected prior to the initial formation during the boildown. These solids precipitated while they sat at hot cell ambient temperature and in the 18. 0 °C water bath. Analysis of boil down test samples indicated that natrophosphate (Na7{sub 3}F(PO{sub 4}){sub 2}{centerdot} 19 H{sub 2}O) and kogarkoite (Na3FS04) accounted for a majority of the initial solids (~80% of the

  12. Refrigerating fluids; Fluides frigorigenes

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    1999-03-01

    Refrigerating fluids are experiencing a real revolution since few years. CFCs with their destructive effect on the ozone layer are now prohibited while HCFCs will be progressively eliminated and replaced by HFCs. However, HFCs can contribute to the increase of the greenhouse effect. The solutions proposed by thermal engineering professionals consist in the confinement of air-conditioning installations (elimination of recurrent leaks) and in the improvement of installations efficiency. HCFC fluids like the R 22 are still widely used in air-conditioning but they are supposed to be replaced by HFC fluids like the R 134a, the R 407C or the R 410A. This short paper gives a brief presentation of these fluids and of their chemical characteristics. (J.S.)

  13. Optimization of a high-throughput CTAB-based protocol for the extraction of qPCR-grade DNA from rumen fluid, plant and bacterial pure cultures.

    Science.gov (United States)

    Minas, Konstantinos; McEwan, Neil R; Newbold, Charles Jamie; Scott, Karen P

    2011-12-01

    The quality and yield of extracted DNA are critical for the majority of downstream applications in molecular biology. Moreover, molecular techniques such as quantitative real-time PCR (qPCR) are becoming increasingly widespread; thus, validation and cross-laboratory comparison of data require standardization of upstream experimental procedures. DNA extraction methods depend on the type and size of starting material(s) used. As such, the extraction of template DNA is arguably the most significant variable when cross-comparing data from different laboratories. Here, we describe a reliable, inexpensive and rapid method of DNA purification that is equally applicable to small or large scale or high-throughput purification of DNA. The protocol relies on a CTAB-based buffer for cell lysis and further purification of DNA with phenol : chloroform : isoamyl alcohol. The protocol has been used successfully for DNA purification from rumen fluid and plant cells. Moreover, after slight alterations, the same protocol was used for large-scale extraction of DNA from pure cultures of Gram-positive and Gram-negative bacteria. The yield of the DNA obtained with this method exceeded that from the same samples using commercial kits, and the quality was confirmed by successful qPCR applications.

  14. Experimental prestorage filtration removes antibodies and decreases lipids in RBC supernatants mitigating TRALI in vivo.

    Science.gov (United States)

    Silliman, Christopher C; Kelher, Marguerite R; Khan, Samina Y; LaSarre, Monica; West, F Bernadette; Land, Kevin J; Mish, Barbara; Ceriano, Linda; Sowemimo-Coker, Samuel

    2014-05-29

    Transfusion-related acute lung injury (TRALI) remains a significant cause of transfusion-related mortality with red cell transfusion. We hypothesize that prestorage filtration may reduce proinflammatory activity in the red blood cell (RBC) supernatant and prevent TRALI. Filters were manufactured for both small volumes and RBC units. Plasma containing antibodies to human lymphocyte antigen (HLA)-A2 or human neutrophil antigen (HNA)-3a was filtered, and immunoglobulins and specific HNA-3a and HLA-2a neutrophil (PMN) priming activity were measured. Antibodies to OX27 were added to plasma, and filtration was evaluated in a 2-event animal model of TRALI. RBC units from 31 donors known to have antibodies against HLA antigens and from 16 antibody-negative controls were filtered. Furthermore, 4 RBC units were drawn and underwent standard leukoreduction. Immunoglobulins, HLA antibodies, PMN priming activity, and the ability to induce TRALI in an animal model were measured. Small-volume filtration of plasma removed >96% of IgG, antibodies to HLA-A2 and HNA-3a, and their respective priming activity, as well as mitigating antibody-mediated in vivo TRALI. In RBC units, experimental filtration removed antibodies to HLA antigens and inhibited the accumulation of lipid priming activity and lipid-mediated TRALI. We conclude that filtration removes proinflammatory activity and the ability to induce TRALI from RBCs and may represent a TRALI mitigation step.

  15. Multistage aqueous two-phase extraction of a monoclonal antibody from cell supernatant.

    Science.gov (United States)

    Muendges, Jan; Zalesko, Alexej; Górak, Andrzej; Zeiner, Tim

    2015-01-01

    This article presents results of continuous multistage aqueous two-phase extraction of an immunoglobulin G1 from cell supernatant in a mixer-settler unit. An aqueous two-phase system consisting of polyethylene glycol 2000, phosphate salt, and water was applied without and with sodium chloride (NaCl). Influences of different parameters such as throughput, phase ratio, and stage number on the extraction performance were analyzed. For systems without NaCl, the extraction was carried out as a washing step. An increase of stage number from one to five stages enabled to increase the immunoglobulin G1 purity from 11.8 to 32.6% at a yield of nearly 90%. Furthermore, a reduction of product phase volume due to a higher phase ratio led to an increase of purity from 20.8 to 29.6% in a three-stage countercurrent extraction. For experiments with NaCl moderate partitioning conditions were adjusted by adding 8 wt% NaCl. In that case, the extraction was carried out as a stripping step.

  16. Discovery of Novel Secreted Virulence Factors from Salmonella enterica Serovar Typhimurium by Proteomic Analysis of Culture Supernatants

    Energy Technology Data Exchange (ETDEWEB)

    Niemann, George; Brown, Roslyn N.; Gustin, Jean K.; Stufkens, Afke; Shaikh-Kidwai, Afshan S.; Li, Jie; McDermott, Jason E.; Brewer, Heather M.; Schepmoes, Athena A.; Smith, Richard D.; Adkins, Joshua N.; Heffron, Fred

    2011-01-01

    The intracellular pathogen Salmonella enterica serovar Typhimurium is a leading cause of acute gastroenteritis in the world. This pathogen has two type-III secretion systems (TTSS) necessary for virulence that are encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) and are expressed during extracellular or intracellular infectious states, respectively, to deliver virulence factors (effectors) to the host cell cytoplasm. While many have been identified and at least partially characterized, the full repertoire of effectors has not been catalogued. In this mass spectrometry-based proteomics study, we identified effector proteins secreted under minimal acidic medium growth conditions that induced the SPI-2 TTSS and its effectors, and compared the secretome from the parent strain to the secretome from strains missing either essential (SsaK) or regulatory components (SsaL) of the SPI-2 secretion apparatus. We identified 75% of the known TTSS effector repertoire. Excluding translocon components, 95% of the known effectors were biased for identification in the ssaL mutant background, which demonstrated that SsaL regulates SPI-2 type III secretion. To confirm secretion to animal cells, we made translational fusions of several of the best candidates to the calmodulin-dependent adenylate cyclase of Bordetella pertussis and assayed cAMP levels of infected J774 macrophage-like cells. From these infected cells we identified six new TTSS effectors and two others that are secreted independent of TTSS. Our results substantiate reports of additional secretion systems encoded by Salmonella other than TTSS.

  17. Separation of active laccases from Pleurotus sapidus culture supernatant using aqueous two-phase systems in centrifugal partition chromatography.

    Science.gov (United States)

    Schwienheer, C; Prinz, A; Zeiner, T; Merz, J

    2015-10-01

    For the production of bio active compounds, e.g., active enzymes or antibodies, a conserved purification process with a minimum loss of active compounds is necessary. In centrifugal partition chromatography (CPC), the separation effect is based on the different distribution of the components to be separated between two immiscible liquid phases. Thereby, one liquid phase is kept stationary in chambers by a centrifugal field and the mobile phase is pumped through via connecting ducts. Aqueous two phase systems (ATPS) are known to provide benign conditions for biochemical products and seem to be promising when used in CPC for purification tasks. However, it is not known if active biochemical compounds can "survive" the conditions in a CPC where strong shear forces can occur due to the two-phasic flow under centrifugal forces. Therefore, this aspect has been faced within this study by the separation of active laccases from a fermentation broth of Pleurotus sapidus. After selecting a suitable ATPS and operating conditions, the activity yield was calculated and the preservation of the active enzymes could be observed. Therefore, CPC could be shown as potentially suitable for the purification of bio-active compounds.

  18. Discovery of novel secreted virulence factors from Salmonella enterica serovar Typhimurium by proteomic analysis of culture supernatants.

    Science.gov (United States)

    Niemann, George S; Brown, Roslyn N; Gustin, Jean K; Stufkens, Afke; Shaikh-Kidwai, Afshan S; Li, Jie; McDermott, Jason E; Brewer, Heather M; Schepmoes, Athena; Smith, Richard D; Adkins, Joshua N; Heffron, Fred

    2011-01-01

    Salmonella enterica serovar Typhimurium is a leading cause of acute gastroenteritis throughout the world. This pathogen has two type III secretion systems (TTSS) encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) that deliver virulence factors (effectors) to the host cell cytoplasm and are required for virulence. While many effectors have been identified and at least partially characterized, the full repertoire of effectors has not been catalogued. In this proteomic study, we identified effector proteins secreted into defined minimal medium designed to induce expression of the SPI-2 TTSS and its effectors. We compared the secretomes of the parent strain to those of strains missing essential (ssaK::cat) or regulatory (ΔssaL) components of the SPI-2 TTSS. We identified 20 known SPI-2 effectors. Excluding the translocon components SseBCD, all SPI-2 effectors were biased for identification in the ΔssaL mutant, substantiating the regulatory role of SsaL in TTS. To identify novel effector proteins, we coupled our secretome data with a machine learning algorithm (SIEVE, SVM-based identification and evaluation of virulence effectors) and selected 12 candidate proteins for further characterization. Using CyaA' reporter fusions, we identified six novel type III effectors and two additional proteins that were secreted into J774 macrophages independently of a TTSS. To assess their roles in virulence, we constructed nonpolar deletions and performed a competitive index analysis from intraperitoneally infected 129/SvJ mice. Six mutants were significantly attenuated for spleen colonization. Our results also suggest that non-type III secretion mechanisms are required for full Salmonella virulence.

  19. Reverse transcriptase activity in chicken embryo fibroblast culture supernatants is associated with particles containing endogenous avian retrovirus EAV-0 RNA.

    OpenAIRE

    Weissmahr, R N; Schüpbach, J; Böni, J

    1997-01-01

    We have recently shown that live attenuated virus vaccines produced on chicken-derived cells contain low levels of particle-associated reverse transcriptase (RT). In both virus and corresponding control harvests produced on chicken embryo fibroblasts, these activities were present at significantly higher concentrations than in the vaccines. In order to identify the putative retrovirus sequence responsible for this activity, a novel method for the selective PCR amplification of particle-associ...

  20. Detecting pM concentrations of prostaglandins in cell culture supernatants by capillary SCX-LC-MS/MS

    DEFF Research Database (Denmark)

    Dahl, Sandra Rinne; Kleiveland, Charlotte Ramstad; Kassem, Moustapha

    2008-01-01

    A highly sensitive, improved online strong cation exchange (SCX)--RP capillary liquid chromatographic (cLC) method with IT mass spectrometric (IT-MS/MS) detection for the simultaneous determination of prostaglandin (PG)A(1), PGD(2), PGE(1), PGE(2), PGF(2alpha), 8-iso-(8i)PGF(2alpha), 6-keto-(6k)P...

  1. Distinct galactofuranose antigens in the cell wall and culture supernatants as a means to differentiate Fusarium from AspergillusspeciesAnnegret

    DEFF Research Database (Denmark)

    Wiedemann, Annegret; Kakoschke, Tamara Katharina; Speth, Cornelia

    2016-01-01

    tDetection of carbohydrate antigens is an important means for diagnosis of invasive fungal infections. Fordiagnosis of systemic Aspergillus infections, galactomannan is commonly used, the core antigenic struc-ture of which consists of chains of several galactofuranose moieties. In this study, we ...

  2. Biological nutrients removal from the supernatant originating from the anaerobic digestion of the organic fraction of municipal solid waste.

    Science.gov (United States)

    Malamis, S; Katsou, E; Di Fabio, S; Bolzonella, D; Fatone, F

    2014-09-01

    This study critically evaluates the biological processes and techniques applied to remove nitrogen and phosphorus from the anaerobic supernatant produced from the treatment of the organic fraction of municipal solid waste (OFMSW) and from its co-digestion with other biodegradable organic waste (BOW) streams. The wide application of anaerobic digestion for the treatment of several organic waste streams results in the production of high quantities of anaerobic effluents. Such effluents are characterized by high nutrient content, because organic and particulate nitrogen and phosphorus are hydrolyzed in the anaerobic digestion process. Consequently, adequate post-treatment is required in order to comply with the existing land application and discharge legislation in the European Union countries. This may include physicochemical and biological processes, with the latter being more advantageous due to their lower cost. Nitrogen removal is accomplished through the conventional nitrification/denitrification, nitritation/denitritation and the complete autotrophic nitrogen removal process; the latter is accomplished by nitritation coupled with the anoxic ammonium oxidation process. As anaerobic digestion effluents are characterized by low COD/TKN ratio, conventional denitrification/nitrification is not an attractive option; short-cut nitrogen removal processes are more promising. Both suspended and attached growth processes have been employed to treat the anaerobic supernatant. Specifically, the sequencing batch reactor, the membrane bioreactor, the conventional activated sludge and the moving bed biofilm reactor processes have been investigated. Physicochemical phosphorus removal via struvite precipitation has been extensively examined. Enhanced biological phosphorus removal from the anaerobic supernatant can take place through the sequencing anaerobic/aerobic process. More recently, denitrifying phosphorus removal via nitrite or nitrate has been explored. The removal of

  3. 利用抗氧化剂促进羊水细胞培养效果的研究%The study of improving the culture results of amniotic fluid cells by adding antioxidants

    Institute of Scientific and Technical Information of China (English)

    丘力功; 蔡小杰; 黄志诚

    2012-01-01

    目的 通过添加抗氧化剂促进羊水细胞的培养效果.方法 分离传代的成纤维型羊水细胞,以生长指数为检测指标,在H配方的基础上利用部分析因设计法和最陡爬坡法对促进羊水细胞生长的抗氧化剂进行筛选和优化.优化后的培养基通过原代羊水细胞培养进行效果验证.结果 添加的维生素E、L-抗坏血酸、α硫辛酸和还原型谷胱甘肽等抗氧化剂皆能促进羊水细胞的生长,且具有协同效应.与H培养基相比,经最陡爬坡法优化的培养基培养的羊水细胞在克隆总数、生长指数等指标都提高3倍以上,分裂相细胞总数增长4倍以上.结论 改良后的羊水细胞培养基培养效果要显著优于H配方培养基,在产前诊断应用中具有推广价值.%Objective To improve the culture results of amniotic fluid by adding antioxidants. Methods Using Fractional factorial designs combined with the steepest ascent path approach, the concentration of antioxidants added in H medium was optimized to increase the growth speed of the passage fibroblast cells from amniotic fluid . And the performance of the optimized culture medium was tested with the growth index, the numbers of clones and metaphase of primary amniotic fluid cells culture. Results Vitamin E, L-ascorbic acid,α-lipoic acid and reduced CSH can all promote the growth of the amniotic fluid cells with synergic effect. Total number of clones,growth index,and total number of metaphase cells from amniotic fluid cells cultured in medium optimized by the steepest ascent path approach are all 3 times higher than those from H medium. Conclusion The optimization method of adding antioxidants in H medium can improve significantly the results of amniotic fluid cells culture and the optimized culture medium is applicable to the prenatal diagnosis.

  4. Upflow anaerobic sludge blanket (UASB) treatment of supernatant of cow manure by thermal pre-treatment.

    Science.gov (United States)

    Yoneyama, Y; Nishii, A; Nishimoto, M; Yamada, N; Suzuki, T

    2006-01-01

    Upflow anaerobic sludge blanket (UASB) methane fermentation treatment of cow manure that was subjected to screw pressing, thermal treatment and subsequent solid-liquid separation was studied. Conducting batch scale tests at temperatures between 140 and 180 degrees C, the optimal temperature for sludge settling and the color suppression was found to be between 160-170 degrees C. UASB treatment was carried out with a supernatant obtained from the thermal treatment at the optimal conditions (170 degrees C for 30 minutes) and polymer-dosed solid-liquid separation. In the UASB treatment with a COD(Cr) loading of 11.7 kg/m3/d and water temperature of 32.2 degrees C, the COD(Cr) level dropped from 16,360 mg/L in raw water to 3,940 mg/L in treated water (COD(Cr), removal rate of 75.9%), and the methane production rate per COD(Cr) was 0.187 Nm3/kg. Using wastewater thermal-treated at the optimal conditions, also a methane fermentation treatment with a continuously stirred tank reactor (CSTR) was conducted (COD(Cr) in raw water: 38,000 mg/L, hydraulic retention time (HRT): 20 days, 35 degrees C). At the COD(Cr) loading of 1.9 kg/m3/d, the methane production rate per COD(Cr), was 0.153 Nm3/kg. This result shows that UASB treatment using thermal pre-treatment provides a COD(Cr), loading of four times or more and a methane production rate of 1.3 times higher than the CSTR treatment.

  5. Boildown Study on Supernatant Liquid Retrieved from AP-107 in May 2010

    Energy Technology Data Exchange (ETDEWEB)

    Callaway, W. S. [Washington River Protection Solutions, LLC, Richland, WA (United States); Page, J. S. [Washington River Protection Solutions, LLC, Richland, WA (United States)

    2013-02-12

    A boildown study was completed on a composite prepared from supernatant liquid grab samples retrieved from tank 241-AP-107 in May of 2010. The composite was a clear, yellow liquid containing no visible solids at hot cell ambient temperatures (25-27 °C). The density of the test composite was 1.216 g/mL at 26.8 °C. The boiling temperature curves generated at three reduced pressures—40-, 60-, and 80 Torr—displayed steadily increasing boiling temperatures with increasing volume reduction with no significant discontinuities. Only minimal foaming was observed after the volume reduction proceeded beyond 50 %WVR (percent waste volume reduction). The bulk densities (D{sub Bulk}{sup 18 °C}) and quantities of settled and centrifuged solids present were measured on samples of the boildown concentrates that were kept at 18 °C for 7-8 days. Estimated values of the bulk densities of the concentrates at 60-Torr boiling temperatures (D{sub Bulk}{sup 60 Torr}) were also calculated. Solids were observed in all boildown concentrates at process temperatures, at hot cell ambient temperatures (25-27 °C), and at 18 °C. The quantity of solids found in the cooled concentrates increased slowly through 50.2 %WVR. The quantity of solids found in concentrates after 54.0 %WVR was noticeably greater. Beyond 54.0 %WVR, the quantity of solids found in cooled concentrates increased dramatically. Analysis of boildown test samples indicated that sodium oxalate and sodium carbonate solids form in cooled concentrates after volume reduction of 8.4 %WVR or less. The major contributors to the large increase in the quantity of solids found in concentrates after 54 %WVR were sodium nitrate and sodium carbonate.

  6. Amniotic fluid

    Science.gov (United States)

    ... carefully. Removing a sample of the fluid through amniocentesis can provide information about the sex, health, and development of the fetus. Images Amniocentesis Amniotic fluid Polyhydramnios Amniotic fluid References Cunningham FG, ...

  7. High level expression of Acidothermus cellulolyticus β-1, 4-endoglucanase in transgenic rice enhances the hydrolysis of its straw by cultured cow gastric fluid

    Energy Technology Data Exchange (ETDEWEB)

    Chou, Hong L.; Dai, Ziyu; Hsieh, Chia W.; Ku, Maurice S.

    2011-12-10

    Large-scale production of effective cellulose hydrolytic enzymes is the key to the bioconversion of agricultural residues to ethanol. The goal of this study was to develop a rice plant as a bioreactor for the large-scale production of cellulose hydrolytic enzymes via genetic transformation, and to simultaneously improve rice straw as an efficient biomass feedstock for conversion of cellulose to glucose. In this study, the cellulose hydrolytic enzyme {beta}-1, 4-endoglucanase (E1) from the thermophilic bacterium Acidothermus cellulolyticus was overexpressed in rice through Agrobacterium-mediated transformation. The expression of the bacterial gene in rice was driven by the constitutive Mac promoter, a hybrid promoter of Ti plasmid mannopine synthetase promoter and cauliflower mosaic virus 35S promoter enhancer with the signal peptide of tobacco pathogenesis-related protein for targeting the protein to the apoplastic compartment for storage. A total of 52 transgenic rice plants from six independent lines expressing the bacterial enzyme were obtained, which expressed the gene at high levels with a normal phenotype. The specific activities of E1 in the leaves of the highest expressing transgenic rice lines were about 20 fold higher than those of various transgenic plants obtained in previous studies and the protein amounts accounted for up to 6.1% of the total leaf soluble protein. Zymogram and temperature-dependent activity analyses demonstrated the thermostability of the enzyme and its substrate specificity against cellulose, and a simple heat treatment can be used to purify the protein. In addition, hydrolysis of transgenic rice straw with cultured cow gastric fluid yielded almost twice more reducing sugars than wild type straw. Taken together, these data suggest that transgenic rice can effectively serve as a bioreactor for large-scale production of active, thermostable cellulose hydrolytic enzymes. As a feedstock, direct expression of large amount of cellulases in

  8. Anti-adherence potential of Enterococcus durans cells and its cell-free supernatant on plastic and stainless steel against foodborne pathogens.

    Science.gov (United States)

    Amel, Ait Meddour; Farida, Bendali; Djamila, Sadoun

    2015-07-01

    It is demonstrated that numerous bacteria are able to attach to surfaces of equipment used for food handling or processing. In this study, a strain of Enterococcus durans, originally isolated from a milking machine surface, was firstly studied for its biofilm formation potential on plastic and stainless steel supports. The strain was found to be a biofilm producer either at 25, 30 or 37 °C on polystyrene microtitre plates, with a best adherence level observed at 25 °C. En. durans showed a strong adhesion to stainless steel AISI-304. Antibacterial and anti-adherence activities of En. durans were tested against four foodborne pathogens (Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853 and Listeria innocua CLIP 74915) which were shown as biofilm producers on both plastic and stainless steel. En. durans cells and cell-free culture supernatant showed a significant (P < 0.05) inhibition potential of the pathogens either on solid media or in broth co-cultures. Characterization of the antibacterial substances indicated their proteinaceous nature which assigned them most probably to bacteriocins group.

  9. Protocol for Identifying the Presence of and Understanding the Nature of Soluble, Non-pertechnetate Technetium in Hanford Tank Supernatants

    Energy Technology Data Exchange (ETDEWEB)

    Rapko, Brian M.

    2014-02-27

    The objective of this report is to propose a method to evaluate the presence and extent of soluble, non-pertechnetate Tc in Hanford tank supernatants as well as methods that might be used to gain insight as to the nature of the specie(s) that make up this fraction. This study will then provide a recommendation as to the preferred approach for identifying and quantifying the presence of Hanford tank supernatant-soluble, non-pertechnetate, technetium. The recommendation will also describe an approach to address the issue of whether inductively coupled plasma mass spectrometry (ICP-MS) analysis, which is useful as a monitoring tool for Tc, may be confounded by the presence of other mass 99 species.

  10. Laboratory bioassay for assessing the effects of sludge supernatant on plant growth and vesicular-arbuscular mycorrhiza formation

    Energy Technology Data Exchange (ETDEWEB)

    Bohn, K.S.; Liberta, A.E.

    1982-12-01

    A laboratory bioassay is described for assessing the effects of sludge supernatant on juvenile corn growth and the ability of vesicular-arbuscular (VA) mycorrhizal fungi, indigenous to coal spoil, to form mycorrhizae. The bioassay demonstrated that application rates can be identified that have the potential to promote increased plant dry weight without suppressing the formation of VA mycorrhizae in a plant's root system.

  11. Facile purification of Escherichia coli expressed tag-free recombinant human tumor necrosis factor alpha from supernatant.

    Science.gov (United States)

    Zhang, Chun; Liu, Yongdong; Zhao, Dawei; Li, Xiunan; Yu, Rong; Su, Zhiguo

    2014-03-01

    Fusing affinity tag at N-terminus was reported to decrease the biological activity of the recombinant human tumor necrosis factor alpha. Although preparation of tag-free rhTNF-α has already been achieved, the processes were yet laborious, especially in large scale. In this paper, tag-free rhTNF-α was almost equally synthesized by Escherichia coli in both soluble and insoluble forms. A two-step ion exchange chromatography, DEAE-Sepharose combined with CM-Sepharose, was developed to purify the soluble specie from supernatant after cell lysis. Native PAGE and HP-SEC showed the rhTNF-α extracted from supernatant existed in a homogeneous form. HP-SAX and SDS-PAGE analysis demonstrated the purity of the final fraction was over 98% with a very high recovery of 75%. Circular dichroism spectrum demonstrated that β-sheet structure was dominant and fluorescence analysis suggested no dramatic exposure of aromatic amino acid residues on the protein surface. Bioassay indicated that purified rhTNF-α was biologically active with a specific activity of approximately 2.0×10(7)U/mg. All these results suggested that this two-step ion exchange chromatography is efficient for preparation of biologically active tag-free rhTNF-α from supernatant.

  12. Microflora of the seminal fluid of healthy men and men suffering from chronic prostatitis syndrome.

    Science.gov (United States)

    Ivanov, Iuri B; Kuzmin, Michael D; Gritsenko, Viktor A

    2009-10-01

    Chronic prostatitis syndrome (CPS) is a common urologic condition that many clinicians find difficult to diagnose and treat effectively. The information about the composition of the flora of the seminal fluid in healthy men and patients with CPS is limited. The aim of this study was to define the microbial communities present in the seminal fluid of healthy men and patients with CPS and at in vitro detection of decomplementary activity (DCA) phenotypes of isolates along with their comparison with isolates from patients with or without CPS. The bacteriological study was carried out to 48 healthy men and 60 men with CPS. Culture specimens were spread on various selective media. Bacterial DCA was tested by measuring the decrease in complement activity (CH(50)) under the influence of culture supernatants. The most common isolates in both groups were coryneforms, lactobacilli, coagulase-negative staphylococci, micrococci and streptococci. Enterobacteriaceae, enterococci and Staphylococcus aureus were isolated only from the CPS group. The organisms from seminal fluid of healthy men exerted DCA at 3.56 +/- 2.15; 2.47 +/- 1.23 and 4.36 +/- 2.2 anti-CH(50) for staphylococci, micrococci and diphtheroids respectively. The DCA of staphylococci, coryneforms, Enterobacteriaceae, enterococci and micrococci from CPS group were 12.8 +/- 2.1 (p 0.05), 16.8 +/- 2.1, 7.2 +/- 1.9 and 11.6 +/- 3.3 (p < 0.05) anti-CH(50) respectively. The data obtained in this study testify the microecological disorders in microbiota of seminal fluid in CPS.

  13. Challenges of culturing human norovirus in three-dimensional organoid intestinal cell culture models.

    Directory of Open Access Journals (Sweden)

    Efstathia Papafragkou

    Full Text Available Human noroviruses are the most common cause of acute gastroenteritis worldwide. Recently, cell culture systems have been described using either human embryonic intestinal epithelial cells (Int-407 or human epithelial colorectal adenocarcinoma cells (Caco-2 growing on collagen-I porous micro carrier beads in a rotating bioreactor under conditions of physiological fluid shear. Here, we describe the efforts from two independent laboratories to implement this three dimensional (3D cell culture system for the replication of norovirus. Int-407 and Caco-2 were grown in a rotating bioreactor for up to 28 days. Prior to infection, cells were screened for the presence of microvilli by electron microscopy and stained for junction proteins (zonula occludens-1, claudin-1, and β-catenin. Differentiated 3D cells were transferred to 24-well plates and infected with bacteria-free filtrates of various norovirus genotypes (GI.1, GI.3, GI.8, GII.2, GII.4, GII.7, and GII.8. At 12 h, 24 h, and 48 h post inoculation, viral RNA from both cells and supernatants were collected and analyzed for norovirus RNA by real-time reverse transcription PCR. Despite observations of high expression of junction proteins and microvilli development in stained thin sections, our data suggest no significant increase in viral titer based on norovirus RNA copy number during the first 48 h after inoculation for the different samples and virus culture conditions tested. Our combined efforts demonstrate that 3D cell culture models using Int-407 or Caco-2 cells do not support norovirus replication and highlight the complexity and difficulty of developing a reproducible in vitro cell culture system for human norovirus.

  14. Fluid Mechanics.

    Science.gov (United States)

    Drazin, Philip

    1987-01-01

    Outlines the contents of Volume II of "Principia" by Sir Isaac Newton. Reviews the contributions of subsequent scientists to the physics of fluid dynamics. Discusses the treatment of fluid mechanics in physics curricula. Highlights a few of the problems of modern research in fluid dynamics. Shows that problems still remain. (CW)

  15. Fluid Interfaces

    DEFF Research Database (Denmark)

    Hansen, Klaus Marius

    2001-01-01

    Fluid interaction, interaction by the user with the system that causes few breakdowns, is essential to many user interfaces. We present two concrete software systems that try to support fluid interaction for different work practices. Furthermore, we present specificity, generality, and minimality...... as design goals for fluid interfaces....

  16. Fluid Mechanics.

    Science.gov (United States)

    Drazin, Philip

    1987-01-01

    Outlines the contents of Volume II of "Principia" by Sir Isaac Newton. Reviews the contributions of subsequent scientists to the physics of fluid dynamics. Discusses the treatment of fluid mechanics in physics curricula. Highlights a few of the problems of modern research in fluid dynamics. Shows that problems still remain. (CW)

  17. Diagnosing feline infectious peritonitis using the Sysmex XT-2000iV based on frozen supernatants from cavitary effusions.

    Science.gov (United States)

    Stranieri, Angelica; Paltrinieri, Saverio; Giordano, Alessia

    2017-02-01

    The delta total nucleated cells (ΔTNC) measurement with the Sysmex XT-2000iV (Sysmex Europe, Norderstedt, Germany) has high diagnostic accuracy on effusions in feline infectious peritonitis (FIP) cases, but the test can be performed only on fresh samples. We evaluated whether supernatants from effusions retain the ability to induce cell clumping and assessed the diagnostic accuracy of this modified ΔTNC method. Effusions were collected from FIP cats ( n = 19) and from cats with other diseases ( n = 15). ΔTNC was measured on fresh samples and on frozen-thawed supernatants after the addition of feline blood at 1:10 dilution. Diagnostic accuracy was assessed at the cutoffs of suggestive of FIP (ΔTNC = 1.7) and consistent with FIP (ΔTNC = 3.4). The influence of the protein content, number of added cells, and magnitude of dilution were also investigated. Specificity and positive predictive value were 100% for both the methods. Sensitivity and negative predictive value were higher for the modified ΔTNC (84.2% and 83.3%, respectively, at the cutoff of 1.7; 78.9% and 78.9%, respectively, at the cutoff of 3.4) than for the ΔTNC on fresh samples (78.6% and 81.3%, respectively, at the cutoff of 1.7; 57.1% and 68.4%, respectively, at the cutoff of 3.4). Protein content, total cell count of the added blood, and magnitude of dilutions did not influence the results. Supernatants of frozen effusions from FIP cats retain the ability to induce cell clumping, thus the modified ΔTNC measurement is a reliable tool to diagnose FIP on samples that cannot be analyzed immediately.

  18. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    Directory of Open Access Journals (Sweden)

    Ronaldo Costa Argôlo-Filho

    2014-05-01

    Full Text Available Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt toxins in culture supernatants (SN have biocontrol potential (e.g., Vip3A, Cry1I, Sip1, whereas others are unwanted (β-exotoxins, as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01 was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  19. Requirement of simultaneous assessment of crystal- and supernatant-related entomotoxic activities of Bacillus thuringiensis strains for biocontrol-product development.

    Science.gov (United States)

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-05-20

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 10(7) endospores mL(-1) caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems.

  20. Inhibition of biofilm development and spoilage potential of Shewanella baltica by quorum sensing signal in cell-free supernatant from Pseudomonas fluorescens.

    Science.gov (United States)

    Zhao, Aifei; Zhu, Junli; Ye, Xiaofeng; Ge, Yangyang; Li, Jianrong

    2016-08-02

    The objective of this study was to in vitro evaluate the effect of a cell-free supernatant (CFS) containing quorum sensing (QS) signal of Pseudomonas fluorescens on the growth, biofilm development and spoilage potential of Shewanella baltica, and preliminarily assess the interactive influences of various chemically synthesized autoinducers on spoilage phenotypes of S. baltica. PF01 strain isolated from spoiled Pseudosciaen crocea was identified P. fluorescens. The addition of 25% and 50% CFS to S. baltica culture had no effect on the growth rate during the lag and exponential phase, however, caused cell decline during the stationary phase. The presence of CFS from P. fluorescens significantly inhibited biofilm development, and greatly decreased the production of trimethylamine (TMA) and biogenic amino in S. baltica. Various signal molecules of QS in the CFS of P. fluorescens culture were detected, including seven N-acyl-l-homoserine lactones (AHLs), autoinducer-2 (AI-2) and two diketopiperazines (DKPs). Exogenous supplement of synthesized seven AHLs containing in the CFS decreased biofilm formation and TMA production in S. baltica, while exposure to exogenous cyclo-(l-Pro-l-Leu) was showed to promote spoilage potential, which revealed that S. baltica also sense the two QS molecules. Furthermore, the stimulating effect of cyclo-(l-Pro-l-Leu) was affected when AHL was simultaneously added, suggesting that the inhibitory activity of spoilage phenotypes in S. baltica might be attributed to a competitive effect of these QS compounds in the CFS of P. fluorescens. The present studies provide a good basis for future research on the role of QS in the regulation of spoilage microbial flora.

  1. Requirement of Simultaneous Assessment of Crystal- and Supernatant-Related Entomotoxic Activities of Bacillus thuringiensis Strains for Biocontrol-Product Development

    Science.gov (United States)

    Argôlo-Filho, Ronaldo Costa; Costa, Robson Luz; Pinheiro, Daniele Heloisa; Corrêa, Fábio Mathias; Valicente, Fernando Hercos; Pomella, Alan William Vilela; Loguercio, Leandro Lopes

    2014-01-01

    Bioinsecticides with lower concentrations of endospores/crystals and without loss of efficiency are economically advantageous for pest biocontrol. In addition to Cry proteins, other Bacillus thuringiensis (Bt) toxins in culture supernatants (SN) have biocontrol potential (e.g., Vip3A, Cry1I, Sip1), whereas others are unwanted (β-exotoxins), as they display widespread toxicity across taxa. A strain simultaneously providing distinct toxin activities in crystals and SN would be desirable for bioinsecticides development; however, strains secreting β-exotoxins should be discarded, independently of other useful entomotoxins. Entomotoxicity of crystals and SN from a Brazilian Bt tolworthi strain (Btt01) was tested against Spodoptera frugiperda to assess the potential for biocontrol-product development based on more than one type of toxin/activity. Tests showed that 107 endospores mL−1 caused >80% of larvae mortality, suggesting Btt01 may be used in similar concentrations as those of other Bt-based biopesticides. When it was applied to cornfields, a significant 60% reduction of larvae infestation was observed. However, bioassays with Btt01 SN revealed a thermostable toxic activity. Physicochemical characterization strongly suggests the presence of unwanted β-exotoxins, with isolate-specific temporal variation in its secretion. Knowledge of the temporal pattern of secretion/activity in culture for all forms of toxins produced by a single strain is required to both detect useful activities and avoid the potential lack of identification of undesirable toxins. These findings are discussed in the contexts of commercial Bt product development, advantages of multiple-activity strains, and care and handling recommended for large-scale fermentation systems. PMID:24854738

  2. Modern embalming, circulation of fluids, and the voyage through the human arterial system: Carl L. Barnes and the culture of immortality in America.

    Science.gov (United States)

    Podgorny, Irina

    2011-01-01

    By considering the work of American embalmer, lawyer, and physician Carl Lewis Barnes (1872-1927), this paper analyzes the emergence of modern embalming in America. Barnes experimented with and exhibited the techniques by which embalming fluids travelled into the most remote cavities of the human body. In this sense, modem embalmers based their skills and methods on experimental medicine, turning the anatomy of blood vessels, physiology of circulation, and composition of blood into a circuit that allowed embalming fluids to move throughout the corpse. Embalmers in the late 19th century took ownership of the laws of hydrodynamics and the physiology of blood circulation to market their fluids and equipment, thus playing the role of physiologists of death, performing and demonstrating physiological experiments with dead bodies.

  3. Analysis of tank 4 (FTF-4-15-22, 23) surface and subsurface supernatant samples in support of enrichment control, corrosion control and evaporator feed qualification programs

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-09-09

    This report provides the results of analyses on Savannah River Site Tank 4 surface and subsurface supernatant liquid samples in support of the Enrichment Control Program (ECP), the Corrosion Control Program (CCP) and the Evaporator Feed Qualification (EFQ) Program. The purpose of the ECP sample taken from Tank 4 in August 2015 was to determine if the supernatant liquid would be “acceptable feed” to the 2H and 3H evaporator systems.

  4. Selection for autochthonous bifidobacteial isolates adapted to simulated gastrointestinal fluid

    Directory of Open Access Journals (Sweden)

    H Jamalifar

    2010-03-01

    Full Text Available "nBackground and the purpose of the study: Bifidobacterial strains are excessively sensitive to acidic conditions and this can affect their living ability in the stomach and fermented foods, and as a result, restrict their use as live probiotic cultures. The aim of the present study was to obtain bifidobacterial isolates with augmented tolerance to simulated gastrointestinal condition using cross-protection method. "nMethods: Individual bifidobacterial strains were treated in acidic environment and also in media containing bile salts and NaCl. Viability of the acid and acid-bile-NaCl tolerant isolates was further examined in simulated gastric and small intestine by subsequent incubation of the probiotic bacteria in the corresponding media for 120 min. Antipathogenic activities of the adapted isolates were compared with those of the original strains. "nResults and major conclusion: The acid and acid-bile-NaCl adapted isolates showed improved viabilities significantly (p<0.05 in simulated gastric fluid compared to their parent strains. The levels of reduction in bacterial count (Log cfu/ml of the acid and acid-bile-NaCl adapted isolates obtained in simulated gastric fluid ranged from 0.64-3.06 and 0.36-2.43 logarithmic units after 120 min of incubation. There was no significant difference between the viability of the acid-bile-NaCl-tolerant isolates and the original strains in simulated small intestinal condition except for Bifidobacterium adolescentis (p<0.05. The presence of 15 ml of supernatants of acid-bile-NaCl-adapted isolates and also those of the initial Bifidobacterium strains inhibited pathogenic bacterial growth for 24 hrs. Probiotic bacteria with improved ability to survive in harsh gastrointestinal environment could be obtained by subsequent treatment of the strains in acid, bile salts and NaCl environments.

  5. Sphingolipid metabolism correlates with cerebrospinal fluid Beta amyloid levels in Alzheimer's disease.

    Directory of Open Access Journals (Sweden)

    Alfred N Fonteh

    Full Text Available Sphingolipids are important in many brain functions but their role in Alzheimer's disease (AD is not completely defined. A major limit is availability of fresh brain tissue with defined AD pathology. The discovery that cerebrospinal fluid (CSF contains abundant nanoparticles that include synaptic vesicles and large dense core vesicles offer an accessible sample to study these organelles, while the supernatant fluid allows study of brain interstitial metabolism. Our objective was to characterize sphingolipids in nanoparticles representative of membrane vesicle metabolism, and in supernatant fluid representative of interstitial metabolism from study participants with varying levels of cognitive dysfunction. We recently described the recruitment, diagnosis, and CSF collection from cognitively normal or impaired study participants. Using liquid chromatography tandem mass spectrometry, we report that cognitively normal participants had measureable levels of sphingomyelin, ceramide, and dihydroceramide species, but that their distribution differed between nanoparticles and supernatant fluid, and further differed in those with cognitive impairment. In CSF from AD compared with cognitively normal participants: a total sphingomyelin levels were lower in nanoparticles and supernatant fluid; b levels of ceramide species were lower in nanoparticles and higher in supernatant fluid; c three sphingomyelin species were reduced in the nanoparticle fraction. Moreover, three sphingomyelin species in the nanoparticle fraction were lower in mild cognitive impairment compared with cognitively normal participants. The activity of acid, but not neutral sphingomyelinase was significantly reduced in the CSF from AD participants. The reduction in acid sphingomylinase in CSF from AD participants was independent of depression and psychotropic medications. Acid sphingomyelinase activity positively correlated with amyloid β42 concentration in CSF from cognitively normal but

  6. The Relation between Fluid Intelligence and the General Factor as a Function of Cultural Background: A Test of Cattell's Investment Theory

    Science.gov (United States)

    Kvist, Ann Valentin; Gustafsson, Jan-Eric

    2008-01-01

    According to Cattell's [Cattell, R.B. (1987). "Intelligence: Its structure, growth and action." New York: North-Holland.] Investment theory individual differences in acquisition of knowledge and skills are partly the result of investment of Fluid Intelligence ("Gf") in learning situations demanding insights in complex…

  7. The Relation between Fluid Intelligence and the General Factor as a Function of Cultural Background: A Test of Cattell's Investment Theory

    Science.gov (United States)

    Kvist, Ann Valentin; Gustafsson, Jan-Eric

    2008-01-01

    According to Cattell's [Cattell, R.B. (1987). "Intelligence: Its structure, growth and action." New York: North-Holland.] Investment theory individual differences in acquisition of knowledge and skills are partly the result of investment of Fluid Intelligence ("Gf") in learning situations demanding insights in complex…

  8. Use of Bacillus thuringiensis supernatant from a fermentation process to improve bioremediation of chlorpyrifos in contaminated soils.

    Science.gov (United States)

    Aceves-Diez, Angel E; Estrada-Castañeda, Kelly J; Castañeda-Sandoval, Laura M

    2015-07-01

    The aim of this research was to investigate the potential of a nutrient-rich organic waste, namely the cell-free supernatant of Bacillus thuringiensis (BtS) gathered from fermentation, as a biostimulating agent to improve and sustain microbial populations and their enzymatic activities, thereby assisting in the bioremediation of chlorpyrifos-contaminated soil at a high dose (70 mg kg(-1)). Experiments were performed for up to 80 d. Chlorpyrifos degradation and its major metabolic product, 3,5,6-trichloro-2-pyridinol (TCP), were quantified by high-performance liquid chromatography (HPLC); total microbial populations were enumerated by direct counts in specific medium; and fluorescein diacetate (FDA) hydrolysis was measured as an index of soil microbial activity. Throughout the experiment, there was higher chlorpyrifos degradation in soil supplemented with BtS (83.1%) as compared to non-supplemented soil. TCP formation and degradation occurred in all soils, but the greatest degradation (30.34%) was observed in soil supplemented with BtS. The total microbial populations were significantly improved by supplementation with BtS. The application of chlorpyrifos to soil inhibited the enzymatic activity; however, this negative effect was counteracted by BtS, inducing an increase of approximately 16% in FDA hydrolysis. These results demonstrate the potential of B. thuringiensis supernatant as a suitable biostimulation agent for enhancing chlorpyrifos and TCP biodegradation in chlorpyrifos-contaminated soils.

  9. A combination turbidity and supernatant microplate assay to rank-order the supersaturation limits of early drug candidates.

    Science.gov (United States)

    Morrison, John S; Nophsker, Michelle J; Haskell, Roy J

    2014-10-01

    A unique opportunity exists at the drug discovery stage to overcome inherently poor solubility by selecting drug candidates with superior supersaturation propensity. Existing supersaturation assays compare either precipitation-resistant or precipitation-inhibiting excipients, or higher-energy polymorphic forms, but not multiple compounds or multiple concentrations. Furthermore, these assays lack sufficient throughput and compound conservation necessary for implementation in the discovery environment. A microplate-based combination turbidity and supernatant concentration assay was therefore developed to determine the extent to which different compounds remain in solution as a function of applied concentration in biorelevant media over a specific period of time. Dimethyl sulfoxide stock solutions at multiple concentrations of four poorly soluble, weak base compounds (Dipyridamole, Ketoconazole, Albendazole, and Cinnarizine) were diluted with pH 6.5 buffer as well as FaSSIF. All samples were monitored for precipitation by turbidity at 600 nm over 1 h and the final supernatant concentrations were measured. The maximum supersaturation ratio was calculated from the supersaturation limit and the equilibrium solubility in each media. Compounds were rank-ordered by supersaturation ratio: Ketoconazole > Dipyridamole > Cinnarizine ∼ Albendazole. These in vitro results correlated well with oral AUC ratios from published in vivo pH effect studies, thereby confirming the validity of this approach. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  10. Differential diagnosis of amyotrophic lateral sclerosis from Guillain-Barré syndrome by quantitative determination of TDP-43 in cerebrospinal fluid.

    Science.gov (United States)

    Hosokawa, Masato; Arai, Tetsuaki; Yamashita, Makiko; Tsuji, Hiroshi; Nonaka, Takashi; Masuda-Suzukake, Masami; Tamaoka, Akira; Hasegawa, Masato; Akiyama, Haruhiko

    2014-05-01

    The aim of this study was to investigate whether an increased level of TAR DNA-binding protein 43 (TDP-43) in the cerebrospinal fluid (CSF) could be a biomarker for amyotrophic lateral sclerosis (ALS) and facilitate differential diagnosis of ALS from peripheral motor neuropathy. TDP-43 is the major constituent of neuronal and glial inclusions that neuropathologically characterize both ALS and tau-negative frontotemporal lobar degeneration. Recent discoveries of various missense mutations in the TDP-43 gene in familial ALS indicate a pivotal role of the aberrant accumulation of TDP-43 in neurodegeneration. Increased TDP-43 in the CSF could be a hallmark of ALS and other TDP-43 proteinopathy. Sandwich enzyme-linked immunosorbent assay (ELISA) was established to measure the concentration of TDP-43 in biological fluids. Culture supernatants of cells transfected with various TDP-43 constructs were used to confirm that the ELISA detected TDP-43. TDP-43 in the culture supernatant of TDP-43 transfected cells was detected by immunoprecipitation with subsequent immunoblotting and concentrations were successfully measured by sandwich ELISA. We then measured TDP-43 concentrations in the CSF of patients with ALS and Guillain-Barré syndrome (GBS). TDP-43 concentrations in CSF were significantly higher in ALS than in GBS (p = 0.016). The sensitivity of the diagnostic test was 71.4% and the specificity was 84.6%. Quantitative determination of TDP-43 concentrations in the CSF by sandwich ELISA is a potential laboratory test for differentiating ALS from peripheral motor neuropathies such as GBS.

  11. Microfluidic Cell Culture Device

    Science.gov (United States)

    Takayama, Shuichi (Inventor); Cabrera, Lourdes Marcella (Inventor); Heo, Yun Seok (Inventor); Smith, Gary Daniel (Inventor)

    2014-01-01

    Microfluidic devices for cell culturing and methods for using the same are disclosed. One device includes a substrate and membrane. The substrate includes a reservoir in fluid communication with a passage. A bio-compatible fluid may be added to the reservoir and passage. The reservoir is configured to receive and retain at least a portion of a cell mass. The membrane acts as a barrier to evaporation of the bio-compatible fluid from the passage. A cover fluid may be added to cover the bio-compatible fluid to prevent evaporation of the bio-compatible fluid.

  12. Fluid mechanics

    CERN Document Server

    Kundu, Pijush K; Dowling, David R

    2011-01-01

    Fluid mechanics, the study of how fluids behave and interact under various forces and in various applied situations-whether in the liquid or gaseous state or both-is introduced and comprehensively covered in this widely adopted text. Revised and updated by Dr. David Dowling, Fluid Mechanics, 5e is suitable for both a first or second course in fluid mechanics at the graduate or advanced undergraduate level. Along with more than 100 new figures, the text has been reorganized and consolidated to provide a better flow and more cohesion of topics.Changes made to the

  13. Cloned, CD117 selected human amniotic fluid stem cells are capable of modulating the immune response.

    Directory of Open Access Journals (Sweden)

    Emily C Moorefield

    Full Text Available Amniotic fluid stem (AFS cells are broadly multipotent, can be expanded extensively in culture, are not tumorigenic and can be readily cryopreserved for cell banking. Mesenchymal stem cells (MSC show immunomodulatory activity and secrete a wide spectrum of cytokines and chemokines that suppress inflammatory responses, block mixed lymphocyte reactions (MLR and other immune reactions, and have proven therapeutic against conditions such as graft-versus-host disease. AFS cells resemble MSCs in many respects including surface marker expression and differentiation potential. We therefore hypothesized that AFS cells may exhibit similar immunomodulatory capabilities. We present data to demonstrate that direct contact with AFS cells inhibits lymphocyte activation. In addition, we show that cell-free supernatants derived from AFS cells primed with total blood monocytes or IL-1β, a cytokine released by monocytes and essential in mediation of the inflammatory response, also inhibited lymphocyte activation. Further investigation of AFS cell-free supernatants by protein array revealed secretion of multiple factors in common with MSCs that are known to be involved in immune regulation including growth related oncogene (GRO and monocyte chemotactic protein (MCP family members as well as interleukin-6 (IL-6. AFS cells activated by PBMCs released several additional cytokines as compared to BM-MSCs, including macrophage inflammatory protein-3α (MIP-3α, MIP-1α and Activin. AFS cells also released higher levels of MCP-1 and lower levels of MCP-2 compared to BM-MSCs in response to IL-1β activation. This suggests that there may be some AFS-specific mechanisms of inhibition of lymphocyte activation. Our results indicate that AFS cells are able to suppress inflammatory responses in vitro and that soluble factors are an essential component in the communication between lymphocytes and AFS cells. Their extensive self-renewal capacity, possibility for banking and

  14. Fluid Dynamics

    DEFF Research Database (Denmark)

    Brorsen, Michael

    These lecture notes are intended mainly for the 7th semester course "Fluid Dynamics" offered by the Study Committee on Civil Engineering, Aalborg University.......These lecture notes are intended mainly for the 7th semester course "Fluid Dynamics" offered by the Study Committee on Civil Engineering, Aalborg University....

  15. Lipopolysaccharide-Induced Profiles of Cytokine, Chemokine, and Growth Factors Produced by Human Decidual Cells Are Altered by Lactobacillus rhamnosus GR-1 Supernatant.

    Science.gov (United States)

    Li, Wei; Yang, Siwen; Kim, Sung O; Reid, Gregor; Challis, John R G; Bocking, Alan D

    2014-07-01

    The aim of this study was to assess the effects of bacterial lipopolysaccharide (LPS) and Lactobacillus rhamnosus GR-1 supernatant (GR-1SN) on secretion profiles of cytokines, chemokines, and growth factors from primary cultures of human decidual cells. Lipopolysaccharide significantly increased the output of proinflammatory cytokines (interleukin [IL]-1B, IL-2, IL-6, IL-12p70, IL-15, IL-17A, interferon gamma [IFN-γ], and tumor necrosis factor [TNF]); anti-inflammatory cytokines (IL-1RN, IL-4, IL-9, and IL-10); chemokines (IL-8, eotaxin, IFN-inducible protein 10 [IP-10], monocyte chemoattractant protein 1 [MCP-1], macrophage inflammatory protein-1α [MIP-1α], macrophage inflammatory protein-1β [MIP-1β], and regulated on activation normal T cell expressed and secreted [RANTES]); and growth factors (granulocyte colony-stimulating factor [CSF] 3, CSF-2, and vascular endothelial growth factor A [VEGFA]). Lactobacillus rhamnosus GR-1SN alone significantly increased CSF-3, MIP-1α MIP-1β, and RANTES but decreased IL-15 and IP-10 output. The GR-1SN also significantly or partially reduced LPS-induced proinflammatory cytokines TNF, IFN-γ, IL-1β, IL-2 IL-6, IL-12p70, IL-15, IL-17, and IP-10; partially reduced LPS-induced anti-inflammatory cytokines IL-1RN, IL-4 and IL-10, and LPS-induced VEGFA output but did not affect CSF-3, MIP-1α, MIP-1β, MCP-1, IL-8, and IL-9. Our results demonstrate that GR-1SN attenuates the inflammatory responses to LPS by human decidual cells, suggesting its potential role in ameliorating intrauterine infection.

  16. Analysis of tank 7 surface supernatant sample (FTF-7-15-26) in support of corrosion control program

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-10-01

    This report provides the results of analyses on Savannah River Site Tank 7 surface supernatant liquid sample in support of the Corrosion Control Program (CCP). The measured nitrate, nitrite and free-hydroxide concentrations for the Tank 7 surface sample averaged, 3.74E-01 ± 1.88E-03, 4.17E-01 ± 9.01E-03 and 0.602 ± 0.005 M, respectively. The Tank 7 surface cesium-137, sodium and silicon concentrations were, respectively, 3.99E+08, ± 3.25E+06 dpm/mL, 2.78 M and <3.10 mg/L. The measured aluminum concentration in the Tank 7 surface sample averaged 0.11 M.

  17. Mescaline-induced changes of brain-cortex ribosomes. Mescaline demethylase activity of brain-cortex soluble supernatant.

    Science.gov (United States)

    Datta, R K; Ghosh, J J

    1977-02-01

    Brain-cortex slices demethylate mescaline and p-methoxyacetanilide, a reference O-demethylating substrate, though the rate of demethylation of mescaline is about one third that of the reference substrate. The demethylase activity is localized mostly in the soluble supernatant (105 000 x g). It is purified 47-fold with respect to the demethylation of mescaline by ammonium sulfate precipitation and DEAE cellulose chromatography. The partially purified demethylase, which is stable for 3-5 days at -5 degrees C in the presence of dithiothreitol and glutathione and is inhibited by p-chloromercuribenzoate, has maximal activity at pH between 7.2 and 8.0. It demethylates mescaline into 3,4-dimethoxy-5-hydroxyphenethylamine and 3,5-dimethoxy-4-hydroxyphenethylamine and some unidentified derivatives.

  18. Use of quantitative 16S rRNA PCR to determine bacterial load does not augment conventional cerebrospinal fluid (CSF) cultures among children undergoing treatment for CSF shunt infection☆,☆☆

    Science.gov (United States)

    Simon, Tamara D.; Van Yserloo, Brian; Nelson, Kevin; Gillespie, David; Jensen, Randy; McAllister, James P.; Riva-Cambrin, Jay; Stockmann, Chris; Daly, Judy A.; Blaschke, Anne J.

    2013-01-01

    The aim of this study was to develop a quantitative 16S rRNA assay for determination of bacterial nucleic acid load in cerebrospinal fluid (CSF) shunt infection and to compare quantitative 16S rRNA polymerase chain reaction (PCR) findings to those of conventional bacterial culture in patients treated for CSF shunt infection. We developed a quantitative 16S rRNA PCR assay that detected bacterial load across a range of 2.5 × 109 down to 2.5 × 104 16S copies/mL CSF under experimental conditions for numerous Gram-positive and Gram-negative organisms. However, when applied to archived CSF samples from 25 shunt infection episodes, correlations between positive bacterial culture and 16S rRNA levels were seen in only half of infections, and 16S rRNA levels dropped precipitously after an initial peak on the first day of sample collection. Bacterial load measured using 16S rRNA PCR does not provide sufficient information beyond bacterial culture to inform CSF shunt infection treatment. PMID:23953744

  19. Production of immunoglobulins in gingival tissue explant cultures from juvenile periodontitis patients

    Energy Technology Data Exchange (ETDEWEB)

    Hall, E.R.; Falkler, W.A. Jr.; Suzuki, J.B. (Univ. of Maryland Dental School, Baltimore (USA))

    1990-10-01

    B lymphocytes and plasma cells are histologically observed in granulomatous periodontal tissues of juvenile periodontitis (JP) patients. Local immune processes may participate in protective or immunopathologic roles in the pathogenesis of this disease. An in vitro explant culture system was utilized to demonstrate the production of immunoglobulins by diseased JP tissues. Immunodiffusion studies using goat anti-human gamma, alpha, or mu chain serum revealed IgG to be the major immunoglobulin present in 92% of the day 1 supernatant fluids (SF) of the 47 JP gingival tissue explant cultures. IgA was present in 15% of the SF; however, no IgM was detected. Staph Protein A isolated 14C-labeled IgG from the SF, when allowed to react with goat anti-human gamma chain serum, formed lines of precipitation. Positive autoradiographs confirmed the biosynthesis of IgG by the explant cultures. The in vitro gingival tissue explant culture system described provides a useful model for the study of localized immunoglobulins produced by diseased tissues of JP patients.

  20. Fluid dynamics of dilatant fluid

    DEFF Research Database (Denmark)

    Nakanishi, Hiizu; Nagahiro, Shin-ichiro; Mitarai, Namiko

    2012-01-01

    A dense mixture of granules and liquid often shows a severe shear thickening and is called a dilatant fluid. We construct a fluid dynamics model for the dilatant fluid by introducing a phenomenological state variable for a local state of dispersed particles. With simple assumptions for an equation...... of the state variable, we demonstrate that the model can describe basic features of the dilatant fluid such as the stress-shear rate curve that represents discontinuous severe shear thickening, hysteresis upon changing shear rate, and instantaneous hardening upon external impact. An analysis of the model...... reveals that the shear thickening fluid shows an instability in a shear flow for some regime and exhibits the shear thickening oscillation (i.e., the oscillatory shear flow alternating between the thickened and the relaxed states). The results of numerical simulations are presented for one- and two...

  1. Beta-carotene-rich carotenoid-protein preparation and exopolysaccharide production by Rhodotorula rubra GED8 grown with a yogurt starter culture.

    Science.gov (United States)

    Frengova, Ginka I; Simova, Emilina D; Beshkova, Dora M

    2006-01-01

    The underlying method for obtaining a beta-carotene-rich carotenoid-protein preparation and exopolysaccharides is the associated cultivation of the carotenoid-synthesizing lactose-negative yeast strain Rhodotorula rubra GED8 with the yogurt starter culture (Lactobacillus bulgaricus 2-11 + Streptococcus thermophilus 15HA) in whey ultrafiltrate (45 g lactose/l) with a maximum carotenoid yield of 13.37 mg/l culture fluid on the 4.5th day. The chemical composition of the carotenoid-protein preparation has been identified. The respective carotenoid and protein content is 497.4 microg/g dry cells and 50.3% per dry weight, respectively. An important characteristic of the carotenoid composition is the high percentage (51.1%) of beta-carotene (a carotenoid pigment with the highest provitamin A activity) as compared to 12.9% and 33.7%, respectively, for the other two individual pigments--torulene and torularhodin. Exopolysaccharides (12.8 g/l) synthesized by the yeast and lactic acid cultures, identified as acid biopolymers containing 7.2% glucuronic acid, were isolated in the cell-free supernatant. Mannose, produced exclusively by the yeast, predominated in the neutral carbohydrate biopolymer component (76%). The mixed cultivation of R. rubra GED8 with the yogurt starter (L. bulgaricus 2-11 + S. thermophilus 15HA) in ultrafiltrate under conditions of intracellular production of maximum amount of carotenoids and exopolysaccharides synthesis enables combined utilization of the culture fluid from the fermentation process.

  2. Fluid dynamics

    CERN Document Server

    Bernard, Peter S

    2015-01-01

    This book presents a focused, readable account of the principal physical and mathematical ideas at the heart of fluid dynamics. Graduate students in engineering, applied math, and physics who are taking their first graduate course in fluids will find this book invaluable in providing the background in physics and mathematics necessary to pursue advanced study. The book includes a detailed derivation of the Navier-Stokes and energy equations, followed by many examples of their use in studying the dynamics of fluid flows. Modern tensor analysis is used to simplify the mathematical derivations, thus allowing a clearer view of the physics. Peter Bernard also covers the motivation behind many fundamental concepts such as Bernoulli's equation and the stream function. Many exercises are designed with a view toward using MATLAB or its equivalent to simplify and extend the analysis of fluid motion including developing flow simulations based on techniques described in the book.

  3. R fluids

    Directory of Open Access Journals (Sweden)

    Caimmi R.

    2008-01-01

    Full Text Available A theory of collisionless fluids is developed in a unified picture, where nonrotating (Ωf1 = Ωf2 = Ωf3 = 0 figures with some given random velocity component distributions, and rotating (Ωf1 = Ωf2 = Ωf3 figures with a different random velocity component distributions, make adjoint configurations to the same system. R fluids are defined as ideal, self-gravitating fluids satisfying the virial theorem assumptions, in presence of systematic rotation around each of the principal axes of inertia. To this aim, mean and rms angular velocities and mean and rms tangential velocity components are expressed, by weighting on the moment of inertia and the mass, respectively. The figure rotation is defined as the mean angular velocity, weighted on the moment of inertia, with respect to a selected axis. The generalized tensor virial equations (Caimmi and Marmo 2005 are formulated for R fluids and further attention is devoted to axisymmetric configurations where, for selected coordinate axes, a variation in figure rotation has to be counterbalanced by a variation in anisotropy excess and vice versa. A microscopical analysis of systematic and random motions is performed under a few general hypotheses, by reversing the sign of tangential or axial velocity components of an assigned fraction of particles, leaving the distribution function and other parameters unchanged (Meza 2002. The application of the reversion process to tangential velocity components is found to imply the conversion of random motion rotation kinetic energy into systematic motion rotation kinetic energy. The application of the reversion process to axial velocity components is found to imply the conversion of random motion translation kinetic energy into systematic motion translation kinetic energy, and the loss related to a change of reference frame is expressed in terms of systematic motion (imaginary rotation kinetic energy. A number of special situations are investigated in greater

  4. Determining antioxidant activities of lactobacilli cell-free supernatants by cellular antioxidant assay: a comparison with traditional methods.

    Directory of Open Access Journals (Sweden)

    Jiali Xing

    Full Text Available Antioxidant activity of lactic acid bacteria is associated with multiple health-protective effects. Traditional indexes of chemical antioxidant activities poorly reflect the antioxidant effects of these bacteria in vivo. Cellular antioxidant activity (CAA assay was used in this study to determine the antioxidant activity of cell-free supernatants (CFSs of 10 Lactobacillus strains. The performance of the CAA assay was compared with that of four chemical antioxidant activity assays, namely, DPPH radical scavenging, hydroxyl radical scavenging (HRS, reducing power (RP, and inhibition of linoleic acid peroxidation (ILAP. Results of the CAA assay were associated with those of DPPH and ILAP assays, but not with those of RP and HRS assays. The inter- and intra-specific antioxidant activities of CFS were characterized by chemical and CAA assays. L. rhamnosus CCFM 1107 displayed a high antioxidative effect similar to positive control L. rhamnosus GG ATCC 53103 in all of the assays. The CAA assay is a potential method for the detection of antioxidant activities of lactobacilli CFSs.

  5. MicroRNA Expression in Salivary Supernatant of Patients with Pancreatic Cancer and Its Relationship with ZHENG

    Directory of Open Access Journals (Sweden)

    Song Gao

    2014-01-01

    Full Text Available In traditional Chinese medicine (TCM, diagnosis and prescriptions are based on the signs and symptoms which are recognized as ZHENG. The cornerstone of TCM is to differentially treat one ZHENG from others, which is also known as syndrome differentiation, and this relies on the gathering of clinical information through inspection, auscultation and olfaction, inquiry, and palpation. However, the biomolecular basis of the ZHENG remains unclear. In this study, the expressions of 384 cancer-related miRNAs in salivary supernatant of patients with pancreatic cancer were assessed by miRNA polymerase chain reaction (PCR array, and the different expression patterns of miRNA in three different groups of ZHENG were studied with use of real-time quantitative PCR (qRT-PCR. Some miRNAs were found to be specifically expressed in some ZHENGs, for instance, miR-17, miR-21, and miR-181b in Shi-Re ZHENG and miR-196a in Pi-Xu ZHENG. This indicates that these miRNAs may play important roles in different ZHENG condition. Therefore, this study to some extent revealed the molecular basis of TCM ZHENG in pancreatic cancer.

  6. R Fluids

    Directory of Open Access Journals (Sweden)

    Caimmi, R.

    2008-06-01

    Full Text Available A theory of collisionless fluids is developed in a unified picture, where nonrotating $(widetilde{Omega_1}=widetilde{Omega_2}= widetilde{Omega_3}=0$ figures with some given random velocity component distributions, and rotating $(widetilde{Omega_1} ewidetilde{Omega_2} e widetilde{Omega_3} $ figures with a different random velocity component distributions, make adjoint configurations to the same system. R fluids are defined as ideal, self-gravitating fluids satisfying the virial theorem assumptions, in presence of systematic rotation around each of the principal axes of inertia. To this aim, mean and rms angular velocities and mean and rms tangential velocity components are expressed, by weighting on the moment of inertia and the mass, respectively. The figure rotation is defined as the mean angular velocity, weighted on the moment of inertia, with respectto a selected axis. The generalized tensor virial equations (Caimmi and Marmo 2005 are formulated for R fluidsand further attention is devoted to axisymmetric configurations where, for selected coordinateaxes, a variation in figure rotation has to be counterbalanced by a variation in anisotropy excess and viceversa. A microscopical analysis of systematic and random motions is performed under a fewgeneral hypotheses, by reversing the sign of tangential or axial velocity components of anassigned fraction of particles, leaving the distribution function and other parametersunchanged (Meza 2002. The application of the reversion process to tangential velocitycomponents is found to imply the conversion of random motion rotation kinetic energy intosystematic motion rotation kinetic energy. The application ofthe reversion process to axial velocity components is found to imply the conversionof random motion translation kinetic energy into systematic motion translation kinetic energy, and theloss related to a change of reference frame is expressed in terms of systematic motion (imaginary rotation kinetic

  7. Analysis of tank 39H (HTF-39-15-61, 62) surface and subsurface supernatant samples in support of corrosion control program

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-08-19

    This report provides the results of analyses on Tanks 39H surface and subsurface supernatant liquid samples in support of the Corrosion Control Program. Analyses included warm acid strike preparation followed by analysis for silicon, aluminum, and sodium and water dilution preparation followed by analysis for anions. Other reported analytical results include analyses results for uranium, Pu-241 and Pu-239.

  8. Evaluation of the Antioxidative, Antibacterial, and Anti-Inflammatory Effects of the Aloe Fermentation Supernatant Containing Lactobacillus plantarum HM218749.1

    Directory of Open Access Journals (Sweden)

    Meixiu Jiang

    2016-01-01

    Full Text Available Little work is done to develop Aloe vera (AV using probiotics. To explore the potential benefits, the antioxidant effects and the antibacterial effects on foodborne pathogens of Aloe fermentation supernatant were evaluated in vitro. Our results indicated that the Aloe fermentation supernatant fermented by Lactobacillus plantarum HM218749.1 had very strong scavenging capacities of the DPPH (86%, O2•- (85%, OH• (76%, and Fe2+ chelation (82% and reducing powers (242.5 mg/L, and the inhibition zones for Salmonella typhimurium, Salmonella enteritidis, Shigella flexneri, Escherichia coli, Listeria monocytogenes, S. dysenteriae 301, Staphylococcus aureus Cowan1, and Propionibacterium acnes were 16, 15, 19, 20, 21, 20, and 27 mm. Moreover, the low concentration of Aloe fermentation supernatant had significantly reduced the production of IL-1β, TNF-α, and IL-6 in both mRNA and protein levels (P<0.01. Therefore, the Aloe fermentation supernatant can be used as functional beverage or cosmetic ingredients to guard human intestinal health, delaying senescence, and prevent chronic diseases.

  9. Evaluation of the Antioxidative, Antibacterial, and Anti-Inflammatory Effects of the Aloe Fermentation Supernatant Containing Lactobacillus plantarum HM218749.1

    Science.gov (United States)

    Deng, Kan; Jiang, Chunling; Fu, Mingui; Guo, Chunlan; Wang, Xiaolei; Wang, Xin; Meng, Fanjing; Yang, Shaoguo; Deng, Keyu

    2016-01-01

    Little work is done to develop Aloe vera (AV) using probiotics. To explore the potential benefits, the antioxidant effects and the antibacterial effects on foodborne pathogens of Aloe fermentation supernatant were evaluated in vitro. Our results indicated that the Aloe fermentation supernatant fermented by Lactobacillus plantarum HM218749.1 had very strong scavenging capacities of the DPPH (86%), O2•− (85%), •OH (76%), and Fe2+ chelation (82%) and reducing powers (242.5 mg/L), and the inhibition zones for Salmonella typhimurium, Salmonella enteritidis, Shigella flexneri, Escherichia coli, Listeria monocytogenes, S. dysenteriae 301, Staphylococcus aureus Cowan1, and Propionibacterium acnes were 16, 15, 19, 20, 21, 20, and 27 mm. Moreover, the low concentration of Aloe fermentation supernatant had significantly reduced the production of IL-1β, TNF-α, and IL-6 in both mRNA and protein levels (P < 0.01). Therefore, the Aloe fermentation supernatant can be used as functional beverage or cosmetic ingredients to guard human intestinal health, delaying senescence, and prevent chronic diseases. PMID:27493450

  10. [The comparative characteristics of antibacterial properties of the peptides of the active site of GM-CSF, and substances delivered from supernatants of hematopoietic progenitor CD34+45- cells].

    Science.gov (United States)

    Zurochka, A V; Zurochka, V A; Kostolomova, E G; Dobrynina, M A; Sykhoveĭ, Iu G; Gritsenko, V A

    2012-01-01

    The antibacterial activity of synthetic peptides of the active site of GM-CSF and supernatants of CD34+45- hematopoietic progenitor cells has been investigated GM-CSF peptides and cell supernatants were found to possess pronounced antibacterial activity, at that a combination of these substances has a more pronounced activity in comparison with the single substances. Possible mechanisms of the identified effects of synthetic peptides and substances from the supernatants of CD34+5- cells are discussed.

  11. Fluid Shifts

    Science.gov (United States)

    Stenger, M. B.; Hargens, A. R.; Dulchavsky, S. A.; Arbeille, P.; Danielson, R. W.; Ebert, D. J.; Garcia, K. M.; Johnston, S. L.; Laurie, S. S.; Lee, S. M. C.; Liu, J.; Macias, B.; Martin, D. S.; Minkoff, L.; Ploutz-Snyder, R.; Ribeiro, L. C.; Sargsyan, A.; Smith, S. M.

    2017-01-01

    Introduction. NASA's Human Research Program is focused on addressing health risks associated with long-duration missions on the International Space Station (ISS) and future exploration-class missions beyond low Earth orbit. Visual acuity changes observed after short-duration missions were largely transient, but now more than 50 percent of ISS astronauts have experienced more profound, chronic changes with objective structural findings such as optic disc edema, globe flattening and choroidal folds. These structural and functional changes are referred to as the visual impairment and intracranial pressure (VIIP) syndrome. Development of VIIP symptoms may be related to elevated intracranial pressure (ICP) secondary to spaceflight-induced cephalad fluid shifts, but this hypothesis has not been tested. The purpose of this study is to characterize fluid distribution and compartmentalization associated with long-duration spaceflight and to determine if a relation exists with vision changes and other elements of the VIIP syndrome. We also seek to determine whether the magnitude of fluid shifts during spaceflight, as well as any VIIP-related effects of those shifts, are predicted by the crewmember's pre-flight status and responses to acute hemodynamic manipulations, specifically posture changes and lower body negative pressure. Methods. We will examine a variety of physiologic variables in 10 long-duration ISS crewmembers using the test conditions and timeline presented in the figure below. Measures include: (1) fluid compartmentalization (total body water by D2O, extracellular fluid by NaBr, intracellular fluid by calculation, plasma volume by CO rebreathe, interstitial fluid by calculation); (2) forehead/eyelids, tibia, and calcaneus tissue thickness (by ultrasound); (3) vascular dimensions by ultrasound (jugular veins, cerebral and carotid arteries, vertebral arteries and veins, portal vein); (4) vascular dynamics by MRI (head/neck blood flow, cerebrospinal fluid

  12. Fluid dynamics

    CERN Document Server

    Ruban, Anatoly I

    This is the first book in a four-part series designed to give a comprehensive and coherent description of Fluid Dynamics, starting with chapters on classical theory suitable for an introductory undergraduate lecture course, and then progressing through more advanced material up to the level of modern research in the field. The present Part 1 consists of four chapters. Chapter 1 begins with a discussion of Continuum Hypothesis, which is followed by an introduction to macroscopic functions, the velocity vector, pressure, density, and enthalpy. We then analyse the forces acting inside a fluid, and deduce the Navier-Stokes equations for incompressible and compressible fluids in Cartesian and curvilinear coordinates. In Chapter 2 we study the properties of a number of flows that are presented by the so-called exact solutions of the Navier-Stokes equations, including the Couette flow between two parallel plates, Hagen-Poiseuille flow through a pipe, and Karman flow above an infinite rotating disk. Chapter 3 is d...

  13. Selective extraction of trace levels of polychlorinated and polybrominated contaminants by supercritical fluid-solid-phase microextraction and determination by gas chromatography/mass spectrometry. Application to aquaculture fish feed and cultured marine species.

    Science.gov (United States)

    Rodil, R; Carro, A M; Lorenzo, R A; Cela Torrijos, R

    2005-04-01

    The persistence, ubiquity, and toxicity of polyhalogenated compounds, together with their presence in fish feed, make it necessary to monitor these organic pollutants in the routine quality assurance programs of aquaculture activities, as this food chain is a source of these toxic compounds for human consumers. A new approach based on simultaneous supercritical fluid extraction-sample cleanup, followed by solid-phase microextraction-gas chromatography/mass spectrometry (SFE-SPME-GC/MS/MS) has been developed as an advantageous analytical tool for the determination of 15 organohalogenated compounds (including pesticides, polychlorinated and polybrominated biphenyls, and polybrominated diphenyl ethers) in aquaculture feed at very low levels. The influence of several parameters in the efficiency of the SPE/SPME combination was systematically investigated by chemometric approaches. In the optimal conditions, the developed procedure provides an excellent linearity, detection, and quantification limits (below 10 pg/g) for most of the analytes investigated, being at the same time advantageous in terms of rapidity, convenience, and avoiding the need of toxic organic solvents. The procedure was applied to the analysis of aquaculture feed and cultured marine species and tested for accuracy against IAEA 406 reference material.

  14. Genome Wide Expression of Amniotic Fluid Stem Cells in Early and Late Passage during in Vitro Culture%羊水干细胞体外培养早期和后期基因表达谱分析

    Institute of Scientific and Technical Information of China (English)

    焦扬; 刘建军; 冯国华; 于明; 赵洪波

    2013-01-01

    Objective This study was purposed to investigate the changes of biological properties and expression patterns of the amniotic fluid stem cells (AFSCs) during in vitro culture.Methods The gene chip data of amniotic fluid stem cells were obtained from GEO database and statistically analyzed using R and Bioconductor to identify the differentially expressed genes,then do the Gene Ontology analysis and KEGG pathway analysis.Results We did not find differentially expressed genes by linear model using limma package,but 495 differentially expressed probes were identified by RankProd,including 217 up-regulated and 278 down-regulated probes.Further analysis with Gene Ontology functional categories showed that the up-regulated genes were concentrated in those related to collagen fibril organization,extracellular matrix organization,extracellular structure organization,skeletal system development,cell adhesion and biological adhesion and the down-regulated probes in late passage were associated with nuclear division,mitosis,and cell cycle.The up-regulated pathway was ECM-receptor interaction and focal adhesion.Cell cycle,cytokine-cytokine receptor interaction,p53 signaling pathway and oocyte meiosis were down-regulated.Conclusion AFSCs maintain their genome-wide expression profile during in-vitro culture.%目的 探讨羊水干细胞(amniotic fluid stem cells,AFSCs)体外培养早期和后期全基因组表达规律.方法 在公共基因芯片数据库GEO中找到羊水干细胞相关的基因芯片数据,使用R和Bioconductor软件包对其进行统计学分析,筛选差异表达基因,并进行GO分析和KEGG通路分析.结果 通过线性模型没有找到差异表达基因,而通过秩积法找到217个上调探针,278个下调探针.对这些差异表达基因进行功能富集,上调的生物过程是胶原纤维组织相关过程、细胞外基质组织、胞外结构组织、骨骼系统发育、细胞粘附等.下调过程最显著的是核分裂过程,有丝分

  15. Culture medium for amylase production by toxigenic fungi

    Directory of Open Access Journals (Sweden)

    Figueira Edson Luiz Zangrando

    2000-01-01

    Full Text Available Mycelial growth and amylase production by a mycotoxigenic strain of Fusarium moniliforme and Aspergillus flavus were evaluated in a culture medium containing starch, glycerol, wheat bran or corn. With emphasis on corn, different fractions composed by germ, degermed seed, starch, milky stage corn and the respective starch or supernatant fraction were analyzed for F. moniliforme growth . The medium composed of milky stage corn supernatant promoted the best mycelial growth (p<0.05, and it was used to prepare amylase production medium in the next step. The medium composed with 2% ground corn in milky stage corn supernatant (350g of milky stage corn blended with 250mL water and centrifuged promoted the highest amylase production, which was at the 10th day of fermentation, both for F. moniliforme (42.32U/mL and A. flavus (4,745.54U/mL.

  16. Pleural Fluid Analysis Test

    Science.gov (United States)

    ... Home Visit Global Sites Search Help? Pleural Fluid Analysis Share this page: Was this page helpful? Formal name: Pleural Fluid Analysis Related tests: Pericardial Fluid Analysis , Peritoneal Fluid Analysis , ...

  17. Significance of amniotic fluid cells culture in the prenatal diagnosis for chromosomal diseases%羊水细胞培养用于染色体病产前诊断分析

    Institute of Scientific and Technical Information of China (English)

    刘慈; 刘义辉; 刘学军; 王振海; 张晓艳; 辛虹

    2014-01-01

    Objective To investigate the feasibility and necessity of amniotic fluid cells culture in the prenatal diagnosis for chromosomal diseases in order to prevent the birth of fetus with chromosomal diseases .Methods The 293 specimens of amniotic cell obtained by amniocentesis from 293 pregnant women with prenatal diagnosis indications were cultured and analyzed .Results Among the 293 specimens of amniotic cell ,14 cases of chromosome abnormality were found , with the chromosome abnormality rate being 4.78%,including 9 cases of chromosome number abnormality and 5 cases of chromosomal structure abnormality .The chromosome abnormality rates were different in different groups with different prenatal diagnosis indications ,in which the chromosome abnormality rates were the highest in the group with one party of the couple with chromosome abnormality .Conclusion The amniotic cell culture and chromosome karyotype analysis are the effective means in prenatal diagnosis for chromosomal diseases in the pregnant women with prenatal diagnosis indications at mid trimester of pregnancy, moreover, the combined detection of serological screening , chromosome examination and ultrasonic inspection plays an important role in the prevention of child birth defects .%目的:探讨羊水细胞培养对孕中期孕妇进行产前诊断的可行性及必要性,防止染色体病患儿的出生。方法对293例有产前诊断指征的孕妇进行羊膜腔穿刺,采取羊水细胞培养,制备中期染色体,分析胎儿核型,进行产前诊断。结果发现染色体异常14例,异常率4.78%,其中数目异常9例,结构异常5例,不同产前诊断指征分组中的异常率不同,以夫妇一方为染色体异常组异常率最高。结论妊娠中期对有产前诊断指征的孕妇进行羊水细胞培养染色体核型分析是产前诊断的重要手段,而血清学筛查,染色体检查和超声检查三者相互结合对于预防出生缺陷意义重大。

  18. Analysis of tank 51H (HTF-51-15-77) subsurface supernatant sample in support of enrichment and corrosion control programs

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-08-18

    This report provides the results of analyses on Tank 51H subsurface supernatant liquid sample in support of the Enrichment Control Program (ECP) and the Corrosion Control Program (CCP). The purpose of the ECP sample taken from Tank 51H in early June was to determine if the later decants would be “acceptable feed” to the 2H and 3H evaporator systems.

  19. Analysis of tank 51H (HTF-51-15-77) subsurface supernatant sample in support of enrichment and corrosion control programs

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-08-18

    This report provides the results of analyses on Tank 51H subsurface supernatant liquid sample in support of the Enrichment Control Program (ECP) and the Corrosion Control Program (CCP).The purpose of the ECP sample taken from Tank 51H in early June was to determine if the later decants would be “acceptable feed” to the 2H and 3H evaporator systems.

  20. 热损伤角质形成细胞培养上清液对成纤维细胞生物学行为的影响%Effect of heat injured keratinocytes supernatant on biological behavior of fibroblasts

    Institute of Scientific and Technical Information of China (English)

    白晓智; 胡大海; 张万福; 张战凤; 石继红; 蔡维霞; 朱华宇; 朱雄翔; 汤朝武

    2010-01-01

    Objective To observe the effect of the supernatant of heat injured keratinocytes (KC) on biological behavior of the dermal fibroblasts ( Fb). Methods Human dermal Fb were isolated and cultured. A model of heat injured KC ( HaCaT) was reproduced in vitro. Supernatant of normal KC and the supernatant of KC culture 12 hours after heat injury were collected and diluted with non-serum DMEM in 1:1 volume ratio to make normal KC conditioned medium ( NKCM ) and heat injury KC conditioned medium ( HKCM) respectively. Fb was respectively treated with non-serum DMEM and 2 kinds of conditioned medium. (1) The proliferation of Fb was detected with MTT method at post culture hour (PCH) 12, 24, 36, 48. (2) The apoptosis of Fb was determined by flow cytometry at PCH 12 ( Fb were heat injured in advance; Fb without heat treatment was used as control). (3) At PCH 24, expression of a-SMA in Fb cytoplasm was determined with immunofluorescence method; expression of a-SMA mRNA in Fb was determined with real-time quantified PCR. Data were processed with one-way analysis of variance, and pairwise comparison among groups with LSD-t test. Results (1) The proliferation of Fb: the absorbance value of Fb cultured with HKCM at PCH 12, 24, 36, 48 wa9 respectively higher than that of Fb cultured with non-serum DMEM ( with t value respectively 1. 89, 2. 35 , 2.02, 1.94, and P values all below 0. 01). There were significant statistical differences between the absorbance values of Fb cultured with HKCM and those of Fb cultured with 1MKCM at PCH 12, 24, and 48 (at PCH 12, t = 1. 83, P <0. 01; at PCH 24,t =2.91, P < 0. 05 ; at PCH 48 , t = 1. 83 , P < 0.05). (2 ) Apoptosis of Fb cultured with HKCM was diminished as compared with that of Fb cultured with NKCM and of Fb without treatment ( t =3.31 , P <0.05; t = 1.47, P < 0.01). (3) The expression of α-SMA (red fluorescence) in Fb cultured with non-serum DMEM or NKCM was less as seen under fluorescence scope, and it was obviously increased in Fb

  1. Activation of resting human B cells by helper T-cell clone supernatant: characterization of a human B-cell-activating factor.

    Science.gov (United States)

    Diu, A; Gougeon, M L; Moreau, J L; Reinherz, E L; Thèze, J

    1987-12-01

    The effects of helper T-cell clone supernatants on resting human B cells were investigated. Four different helper T-cell clones (two T4+ and two T8+) were stimulated by anti-T3 monoclonal antibodies on Sepharose beads or anti-T11(2) plus anti-T11(3) monoclonal antibodies. The supernatants from these activated clones induced the proliferation of highly purified resting B lymphocytes from the peripheral blood. The B cells exhibited a cell size and a surface-antigen pattern (4F2 antigen and transferrin receptor) of phase G0 B cells, and they were functionally resting. In response to T-cell supernatants a large fraction of the B cells enlarged and expressed 4F2 antigens and transferrin receptors. In gel filtration, the corresponding activity migrated with an apparent Mr of 12,000-15,000. Our findings strongly support the existence of a human B-cell-activating factor acting on resting B cells and causing them to enter phase G1 of the cell cycle.

  2. [Induced sputum supernatant prostaglandin E2 during oral aspirin challenge of asthmatic patients with and without aspirin hypersensitivity and healthy controls--pilot study].

    Science.gov (United States)

    Ignacak, Maria; Celejewska-Wójcik, Natalia; Wójcik, Krzysztof; Sałapa, Kinga; Konduracka, Ewa; Sanak, Marek; Tyrak, Katarzyna; Sładek, Krzysztof; Musiał, Jacek; Mastalerz, Lucyna

    2016-01-01

    The aim of this pilot study was to evaluate changes in the concentration of prostaglandin E2 (PGE2) in induced sputum supernatant in 3 groups: sub- jects with NSAID-exacerbated respira- tory disease (NERD), aspirin tolerant asthma (ATA) and healthy controls (HC), before and after oral aspirin chal- lenge test. The study was conducted in the years 2014-2015 at the Clinical Department of the Pulmonology Clinic at the University Hospital in Cracow. 43 patients were enrolled in the study (NERD - n = 15, ATA - n = 15 and HC - n = 13). All of them underwent a placebo-controlled oral aspirin challenge. Sputum was induced 24 hours before the challenge and immediately after the test. Induced sputum was processed in order to obtain cystospin slides to depict inflammatory cell patterns and supernatants, in which PGE2 was measured. The concentration of PGE2 was determined using mass spectrometry coupled with gas chromatography (gas chromatography/mass spectrometry - GC/MS). After aspirin challenge, the concentration of PGE2 in induced sputum supernatant decreased in both asthmatics hypersensitive to aspirin (p = 0.01) and those who tolerated aspirin well (p = 0.17). The change in the healthy control group was not statistically significant. These results support the cyclooxygenase theory of PGE2 inhibition by aspirin. However, the mechanism of bronchoconstriction after aspirin administration alone in patients with NSAID-exacerbated respiratory disease remains unclear.

  3. Production and assay of cellulolytic enzyme activity of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya abbatoir, Indonesia

    Directory of Open Access Journals (Sweden)

    W. P. Lokapirnasari

    2015-03-01

    Full Text Available Aim: This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4-β-D-glucanase, exo-(1,4-β-Dglucanase, and β-glucosidase, at optimum temperature and optimum pH of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia. Materials and Methods: To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase, and β-glucosidase. Results: Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5. Conclusion: E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4-β-D-glucanase, exo-(1,4-β-D-glucanase and β-glucosidase.

  4. Fast and simple online sample preparation coupled with capillary LC-MS/MS for determination of prostaglandins in cell culture supernatants

    DEFF Research Database (Denmark)

    Rinne, Sandra; Ramstad Kleiveland, Charlotte; Kassem, Moustapha

    2007-01-01

    An online 2-D strong cation exchange (SCX)-RP capillary liquid chromatographic (cLC) method with IT mass spectrometric (IT-MS/MS) detection for the simultaneous determination of prostaglandin (PG) A(1), PGD(2), PGE(1), PGE(2), PGF(2a), 6-keto-(6k)PGF(1a), and 15-Delta(12,14)-deoxy-PGJ(2) (15dPGJ(...

  5. High-throughput screening techniques for rapid PEG-based precipitation of IgG4 mAb from clarified cell culture supernatant.

    Science.gov (United States)

    Knevelman, Carol; Davies, Jim; Allen, Lee; Titchener-Hooker, Nigel J

    2010-01-01

    Locating optimal protein precipitation conditions for complex biological feed materials is problematic. This article describes the application of a series of high-throughput platforms for the rapid identification and selection of conditions for the precipitation of an IgG(4) monoclonal antibody (mAb) from a complex feedstock using only microliter quantities of material. The approach uses 96-microwell filter plates combined with high-throughput analytical methods and a method for well volume determination for product quantification. The low material, time and resource requirements facilitated the use of a full factorial Design of Experiments (DoE) for the rapid investigation into how critical parameters impact the IgG(4) precipitation. To aid the DoE, a set of preliminary range-finding studies were conducted first. Data collected through this approach describing Polyethylene Glycol (PEG) precipitation of the IgG(4) as a function of mAb concentration, precipitant concentration, and pH are presented. Response surface diagrams were used to explore interactions between parameters and to inform selection of the most favorable conditions for maximum yield and purification. PEG concentrations required for maximum yield and purity were dependant on the IgG(4) concentration; however, concentrations of 14 to 20% w/v, pH 6.5, gave optimal levels of yield and purity. Application of the high-throughput approach enabled 1,155 conditions to be examined with less than 1 g of material. The level of insights gained over such a short time frame is indicative of the power of microwell experimentation in allowing the rapid identification of appropriate processing conditions for key bioprocess operations. Copyright 2009 American Institute of Chemical Engineers

  6. Screening for epitope specificity directly on culture supernatants in the early phase of monoclonal antibody production by an ELISA with biotin-labeled antigen

    DEFF Research Database (Denmark)

    Andersen, Ditte C; Jensen, Charlotte H; Gregersen, Annemette

    2004-01-01

    This report describes an assay for comparison of epitope specificity in groups of monoclonal antibodies against a given antigen. The only prerequisite is the biotin-labeled antigen. One of the monoclonal antibodies is captured onto a plastic surface via a rabbit anti-mouse Ig, and the other...... preincubated with biotinylated antigen. When the two antibodies react with the same epitope subsequent binding of the biotin-labeled antigen is abolished (inhibition). In the cases where no inhibition was observed, the two antibodies were considered to react with distinct, independent epitopes. The obvious...

  7. Toxigenicity testing of clinical isolates of non-typhoidal salmonellae in Vero cell culture & Caenorhabditis elegans model.

    Science.gov (United States)

    Jesudason, Mary V; V Balaji, V; Densibai, Shoba

    2006-06-01

    The non-typhoidal salmonellae (NTS) are recognized agents of gastroenteritis worldwide. Some of the NTS do not produces cytotoxic changes in tissue culture and not much is known about the endotoxicity of the clinical isolates of NTS (mostly Salmonella enterica serotype Typhimurium and Salmonella enterica serotype Enteritidis). We examined the exotoxic (cytotoxin) and endotoxic activity of clinical isolates of NTS in two assay models namely Vero cell culture and the nematode, Caenorhabditis elegans. Bacteria-free culture supernatants of 40 isolates NTS were tested in 96 well microtitre plate containing confluent monolayers of Vero cells. For the effects on C. elegans, the worms were exposed to bacteria free culture supernatants in 24 well microtitre plate for 24 h and then transferred to OP50 Escherichia coli lawn culture. The endotoxic activity of the live bacterium was studied by feeding the worms in the lawn culture of NTS separately. No cytopathic effect was observed with NTS tested in Vero cell culture assay. Likewise, the worms exposed to the bacteria-free culture supernatants were found active up to 7 days. In the co-culture killing assay, worms were found dead with characteristic stiff and straight appearance by 16(th) day. The worms were alive up to 21 days in OP50 E. coli. Bacteria-free culture supernatants did not have any deleterious effect on worms or in Vero cell culture, suggesting that there is no soluble toxic factor (diffusible toxin) in the culture supernatants. However, live NTS were found to be lethal to the worms; indicating that direct interaction between viable NTS and C. elegans is necessary for killing.

  8. Effects of human promyelocytic leukemia HL-60 cel supernatant on differetiation of mononulear cel s from human umbilical cord blood%急性早幼粒白血病细胞培养上清液对人脐带血单个核细胞分化的影响

    Institute of Scientific and Technical Information of China (English)

    郑毅; 方宁; 陈代雄

    2013-01-01

    目的白血病细胞能够通过自分泌方式分泌细胞因子而作用于自身,使细胞大量的增殖而不分化;白血病细胞所分泌的细胞因子也能作用于人脐带血(UCB)单个核细胞(MNCs)中的CD34+/CD133+细胞亚群使之增殖而抑制其分化。关于急性早幼粒白血病细胞(HL-60细胞)培养上清液能否使UCB- MNCs分化尚未见报道,本实验主要目的是观察HL-60细胞培养上清液对人UCB-MNCs分化的影响。方法实验分为①有HL-60细胞培养上清液+StemSpan® SFEM组;②StemSpan® SFEM组;③HL-60细胞培养上清液+高糖(HG)-DMEM组;④HG-DMEM组;⑤HL-60细胞培养上清液+低糖(LG)-DMEM组;⑥LG-DMEM组。收分离的人UCB-MNCs按2.5×106/瓶分别接种于6组培养液中,37℃,5%CO2,饱和湿度培养,于培养前和培养12天用流式细胞仪(FCM)分析表型CD34,CD90和CD166的变化。结果各实验组CD34,CD34/CD90和CD166阳性细胞百分比均较培养前升高。结论%Objective:Leukemic cel s can excrete cytokines by an autocrine way, which contribute to its large-scale proliferation, and the cytokines may also contribute to the proliferations of CD34+/CD133+ subpopulation of UCB-MNCs too. To the effect of human promyelocytic leukemia HL-60 cel supernatant on the differentiation of UCB-MNCs, there is not related report so far. The aim of this study is to investigate effects of HL-60 cel supernatant on differentiations of mononuclear cel s from human umbilical cord blood. Methods:Six various cultural system are listed as fol ows: ① HL-60 cel supernatant+ StemSpan–SFEM; ② StemSpan–SFEM; ③ HL-60 cel supernatant+ HG-DMEM; ④ HG-DMEM; ⑤HL-60 cel supernatant LG-DMEM; ⑥ LG-DMEM. 2.50×106 of UCB-MNCs per bottle with the above each cultural system is incubated at 37℃ in a 5%CO2, saturation humidity of incubator, the changes of phenotype of CD34,CD90 and CD166 analyzed by FCM after 12 days. Results: Positive cel s with CD34+, CD166+ and CD34+/CD

  9. Thermophysical Properties of Fluids and Fluid Mixtures

    Energy Technology Data Exchange (ETDEWEB)

    Sengers, Jan V.; Anisimov, Mikhail A.

    2004-05-03

    The major goal of the project was to study the effect of critical fluctuations on the thermophysical properties and phase behavior of fluids and fluid mixtures. Long-range fluctuations appear because of the presence of critical phase transitions. A global theory of critical fluctuations was developed and applied to represent thermodynamic properties and transport properties of molecular fluids and fluid mixtures. In the second phase of the project, the theory was extended to deal with critical fluctuations in complex fluids such as polymer solutions and electrolyte solutions. The theoretical predictions have been confirmed by computer simulations and by light-scattering experiments. Fluctuations in fluids in nonequilibrium states have also been investigated.

  10. Primary culture of Rhodnius prolixus (Hemiptera: Reduviidae salivary gland cells

    Directory of Open Access Journals (Sweden)

    Fernanda F Rocha

    2010-03-01

    Full Text Available In the present paper, we developed a primary culture of Rhodnius prolixus salivary gland and main salivary canal cells. Cells remained viable in culture for 30 days. Three types of cells were indentified in the salivary gland cultures, with binuclear cells being the most abundant. The supernatants of salivary cultures contained mainly 16-24 kDa proteins and presented anticoagulant and apyrase activities. Secretion vesicles were observed budding from the cellular monolayer of the main salivary canal cells. These results indicate that R. prolixus salivary proteins may be produced in vitro and suggest that the main salivary canal may have a possible secretory role.

  11. Determination of Sr{sup 90}, Sb{sup 125}, Cs{sup 137} and absolute beta activities in First Cycle Supernatent

    Energy Technology Data Exchange (ETDEWEB)

    Helmholz, H.R.

    1954-03-15

    This report discusses procedures have been established for determining the absolute activities of Sr{sup 90}, Sb{sup 125}, Cs{sup 137}, and gross beta in First Cycle Supernatant. These procedures are also valid for other samples such as scavenged Radioactive Waste (RAW), the primary restriction being that the age of the material be at least 250 days so that any Sr{sup 89} activity (53 day half-life) is negligible. With the exception of the gross beta analysis the results are believed to be accurate to within 10%.

  12. Rapid method for identification of group B streptococci in neonatal blood cultures.

    OpenAIRE

    Holmes, R. L.; Harada, W A

    1981-01-01

    A rapid technique used for the identification of Streptococcus agalactiae, Lancefield group B, from the blood cultures of two neonatal infants is reported. The method utilized the Phadebact Streptococcus Test System (Pharmacia Diagnostics, Piscataway, N.J.) and the supernatant from 13- and 14-h blood cultures. Additional studies with simulated neonatal blood cultures revealed that this method was reproducible. Additional studies also revealed that some non-specific agglutination did occur, wh...

  13. Analysis of Tank 38H (HTF-38-15-47, 49) and Tank 43H (HTF-43-15-51, 53) surface and subsurface supernatant samples in support of enrichment and corrosion control programs

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-06-30

    This report provides the results of analyses on Tanks 38H and 43H surface and subsurface supernatant liquid samples in support of the Enrichment Control Program (ECP) and the Corrosion Control Program (CCP).

  14. Analysis of Tank 38H (HTF-38-14-150, 151) and Tank 43H (HTF- 43-14-152, 53) Surface and Subsurface Supernatant Samples in Support of Enrichment Control, Corrosion Control and Sodium Aluminosilicate Formation Potential Programs

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-01-14

    This report provides the results of analyses on Tanks 38H and 43H surface and subsurface supernatant liquid samples in support of the Enrichment Control Program (ECP), the Corrosion Control Program and Sodium Aluminosilicate Formation Potential in the Evaporator.

  15. Robotic art, culture and cultural imagination

    DEFF Research Database (Denmark)

    Romic, Bojana

    2015-01-01

    In the recent years there has been an ongoing debate about the notion and (possible) function of robotic culture; with the development of the new generation of drones, as well as the advancement in social and health robotics, questions about robot culture seem to open a variety of discussions...... – beginning with the inquiry of how can we grasp the notion of culture, as an umbrella term for the range of habitual and technological practices, in relation to equally heterogeneous field of robotics. This article aims to accentuate the importance of cultural imagination of robots in situating the robotic...... research, observing it 'as a mixed register of fantasy and an actual practice' (Kakoudaki, 2007). The emphasis will be put on the robotic art which, I argue, is in the fluid state of exchange with other areas of robotic research, equally benefiting from the larger context of cultural imagination of robots...

  16. Differentiation efficiency of human umbilical cord mesenchymal stem cells into hepatocytes under two kinds of liver homogenate supernatants: a comparative study%两种肝组织匀浆上清液诱导人脐带间充质干细胞向肝细胞分化的比较

    Institute of Scientific and Technical Information of China (English)

    闫成; 薛改; 吴丽颖; 刘建芳; 侯艳宁

    2015-01-01

    BACKGROUND:Previous studies have demonstrated that normal rat liver homogenate supernatant can induce human umbilical cord mesenchymal stem cels to differentiate into hepatocyte-like cels with partial hepatocyte functions. However, whether fibrotic liver homogenate supernatant can work or how the inducing effect is remains unclear. OBJECTIVE:To investigate the differentiation potential of human umbilical cord mesenchymal stem cels into hepatocytes under the normal liver and fibrotic liver microenvironment in vitro. METHODS:Liver fibrosis was induced in the SD rats by repeated intraperitoneal injections of 3% thioacetamide at a dose of 200 mg/kg body mass, twice a week for 4 weeks, and then fibrotic liver tissues and normal liver tissues were used to prepare liver homogenate supernatants. Passage 3 human umbilical cord mesenchymal stem cels were used and divided into standard control group (cels were cultured in DMEM/F12 with 10% fetal bovine serum), fibrotic liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 50 g/L fibrotic liver homogenate supernatants), normal liver homogenate supernatants group (cels were cultured in DMEM/F12 with 10% fetal bovine serum and 100 g/L normal liver homogenate supernatants). The morphological changes of the cels in each group were recorded under inverted microscope; the protein levels of CK18, AFP, CYP3A4, CYP2E1, CYP2D6 and TPH2 were evaluated using western blot assay. Furthermore, the concentration of albumin in the cels was measured. RESULTS AND CONCLUSION:After a 7-day inducement, the stem cels in liver homogenate supernatants groups lost their fusiform shape and changed into hepatocyte-like cels with the morphous of round shape. Compared with the standard control group, the hepatocyte-like cels in the two liver homogenate supernatants groups exhibited human hepatocyte biomarkers, CK18 and AFP. The standard control group cels could express a little amount of CYP2E1, while cels in

  17. Impact of Cell-free Supernatant of Lactic Acid Bacteria on Putrescine and Other Polyamine Formation by Foodborne Pathogens in Ornithine Decarboxylase Broth.

    Science.gov (United States)

    Ozogul, Fatih; Tabanelli, Giulia; Toy, Nurten; Gardini, Fausto

    2015-06-24

    Conversion of ornithine to putrescine by Salmonella Paratyphi A, Listeria monocytogenes, Staphylococcus aureus, and Escherichia coli was investigated in ornithine decarboxylase broth (ODB) using cell-free supernatants (CFSs) obtained from Leuconostoc mesenterodies subsp. cremoris, Pediococcus acidilactici, Lactococcus lactis subsp. lactis, Streptococcus thermophilus. Two groups of cell-free supernatants (25 or 50%) and control (only ODB) were prepared to investigate putrescine (PUT) and other polyamine formation by foodborne pathogens (FBPs). Significant differences (p < 0.05) were observed among the species for each amine. All of the CFSs reduced the formation of PUT by ≥65%. The production of cadaverine (CAD) was scarcely affected by the presence of CFSs, with the exception of the samples inoculated with L. monocytogenes. The variation in polyamine was found with respect to the control samples. Spermidine (SPD) was produced in lower amount in many samples, especially in Gram-negative FBPs, whereas spermine (SPN) increased drastically in the major part of the samples concerning the control. Histamine (HIS) was characterized by a marked concentration decrease in all of the samples, and tyramine (TYR) was accumulated in very low concentrations in the controls. Therefore, the ability of bacteria to produce certain biogenic amines such as HIS, TYR, PUT, and CAD has been studied to assess their risk and prevent their formation in food products. The results obtained from this study concluded that the lactic acid bacteria (LAB) strains with non-decarboxylase activity are capable of avoiding or limiting biogenic amine formation by FBP.

  18. Effect of phenobarbital pretreatment on benzene biotransformation in the rat. Pt. 2. 9. 000 g supernatant and isolated perfused liver versus living rat

    Energy Technology Data Exchange (ETDEWEB)

    Gut, I.; Hatle, K.; Zizkova, L.

    1981-03-01

    Factors responsible for different quantitative effect of phenobarbital (PB) pretreatment on benzene metabolism to phenol in vivo and in vitro were studied in male Wistar rats. A more than 4-fold increase of benzene metabolism was observed with 9,000 g supernatant of liver homogenate, 2.8- to 4-fold increase with isolated perfused liver; phenol formation in vivo after oral benzene was increased by PB 2-fold, but only shortly following benzene administration and the enhancement rapidly diminished to 1.15-fold increase in the total excreted phenol. Benzene concentrations in 9,000 g supernatant incubations were 2 mM, those with isolated perfused livers were up to 4 mM, but those in blood in vivo were below 0.3 mM; the effect of PB induction in vivo disappeared along with decreasing benzene and increasing phenol blood concentrations which surpassed benzene 2-3 h after oral benzene administration. The effect of benzene concentration on the manifestation of PB induction is also supported by almost a 2-fold increased phenol formation in PB rats over controls in vivo after repeated administration of benzene. The elimination of radioactive metabolites of orally administered benzene-/sup 14/C, in urine was markedly inhibited by intraperitoneal administration of phenol, but not by pyrocatechol, resorcinol or hydroquinol suggesting that phenol might inhibit benzene metabolism in vivo especially when its concentration exceeds that of benzene.

  19. [Diagnosis of amniotic fluid embolism with blood samples by liquid-based cytology technique].

    Science.gov (United States)

    Liu, Bao-qin; Deng, Jian-qiang; Hou, An-chao; Cai, Ji-feng

    2014-12-01

    To establish the diagnosis of amniotic fluid embolism with blood samples by liquid-based cytology technique and to study the validity of method. The blood samples were collected from patients who suffered from amniotic fluid embolism. The components of amniotic fluid in blood samples were examined with blood smear by two direct smear methods (supernatant smear, sediment smear) and two liquid-based cytology methods (automatic smear, manual smear). The positive detection rate of each method was calculated. The positive detection rates of two liquid-based cytology methods (84.6% and 92.3%, respectively) were much higher than those of two direct methods (53.8% and 61.5%, respectively). The liquid-based cytology technique could improve the positive detection rate of amniotic fluid embolism.

  20. Auxillary Fluid Flowmeter

    DEFF Research Database (Denmark)

    RezaNejad Gatabi, Javad; Forouzbakhsh, Farshid; Ebrahimi Darkhaneh, Hadi

    2010-01-01

    and with measuring its travel time between two different positions, its velocity could be calculated. Given the velocity of the auxiliary fluid, the velocity of the main fluid could be calculated. Using this technique, it is possible to measure the velocity of any kind of fluids, if an appropriate auxiliary fluid...

  1. Fluid mechanics in fluids at rest.

    Science.gov (United States)

    Brenner, Howard

    2012-07-01

    Using readily available experimental thermophoretic particle-velocity data it is shown, contrary to current teachings, that for the case of compressible flows independent dye- and particle-tracer velocity measurements of the local fluid velocity at a point in a flowing fluid do not generally result in the same fluid velocity measure. Rather, tracer-velocity equality holds only for incompressible flows. For compressible fluids, each type of tracer is shown to monitor a fundamentally different fluid velocity, with (i) a dye (or any other such molecular-tagging scheme) measuring the fluid's mass velocity v appearing in the continuity equation and (ii) a small, physicochemically and thermally inert, macroscopic (i.e., non-Brownian), solid particle measuring the fluid's volume velocity v(v). The term "compressibility" as used here includes not only pressure effects on density, but also temperature effects thereon. (For example, owing to a liquid's generally nonzero isobaric coefficient of thermal expansion, nonisothermal liquid flows are to be regarded as compressible despite the general perception of liquids as being incompressible.) Recognition of the fact that two independent fluid velocities, mass- and volume-based, are formally required to model continuum fluid behavior impacts on the foundations of contemporary (monovelocity) fluid mechanics. Included therein are the Navier-Stokes-Fourier equations, which are now seen to apply only to incompressible fluids (a fact well-known, empirically, to experimental gas kineticists). The findings of a difference in tracer velocities heralds the introduction into fluid mechanics of a general bipartite theory of fluid mechanics, bivelocity hydrodynamics [Brenner, Int. J. Eng. Sci. 54, 67 (2012)], differing from conventional hydrodynamics in situations entailing compressible flows and reducing to conventional hydrodynamics when the flow is incompressible, while being applicable to both liquids and gases.

  2. Videotapes and Movies on Fluid Dynamics and Fluid Machines

    OpenAIRE

    Carr, Bobbie; Young, Virginia E.

    1996-01-01

    Chapter 17 of Handbook of Fluid Dynamics and Fluid Machinery: Experimental and Computational Fluid Dynamics, Volume 11. A list of videorecordings and 16mm motion pictures about Fluid Dynamics and Fluid Machines.

  3. Cytopathogenicity of Naegleria for cultured neuroblastoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Fulford, D.E.

    1985-01-01

    The cytopathic activity of live Naegleria amoebae and cell-free lysates of Naegleria for B-103 rat neuroblastoma cells was investigated using a /sup 51/Cr release assay. Live amoebae and cell-free lysates of N. fowleri, N. australiensis, N. lovaniensis, and N. gruberi all induced sufficient damage to radiolabeled B-103 cells to cause a significant release of chromium. The cytotoxic activity present in the cell-free lysates of N. fowleri can be recovered in the supernatant fluid following centrifugation at 100,000xg and precipitation of the 100,000xg supernatant fluid with ammonium sulfate. Initial characterization of the cytotoxic factor indicates that it is a heat labile, pH sensitive, soluble protein. The cytotoxic activity is abolished by either extraction, unaffected by repeated freeze-thawing, and is not sensitive to inhibitors of proteolytic enzymes. Phospholipase A activity was detected in the cytotoxic ammonium sulfate precipitable material, suggesting that this enzyme activity may have a role in the cytotoxic activity of the cell-free lysates.

  4. Near infrared spectroscopy combined with multivariate analysis for monitoring the ethanol precipitation process of fraction I+II+III supernatant in human albumin separation.

    Science.gov (United States)

    Li, Can; Wang, Fei; Zang, Lixuan; Zang, Hengchang; Alcalà, Manel; Nie, Lei; Wang, Mingyu; Li, Lian

    2017-03-15

    Nowadays, as a powerful process analytical tool, near infrared spectroscopy (NIRS) has been widely applied in process monitoring. In present work, NIRS combined with multivariate analysis was used to monitor the ethanol precipitation process of fraction I+II+III (FI+II+III) supernatant in human albumin (HA) separation to achieve qualitative and quantitative monitoring at the same time and assure the product's quality. First, a qualitative model was established by using principal component analysis (PCA) with 6 of 8 normal batches samples, and evaluated by the remaining 2 normal batches and 3 abnormal batches. The results showed that the first principal component (PC1) score chart could be successfully used for fault detection and diagnosis. Then, two quantitative models were built with 6 of 8 normal batches to determine the content of the total protein (TP) and HA separately by using partial least squares regression (PLS-R) strategy, and the models were validated by 2 remaining normal batches. The determination coefficient of validation (Rp(2)), root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP) and ratio of performance deviation (RPD) were 0.975, 0.501g/L, 0.465g/L and 5.57 for TP, and 0.969, 0.530g/L, 0.341g/L and 5.47 for HA, respectively. The results showed that the established models could give a rapid and accurate measurement of the content of TP and HA. The results of this study indicated that NIRS is an effective tool and could be successfully used for qualitative and quantitative monitoring the ethanol precipitation process of FI+II+III supernatant simultaneously. This research has significant reference value for assuring the quality and improving the recovery ratio of HA in industrialization scale by using NIRS.

  5. Near infrared spectroscopy combined with multivariate analysis for monitoring the ethanol precipitation process of fraction I + II + III supernatant in human albumin separation

    Science.gov (United States)

    Li, Can; Wang, Fei; Zang, Lixuan; Zang, Hengchang; Alcalà, Manel; Nie, Lei; Wang, Mingyu; Li, Lian

    2017-03-01

    Nowadays, as a powerful process analytical tool, near infrared spectroscopy (NIRS) has been widely applied in process monitoring. In present work, NIRS combined with multivariate analysis was used to monitor the ethanol precipitation process of fraction I + II + III (FI + II + III) supernatant in human albumin (HA) separation to achieve qualitative and quantitative monitoring at the same time and assure the product's quality. First, a qualitative model was established by using principal component analysis (PCA) with 6 of 8 normal batches samples, and evaluated by the remaining 2 normal batches and 3 abnormal batches. The results showed that the first principal component (PC1) score chart could be successfully used for fault detection and diagnosis. Then, two quantitative models were built with 6 of 8 normal batches to determine the content of the total protein (TP) and HA separately by using partial least squares regression (PLS-R) strategy, and the models were validated by 2 remaining normal batches. The determination coefficient of validation (Rp2), root mean square error of cross validation (RMSECV), root mean square error of prediction (RMSEP) and ratio of performance deviation (RPD) were 0.975, 0.501 g/L, 0.465 g/L and 5.57 for TP, and 0.969, 0.530 g/L, 0.341 g/L and 5.47 for HA, respectively. The results showed that the established models could give a rapid and accurate measurement of the content of TP and HA. The results of this study indicated that NIRS is an effective tool and could be successfully used for qualitative and quantitative monitoring the ethanol precipitation process of FI + II + III supernatant simultaneously. This research has significant reference value for assuring the quality and improving the recovery ratio of HA in industrialization scale by using NIRS.

  6. A cytopreparatory method for cerebrospinal fluid in which the cell yield is high and the fluid is saved for chemical analysis.

    Science.gov (United States)

    Tutuarima, J A; Hische, E A; Sylva-Steenland, R M; van der Helm, H J

    1988-01-01

    A method for the concentration of cells from cerebrospinal fluid is described. An adaptation of a commercial cytochamber, consisting of a holder that fixes a disposable chamber directly on a microscope slide, was used. The cells were spun down in a conventional swing-out centrifuge, which was provided with a bucket for the cytochamber system. After removing most of the supernatant with a pipette, the remaining fluid was absorbed by means of a suction device consisting of a disposable pipette tip covered with a piece of Leukopor and filled with Sephadex G10 beads. The method gives a high recovery of cells (90%), together with a good preservation of cell morphology, and leaves about 80% of the fluid available for analysis of the soluble components.

  7. Astrocytes Enhance Streptococcus suis-Glial Cell Interaction in Primary Astrocyte-Microglial Cell Co-Cultures.

    Science.gov (United States)

    Seele, Jana; Nau, Roland; Prajeeth, Chittappen K; Stangel, Martin; Valentin-Weigand, Peter; Seitz, Maren

    2016-06-13

    Streptococcus (S.) suis infections are the most common cause of meningitis in pigs. Moreover, S. suis is a zoonotic pathogen, which can lead to meningitis in humans, mainly in adults. We assume that glial cells may play a crucial role in host-pathogen interactions during S. suis infection of the central nervous system. Glial cells are considered to possess important functions during inflammation and injury of the brain in bacterial meningitis. In the present study, we established primary astrocyte-microglial cell co-cultures to investigate interactions of S. suis with glial cells. For this purpose, microglial cells and astrocytes were isolated from new-born mouse brains and characterized by flow cytometry, followed by the establishment of astrocyte and microglial cell mono-cultures as well as astrocyte-microglial cell co-cultures. In addition, we prepared microglial cell mono-cultures co-incubated with uninfected astrocyte mono-culture supernatants and astrocyte mono-cultures co-incubated with uninfected microglial cell mono-culture supernatants. After infection of the different cell cultures with S. suis, bacteria-cell association was mainly observed with microglial cells and most prominently with a non-encapsulated mutant of S. suis. A time-dependent induction of NO release was found only in the co-cultures and after co-incubation of microglial cells with uninfected supernatants of astrocyte mono-cultures mainly after infection with the capsular mutant. Only moderate cytotoxic effects were found in co-cultured glial cells after infection with S. suis. Taken together, astrocytes and astrocyte supernatants increased interaction of microglial cells with S. suis. Astrocyte-microglial cell co-cultures are suitable to study S. suis infections and bacteria-cell association as well as NO release by microglial cells was enhanced in the presence of astrocytes.

  8. Enhancing the culturability of bacteria from the gastrointestinal tract of farmed adult turbot Scophthalmus maximus

    Science.gov (United States)

    Xing, Mengxin; Hou, Zhanhui; Qu, Yanmei; Liu, Bin

    2014-03-01

    Eighteen agar media were tested for the culture of gut-associated bacteria from farmed adult turbot ( Scophthalmus maximus), including 16 agar media with or without 1% gastrointestinal (GI) supernatant, or with 2% or 4% GI supernatant. A total of 1 711 colonies were analyzed and 24 operational taxonomic units (OTUs) were identified. The greatest bacterial diversity was isolated on Zobell 2216E/Zobell 2216E+ agar media, whereas MRS/MRS+ agar media produced a low diversity of colonies. Agar media with GI supernatant (1%, 2%, or 4%) showed increased diversity and yielded different profiles of OTUs from the corresponding original media, suggesting that GI supernatant provides substances that enhance the culture efficiency of bacteria from the turbot GI tract. The large majority of the colonies (82%) were γ-Proteobacteria, whereas 15.6% and 2.4% of colonies were Firmicutes and Actinobacteria, respectively. At the genus level, 49.4% of all colonies were assigned to Vibrio. Other potential pathogens, including Pseudomonas, Photobacterium, and Enterobacter, and potential probiotics, including Bacillus, Paenibacillus, and Pseudomonas, were also isolated on agar media. Most cultured bacteria belonged to species that were first described in the turbot GI tract. The impact of these species on turbot physiology and health should be investigated further.

  9. Synovial fluid analysis

    Science.gov (United States)

    ... bursae (fluid-filled sacs in the joints), and tendon sheaths. After the joint area is cleaned, the ... HS. Synovial fluid analysis, synovial biopsy, and synovial pathology. In: Firestein GS, Budd RC, Gabriel SE, McInnes ...

  10. Amniotic fluid (image)

    Science.gov (United States)

    Amniotic fluid surrounds the growing fetus in the womb and protects the fetus from injury and temperature changes. ... of fetal movement and permits musculoskeletal development. The amniotic fluid can be withdrawn in a procedure called amniocentsis ...

  11. Pericardial Fluid Analysis

    Science.gov (United States)

    ... help diagnose the cause of inflammation of the pericardium (pericarditis) and/or fluid accumulation around the heart ( ... pressure within blood vessels or inflammation of the pericardium. An initial set of tests, including fluid protein ...

  12. Pericardial fluid Gram stain

    Science.gov (United States)

    ... staining a sample of fluid taken from the pericardium. This is the sac surrounding the heart to ... sample of fluid will be taken from the pericardium. This is done through a procedure called pericardiocentesis . ...

  13. Lectures on fluid mechanics

    CERN Document Server

    Shinbrot, Marvin

    2012-01-01

    Readable and user-friendly, this high-level introduction explores the derivation of the equations of fluid motion from statistical mechanics, classical theory, and a portion of the modern mathematical theory of viscous, incompressible fluids. 1973 edition.

  14. Electric fluid pump

    Science.gov (United States)

    Van Dam, Jeremy Daniel; Turnquist, Norman Arnold; Raminosoa, Tsarafidy; Shah, Manoj Ramprasad; Shen, Xiaochun

    2015-09-29

    An electric machine is presented. The electric machine includes a hollow rotor; and a stator disposed within the hollow rotor, the stator defining a flow channel. The hollow rotor includes a first end portion defining a fluid inlet, a second end portion defining a fluid outlet; the fluid inlet, the fluid outlet, and the flow channel of the stator being configured to allow passage of a fluid from the fluid inlet to the fluid outlet via the flow channel; and wherein the hollow rotor is characterized by a largest cross-sectional area of hollow rotor, and wherein the flow channel is characterized by a smallest cross-sectional area of the flow channel, wherein the smallest cross-sectional area of the flow channel is at least about 25% of the largest cross-sectional area of the hollow rotor. An electric fluid pump and a power generation system are also presented.

  15. Fluid force transducer

    Science.gov (United States)

    Jendrzejczyk, Joseph A.

    1982-01-01

    An electrical fluid force transducer for measuring the magnitude and direction of fluid forces caused by lateral fluid flow, includes a movable sleeve which is deflectable in response to the movement of fluid, and a rod fixed to the sleeve to translate forces applied to the sleeve to strain gauges attached to the rod, the strain gauges being connected in a bridge circuit arrangement enabling generation of a signal output indicative of the magnitude and direction of the force applied to the sleeve.

  16. Characteristics of HCV replication and expression in a cultured human liver carcinoma cell line in vitro

    Institute of Scientific and Technical Information of China (English)

    SONG Zhi-qing; HAO Fei; MIN Feng; LIU Dao-jian

    2001-01-01

    Objective: To establish a cell culture system to support HCV long-term replication in vitro. Methods: A human hepatoma cell line 7721 was tested for its susceptibility to HCV by incubating with a serum from chronic hepatitis C patient. Cells and supernatant of the culture medium were harvested at various time-phases during the culturing periods. The presence of HCV RNA, the expression of HCV antigens in cells and/or supematant were examined with RT-PCR, in situ hybridization and immunohistochemistry respectively. Results: It was found that the intracellular HCV RNA was first detected on the 2nd day after culture, and then could be intermittently detected in both cells and supernatant over a period of at least 3 months after culture. HCV NS3, CP10 antigens were expressed in the cells. The fresh cells could be infected with the supernatant from cultured infected cells and the transmission of viral genome from HCV-infected 7721 cells to peripheral blood mononuclear cells (PBMCs) was also observed. Conclusion: Our findings suggest that the human liver carcinoma cell line7721 is not only susceptible to HCV but also can support its long replication in vitro. This cell line with HCV infection in vitro can serve as a useful tool for the study of the mechanism of HCV infection and replication, the evaluation of antiviral agents, and the primary selection of neutralization assays and HCV vaccine development.

  17. Fluid and particle mechanics

    CERN Document Server

    Michell, S J

    2013-01-01

    Fluid and Particle Mechanics provides information pertinent to hydraulics or fluid mechanics. This book discusses the properties and behavior of liquids and gases in motion and at rest. Organized into nine chapters, this book begins with an overview of the science of fluid mechanics that is subdivided accordingly into two main branches, namely, fluid statics and fluid dynamics. This text then examines the flowmeter devices used for the measurement of flow of liquids and gases. Other chapters consider the principle of resistance in open channel flow, which is based on improper application of th

  18. Catalase expression is modulated by vancomycin and ciprofloxacin and influences the formation of free radicals in Staphylococcus aureus cultures

    DEFF Research Database (Denmark)

    Wang, Ying; Hougaard, Anni Bygvrå; Paulander, Wilhelm Erik Axel

    2015-01-01

    formation in the human pathogen Staphylococcus aureus treated with a bactericidal antibiotic, vancomycin or ciprofloxacin. While we were unable to detect ESR signals in bacterial cells, hydroxyl radicals were observed in the supernatant of bacterial cell cultures. Surprisingly, the strongest signal...

  19. Identification and localization of a soluble antigen, Ag2, of 136 kDa from Plasmodium falciparum in vitro cultures

    DEFF Research Database (Denmark)

    Jakobsen, P H; Grellier, P; Theander, T G

    1991-01-01

    The soluble antigens, antigen 2 (Ag2) and antigen 6 (Ag6), were copurified from supernatants of P. falciparum in vitro cultures by affinity chromatography and Fast Protein Liquid Chromatography. Rabbit antibodies to Ag2 were raised and characterized by crossed immunoelectrophoresis. Ag2 appeared ...... vacuole and in clefts in the infected erythrocyte cytoplasma as shown by immunogold electron microscopy....

  20. Fluid cooled electrical assembly

    Science.gov (United States)

    Rinehart, Lawrence E.; Romero, Guillermo L.

    2007-02-06

    A heat producing, fluid cooled assembly that includes a housing made of liquid-impermeable material, which defines a fluid inlet and a fluid outlet and an opening. Also included is an electrical package having a set of semiconductor electrical devices supported on a substrate and the second major surface is a heat sink adapted to express heat generated from the electrical apparatus and wherein the second major surface defines a rim that is fit to the opening. Further, the housing is constructed so that as fluid travels from the fluid inlet to the fluid outlet it is constrained to flow past the opening thereby placing the fluid in contact with the heat sink.

  1. Spinning fluids reactor

    Science.gov (United States)

    Miller, Jan D; Hupka, Jan; Aranowski, Robert

    2012-11-20

    A spinning fluids reactor, includes a reactor body (24) having a circular cross-section and a fluid contactor screen (26) within the reactor body (24). The fluid contactor screen (26) having a plurality of apertures and a circular cross-section concentric with the reactor body (24) for a length thus forming an inner volume (28) bound by the fluid contactor screen (26) and an outer volume (30) bound by the reactor body (24) and the fluid contactor screen (26). A primary inlet (20) can be operatively connected to the reactor body (24) and can be configured to produce flow-through first spinning flow of a first fluid within the inner volume (28). A secondary inlet (22) can similarly be operatively connected to the reactor body (24) and can be configured to produce a second flow of a second fluid within the outer volume (30) which is optionally spinning.

  2. Inflammatory cytokine detection in adenotonsill and peripheral blood mononuclear cells- culture in adenotonsillectomy patients: a comparative study

    Directory of Open Access Journals (Sweden)

    Farhadi M

    2013-04-01

    Full Text Available Background: Tonsils and adenoid hypertrophy is a major respiratory symptom in children which is partly due to recruitment of inflammatory cells in upper airway lymph nodes as a result of the effects of synthesis and release of different inflammatory cytokines. It seems that infections play role in concert with these cytokines leading to tonsilar hypertrophy and other pathologic consequences. It is proposed that cellular infiltrate of tonsils and adenoids may secrete different quantities of these cytokines compared with peripheral blood mononuclear cells (PBMC cultures.Methods: Among patients who were admitted for adenotonsillectomy to the ENT ward, 37 patients, under 1-12 years old patients with fulfill criteria selected to include the study. Excised adenoid and tonsils cultured and inflammatory cytokines Interferon-γ (INF-γ, Interlukine-1 (IL-1, IL-6, IL-8 and tumor necrosis factor-α (TNF-α measured in cellular culture supernatant. The same cytokines measured in PBMC cultures.Results: The data shows that there is a significant difference between IFN-γ and IL-8 amounts in adenoid tissue culture supernatant and PBMC culture of our patients. Furth-ermore, the amounts of IFN-γ, IL-1 and IL-8 showed considerable difference between tonsilar tissue culture supernatant and PBMC culture of these patients. Although there is a significant correlation between IL-6 amounts in tissue culture supernatant and PBMC culture (P=0.02, the respective data for TNF is only almost significant.Conclusion: Inflammatory cytokines may have significant role in the early provoke of inflammation occurred in hypertrophied tonsils and adenoid. The majority of these cyt-okines increase the expression of adhesion molecules on epithelial cells and influence the recruitment of leucocytes and inflamed tonsils. On the other hand lack of sufficient cytokine release may lead to persistent infections and may cause chronic inflammation and hypertrophied tissue.

  3. Diminished exoproteome of Frankia spp. in culture and symbiosis.

    Science.gov (United States)

    Mastronunzio, J E; Huang, Y; Benson, D R

    2009-11-01

    Frankia species are the most geographically widespread gram-positive plant symbionts, carrying out N(2) fixation in root nodules of trees and woody shrubs called actinorhizal plants. Taking advantage of the sequencing of three Frankia genomes, proteomics techniques were used to investigate the population of extracellular proteins (the exoproteome) from Frankia, some of which potentially mediate host-microbe interactions. Initial two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of culture supernatants indicated that cytoplasmic proteins appeared in supernatants as cells aged, likely because older hyphae lyse in this slow-growing filamentous actinomycete. Using liquid chromatography coupled to tandem mass spectrometry to identify peptides, 38 proteins were identified in the culture supernatant of Frankia sp. strain CcI3, but only three had predicted export signal peptides. In symbiotic cells, 42 signal peptide-containing proteins were detected from strain CcI3 in Casuarina cunninghamiana and Casuarina glauca root nodules, while 73 and 53 putative secreted proteins containing signal peptides were identified from Frankia strains in field-collected root nodules of Alnus incana and Elaeagnus angustifolia, respectively. Solute-binding proteins were the most commonly identified secreted proteins in symbiosis, particularly those predicted to bind branched-chain amino acids and peptides. These direct proteomics results complement a previous bioinformatics study that predicted few secreted hydrolytic enzymes in the Frankia proteome and provide direct evidence that the symbiosis succeeds partly, if not largely, because of a benign relationship.

  4. Analysis of tank 39H (HTF-39-15-61, 62) surface and subsurface supernatant samples in support of corrosion control program

    Energy Technology Data Exchange (ETDEWEB)

    Oji, L. N. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2015-08-01

    This report provides the results of analyses on Tanks 39H surface and subsurface supernatant liquid samples in support of the Corrosion Control Program. Analyses included warm acid strike preparation followed by analysis for silicon, aluminum, and sodium and water dilution preparation followed by analysis for anions. Other reported analytical results include analyses results for uranium, Pu-241 and Pu-239. The measured sodium concentration averaged, respectively, 4.28E+00 ± 9.30E-02 M and 4.32E+00 ± 1.076E-01 M in the Tank 39H surface sample and Tank 39H subsurface sample. In general, the nitrate, nitrite, free-OH and specific gravity of the Tank 39H surface and subsurface samples were all about the same in magnitude, respectively, averaging 1.98 M, 0.314 M, 1.26 M and 1.24. The measured silicon concentration for the Tank 39H surface and subsurface samples were, respectively, 3.84E+01± 5.51E+00 and 4.14E+01± 1.17E+00 mg/L. Based on the uranium, Pu-241 and Pu-239 concentrations, the calculated U-235 equivalent is 21.41 wt% for the surface sample and 21.32 wt% for the subsurface sample.

  5. Bio-chemical process for nitrogen and phosphorus removal by draining out anaerobic rich phosphate supernatant in ERP-SBR system

    Institute of Scientific and Technical Information of China (English)

    JIFangying; XUXiaoyi; LUOGuyuan

    2003-01-01

    The method of fixed phosphate coming from anaerobic reactor by the auxiliary chemical process is applied in External Recycle Process-SBR (ERP-SBR). This process changes the model of draining out activated sludge in the traditional biological phosphorus removal system to discharge anaerobic poly-phosphate supernatant. This process eliminates the contradiction of control for Solid Removal Time (SRT) in process of biological nitrogen and phosphorus removal. It can obtain high removal efficiency of nitrogen(N) and phosphorus(P) in longer SRT. Experiment results show that: when SRT=50 ~ 80 d, TN=28.6~ 58.3 mg/L, TP=5.5~ 13.5 mg/L in influent, COD≤ 34mg/L, TN≤ 6.02 mg/L, PO43-≤0.23 mg/L in effluent. The amount of lime is only 5% of traditional methods. The phosphorus content in the chemical sludge is 12 %~15 % and the recycle of phosphorus can be realized easily.

  6. Isolation of monoclonal antibody from a Chinese hamster ovary supernatant. II: dynamics of the integrated separation on ion exchange and hydrophobic interaction chromatography media.

    Science.gov (United States)

    Marek, Wojciech; Muca, Renata; Woś, Sylwia; Piątkowski, Wojciech; Antos, Dorota

    2013-08-30

    Dynamics of the purification process of a CHO derived monoclonal antibody by ion exchange chromatography (IEC), hydrophobic interaction chromatography (HIC) and their integration has been investigated. To quantify the adsorption behavior of the target protein (IgG1) and impurities contained in the supernatant, their elution course on IEC and HIC columns has been analyzed versus pH and/or the salt concentration in the mobile phase. A short-cut method has been proposed for mathematical modeling and determining underlying kinetic and thermodynamic parameters. The accuracy of the model predictions has been verified by comparing the simulated and experimental band profiles recorded in both chromatographic processes. After verification, the model was used to optimize operating conditions for the column loading and chromatographic elution in the integrated process IEC/HIC. Two alternative loading techniques based on the upstream and downstream feed dilution were taken into account in the optimization routine. In the first one the feed stream was diluted with the loading buffer prior to the column loading, while in the latter one the feed dilution was realized inside the column using the multiple-injection technique. It was shown that the downstream dilution allowed significant reduction of the contact time between the protein and the loading buffer.

  7. Bicontinuous Fluid Structure with Low Cohesive Energy: Molecular Basis for Exceptionally Low Interfacial Tension of Complex Coacervate Fluids.

    Science.gov (United States)

    Huang, Kuo-Ying; Yoo, Hee Young; Jho, YongSeok; Han, Songi; Hwang, Dong Soo

    2016-05-24

    An exceptionally low interfacial tension of a dense fluid of concentrated polyelectrolyte complexes, phase-separated from a biphasic fluid known as complex coacervates, represents a unique and highly sought-after materials property that inspires novel applications from superior coating to wet adhesion. Despite extensive studies and broad interest, the molecular and structural bases for the unique properties of complex coacervates are unclear. Here, a microphase-separated complex coacervate fluid generated by mixing a recombinant mussel foot protein-1 (mfp-1) as the polycation and hyaluronic acid (HA) as the polyanion at stoichiometric ratios was macroscopically phase-separated into a dense complex coacervate and a dilute supernatant phase to enable separate characterization of the two fluid phases. Surprisingly, despite up to 4 orders of magnitude differing density of the polyelectrolytes, the diffusivity of water in these two phases was found to be indistinguishable. The presence of unbound, bulk-like, water in the dense fluid can be reconciled with a water population that is only weakly perturbed by the polyelectrolyte interface and network. This hypothesis was experimentally validated by cryo-TEM of the macroscopically phase-separated dense complex coacervate phase that was found to be a bicontinuous and biphasic nanostructured network, in which one of the phases was confirmed by staining techniques to be water and the other polyelectrolyte complexes. We conclude that a weak cohesive energy between water-water and water-polyelectrolytes manifests itself in a bicontinuous network, and is responsible for the exceptionally low interfacial energy of this complex fluid phase with respect to virtually any surface within an aqueous medium.

  8. Exosomal miRNAs from Peritoneum Lavage Fluid as Potential Prognostic Biomarkers of Peritoneal Metastasis in Gastric Cancer.

    Science.gov (United States)

    Tokuhisa, Motohiko; Ichikawa, Yasushi; Kosaka, Nobuyoshi; Ochiya, Takahiro; Yashiro, Masakazu; Hirakawa, Kosei; Kosaka, Takashi; Makino, Hirochika; Akiyama, Hirotoshi; Kunisaki, Chikara; Endo, Itaru

    2015-01-01

    Peritoneal metastasis is the most frequent type of recurrence in patients with gastric cancer (GC) and is associated with poor prognosis. Peritoneal lavage cytology, used to evaluate the risk of peritoneal metastasis, has low sensitivity. Here, we assessed the diagnostic potential of exosomal miRNA profiles in peritoneal fluid for the prediction of peritoneal dissemination in GC. Total RNA was extracted from exosomes isolated from six gastric malignant ascites (MA) samples, 24 peritoneal lavage fluid (PLF) samples, and culture supernatants (CM) of two human gastric carcinoma cell lines that differ in their potential for peritoneal metastasis. Expression of exosomal miRNAs was evaluated with Agilent Human miRNA microarrays and quantitative reverse transcription polymerase chain reaction (qRT-PCR). The microarray analysis indicated a low variability in the number and signal intensity of miRNAs detected among the samples. In the six MA fluids, miR-21 showed the highest signal intensity. We identified five miRNAs (miR-1225-5p, miR-320c, miR-1202, miR-1207-5p, and miR-4270) with high expression in MA samples, the PLF of serosa-invasive GC, and the CM of a highly metastatic GC cell line; these candidate miRNA species appear to be related to peritoneal dissemination. Differential expression of miR-21, miR-320c, and miR-1225-5p was validated in the PLF of serosa-invasive and non-invasive GC by qRT-PCR and miR-21 and miR-1225-5p were confirmed to be associated with serosal invasion in GC. PLF can be used to profile the expression of exosomal miRNAs. Our findings suggest that miR-21 and miR-1225-5p may serve as biomarkers of peritoneal recurrence after curative GC resection, thus providing a novel approach to early diagnosis of peritoneal dissemination of GC.

  9. Exosomal miRNAs from Peritoneum Lavage Fluid as Potential Prognostic Biomarkers of Peritoneal Metastasis in Gastric Cancer.

    Directory of Open Access Journals (Sweden)

    Motohiko Tokuhisa

    Full Text Available Peritoneal metastasis is the most frequent type of recurrence in patients with gastric cancer (GC and is associated with poor prognosis. Peritoneal lavage cytology, used to evaluate the risk of peritoneal metastasis, has low sensitivity. Here, we assessed the diagnostic potential of exosomal miRNA profiles in peritoneal fluid for the prediction of peritoneal dissemination in GC. Total RNA was extracted from exosomes isolated from six gastric malignant ascites (MA samples, 24 peritoneal lavage fluid (PLF samples, and culture supernatants (CM of two human gastric carcinoma cell lines that differ in their potential for peritoneal metastasis. Expression of exosomal miRNAs was evaluated with Agilent Human miRNA microarrays and quantitative reverse transcription polymerase chain reaction (qRT-PCR. The microarray analysis indicated a low variability in the number and signal intensity of miRNAs detected among the samples. In the six MA fluids, miR-21 showed the highest signal intensity. We identified five miRNAs (miR-1225-5p, miR-320c, miR-1202, miR-1207-5p, and miR-4270 with high expression in MA samples, the PLF of serosa-invasive GC, and the CM of a highly metastatic GC cell line; these candidate miRNA species appear to be related to peritoneal dissemination. Differential expression of miR-21, miR-320c, and miR-1225-5p was validated in the PLF of serosa-invasive and non-invasive GC by qRT-PCR and miR-21 and miR-1225-5p were confirmed to be associated with serosal invasion in GC. PLF can be used to profile the expression of exosomal miRNAs. Our findings suggest that miR-21 and miR-1225-5p may serve as biomarkers of peritoneal recurrence after curative GC resection, thus providing a novel approach to early diagnosis of peritoneal dissemination of GC.

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

    Science.gov (United States)

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

    1998-01-01

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

  11. Metalworking and machining fluids

    Science.gov (United States)

    Erdemir, Ali; Sykora, Frank; Dorbeck, Mark

    2010-10-12

    Improved boron-based metal working and machining fluids. Boric acid and boron-based additives that, when mixed with certain carrier fluids, such as water, cellulose and/or cellulose derivatives, polyhydric alcohol, polyalkylene glycol, polyvinyl alcohol, starch, dextrin, in solid and/or solvated forms result in improved metalworking and machining of metallic work pieces. Fluids manufactured with boric acid or boron-based additives effectively reduce friction, prevent galling and severe wear problems on cutting and forming tools.

  12. Protoscoleces and hydatid fluid promote expression of TGF-β from mouse spleen cells in vitro%原头蚴及囊液促体外培养小鼠脾细胞TGF-β表达的研究

    Institute of Scientific and Technical Information of China (English)

    陈小林; 印双红; 吴向未; 陈雪玲

    2013-01-01

    目的 观察细粒棘球蚴囊液及原头蚴对体外培养BABL/c小鼠脾细胞TGF-β表达的影响. 方法 取BABL/c小鼠脾细胞,制成细胞悬液,加入不同浓度囊液/原头蚴共培养72 h,用ELISA检测上清液TGF-β含量;同时收集细胞并提取mRNA,用RT-PCR检测TGF-βmRNA的相对表达量.试验以PBS与脾细胞混合培养组为对照. 结果 ELISA检测囊液组和原头蚴组TGF-β分泌均增加,与对照组相比差异有统计学意义(P<0.05);RT-PCR检测囊液组和原头蚴不同浓度组及对照组TGF-β mRNA的相对表达量,差异均有统计学意义(P<0.05),但囊液/原头蚴不同浓度组间差异无统计学意义(P>0.05). 结论 原头蚴和囊液均能刺激体外培养小鼠脾细胞TGF-β表达增加,提示原头蚴和囊液中含有能促使宿主产生TGF-β的抗原物质,这些抗原物质在包虫病的免疫逃避中发挥一定作用.%Objective To observe the effect of treatment with Echinococcus granulosus fluid/protoscoleces on TGF-β expression in mouse spleen cells in vitro.Methods After spleen cells from BABL/c mice were co-cultured with different concentrations of hydatid fluid or protoscoleces,the level of TGF-β in the co-culture supernatant were measured using ELISA.mRNA extracted from cells was collected and the relative level of expression of TGF-β mRNA was detected using RT-PCR.Spleen cells were co-cultured with PBS to serve as a control.Results ELISA indicated that,compared to the controls,cells co-cultured with hydatid fluid and protoscoleces had increased secretion of TGF-β; the difference in secretion was statistically significant (P<0.05).RT-PCR to detect the relative level of expression of TGF-β mRNA indicated that,compared to the controls,cells co-cultured with hydatid fluid and protoscoleces had significantly increased expression (P<0.05),but this increased expression did not differ significantly with different concentrations of hydatid fluid or protoscoleces

  13. Electrorheological fluids and methods

    Energy Technology Data Exchange (ETDEWEB)

    Green, Peter F.; McIntyre, Ernest C.

    2015-06-02

    Electrorheological fluids and methods include changes in liquid-like materials that can flow like milk and subsequently form solid-like structures under applied electric fields; e.g., about 1 kV/mm. Such fluids can be used in various ways as smart suspensions, including uses in automotive, defense, and civil engineering applications. Electrorheological fluids and methods include one or more polar molecule substituted polyhedral silsesquioxanes (e.g., sulfonated polyhedral silsesquioxanes) and one or more oils (e.g., silicone oil), where the fluid can be subjected to an electric field.

  14. The Fluids RAP

    Science.gov (United States)

    Nedyalkov, Ivaylo

    2016-11-01

    After fifteen years of experience in rap, and ten in fluid mechanics, "I am coming here with high-Reynolds-number stamina; I can beat these rap folks whose flows are... laminar." The rap relates fluid flows to rap flows. The fluid concepts presented in the song have varying complexity and the listeners/viewers will be encouraged to read the explanations on a site dedicated to the rap. The music video will provide an opportunity to share high-quality fluid visualizations with a general audience. This talk will present the rap lyrics, the vision for the video, and the strategy for outreach. Suggestions and comments will be welcomed.

  15. Theoretical Fluid Dynamics

    CERN Document Server

    Shivamoggi, Bhimsen K

    1998-01-01

    "Although there are many texts and monographs on fluid dynamics, I do not know of any which is as comprehensive as the present book. It surveys nearly the entire field of classical fluid dynamics in an advanced, compact, and clear manner, and discusses the various conceptual and analytical models of fluid flow." - Foundations of Physics on the first edition. Theoretical Fluid Dynamics functions equally well as a graduate-level text and a professional reference. Steering a middle course between the empiricism of engineering and the abstractions of pure mathematics, the author focuses

  16. Fluid dynamics transactions

    CERN Document Server

    Fiszdon, W

    1965-01-01

    Fluid Dynamics Transactions, Volume 2 compiles 46 papers on fluid dynamics, a subdiscipline of fluid mechanics that deals with fluid flow. The topics discussed in this book include developments in interference theory for aeronautical applications; diffusion from sources in a turbulent boundary layer; unsteady motion of a finite wing span in a compressible medium; and wall pressure covariance and comparison with experiment. The certain classes of non-stationary axially symmetric flows in magneto-gas-dynamics; description of the phenomenon of secondary flows in curved channels by means of co

  17. Organ culture system as a means to detect celiac disease.

    Science.gov (United States)

    Picarelli, Antonio; Libanori, Valerio; De Nitto, Daniela; Saponara, Annarita; Di Tola, Marco; Donato, Giuseppe

    2010-01-01

    Anti-endomysial and anti-transglutaminase antibodies can be produced in vitro by the intestinal mucosa of celiac disease (CD) patients in clinical remission, when the culture is performed in the presence of gliadin peptides. Our aim was to use this organ culture system as a means to detect the pathognomonic antibodies of celiac disease (CD) in the culture supernatants. Organ culture was performed in the presence of three different activators to evaluate which one induced the strongest antibody response in intestinal mucosa from patients in clinical remission of CD. Our data confirm the high efficiency of synthetic peptide 31-43 as a specific immunological activator in CD and demonstrate its capability to stimulate production/secretion of CD-specific antibodies. We envision that this organ culture system may prove to be useful as a new technique for CD diagnosis.

  18. [Simultaneous determination of guanidinoacetate, creatine and creatinine by liquid chromatography-tandem mass spectrometry: a diagnostic tool for creatine deficiency syndromes in body fluids and a perspective use on cultured fibroblasts].

    Science.gov (United States)

    Van Noolen, Laetitia; Monneret, Denis; Ducros, Véronique; Corne, Christelle; Lunardi, Joël; Faure, Patrice

    2013-01-01

    Guanidinoacetate (GAA) and creatine (Cr) are creatine deficiency syndromes (CDS) biochemical markers. We describe a liquid chromatography - tandem mass spectrometry method (LC/MSMS) performing simultaneous analysis of GAA, Cr and creatinine (Crn). Study of Cr uptake by fibroblasts for Cr transporter defect diagnosis is also assessed. The three butylated compounds were separated by liquid chromatography and MSMS quantification was achieved by isotopic dilution with electrospray positive ion mode. Linearity was demonstrated from 0 to 600, 675 and 4500 μmol/L and limit of quantification was 0.1, 0.04 and 0.9 μmol/L for GAA, Cr, and Crn respectively. Intra- and inter-assay precision for each analyte was better than 11%, and standard recoveries ranged from 83 to 109%. Reference values in cerebrospinal fluid samples for subjects ≥14 years were also established for GAA and Cr. Five fibroblast cell lines were used for Cr uptake study. Cr uptake by fibroblasts increased with the Cr media concentrations and was significantly inhibited by 3-guanidinopropionate (500 μmol/L), a Cr transporter inhibitor (96h incubation, [Cr media] = 25 μmol/L, p<0.05). A reliable LC/MSMS method for the diagnosis of CDS was developed in different biological fluids. Finally, results of the Cr uptake study reinforce the interest of this technique to diagnose Cr transporter deficiencies.

  19. Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

    Science.gov (United States)

    Reisinger, Nicole; Schaumberger, Simone; Nagl, Veronika; Hessenberger, Sabine; Schatzmayr, Gerd

    2015-01-01

    Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS), and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL) on cell viability of isolated epidermal and dermal hoof cells as well as on the tissue integrity of hoof explants. Furthermore, glucose, acetic acid, lactic acid, and propionic acid concentrations in explant supernatants were measured to evaluate the energy metabolism in the hoof tissue. LPS did not exhibit cytotoxic effects on epidermal or dermal cells. Force required to separate LPS treated hoof explants decreased in a concentration dependent manner. Specifically, explants incubated with 10 and 100 μg/mL needed significantly less force to separate compared to control explants. Lactic acid concentrations were significantly decreased in explants incubated with 5, 10, or 100 μg/mL LPS, while glucose, acetic acid and propionic acid concentrations were unaffected by LPS treatment. Our study indicates that LPS has no cytotoxic effect on epidermal and dermal cells isolated from hoof tissue, but impairs integrity of hoof explants. In addition, LPS led to an alteration of the lactic acid production in the lamellar tissue. Since our data highlight that LPS can affect the integrity of the equine hoof tissue in vitro, endotoxins should be further explored for their contribution to facilitate the development of laminitis.

  20. Concentration Dependent Influence of Lipopolysaccharides on Separation of Hoof Explants and Supernatant Lactic Acid Concentration in an Ex Vivo/In Vitro Laminitis Model.

    Directory of Open Access Journals (Sweden)

    Nicole Reisinger

    Full Text Available Laminitis is one of the most common diseases in horses. It is not only painful for the animal, but also has a significant financial impact on the equine industry. This multifactorial disease affects the connective tissue of the hoof. However, the pathogenesis of laminitis is still not fully understood. Endotoxins, also known as lipopolysaccharides (LPS, and bacterial exotoxins seem to play an important role during the development of laminitis. The aim of our study was to investigate the effect of increasing LPS concentrations (0, 2.5, 5, 10, and 100 μg/mL on cell viability of isolated epidermal and dermal hoof cells as well as on the tissue integrity of hoof explants. Furthermore, glucose, acetic acid, lactic acid, and propionic acid concentrations in explant supernatants were measured to evaluate the energy metabolism in the hoof tissue. LPS did not exhibit cytotoxic effects on epidermal or dermal cells. Force required to separate LPS treated hoof explants decreased in a concentration dependent manner. Specifically, explants incubated with 10 and 100 μg/mL needed significantly less force to separate compared to control explants. Lactic acid concentrations were significantly decreased in explants incubated with 5, 10, or 100 μg/mL LPS, while glucose, acetic acid and propionic acid concentrations were unaffected by LPS treatment. Our study indicates that LPS has no cytotoxic effect on epidermal and dermal cells isolated from hoof tissue, but impairs integrity of hoof explants. In addition, LPS led to an alteration of the lactic acid production in the lamellar tissue. Since our data highlight that LPS can affect the integrity of the equine hoof tissue in vitro, endotoxins should be further explored for their contribution to facilitate the development of laminitis.

  1. Lactobacillus rhamnosus GG supernatant promotes intestinal barrier function, balances Treg and TH17 cells and ameliorates hepatic injury in a mouse model of chronic-binge alcohol feeding.

    Science.gov (United States)

    Chen, Rui-Cong; Xu, Lan-Man; Du, Shan-Jie; Huang, Si-Si; Wu, He; Dong, Jia-Jia; Huang, Jian-Rong; Wang, Xiao-Dong; Feng, Wen-Ke; Chen, Yong-Ping

    2016-01-22

    Impaired intestinal barrier function plays a critical role in alcohol-induced hepatic injury, and the subsequent excessive absorbed endotoxin and bacterial translocation activate the immune response that aggravates the liver injury. Lactobacillus rhamnosus GG supernatant (LGG-s) has been suggested to improve intestinal barrier function and alleviate the liver injury induced by chronic and binge alcohol consumption, but the underlying mechanisms are still not clear. In this study, chronic-binge alcohol fed model was used to determine the effects of LGG-s on the prevention of alcoholic liver disease in C57BL/6 mice and investigate underlying mechanisms. Mice were fed Lieber-DeCarli diet containing 5% alcohol for 10 days, and one dose of alcohol was gavaged on Day 11. In one group, LGG-s was supplemented along with alcohol. Control mice were fed isocaloric diet. Nine hours later the mice were sacrificed for analysis. Chronic-binge alcohol exposure induced an elevation in liver enzymes, steatosis and morphology changes, while LGG-s supplementation attenuated these changes. Treatment with LGG-s significantly improved intestinal barrier function reflected by increased mRNA expression of tight junction (TJ) proteins and villus-crypt histology in ileum, and decreased Escherichia coli (E. coli) protein level in liver. Importantly, flow cytometry analysis showed that alcohol reduced Treg cell population while increased TH17 cell population as well as IL-17 secretion, which was reversed by LGG-s administration. In conclusion, our findings indicate that LGG-s is effective in preventing chronic-binge alcohol exposure-induced liver injury and shed a light on the importance of the balance of Treg and TH17 cells in the role of LGG-s application.

  2. Space Station fluid management logistics

    Science.gov (United States)

    Dominick, Sam M.

    1990-01-01

    Viewgraphs and discussion on space station fluid management logistics are presented. Topics covered include: fluid management logistics - issues for Space Station Freedom evolution; current fluid logistics approach; evolution of Space Station Freedom fluid resupply; launch vehicle evolution; ELV logistics system approach; logistics carrier configuration; expendable fluid/propellant carrier description; fluid carrier design concept; logistics carrier orbital operations; carrier operations at space station; summary/status of orbital fluid transfer techniques; Soviet progress tanker system; and Soviet propellant resupply system observations.

  3. Amniotic Fluid Glucose Concentration: A Marker for Infection in Preterm Labor and Preterm Premature Rupture of Membranes

    Directory of Open Access Journals (Sweden)

    Gary A. Dildy

    1994-01-01

    Full Text Available Amniotic fluid Gram stain and culture have been utilized as laboratory tests of microbial invasion of the amniotic cavity. The Gram stain of amniotic fluid has a low sensitivity in the detection of clinical infection or microbial invasion of the amniotic cavity, and amniotic fluid culture results are not immediately available for management decisions. Glucose concentration is used to diagnose infection in other sites such as cerebrospinal fluid.

  4. Urine culture

    Science.gov (United States)

    Culture and sensitivity - urine ... when urinating. You also may have a urine culture after you have been treated for an infection. ... when bacteria or yeast are found in the culture. This likely means that you have a urinary ...

  5. Dynamics of Complex Fluid-Fluid Interfaces

    NARCIS (Netherlands)

    Sagis, L.M.C.

    2016-01-01

    This chapter presents an overview of recent progress in modelling the behaviour of complex fluid–fluid interfaces with non-equilibrium thermodynamics. We will limit ourselves to frameworks employing the Gibbs dividing surface model, and start with a general discussion of the surface excess variables

  6. Safeguards Culture

    Energy Technology Data Exchange (ETDEWEB)

    Frazar, Sarah L.; Mladineo, Stephen V.

    2012-07-01

    The concepts of nuclear safety and security culture are well established; however, a common understanding of safeguards culture is not internationally recognized. Supported by the National Nuclear Security Administration, the authors prepared this report, an analysis of the concept of safeguards culture, and gauged its value to the safeguards community. The authors explored distinctions between safeguards culture, safeguards compliance, and safeguards performance, and evaluated synergies and differences between safeguards culture and safety/security culture. The report concludes with suggested next steps.

  7. Fluid loading responsiveness

    NARCIS (Netherlands)

    Geerts, Bart

    2011-01-01

    Patients in the intensive care unit (ICU) and in the peri-operative phase are dependent on physicians and nurses for their fluid intake. Volume status optimization is required to maximize oxygen delivery to vital organs. Unnecessary fluid administration can, however, lead to general and pulmonary

  8. Fluid loading responsiveness

    NARCIS (Netherlands)

    Geerts, Bart

    2011-01-01

    Patients in the intensive care unit (ICU) and in the peri-operative phase are dependent on physicians and nurses for their fluid intake. Volume status optimization is required to maximize oxygen delivery to vital organs. Unnecessary fluid administration can, however, lead to general and pulmonary oe

  9. Applications of fluid dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Round, G.R.; Garg, V.K.

    1986-01-01

    This book describes flexible and practical approach to learning the basics of fluid dynamics. Each chapter is a self-contained work session and includes a fluid dynamics concept, an explanation of the principles involved, an illustration of their application and references on where more detailed discussions can be found.

  10. Fluid loading responsiveness

    NARCIS (Netherlands)

    Geerts, Bart

    2011-01-01

    Patients in the intensive care unit (ICU) and in the peri-operative phase are dependent on physicians and nurses for their fluid intake. Volume status optimization is required to maximize oxygen delivery to vital organs. Unnecessary fluid administration can, however, lead to general and pulmonary oe

  11. Organizational Culture

    Directory of Open Access Journals (Sweden)

    Adrian HUDREA

    2006-02-01

    Full Text Available Cultural orientations of an organization can be its greatest strength, providing the basis for problem solving, cooperation, and communication. Culture, however, can also inhibit needed changes. Cultural changes typically happen slowly – but without cultural change, many other organizational changes are doomed to fail. The dominant culture of an organization is a major contributor to its success. But, of course, no organizational culture is purely one type or another. And the existence of secondary cultures can provide the basis for change. Therefore, organizations need to understand the cultural environments and values.

  12. Fluid blade disablement tool

    Energy Technology Data Exchange (ETDEWEB)

    Jakaboski, Juan-Carlos [Albuquerque, NM; Hughs, Chance G [Albuquerque, NM; Todd, Steven N [Rio Rancho, NM

    2012-01-10

    A fluid blade disablement (FBD) tool that forms both a focused fluid projectile that resembles a blade, which can provide precision penetration of a barrier wall, and a broad fluid projectile that functions substantially like a hammer, which can produce general disruption of structures behind the barrier wall. Embodiments of the FBD tool comprise a container capable of holding fluid, an explosive assembly which is positioned within the container and which comprises an explosive holder and explosive, and a means for detonating. The container has a concavity on the side adjacent to the exposed surface of the explosive. The position of the concavity relative to the explosive and its construction of materials with thicknesses that facilitate inversion and/or rupture of the concavity wall enable the formation of a sharp and coherent blade of fluid advancing ahead of the detonation gases.

  13. Amniotic fluid water dynamics.

    Science.gov (United States)

    Beall, M H; van den Wijngaard, J P H M; van Gemert, M J C; Ross, M G

    2007-01-01

    Water arrives in the mammalian gestation from the maternal circulation across the placenta. It then circulates between the fetal water compartments, including the fetal body compartments, the placenta and the amniotic fluid. Amniotic fluid is created by the flow of fluid from the fetal lung and bladder. A major pathway for amniotic fluid resorption is fetal swallowing; however in many cases the amounts of fluid produced and absorbed do not balance. A second resorption pathway, the intramembranous pathway (across the amnion to the fetal circulation), has been proposed to explain the maintenance of normal amniotic fluid volume. Amniotic fluid volume is thus a function both of the amount of water transferred to the gestation across the placental membrane, and the flux of water across the amnion. Membrane water flux is a function of the water permeability of the membrane; available data suggests that the amnion is the structure limiting intramembranous water flow. In the placenta, the syncytiotrophoblast is likely to be responsible for limiting water flow across the placenta. In human tissues, placental trophoblast membrane permeability increases with gestational age, suggesting a mechanism for the increased water flow necessary in late gestation. Membrane water flow can be driven by both hydrostatic and osmotic forces. Changes in both osmotic/oncotic and hydrostatic forces in the placenta my alter maternal-fetal water flow. A normal amniotic fluid volume is critical for normal fetal growth and development. The study of amniotic fluid volume regulation may yield important insights into the mechanisms used by the fetus to maintain water homeostasis. Knowledge of these mechanisms may allow novel treatments for amniotic fluid volume abnormalities with resultant improvement in clinical outcome.

  14. The method used to culture host cells (Sf9 cells) can affect the qualities of baculovirus budding particles expressing recombinant proteins.

    Science.gov (United States)

    Hattori, Tomomi; Nakanishi, Kohei; Mori, Takaaki; Tomita, Masahiro; Tsumoto, Kanta

    2016-01-01

    Budded virus (BV) particles of baculovirus (Autographa californica nucleopolyhedrovirus, AcNPV) are harvested from the supernatant of liquid culture of Sf9 host cells by ultracentrifugation. Using polyacrylamide gel electrophoresis, Western blot and transmission electron microscopy (TEM) of BV samples fractionated closely by sucrose density gradient centrifugation, we observed that BVs exhibited different qualities depending on whether they had been harvested from the supernatant from a standing (static), shaking (suspension), or standing/shaking (pre-/post-infection) culture of Sf9 cells. The amount of BV protein apparently increased in the order of standing, standing/shaking, and shaking procedure, and the yield of intact particles showed an opposite trend. TEM observation clearly showed that appropriate fractions of the standing and standing/shaking cultures contained more intact BV particles than those from the shaking culture. These results suggest that the qualities of recombinant BV particles may be related to the culture conditions of the host cells.

  15. Identification and localization of a soluble antigen, Ag2, of 136 kDa from Plasmodium falciparum in vitro cultures

    DEFF Research Database (Denmark)

    Jakobsen, P H; Grellier, P; Theander, T G

    1991-01-01

    The soluble antigens, antigen 2 (Ag2) and antigen 6 (Ag6), were copurified from supernatants of P. falciparum in vitro cultures by affinity chromatography and Fast Protein Liquid Chromatography. Rabbit antibodies to Ag2 were raised and characterized by crossed immunoelectrophoresis. Ag2 appeared ...... vacuole and in clefts in the infected erythrocyte cytoplasma as shown by immunogold electron microscopy....

  16. Micromachined Fluid Inertial Sensors

    Directory of Open Access Journals (Sweden)

    Shiqiang Liu

    2017-02-01

    Full Text Available Micromachined fluid inertial sensors are an important class of inertial sensors, which mainly includes thermal accelerometers and fluid gyroscopes, which have now been developed since the end of the last century for about 20 years. Compared with conventional silicon or quartz inertial sensors, the fluid inertial sensors use a fluid instead of a solid proof mass as the moving and sensitive element, and thus offer advantages of simple structures, low cost, high shock resistance, and large measurement ranges while the sensitivity and bandwidth are not competitive. Many studies and various designs have been reported in the past two decades. This review firstly introduces the working principles of fluid inertial sensors, followed by the relevant research developments. The micromachined thermal accelerometers based on thermal convection have developed maturely and become commercialized. However, the micromachined fluid gyroscopes, which are based on jet flow or thermal flow, are less mature. The key issues and technologies of the thermal accelerometers, mainly including bandwidth, temperature compensation, monolithic integration of tri-axis accelerometers and strategies for high production yields are also summarized and discussed. For the micromachined fluid gyroscopes, improving integration and sensitivity, reducing thermal errors and cross coupling errors are the issues of most concern.

  17. FRACTURING FLUID CHARACTERIZATION FACILITY

    Energy Technology Data Exchange (ETDEWEB)

    Subhash Shah

    2000-08-01

    Hydraulic fracturing technology has been successfully applied for well stimulation of low and high permeability reservoirs for numerous years. Treatment optimization and improved economics have always been the key to the success and it is more so when the reservoirs under consideration are marginal. Fluids are widely used for the stimulation of wells. The Fracturing Fluid Characterization Facility (FFCF) has been established to provide the accurate prediction of the behavior of complex fracturing fluids under downhole conditions. The primary focus of the facility is to provide valuable insight into the various mechanisms that govern the flow of fracturing fluids and slurries through hydraulically created fractures. During the time between September 30, 1992, and March 31, 2000, the research efforts were devoted to the areas of fluid rheology, proppant transport, proppant flowback, dynamic fluid loss, perforation pressure losses, and frictional pressure losses. In this regard, a unique above-the-ground fracture simulator was designed and constructed at the FFCF, labeled ''The High Pressure Simulator'' (HPS). The FFCF is now available to industry for characterizing and understanding the behavior of complex fluid systems. To better reflect and encompass the broad spectrum of the petroleum industry, the FFCF now operates under a new name of ''The Well Construction Technology Center'' (WCTC). This report documents the summary of the activities performed during 1992-2000 at the FFCF.

  18. Industrial cultures

    DEFF Research Database (Denmark)

    Rasmussen, Lauge Baungaard

    1996-01-01

    The chapter deals with different paradigms andtheories of cultural development. The problem toexplain change and methods to analyse developmentin different cultures are presented and discussed.......The chapter deals with different paradigms andtheories of cultural development. The problem toexplain change and methods to analyse developmentin different cultures are presented and discussed....

  19. Applied fluid mechanics; Mecanique des fluides appliquee

    Energy Technology Data Exchange (ETDEWEB)

    Viollet, P.L.; Chabard, J.P.; Esposito, P.; Laurence, D. [Ecole Nationale des Ponts et Chaussees (ENPC), 75 - Paris (France)]|[Electricite de France (EDF), 75 - Paris (France). Direction des Etudes et Recherches

    2002-07-01

    Computational hydraulics, computational fluid dynamics, and hydro-informatics have invaded virtually all domains of research and application in hydro-science and fluids engineering. To the extent that this invasion has led to improved understanding of complex fluid phenomena and provided a frame of reference for testing and verifying designs and operational schemes, we have all benefited from it. But to the extent that it has shifted attention away from fundamental descriptions and understanding of fluid phenomena, and toward computational and numerical issues, this invasion has left avoid in the scientific and technical literature. This void exists somewhere between student exposure to first principles of solid and fluid mechanics on the one hand, and advanced-student and researcher/practitioner exposure to computational techniques and applications on the other. This new text naturally and refreshingly steps in to fill this void, and as such is a most welcome addition to the literature and to personal and institutional libraries. The text is refreshing in its innovative and careful attention to setting the historical framework of general and specific topics. This is most notable in the first chapter, which very gracefully and efficiently leads the reader through historical developments to contemporary mathematical statements of basic fluid phenomena. Once the authors have established this foundation of fundamental principles, they tie each succeeding chapter back into the introduction with appropriate and supportive historical contexts. Although the text does not shy away from rigorous analytical descriptions of fluid phenomena, it is unique in providing this delightful historical context for each topic. The authors have also made a special effort to tie the chapters together into a unified whole, with ample references forward and back; this is indeed rare, and much appreciated, in a text of multiple authorship. The topics treated and chapter structures reflect

  20. Fundamentals of fluid lubrication

    Science.gov (United States)

    Hamrock, Bernard J.

    1991-01-01

    The aim is to coordinate the topics of design, engineering dynamics, and fluid dynamics in order to aid researchers in the area of fluid film lubrication. The lubrication principles that are covered can serve as a basis for the engineering design of machine elements. The fundamentals of fluid film lubrication are presented clearly so that students that use the book will have confidence in their ability to apply these principles to a wide range of lubrication situations. Some guidance on applying these fundamentals to the solution of engineering problems is also provided.

  1. Physics of Fluids

    OpenAIRE

    2007-01-01

    Periodic motion of three stirrers in a two-dimensional flow can lead to chaotic transport of the surrounding fluid. For certain stirrer motions, the generation of chaos is guaranteed solely by the topology of that motion and continuity of the fluid. Work in this area has focused largely on using physical rods as stirrers, but the theory also applies when the "stirrers" are passive fluid particles. We demonstrate the occurrence of topological chaos for Stokes flow in a two-dimensional lid-driv...

  2. Supercritical fluid extraction

    Science.gov (United States)

    Wai, Chien M.; Laintz, Kenneth

    1994-01-01

    A method of extracting metalloid and metal species from a solid or liquid material by exposing the material to a supercritical fluid solvent containing a chelating agent. The chelating agent forms chelates that are soluble in the supercritical fluid to allow removal of the species from the material. In preferred embodiments, the extraction solvent is supercritical carbon dioxide and the chelating agent is a fluorinated or lipophilic crown ether or fluorinated dithiocarbamate. The method provides an environmentally benign process for removing contaminants from industrial waste without using acids or biologically harmful solvents. The chelate and supercritical fluid can be regenerated, and the contaminant species recovered, to provide an economic, efficient process.

  3. Geophysical fluid flow experiment

    Science.gov (United States)

    Broome, B. G.; Fichtl, G.; Fowlis, W.

    1979-01-01

    The essential fluid flow processes associated with the solar and Jovian atmospheres will be examined in a laboratory experiment scheduled for performance on Spacelab Missions One and Three. The experimental instrumentation required to generate and to record convective fluid flow is described. Details of the optical system configuration, the lens design, and the optical coatings are described. Measurement of thermal gradient fields by schlieren techniques and measurement of fluid flow velocity fields by photochromic dye tracers is achieved with a common optical system which utilizes photographic film for data recording. Generation of the photochromic dye tracers is described, and data annotation of experimental parameters on the film record is discussed.

  4. Synthetic Base Fluids

    Science.gov (United States)

    Brown, M.; Fotheringham, J. D.; Hoyes, T. J.; Mortier, R. M.; Orszulik, S. T.; Randles, S. J.; Stroud, P. M.

    The chemical nature and technology of the main synthetic lubricant base fluids is described, covering polyalphaolefins, alkylated aromatics, gas-to-liquid (GTL) base fluids, polybutenes, aliphatic diesters, polyolesters, polyalkylene glycols or PAGs and phosphate esters.Other synthetic lubricant base oils such as the silicones, borate esters, perfluoroethers and polyphenylene ethers are considered to have restricted applications due to either high cost or performance limitations and are not considered here.Each of the main synthetic base fluids is described for their chemical and physical properties, manufacture and production, their chemistry, key properties, applications and their implications when used in the environment.

  5. Expression and significance of thrombospondin-1 in rats' hippocampal neurons cultured in vitro by fluid percussion injury%凝血酶敏感蛋白-1在液压冲击损伤体外培养大鼠海马神经细胞中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    杨术旺; 梁林; 战丽; 舒清明; 秦晓静; 张永亮; 董化江

    2013-01-01

    Objective To investigate the expression of thrombospondin-1 (TSP-1) in hippocampal neurons of rats cultured in vitro by fluid percussion injury. Methods The primary cultural neuron models of fluid percussion injury were established. The degree of neurons injury was observed by toluidine blue stain. The expression of TSP-1 in neurons was detected by Western blot and immunocytochemical stain. Results After fluid percussion, the results of toluidine blue stain showed that most post-injury neurons were necrotic cells, the cells body were condensed, and nuclei were pycnosis and dark blue. The results of immunocytochemical stain showed that there was some expression of TSP-1 in the neurons of control group, and the expression of TSP-1 and the positive cells increased in injury group. The results of Western blot showed that the optical density value was 0.937 ±0.194 in control group and 2. 318 ±0.495 in injury group, the expression of TSP-1 in the neurons of injury group was significantly stronger than that in control group (P < 0. 01). Conclusion The increased expression of TSP-1 may play an important role in the development of neurons cultured in vitro by fluid percussion injury.%目的 探讨凝血酶敏感蛋白-1(TSP-1)在液压冲击损伤大鼠体外培养海马神经细胞中的表达.方法 建立体外原代培养神经细胞液压冲击损伤模型,采用甲苯胺蓝染色观察神经细胞损伤程度,免疫细胞化学染色和West-ern blot技术检测神经细胞中TSP-1的表达水平.结果 液压冲击后,甲苯胺蓝染色显示,损伤组部分神经细胞胞体浓缩,细胞核固缩,呈深蓝色,多为坏死型损伤细胞.免疫细胞化学染色显示,对照组神经细胞内有一定量的TSP-1表达,损伤组神经细胞内TSP-1表达增多,阳性细胞数目增多.Western blot结果显示,对照组和损伤组光密度值分别为0.937±0.194与2.318±0.495,损伤组神经细胞TSP-1蛋白表达显著强于对照组(P<0.01).结论 体外培养

  6. Combined Analysis of IFN-γ, IL-2, IL-5, IL-10, IL-1RA and MCP-1 in QFT Supernatant Is Useful for Distinguishing Active Tuberculosis from Latent Infection.

    Directory of Open Access Journals (Sweden)

    Maho Suzukawa

    Full Text Available The QuantiFERON®-TB Gold In-Tube test (QFT, an interferon-γ release assay, is used to diagnose Mycobacterium tuberculosis, but its inaccuracy in distinguishing active tuberculosis from latent infection is a major concern. There is thus a need for an easy and accurate tool for achieving that goal in daily clinical settings. This study aimed to identify candidate cytokines for specifically differentiating active tuberculosis from latent infection. Our study population consisted of 31 active TB (tuberculosis patients, 29 LTBI (latent tuberculosis infection patients and 10 healthy control subjects. We assayed for 27 cytokines in QFT supernatants of both specific antigen-stimulated blood samples (TBAg and negative-control samples (Nil. We analyzed their specificities and sensitivities by creating receiver operating characteristic (ROC curves and measuring the area under those curves (AUCs. In TBAg-Nil supernatants, IL-10, IFN-γ, MCP-1 and IL-1RA showed high AUCs of 0.8120, 0.7842, 0.7419 and 0.7375, respectively. Compared with each cytokine alone, combined assay for these top four cytokines showed positive rates in diagnosing active TB, and GDA analysis revealed that MCP-1 and IL-5 are potent in distinguishing active TB from LTBI, with Wilk's lambda = 0.718 (p < 0.001. Furthermore, utilizing the unique characteristic of IL-2 that its TBAg-Nil supernatant levels are higher in LTBI compared to active TB, the difference between IFN-γ and IL-2 showed a large AUC of 0.8910. In summary, besides IFN-γ, IL-2, IL-5, IL-10, IL-1RA and MCP-1 in QFT supernatants may be useful for distinguishing active TB from LTBI. Those cytokines may also help us understand the difference in pathogenesis between active TB and LTBI.

  7. [The phenol turbidity test for measurement of pulmonary surfactants in amniotic fluid--rapid test for fetal lung maturity (author's transl)].

    Science.gov (United States)

    Tatsumi, H; Shimada, N; Kuramoto, R; Mochizuki, Y; Nishizima, M; Arai, M; Osanai, K; Ishihara, K; Goso, K; Hotta, K

    1981-05-01

    A simple and sensitive procedure for the quantitative estimation of pulmonary surfactants in the amniotic fluid is described. The method is based on the formation of turbidity from the amniotic fluid surfactants with phenol. Amniotic fluid drawn through an intrauterine catheter was centrifuged at 2000 rpm for 5 min. One ml of 5% aqueous phenol solution was added to 1.0 ml of the diluted supernatant. Control solution was prepared by addition of 1.0 ml of distilled water to the supernatant, instead of phenol solution. The turbidity was measured spectrophotometrically against control at 340 nm 5 to 10 min after agitation on a Vortex mixer for 5 sec. The turbidity obtained from amniotic fluid with phenol was proportional to the increase in total phospholipids in the fluid measured enzymatically. Moreover, only lecithin in the phospholipids was related to the turbidity formation. Sphyngomyelin, lysolecithin, phosphatidylethanolamin did not produce any turbidity with phenol. The spectrophotometric reading of 87 cases (31-41 weeks) was ranged at 0.18-3.52. Four cases with lower value (0.40) showed neonatal respiratory problem. The phenol turbidity test is more useful for the detection of fetal lung maturity comparing with the generally used shake test.

  8. Peritoneal Fluid Analysis

    Science.gov (United States)

    ... Peritoneal fluid glucose, amylase, tumor markers, bilirubin, creatinine, lactate dehydrogenase (LD) Microscopic examination – may be performed if infection or cancer is suspected; a laboratory professional may use a ...

  9. Polymer Fluid Dynamics.

    Science.gov (United States)

    Bird, R. Byron

    1980-01-01

    Problems in polymer fluid dynamics are described, including development of constitutive equations, rheometry, kinetic theory, flow visualization, heat transfer studies, flows with phase change, two-phase flow, polymer unit operations, and drag reduction. (JN)

  10. Nonpolluting drilling fluid composition

    Energy Technology Data Exchange (ETDEWEB)

    Larson, D.E.; Mocek, C.J.; Mouton, R.J.

    1983-02-22

    Disclosed is a nonpolluting drilling fluid composition. The composition mixture consisting essentially of a concentrate and any nonpolluting oil. The concentrate consists essentially of diethanolamide, a fatty acid, and a imidazoline/amide mixture.

  11. Cervical Cancer Cell Supernatants Induce a Phenotypic Switch from U937-Derived Macrophage-Activated M1 State into M2-Like Suppressor Phenotype with Change in Toll-Like Receptor Profile

    Science.gov (United States)

    Sánchez-Reyes, Karina; Bravo-Cuellar, Alejandro; Hernández-Flores, Georgina; Lerma-Díaz, José Manuel; Jave-Suárez, Luis Felipe; Gómez-Lomelí, Paulina; de Celis, Ruth; Aguilar-Lemarroy, Adriana; Domínguez-Rodríguez, Jorge Ramiro; Ortiz-Lazareno, Pablo Cesar

    2014-01-01

    Cervical cancer (CC) is the second most common cancer among women worldwide. Infection with human papillomavirus (HPV) is the main risk factor for developing CC. Macrophages are important immune effector cells; they can be differentiated into two phenotypes, identified as M1 (classically activated) and M2 (alternatively activated). Macrophage polarization exerts profound effects on the Toll-like receptor (TLR) profile. In this study, we evaluated whether the supernatant of human CC cells HeLa, SiHa, and C-33A induces a shift of M1 macrophage toward M2 macrophage in U937-derived macrophages. Results. The results showed that soluble factors secreted by CC cells induce a change in the immunophenotype of macrophages from macrophage M1 into macrophage M2. U937-derived macrophages M1 released proinflammatory cytokines and nitric oxide; however, when these cells were treated with the supernatant of CC cell lines, we observed a turnover of M1 toward M2. These cells increased CD163 and IL-10 expression. The expression of TLR-3, -7, and -9 is increased when the macrophages were treated with the supernatant of CC cells. Conclusions. Our result strongly suggests that CC cells may, through the secretion of soluble factors, induce a change of immunophenotype M1 into M2 macrophages. PMID:25309919

  12. Cervical Cancer Cell Supernatants Induce a Phenotypic Switch from U937-Derived Macrophage-Activated M1 State into M2-Like Suppressor Phenotype with Change in Toll-Like Receptor Profile

    Directory of Open Access Journals (Sweden)

    Karina Sánchez-Reyes

    2014-01-01

    Full Text Available Cervical cancer (CC is the second most common cancer among women worldwide. Infection with human papillomavirus (HPV is the main risk factor for developing CC. Macrophages are important immune effector cells; they can be differentiated into two phenotypes, identified as M1 (classically activated and M2 (alternatively activated. Macrophage polarization exerts profound effects on the Toll-like receptor (TLR profile. In this study, we evaluated whether the supernatant of human CC cells HeLa, SiHa, and C-33A induces a shift of M1 macrophage toward M2 macrophage in U937-derived macrophages. Results. The results showed that soluble factors secreted by CC cells induce a change in the immunophenotype of macrophages from macrophage M1 into macrophage M2. U937-derived macrophages M1 released proinflammatory cytokines and nitric oxide; however, when these cells were treated with the supernatant of CC cell lines, we observed a turnover of M1 toward M2. These cells increased CD163 and IL-10 expression. The expression of TLR-3, -7, and -9 is increased when the macrophages were treated with the supernatant of CC cells. Conclusions. Our result strongly suggests that CC cells may, through the secretion of soluble factors, induce a change of immunophenotype M1 into M2 macrophages.

  13. [Diagnosis: synovial fluid analysis].

    Science.gov (United States)

    Gallo Vallejo, Francisco Javier; Giner Ruiz, Vicente

    2014-01-01

    Synovial fluid analysis in rheumatological diseases allows a more accurate diagnosis in some entities, mainly infectious and microcrystalline arthritis. Examination of synovial fluid in patients with osteoarthritis is useful if a differential diagnosis will be performed with other processes and to distinguish between inflammatory and non-inflammatory forms. Joint aspiration is a diagnostic and sometimes therapeutic procedure that is available to primary care physicians. Copyright © 2014 Elsevier España, S.L. All rights reserved.

  14. Conventional cerebrospinal fluid scanning

    Energy Technology Data Exchange (ETDEWEB)

    Schicha, H.

    1985-06-01

    Conventional cerebrospinal fluid scanning (CSF scanning) today is mainly carried out in addition to computerized tomography to obtain information about liquor flow kinetics. Especially in patients with communicating obstructive hydrocephalus, CSF scanning is clinically useful for the decision for shunt surgery. In patients with intracranial cysts, CSF scanning can provide information about liquor circulation. Further indications for CSF scanning include the assessment of shunt patency especially in children, as well as the detection and localization of cerebrospinal fluid leaks.

  15. Glycerol production by Oenococcus oeni during sequential and simultaneous cultures with wine yeast strains.

    Science.gov (United States)

    Ale, Cesar E; Farías, Marta E; Strasser de Saad, Ana M; Pasteris, Sergio E

    2014-07-01

    Growth and fermentation patterns of Saccharomyces cerevisiae, Kloeckera apiculata, and Oenococcus oeni strains cultured in grape juice medium were studied. In pure, sequential and simultaneous cultures, the strains reached the stationary growth phase between 2 and 3 days. Pure and mixed K. apiculata and S. cerevisiae cultures used mainly glucose, producing ethanol, organic acids, and 4.0 and 0.1 mM glycerol, respectively. In sequential cultures, O. oeni achieved about 1 log unit at 3 days using mainly fructose and L-malic acid. Highest sugars consumption was detected in K. apiculata supernatants, lactic acid being the major end-product. 8.0 mM glycerol was found in 6-day culture supernatants. In simultaneous cultures, total sugars and L-malic acid were used at 3 days and 98% of ethanol and glycerol were detected. This study represents the first report of the population dynamics and metabolic behavior of yeasts and O. oeni in sequential and simultaneous cultures and contributes to the selection of indigenous strains to design starter cultures for winemaking, also considering the inclusion of K. apiculata. The sequential inoculation of yeasts and O. oeni would enhance glycerol production, which confers desirable organoleptic characteristics to wines, while organic acids levels would not affect their sensory profile.

  16. Fluid Dynamic Verification Experiments on STS-70

    Science.gov (United States)

    Kleis, Stanley J.

    1996-01-01

    Fluid dynamic experiments were flown on STS-70 as phase two of the engineering evaluation of the first bioreactor Engineering Development Unit (EDU#1). The phase one experiments were comparative cell cultures in identical units on earth and onboard STS-70. In phase two, two types of fluid dynamic experiments were performed. Qualitative comparisons of the basic flow patterns were evaluated with the use of 'dye' streaklines formed from alternate injections of either a mild acid or base solution into the external flow loop that was then perfused into the vessel. The presence of Bromothymol Blue in the fluid then caused color changes from yellow to blue or vice versa, indicating the basic fluid motions. This reversible change could be repeated as desired. In the absence of significant density differences in the fluid, the flow patterns in space should be the same as on earth. Video tape records of the flow patterns for a wide range of operating conditions were obtained. The second type of fluid dynamic experiment was the quantitative evaluation of the trajectories of solid beads of various densities and sizes. The beads were introduced into the vessel and the paths recorded on video tape, with the vessel operated at various rotation rates and flow perfusion rates. Because of space limitations, the video camera was placed as close as possible to the vessel, resulting in significant optical distortion. This report describes the analysis methods to obtain comparisons between the in-flight fluid dynamics and numerical models of the flow field. The methods include optical corrections to the video images and calculation of the bead trajectories for given operating conditions and initial bead locations.

  17. The work of cultural transition

    DEFF Research Database (Denmark)

    Ryba, T. V.; Stambulova, N. B.; Ronkainen, Noora J.

    2016-01-01

    In today's uncertain, fluid job market, transnational mobility has intensified. Though the concept of cultural transition is increasingly used in sport and career research, insight into the processes of how individuals produce their own development through work and relationships in shifting...... cultural patterns of meaning remains limited. The transnational industry of sports, in which athletes' psychological adjustment to cultural transitions has implications for both performance and meaningful life, serves as a backdrop for this article. This study applied the life story method to interviews...

  18. Fullerol ionic fluids

    KAUST Repository

    Fernandes, Nikhil

    2010-01-01

    We report for the first time an ionic fluid based on hydroxylated fullerenes (fullerols). The ionic fluid was synthesized by neutralizing the fully protonated fullerol with an amine terminated polyethylene/polypropylene oxide oligomer (Jeffamine®). The ionic fluid was compared to a control synthesized by mixing the partially protonated form (sodium form) of the fullerols with the same oligomeric amine in the same ratio as in the ionic fluids (20 wt% fullerol). In the fullerol fluid the ionic bonding significantly perturbs the thermal transitions and melting/crystallization behavior of the amine. In contrast, both the normalized heat of fusion and crystallization of the amine in the control are similar to those of the neat amine consistent with a physical mixture of the fullerols/amine with minimal interactions. In addition to differences in thermal behavior, the fullerol ionic fluid exhibits a complex viscoelastic behavior intermediate between the neat Jeffamine® (liquid-like) and the control (solid-like). © 2010 The Royal Society of Chemistry.

  19. Oscillating Cell Culture Bioreactor

    Science.gov (United States)

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

    2010-01-01

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

  20. Cultural psychology.

    Science.gov (United States)

    Heine, Steven J; Ruby, Matthew B

    2010-03-01

    Humans are a cultural species, constantly navigating a complex web of culturally bound practices, norms, and worldviews. This article provides a brief overview of the relatively young field of cultural psychology, which investigates the many ways psychology and culture interweave with one another. Highlighting the cultural nature of the human species, it draws upon research on cultural evolution, enculturation, and developmental processes. This review further summarizes a number of cultural differences in how people perceive the self, and the behavioral consequences that follow from these differences, in the domains of internal and external attribution styles, motivations for self-enhancement, approach/avoidance, primary and secondary control, as well as motivations for distinctiveness and conformity. Additionally, the review discusses research on the intersection of culture and emotion, as well as cultural differences in cognition, perception, and reasoning. Copyright © 2010 John Wiley & Sons, Ltd. For further resources related to this article, please visit the WIREs website. Copyright © 2010 John Wiley & Sons, Ltd.

  1. Prenatal diagnosis of Down syndrome using cell-free fetal DNA in amniotic fluid by quantitative fluorescent polymersase chain reaction

    Institute of Scientific and Technical Information of China (English)

    Wu Dan; Chi Hongbin; Shao Minjie; Wu Yao; Jin Hongyan; Wu Baiyan; Qiao Jie

    2014-01-01

    Backgroud Amniotic fluid (AF) supernatant contains cell-free fetal DNA (cffDNA) fragments.This study attempted to take advantage of cffDNA as a new material for prenatal diagnosis,which could be combined with simple quantitative fluorescent polymerase chain reaction (QF-PCR) to provide an ancillary method for the prenatal diagnosis of trisomy 21 syndrome.Methods AF supernatant samples were obtained from 27 women carrying euploid fetuses and 28 women carrying aneuploid fetuses with known cytogenetic karyotypes.Peripheral blood samples of the parents were collected at the same time.Short tandem repeat (STR) fragments on chromosome 21 were amplified by QF-PCR.Fetal condition and the parental source of the extra chromosome could be determined by the STR peaks.Results The sensitivity of the assay for the aneuploid was 93% (26/28; confidence interval,CI:77%-98%) and the specificity was 100% (26/26; CI:88%-100%).The determination rate of the origin of the extra chromosome was 69%.The sensitivity and the specificity of the assay in the euploid were 100% (27/27).Conclusions Trisomy 21 can be prenatally diagnosed by the QF-PCR method in AF supernatant.This karyotype analysis method greatly reduces the requirement for the specimen size.It will be a benefit for early amniocentesis and could avoid pregnancy complications.The method may become an ancillary method for prenatal diagnosis of trisomy 21.

  2. Cultural commons and cultural evolution

    OpenAIRE

    Giangiacomo Bravo

    2010-01-01

    Culture evolves following a process that is akin to biological evolution, although with some significant differences. At the same time culture has often a collective good value for human groups. This paper studies culture in an evolutionary perspective, with a focus on the implications of group definition for the coexistence of different cultures. A model of cultural evolution is presented where agents interacts in an artificial environment. The belonging to a specific memetic group is a majo...

  3. Viscous Flow with Large Fluid-Fluid Interface Displacement

    DEFF Research Database (Denmark)

    Rasmussen, Henrik Koblitz; Hassager, Ole; Saasen, Arild

    1998-01-01

    The arbitrary Lagrange-Euler (ALE) kinematic description has been implemented in a 3D transient finite element program to simulate multiple fluid flows with fluid-fluid interface or surface displacements. The description of fluid interfaces includes variable interfacial tension, and the formulation...

  4. Numerical simulation of orbitally shaken viscous fluids with free surface

    OpenAIRE

    Discacciati, Marco; Hacker, David; Quarteroni, A.; Quinodoz, Samuel; Tissot, Stéphanie; Wurm, M. Florian

    2013-01-01

    Orbitally shaken bioreactors are an emerging alternative to stirred-tank bioreactors for large-scale mam- malian cell culture, but their fluid dynamics is still not well defined. Among the theoretical and practical issues that remain to be resolved, the characterization of the liquid free surface during orbital shaking remains a major challenge because it is an essential aspect of gas transfer and mixing in these reactors. To simulate the fluid behavior and the free surface shape, we develope...

  5. Two-phase cooling fluids; Les fluides frigoporteurs diphasiques

    Energy Technology Data Exchange (ETDEWEB)

    Lallemand, A. [Institut National des Sciences Appliquees (INSA), 69 - Lyon (France)

    1997-12-31

    In the framework of the diminution of heat transfer fluid consumption, the concept of indirect refrigerating circuits, using cooling intermediate fluids, is reviewed and the fluids that are currently used in these systems are described. Two-phase cooling fluids advantages over single-phase fluids are presented with their thermophysical characteristics: solid fraction, two-phase mixture enthalpy, thermal and rheological properties, determination of heat and mass transfer characteristics, and cold storage through ice slurry

  6. Some biochemical constituents in follicular fluid of indigenous cows of Assam

    Directory of Open Access Journals (Sweden)

    S. S. Deka

    2014-11-01

    Full Text Available Aim: Estimation of some biochemical constituents in follicular fluid of non-descript cows of Assam. Materials and Methods: Twenty five pairs of ovaries were collected from local slaughter house and the follicular fluid was aspirated from small (2-4 mm, medium (4-6 mm and large (6-9 mm follicles. Aspirated fluid samples were centrifuged at 4000 rpm for 20 min in a refrigerated centrifuge to remove granulosa cells and other cell debris. Supernatant was used for estimation of glucose, total protein, cholesterol, acid phosphatase and alkaline phosphatase. Data generated in the study were analyzed statiscally by SPSS (version 16.0. SPSS South Asia Pvt. Limited, Kacharakanahalli, Bangalore, 560043. Results: A non significant difference was recorded in case of total protein and cholesterol of follicular fluid of small, medium and large sized follicles of cow. However, the glucose level significantly (p<0.01 increased with the increase of follicular size where as the activity of acid phosphatase and alkaline phosphatase significantly (p<0.01 decreased with increase of size. Conclusion: Certain biochemical constituents and enzyme activities of follicular fluid changes with the growth of follicles in non-descript cows of Assam. The glucose concentration increased with the growth of the follicles while acid phosphatase and alkaline phosphatase levels had an inverse relation with the size of the ovarian follicles.

  7. Oral bacterial DNA findings in pericardial fluid

    Directory of Open Access Journals (Sweden)

    Anne-Mari Louhelainen

    2014-11-01

    Full Text Available Background: We recently reported that large amounts of oral bacterial DNA can be found in thrombus aspirates of myocardial infarction patients. Some case reports describe bacterial findings in pericardial fluid, mostly done with conventional culturing and a few with PCR; in purulent pericarditis, nevertheless, bacterial PCR has not been used as a diagnostic method before. Objective: To find out whether bacterial DNA can be measured in the pericardial fluid and if it correlates with pathologic–anatomic findings linked to cardiovascular diseases. Methods: Twenty-two pericardial aspirates were collected aseptically prior to forensic autopsy at Tampere University Hospital during 2009–2010. Of the autopsies, 10 (45.5% were free of coronary artery disease (CAD, 7 (31.8% had mild and 5 (22.7% had severe CAD. Bacterial DNA amounts were determined using real-time quantitative PCR with specific primers and probes for all bacterial strains associated with endodontic disease (Streptococcus mitis group, Streptococcus anginosus group, Staphylococcus aureus/Staphylococcus epidermidis, Prevotella intermedia, Parvimonas micra and periodontal disease (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, Fusobacterium nucleatus, and Dialister pneumosintes. Results: Of 22 cases, 14 (63.6% were positive for endodontic and 8 (36.4% for periodontal-disease-associated bacteria. Only one case was positive for bacterial culturing. There was a statistically significant association between the relative amount of bacterial DNA in the pericardial fluid and the severity of CAD (p=0.035. Conclusions: Oral bacterial DNA was detectable in pericardial fluid and an association between the severity of CAD and the total amount of bacterial DNA in pericardial fluid was found, suggesting that this kind of measurement might be useful for clinical purposes.

  8. Repellent Culture.

    Science.gov (United States)

    Carroll, Jeffrey

    2001-01-01

    Considers defining "culture," noting how it is difficult to define because those individuals defining it cannot separate themselves from it. Relates these issues to student writing and their writing improvement. Addresses violence in relation to culture. (SG)

  9. Culturing Protozoa.

    Science.gov (United States)

    Stevenson, Paul

    1980-01-01

    Compares various nutrient media, growth conditions, and stock solutions used in culturing protozoa. A hay infusion in Chalkey's solution maintained at a stable temperature is recommended for producing the most dense and diverse cultures. (WB)

  10. Throat Culture

    Science.gov (United States)

    ... products and services. Advertising & Sponsorship: Policy | Opportunities Throat Culture Share this page: Was this page helpful? Collecting | ... treatment | Getting results | see BLOOD SAMPLE Collecting A culture is a test that is often used to ...

  11. Culturing Protozoa.

    Science.gov (United States)

    Stevenson, Paul

    1980-01-01

    Compares various nutrient media, growth conditions, and stock solutions used in culturing protozoa. A hay infusion in Chalkey's solution maintained at a stable temperature is recommended for producing the most dense and diverse cultures. (WB)

  12. Safety culture

    Energy Technology Data Exchange (ETDEWEB)

    Keen, L.J. [Canadian Nuclear Safety Commission, Ottawa, Ontario (Canada)

    2003-07-01

    Safety culture has become a topic of increasing interest for industry and regulators as issues are raised on safety problems around the world. The keys to safety culture are organizational effectiveness, effective communications, organizational learning, and a culture that encourages the identification and resolution of safety issues. The necessity of a strong safety culture places an onus on all of us to continually question whether the safety measures already in place are sufficient, and are being applied. (author)

  13. Boiler using combustible fluid

    Science.gov (United States)

    Baumgartner, H.; Meier, J.G.

    1974-07-03

    A fluid fuel boiler is described comprising a combustion chamber, a cover on the combustion chamber having an opening for introducing a combustion-supporting gaseous fluid through said openings, means to impart rotation to the gaseous fluid about an axis of the combustion chamber, a burner for introducing a fluid fuel into the chamber mixed with the gaseous fluid for combustion thereof, the cover having a generally frustro-conical configuration diverging from the opening toward the interior of the chamber at an angle of between 15/sup 0/ and 55/sup 0/; means defining said combustion chamber having means defining a plurality of axial hot gas flow paths from a downstream portion of the combustion chamber to flow hot gases into an upstream portion of the combustion chamber, and means for diverting some of the hot gas flow along paths in a direction circumferentially of the combustion chamber, with the latter paths being immersed in the water flow path thereby to improve heat transfer and terminating in a gas outlet, the combustion chamber comprising at least one modular element, joined axially to the frustro-conical cover and coaxial therewith. The modular element comprises an inner ring and means of defining the circumferential, radial, and spiral flow paths of the hot gases.

  14. Amniotic fluid embolism

    Directory of Open Access Journals (Sweden)

    Kiranpreet Kaur

    2016-01-01

    Full Text Available Amniotic fluid embolism (AFE is one of the catastrophic complications of pregnancy in which amniotic fluid, fetal cells, hair, or other debris enters into the maternal pulmonary circulation, causing cardiovascular collapse. Etiology largely remains unknown, but may occur in healthy women during labour, during cesarean section, after abnormal vaginal delivery, or during the second trimester of pregnancy. It may also occur up to 48 hours post-delivery. It can also occur during abortion, after abdominal trauma, and during amnio-infusion. The pathophysiology of AFE is not completely understood. Possible historical cause is that any breach of the barrier between maternal blood and amniotic fluid forces the entry of amniotic fluid into the systemic circulation and results in a physical obstruction of the pulmonary circulation. The presenting signs and symptoms of AFE involve many organ systems. Clinical signs and symptoms are acute dyspnea, cough, hypotension, cyanosis, fetal bradycardia, encephalopathy, acute pulmonary hypertension, coagulopathy etc. Besides basic investigations lung scan, serum tryptase levels, serum levels of C3 and C4 complements, zinc coproporphyrin, serum sialyl Tn etc are helpful in establishing the diagnosis. Treatment is mainly supportive, but exchange transfusion, extracorporeal membrane oxygenation, and uterine artery embolization have been tried from time to time. The maternal prognosis after amniotic fluid embolism is very poor though infant survival rate is around 70%.

  15. Cultural clashes

    National Research Council Canada - National Science Library

    2001-01-01

    ... is essentially a cultural, not religious, practice. Siddiq Bazarwala Singapore From Georgina Dubourcq Sir: While I don't disagree with Dr Dalrymple's comments about the way Muslim culture can treat women, I wonder how a culture which condones lap-dancing, pornographic videos and often turns a blind eye to under-age prostitution can really feel superior. I...

  16. Handling Culture

    NARCIS (Netherlands)

    Pieter van Nispen tot Pannerden

    2011-01-01

    The article indicates how companies may prepare for and deal with cultural differences. Because the research base is still rather limited an overall perspective may not be realised. After discussing definitions and concepts of culture, as well as values, cultural differences between states are discu

  17. Handling Culture

    NARCIS (Netherlands)

    Nispen tot Pannerden, P.J.M. van

    2011-01-01

    The article indicates how companies may prepare for and deal with cultural differences. Because the research base is still rather limited an overall perspective may not be realised. After discussing definitions and concepts of culture, as well as values, cultural differences between states are discu

  18. Beyond Culture.

    Science.gov (United States)

    Barron, Daniel D.

    1993-01-01

    Discusses the lack of literature relating to cultural differences and school library media programs and reviews the book "Beyond Culture" by Edward T. Hall. Highlights include the population/environment crisis, cultural literacy, the use of technology, and Marshall McLuhan's idea of the global village. (LRW)

  19. Stochastic interpenetration of fluids

    Energy Technology Data Exchange (ETDEWEB)

    Steinkamp, M.J.; Clark, T.T.; Harlow, F.H.

    1995-11-01

    We describe a spectral approach to the investigation of fluid instability, generalized turbulence, and the interpenetration of fluids across an interface. The technique also applies to a single fluid with large variations in density. Departures of fluctuating velocity components from the local mean are far subsonic, but the mean Mach number can be large. Validity of the description is demonstrated by comparisons with experiments on turbulent mixing due to the late stages of Rayleigh-Taylor instability, when the dynamics become approximately self-similar in response to a constant body force. Generic forms for anisotropic spectral structure are described and used as a basis for deriving spectrally integrated moment equations that can be incorporated into computer codes for scientific and engineering analyses.

  20. Vorticity in holographic fluids

    CERN Document Server

    Caldarelli, Marco M; Petkou, Anastasios C; Petropoulos, P Marios; Pozzoli, Valentina; Siampos, Konstadinos

    2012-01-01

    In view of the recent interest in reproducing holographically various properties of conformal fluids, we review the issue of vorticity in the context of AdS/CFT. Three-dimensional fluids with vorticity require four-dimensional bulk geometries with either angular momentum or nut charge, whose boundary geometries fall into the Papapetrou--Randers class. The boundary fluids emerge in stationary non-dissipative kinematic configurations, which can be cyclonic or vortex flows, evolving in compact or non-compact supports. A rich network of Einstein's solutions arises, naturally connected with three-dimensional Bianchi spaces. We use Fefferman--Graham expansion to handle holographic data from the bulk and discuss the alternative for reversing the process and reconstruct the exact bulk geometries.

  1. The Omega-3 Fatty Acid Docosahexaenoic Acid Modulates Inflammatory Mediator Release in Human Alveolar Cells Exposed to Bronchoalveolar Lavage Fluid of ARDS Patients

    Directory of Open Access Journals (Sweden)

    Paolo Cotogni

    2015-01-01

    Full Text Available Background. This study investigated whether the 1 : 2 ω-3/ω-6 ratio may reduce proinflammatory response in human alveolar cells (A549 exposed to an ex vivo inflammatory stimulus (bronchoalveolar lavage fluid (BALF of acute respiratory distress syndrome (ARDS patients. Methods. We exposed A549 cells to the BALF collected from 12 ARDS patients. After 18 hours, fatty acids (FA were added as docosahexaenoic acid (DHA, ω-3 and arachidonic acid (AA, ω-6 in two ratios (1 : 2 or 1 : 7. 24 hours later, in culture supernatants were evaluated cytokines (TNF-α, IL-6, IL-8, and IL-10 and prostaglandins (PGE2 and PGE3 release. The FA percentage content in A549 membrane phospholipids, content of COX-2, level of PPARγ, and NF-κB binding activity were determined. Results. The 1 : 2 DHA/AA ratio reversed the baseline predominance of ω-6 over ω-3 in the cell membranes (P < 0.001. The proinflammatory cytokine release was reduced by the 1 : 2 ratio (P < 0.01 to <0.001 but was increased by the 1 : 7 ratio (P < 0.01. The 1 : 2 ratio reduced COX-2 and PGE2 (P < 0.001 as well as NF-κB translocation into the nucleus (P < 0.01, while it increased activation of PPARγ and IL-10 release (P < 0.001. Conclusion. This study demonstrated that shifting the FA supply from ω-6 to ω-3 decreased proinflammatory mediator release in human alveolar cells exposed to BALF of ARDS patients.

  2. Cultural Rights and Cultural Diversity

    Institute of Scientific and Technical Information of China (English)

    WANG SIXIN

    2011-01-01

    @@ Culture is a very big concept, big enough almost to comprise all the activities of human beings and the tangible and intangible results caused by human activities.Therefore, it is very difficult to define culture in a few words.

  3. Computational fluid dynamics

    CERN Document Server

    Magoules, Frederic

    2011-01-01

    Exploring new variations of classical methods as well as recent approaches appearing in the field, Computational Fluid Dynamics demonstrates the extensive use of numerical techniques and mathematical models in fluid mechanics. It presents various numerical methods, including finite volume, finite difference, finite element, spectral, smoothed particle hydrodynamics (SPH), mixed-element-volume, and free surface flow.Taking a unified point of view, the book first introduces the basis of finite volume, weighted residual, and spectral approaches. The contributors present the SPH method, a novel ap

  4. Fluids in cosmology

    CERN Document Server

    Cervantes-Cota, Jorge L

    2014-01-01

    We review the role of fluids in cosmology by first introducing them in General Relativity and then applied to a FRW Universe's model. We describe how relativistic and non-relativistic components evolve in the background dynamics. We also introduce scalar fields to show that they are able to yield an inflationary dynamics at very early times (inflation) and late times (quintessence). Then, we proceed to study the thermodynamical properties of the fluids and, lastly, its perturbed kinematics. We make emphasis in the constrictions of parameters by recent cosmological probes.

  5. Computational fluid dynamics

    CERN Document Server

    Blazek, Jiri

    2015-01-01

    Computational Fluid Dynamics: Principles and Applications, Third Edition presents students, engineers, and scientists with all they need to gain a solid understanding of the numerical methods and principles underlying modern computation techniques in fluid dynamics. By providing complete coverage of the essential knowledge required in order to write codes or understand commercial codes, the book gives the reader an overview of fundamentals and solution strategies in the early chapters before moving on to cover the details of different solution techniques. This updated edition includes new

  6. Mechanics of fluid flow

    CERN Document Server

    Basniev, Kaplan S; Chilingar, George V 0

    2012-01-01

    The mechanics of fluid flow is a fundamental engineering discipline explaining both natural phenomena and human-induced processes, and a thorough understanding of it is central to the operations of the oil and gas industry.  This book, written by some of the world's best-known and respected petroleum engineers, covers the concepts, theories, and applications of the mechanics of fluid flow for the veteran engineer working in the field and the student, alike.  It is a must-have for any engineer working in the oil and gas industry.

  7. Bioreactor Studies and Computational Fluid Dynamics

    Science.gov (United States)

    Singh, H.; Hutmacher, D. W.

    The hydrodynamic environment “created” by bioreactors for the culture of a tissue engineered construct (TEC) is known to influence cell migration, proliferation and extra cellular matrix production. However, tissue engineers have looked at bioreactors as black boxes within which TECs are cultured mainly by trial and error, as the complex relationship between the hydrodynamic environment and tissue properties remains elusive, yet is critical to the production of clinically useful tissues. It is well known in the chemical and biotechnology field that a more detailed description of fluid mechanics and nutrient transport within process equipment can be achieved via the use of computational fluid dynamics (CFD) technology. Hence, the coupling of experimental methods and computational simulations forms a synergistic relationship that can potentially yield greater and yet, more cohesive data sets for bioreactor studies. This review aims at discussing the rationale of using CFD in bioreactor studies related to tissue engineering, as fluid flow processes and phenomena have direct implications on cellular response such as migration and/or proliferation. We conclude that CFD should be seen by tissue engineers as an invaluable tool allowing us to analyze and visualize the impact of fluidic forces and stresses on cells and TECs.

  8. 两步法分离和培养人羊水来源胚胎间充质干细胞及其向神经元样细胞诱导分化的实验研究%Isolation of human pluripotent mesenchymal stem cells from second-trimester amniotic fluid using two kinds of culture protocol and their differentiation into neuron-like cells

    Institute of Scientific and Technical Information of China (English)

    冯建勋; 腊晓林; 马艳; 毕晓娟; 温浩

    2009-01-01

    目的 建立两步培养法分离培养人孕中期羊水间充质干细胞(MSCs)的方法,观察羊水MSCs向神经元样细胞的诱导分化.方法 采用改进的两步培养法分离和培养人羊水MSCs,以β-巯基乙醇诱导其向神经元样细胞方向分化;并使用流式细胞术、逆转录-聚合酶链反应(RT-PCR)和免疫细胞化学法研究人羊水MSCs的生物学特性.结果 成功分离、培养和传代出人羊水MSCs.流式细胞术检测人羊水MSCs表达CD44、CD29、CD105,不表达CD45、CD34、人白细胞DR抗原(HLA-DR);免疫荧光法和RT-PCR检测表明,部分人羊水MSCs表达转录因子Oct-4.两步培养法羊水MSCs集落形成率[(15.0±2.3)%]和Oct-4阳性细胞率[(1.2±0.3)%]均高于一步法[分别为(10.0±1.8)%,(0.9±0.2)%,P均<0.05].羊水MSCs经β-巯基乙醇诱导后细胞形态发生改变,伸出突起并呈神经球样改变;免疫细胞化学法检测显示(54.76±3.65)%的细胞表达神经元特异性烯醇化酶(NSE),同时(36.28±4.27)%的细胞表达神经胶质酸性蛋白(GFAP).结论 人羊水中存在MSCs,两步培养法是一种高效、简便、实用而且不干扰常规产前诊断程序的方法.在体外条件下,利用β-巯基乙醇和适宜的培养液可使羊水MSCs定向分化为神经元.%Objective To isolate mesenchymal stem cells (MSCs) from second-trimester amniotic fluid using an improved two-stage culture protocol, and to induce these MSCs into neuron-like cells. Methods An improved two-stage culture protocol for MSCs from amniotic fluid was developed. MSCs from amniotic fluid were induced to differentiate with β-mercaptoethanol into neuron-like cells. Flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry were employed for analysis of the phenotypic characteristics of the cultured MSCs from amniotic fluid. Results MSCs from amniotic fluid were successfully isolated, cultured and enriched without interfering with the routine process of

  9. Culture evolves.

    Science.gov (United States)

    Whiten, Andrew; Hinde, Robert A; Laland, Kevin N; Stringer, Christopher B

    2011-04-12

    Culture pervades human lives and has allowed our species to create niches all around the world and its oceans, in ways quite unlike any other primate. Indeed, our cultural nature appears so distinctive that it is often thought to separate humanity from the rest of nature and the Darwinian forces that shape it. A contrary view arises through the recent discoveries of a diverse range of disciplines, here brought together to illustrate the scope of a burgeoning field of cultural evolution and to facilitate cross-disciplinary fertilization. Each approach emphasizes important linkages between culture and evolutionary biology rather than quarantining one from the other. Recent studies reveal that processes important in cultural transmission are more widespread and significant across the animal kingdom than earlier recognized, with important implications for evolutionary theory. Recent archaeological discoveries have pushed back the origins of human culture to much more ancient times than traditionally thought. These developments suggest previously unidentified continuities between animal and human culture. A third new array of discoveries concerns the later diversification of human cultures, where the operations of Darwinian-like processes are identified, in part, through scientific methods borrowed from biology. Finally, surprising discoveries have been made about the imprint of cultural evolution in the predispositions of human minds for cultural transmission.

  10. Fluid Jet Polishing

    NARCIS (Netherlands)

    Booij, S.M.

    2003-01-01

    The goal of this thesis research was to investigate the possibilities and limitations of the Fluid Jet Polishing (FJP) technique. FJP is a new optical fabrication technique that is capable of making shape corrections and reducing the surface roughness of glass and other materials. The principle of o

  11. Proteomics of body fluids

    NARCIS (Netherlands)

    L.J.M. Dekker (Lennard)

    2007-01-01

    textabstractIn this thesis we present newly developed methods for biomarker discovery. We applied these methods to discover biomarkers of leptomeningeal metastasis (LM) in the cerebrospinal fluid (CSF) from breast cancer patients and in serum from patients with prostate cancer. Early diagnos

  12. Fluid Jet Polishing

    NARCIS (Netherlands)

    Booij, S.M.

    2003-01-01

    The goal of this thesis research was to investigate the possibilities and limitations of the Fluid Jet Polishing (FJP) technique. FJP is a new optical fabrication technique that is capable of making shape corrections and reducing the surface roughness of glass and other materials. The principle of

  13. Orbital Fluid Resupply Assessment

    Science.gov (United States)

    Eberhardt, Ralph N.

    1989-01-01

    Orbital fluid resupply can significantly increase the cost-effectiveness and operational flexibility of spacecraft, satellites, and orbiting platforms and observatories. Reusable tankers are currently being designed for transporting fluids to space. A number of options exist for transporting the fluids and propellant to the space-based user systems. The fluids can be transported to space either in the Shuttle cargo bay or using expendable launch vehicles (ELVs). Resupply can thus be accomplished either from the Shuttle bay, or the tanker can be removed from the Shuttle bay or launched on an ELV and attached to a carrier such as the Orbital Maneuvering Vehicle (OMV) or Orbital Transfer Vehicle (OTV) for transport to the user to be serviced. A third option involves locating the tanker at the space station or an unmanned platform as a quasi-permanent servicing facility or depot which returns to the ground for recycling once its tanks are depleted. Current modular tanker designs for monopropellants, bipropellants, and water for space station propulsion are discussed. Superfluid helium tankers are addressed, including trade-offs in tanker sizes, shapes to fit the range of ELVs currently available, and boil-off losses associated with longer-term (greater than 6-month) space-basing. It is concluded that the mixed fleet approach to on-orbit consumables resupply offers significant advantages to the overall logistics requirements.

  14. Fluid resuscitation in trauma

    Directory of Open Access Journals (Sweden)

    Rudra A

    2006-01-01

    Full Text Available Appropriate fluid replacement is an essential component of trauma fluid resuscitation. Once hemorrhage is controlled, restoration of normovolemia is a priority. In the presence of uncontrolled haemorrhage, aggressive fluid management may be harmful. The crystalloid-colloid debate continues but existing clinical practice is more likely to reflect local biases rather than evidence based medicine. Colloids vary substantially in their pharmacology and pharmacokinetics,and the experimental finding based on one colloid cannot be extrapolated reliably to another. In the initial stages of trauma resuscitation the precise fluid used is probably not important as long as an appropriate volume is given. Later, when the microcirculation is ′leaky′, there may be some advantages to high or medium weight colloids such as hydroxyethyl starch. Hypertonic saline solutions may have some benefit in patients with head injuries. A number of hemoglobin solutions are under development, but one of the most promising of these has been withdrawn recently. It is highly likely that at least one of these solutions will eventually become routine therapy for trauma patient resuscitation. In the meantime, contrary to traditional teaching, recent data suggest that restrictive strategy of red cell transfusion may improve outcome in some critically ill patients.

  15. Amniotic Fluid Embolism

    Science.gov (United States)

    ... embolisms are rare, which makes it difficult to identify risk factors. It's estimated that there are between 1 ... Kramer MS, et al. Amniotic fluid embolism: Incidence, risk factors, and impact on perinatal outcome. BJOG: An International Journal of Obstetrics and Gynaecology. 2012;119:874. Baskett ...

  16. Relativistic viscoelastic fluid mechanics.

    Science.gov (United States)

    Fukuma, Masafumi; Sakatani, Yuho

    2011-08-01

    A detailed study is carried out for the relativistic theory of viscoelasticity which was recently constructed on the basis of Onsager's linear nonequilibrium thermodynamics. After rederiving the theory using a local argument with the entropy current, we show that this theory universally reduces to the standard relativistic Navier-Stokes fluid mechanics in the long time limit. Since effects of elasticity are taken into account, the dynamics at short time scales is modified from that given by the Navier-Stokes equations, so that acausal problems intrinsic to relativistic Navier-Stokes fluids are significantly remedied. We in particular show that the wave equations for the propagation of disturbance around a hydrostatic equilibrium in Minkowski space-time become symmetric hyperbolic for some range of parameters, so that the model is free of acausality problems. This observation suggests that the relativistic viscoelastic model with such parameters can be regarded as a causal completion of relativistic Navier-Stokes fluid mechanics. By adjusting parameters to various values, this theory can treat a wide variety of materials including elastic materials, Maxwell materials, Kelvin-Voigt materials, and (a nonlinearly generalized version of) simplified Israel-Stewart fluids, and thus we expect the theory to be the most universal description of single-component relativistic continuum materials. We also show that the presence of strains and the corresponding change in temperature are naturally unified through the Tolman law in a generally covariant description of continuum mechanics.

  17. Spatial Culture

    DEFF Research Database (Denmark)

    2012-01-01

    – 2006. The essays published here allow us to subdivide the field of spatial culture into five major domains, summarized in the titles of chapters in the book: ”Perception and Strategies: Architecture”, ”Politics and Poetics: Urban Spaces”, ”Movements and Cityscape: Textuality”, ”Crisis and Construction......Spatial Culture – A Humanities Perspective Abstract of introductory essay by Henrik Reeh Secured by alliances between socio-political development and cultural practices, a new field of humanistic studies in spatial culture has developed since the 1990s. To focus on links between urban culture...... and modern society is, however, an intellectual practice which has a much longer history. Already in the 1980s, the debate on the modern and the postmodern cited Paris and Los Angeles as spatio-cultural illustrations of these major philosophical concepts. Earlier, in the history of critical studies, the work...

  18. Fluid transport due to nonlinear fluid-structure interaction

    Energy Technology Data Exchange (ETDEWEB)

    Soendergaard Jensen, J.

    1996-08-01

    This work considers nonlinear fluid-structure interaction for a vibrating pipe containing fluid. Transverse pipe vibrations will force the fluid to move relative to the pipe creating uni-directional fluid flow towards the pipe end. The fluid flow induced affects the damping and the stiffness of the pipe. The behavior of the system in response to lateral resonant base excitation is analyzed numerically mode of vibration seems to be most effective for high mean fluid speed, whereas higher modes of vibration can be used to transport fluid with the same fluid speed but with smaller magnitude of pipe vibrations. The effect of the nonlinear geometrical terms is analyzed and these terms are shown to affect the response for higher modes of vibration. Experimental investigations show good agreement with theoretical predictions. (au) 16 refs.

  19. Culture Shock

    Institute of Scientific and Technical Information of China (English)

    宋文玲

    2004-01-01

    Specialists say that it is not easy to get used to life in a new culture.“Culture shock”is the term these specialists use when talking about the feelings that people have in a new environment.There are three stages of culture shock,say the specialists.In the first stage,the newcomers like their new environment,Then when the fresh experience

  20. Cultural diversity

    OpenAIRE

    Raghavan, Raghu

    2011-01-01

    The concept of cultural diversity has emerged as an influential one having impact on multiple policy and legal instruments especially following the adoption of the UNESCO Convention on the Protection and Promotion of the Diversity of Cultural Expressions in 2005. The discussions on its appropriate implementation are however profoundly fragmented and often laden with political considerations. The present brief paper offers some thoughts on the meaning of cultural diversity and its implementati...

  1. Prolonged effect of fluid flow stress on the proliferative activity of mesothelial cells after abrupt discontinuation of fluid streaming

    Energy Technology Data Exchange (ETDEWEB)

    Aoki, Shigehisa, E-mail: aokis@cc.saga-u.ac.jp [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Ikeda, Satoshi [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Takezawa, Toshiaki [Transgenic Animal Research Center, National Institute of Agrobiological Sciences, Ibaraki (Japan); Kishi, Tomoya [Department of Internal Medicine, Saga University, Saga (Japan); Makino, Junichi [Makino Clinic, Saga (Japan); Uchihashi, Kazuyoshi; Matsunobu, Aki [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan); Noguchi, Mitsuru [Department of Urology, Faculty of Medicine, Saga University, Saga (Japan); Sugihara, Hajime [Department of Physical Therapy, International University of Health and Welfare, Fukuoka (Japan); Toda, Shuji [Department of Pathology and Microbiology, Faculty of Medicine, Saga University, Saga (Japan)

    2011-12-16

    Highlights: Black-Right-Pointing-Pointer Late-onset peritoneal fibrosis leading to EPS remains to be elucidated. Black-Right-Pointing-Pointer Fluid streaming is a potent factor for peritoneal fibrosis in PD. Black-Right-Pointing-Pointer We focused on the prolonged effect of fluid streaming on mesothelial cell kinetics. Black-Right-Pointing-Pointer A history of fluid streaming exposure promoted mesothelial proliferative activity. Black-Right-Pointing-Pointer We have thus identified a potent new factor for late-onset peritoneal fibrosis. -- Abstract: Encapsulating peritoneal sclerosis (EPS) often develops after transfer to hemodialysis and transplantation. Both termination of peritoneal dialysis (PD) and transplantation-related factors are risks implicated in post-PD development of EPS, but the precise mechanism of this late-onset peritoneal fibrosis remains to be elucidated. We previously demonstrated that fluid flow stress induced mesothelial proliferation and epithelial-mesenchymal transition via mitogen-activated protein kinase (MAPK) signaling. Therefore, we speculated that the prolonged bioactive effect of fluid flow stress may affect mesothelial cell kinetics after cessation of fluid streaming. To investigate how long mesothelial cells stay under the bioactive effect brought on by fluid flow stress after removal of the stress, we initially cultured mesothelial cells under fluid flow stress and then cultured the cells under static conditions. Mesothelial cells exposed to fluid flow stress for a certain time showed significantly high proliferative activity compared with static conditions after stoppage of fluid streaming. The expression levels of protein phosphatase 2A, which dephosphorylates MAPK, in mesothelial cells changed with time and showed a biphasic pattern that was dependent on the duration of exposure to fluid flow stress. There were no differences in the fluid flow stress-related bioactive effects on mesothelial cells once a certain time had passed

  2. Skin or nail culture

    Science.gov (United States)

    Mucosal culture; Culture - skin; Culture - mucosal; Nail culture; Culture - fingernail; Fingernail culture ... There, it is placed in a special dish (culture). It is then watched to see if bacteria, ...

  3. Purification and characterization of lectin from humoral fluids of Charybdis feriatus

    Institute of Scientific and Technical Information of China (English)

    DAI Congjie; Wang Guizhong; HE Jianfeng; LI Shaojing; HUANG Huiyang

    2006-01-01

    To search new sources of lectin, an experiment on lectin distribution in humoral serum of crab Charybdisferiatus (in short, CFL) was conducted March, 2002. When adding solid ammonium sulfate into the fluids up to 50% saturation at 4℃, most CFL activity showed precipitates who were then continually extracted by ammonium sulfate of different concentrations. The supernatant, which was called primary CFL fluids, was given a 17.60-fold purification and 45.70% recovery of total activity. Finally, by using Sephadex G-100 column chromatography, the CFL in the primary CFL fluid was highly purified.Compared to the original humoral fluids, the last purified CFL got a 203.90-fold purification and 30.48%recovery of total activity, and demonstrated a single band on SDS-PAGE. In thesame time, the purified CFL was detected for agglutination activity with 7 kinds of animal erythrocytes. Other characterization,such as sugar inhibition, and the effect of temperature on the agglutination activity of CFL were also studied. Our results indicate that agglutination activity of CFL was influenced by sugar and temperature.

  4. Amniotic fluid RNA gene expression profiling provides insights into the phenotype of Turner syndrome.

    Science.gov (United States)

    Massingham, Lauren J; Johnson, Kirby L; Scholl, Thomas M; Slonim, Donna K; Wick, Heather C; Bianchi, Diana W

    2014-09-01

    Turner syndrome is a sex chromosome aneuploidy with characteristic malformations. Amniotic fluid, a complex biological material, could contribute to the understanding of Turner syndrome pathogenesis. In this pilot study, global gene expression analysis of cell-free RNA in amniotic fluid supernatant was utilized to identify specific genes/organ systems that may play a role in Turner syndrome pathophysiology. Cell-free RNA from amniotic fluid of five mid-trimester Turner syndrome fetuses and five euploid female fetuses matched for gestational age was extracted, amplified, and hybridized onto Affymetrix(®) U133 Plus 2.0 arrays. Significantly differentially regulated genes were identified using paired t tests. Biological interpretation was performed using Ingenuity Pathway Analysis and BioGPS gene expression atlas. There were 470 statistically significantly differentially expressed genes identified. They were widely distributed across the genome. XIST was significantly down-regulated (p Turner syndrome transcriptome from other aneuploidies we previously studied. Manual curation of the differentially expressed gene list identified genes of possible pathologic significance, including NFATC3, IGFBP5, and LDLR. Transcriptomic differences in the amniotic fluid of Turner syndrome fetuses are due to genome-wide dysregulation. The hematologic/immune system differences may play a role in early-onset autoimmune dysfunction. Other genes identified with possible pathologic significance are associated with cardiac and skeletal systems, which are known to be affected in females with Turner syndrome. The discovery-driven approach described here may be useful in elucidating novel mechanisms of disease in Turner syndrome.

  5. Suppression of MMP activity in bovine cartilage explants cultures has little if any effect on the release of aggrecanase-derived aggrecan fragments

    DEFF Research Database (Denmark)

    Wang, Bijue; Chen, Pingping; Jensen, Anne-Christine Bay

    2009-01-01

    - and aggrecanase-derived fragments of aggrecan and type II collagen into the supernatant of bovine cartilage explants cultures using neo-epitope specific immunoassays, and to associate the release of these fragments with the activity of proteolytic enzymes using inhibitors. FINDINGS: Bovine cartilage explants were...... cultured in the presence or absence of the catabolic cytokines oncostatin M (OSM) and tumor necrosis factor alpha (TNFalpha). In parallel, explants were co-cultured with protease inhibitors such as GM6001, TIMP1, TIMP2 and TIMP3. Fragments released into the supernatant were determined using a range of neo......-epitope specific immunoassays; (1) sandwich (342)FFGVG-G2 ELISA, (2) competition NITEGE(373)ELISA (3) sandwich G1-NITEGE(373 )ELISA (4) competition (374)ARGSV ELISA, and (5) sandwich (374)ARGSV-G2 ELISA all detecting aggrecan fragments, and (6) sandwich CTX-II ELISA, detecting C-telopeptides of type II collagen...

  6. Microgravity Fluids for Biology, Workshop

    Science.gov (United States)

    Griffin, DeVon; Kohl, Fred; Massa, Gioia D.; Motil, Brian; Parsons-Wingerter, Patricia; Quincy, Charles; Sato, Kevin; Singh, Bhim; Smith, Jeffrey D.; Wheeler, Raymond M.

    2013-01-01

    Microgravity Fluids for Biology represents an intersection of biology and fluid physics that present exciting research challenges to the Space Life and Physical Sciences Division. Solving and managing the transport processes and fluid mechanics in physiological and biological systems and processes are essential for future space exploration and colonization of space by humans. Adequate understanding of the underlying fluid physics and transport mechanisms will provide new, necessary insights and technologies for analyzing and designing biological systems critical to NASAs mission. To enable this mission, the fluid physics discipline needs to work to enhance the understanding of the influence of gravity on the scales and types of fluids (i.e., non-Newtonian) important to biology and life sciences. In turn, biomimetic, bio-inspired and synthetic biology applications based on physiology and biology can enrich the fluid mechanics and transport phenomena capabilities of the microgravity fluid physics community.

  7. Tumor interstitial fluid

    DEFF Research Database (Denmark)

    Gromov, Pavel; Gromova, Irina; Olsen, Charlotta J.

    2013-01-01

    Tumor interstitial fluid (TIF) is a proximal fluid that, in addition to the set of blood soluble phase-borne proteins, holds a subset of aberrantly externalized components, mainly proteins, released by tumor cells and tumor microenvironment through various mechanisms, which include classical...... secretion, non-classical secretion, secretion via exosomes and membrane protein shedding. Consequently, the interstitial aqueous phase of solid tumors is a highly promising resource for the discovery of molecules associated with pathological changes in tissues. Firstly, it allows one to delve deeper...... into the regulatory mechanisms and functions of secretion-related processes in tumor development. Secondly, the anomalous secretion of molecules that is innate to tumors and the tumor microenvironment, being associated with cancer progression, offers a valuable source for biomarker discovery and possible targets...

  8. Electrorheologic fluids; Fluidos electroreologicos

    Energy Technology Data Exchange (ETDEWEB)

    Rejon G, Leonardo; Lopez G, Francisco; Montoya T, Gerardo [Instituto de Investigaciones Electricas, Cuernavaca, Morelos (Mexico); Manero B, Octavio [Instituto de Investigaciones en Materiales, UNAM.(Mexico)

    2003-07-01

    The present article has as an objective to offer a review of the research work made in the Instituto de Investigaciones Electricas (IIE) on the study of the electrorheologic fluids whose flow properties can abruptly change in the presence of an electric field when this is induced by a direct current. The electrorheologic fluids have their main application in the manufacture of self-controlling damping systems. [Spanish] El presente articulo tiene por objetivo ofrecer una resena de los trabajos de investigacion realizados en el Instituto de Investigaciones Electricas (IIE) sobre el estudio de los fluidos electroreologicos cuyas propiedades de flujo pueden cambiar abruptamente en presencia de un campo electrico cuando este es inducido por una corriente directa. Los fluidos electroreologicos tienen su principal aplicacion en la fabricacion de sistemas de amortiguamiento autocontrolables.

  9. COUPLED CHEMOTAXIS FLUID MODEL

    KAUST Repository

    LORZ, ALEXANDER

    2010-06-01

    We consider a model system for the collective behavior of oxygen-driven swimming bacteria in an aquatic fluid. In certain parameter regimes, such suspensions of bacteria feature large-scale convection patterns as a result of the hydrodynamic interaction between bacteria. The presented model consist of a parabolicparabolic chemotaxis system for the oxygen concentration and the bacteria density coupled to an incompressible Stokes equation for the fluid driven by a gravitational force of the heavier bacteria. We show local existence of weak solutions in a bounded domain in d, d = 2, 3 with no-flux boundary condition and in 2 in the case of inhomogeneous Dirichlet conditions for the oxygen. © 2010 World Scientific Publishing Company.

  10. Galilean relativistic fluid mechanics

    CERN Document Server

    Ván, Péter

    2015-01-01

    Single component Galilean-relativistic (nonrelativistic) fluids are treated independently of reference frames. The basic fields are given, their balances, thermodynamic relations and the entropy production is calculated. The usual relative basic fields, the mass, momentum and energy densities, the diffusion current density, the pressure tensor and the heat flux are the time- and spacelike components of the third order mass-momentum-energy density tensor according to a velocity field. The transformation rules of the basic fields are derived and prove that the non-equilibrium thermodynamic background theory, that is the Gibbs relation, extensivity condition and the entropy production is absolute, that is independent of the reference frame and also of the fluid velocity. --- Az egykomponensu Galilei-relativisztikus (azaz nemrelativisztikus) disszipativ folyadekokat vonatkoztatasi rendszertol fuggetlenul targyaljuk. Megadjuk az alapmennyisegeket, ezek merlegeit, a termodinamikai osszefuggeseket es kiszamoljuk az ...

  11. Zinc Determination in Pleural Fluid

    OpenAIRE

    Nazan DEMİR; DEMİR, Yaşar

    2000-01-01

    In this study, an enzymatic zinc determination method was applied to pleural fluid, the basis of which was the regaining of the activity of apo carbonic anhydrase by the zinc present in the sample. The method was used for pleural fluid zinc determination in order to show the application to body fluids other than serum. For this purpose, pleural fluids were obtained from 20 patients and zinc concentrations were determined. Carbonic anhydrase was purified by affinity chromatography from bovine ...

  12. Handbook of hydraulic fluid technology

    CERN Document Server

    Totten, George E

    2011-01-01

    ""The Handbook of Hydraulic Fluid Technology"" serves as the foremost resource for designing hydraulic systems and for selecting hydraulic fluids used in engineering applications. Featuring new illustrations, data tables, as well as practical examples, this second edition is updated with essential information on the latest hydraulic fluids and testing methods. The detailed text facilitates unparalleled understanding of the total hydraulic system, including important hardware, fluid properties, and hydraulic lubricants. Written by worldwide experts, the book also offers a rigorous overview of h

  13. Studies on the Primary Toxic Factor of Bt9816C Culture Supernatant%Bt9816C培养上清中杀虫活性成分的研究

    Institute of Scientific and Technical Information of China (English)

    蔡峻; 关彬; 陈月华; 任改新

    2004-01-01

    Bt9816C的培养上清对棉铃虫和甜菜夜蛾具有很高的杀虫活性.热敏实验表明β-外毒素和Zwittermicin A不是其主要杀虫活性成分.Bt9816C的无晶体株保留有卵磷酯酶C和溶血素活性,但是不再具有vip3A基因,其培养上清也丧失了杀虫活性.SDS-PAGE结果显示Bt9816C培养上清中有Vip3A蛋白特征大小的89 kD的蛋白,而其无晶体株正好缺失了该蛋白.以上结果表明Bt9816C培养上清中主要的杀虫活性成分是Vip3A蛋白.

  14. Cultural probes

    DEFF Research Database (Denmark)

    Madsen, Jacob Østergaard

    2016-01-01

    The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation.......The aim of this study was thus to explore cultural probes (Gaver, Boucher et al. 2004), as a possible methodical approach, supporting knowledge production on situated and contextual aspects of occupation....

  15. Manuscript Cultures

    DEFF Research Database (Denmark)

    What do Mesoamerica, Greece, Byzantium, Island, Chad, Ethiopia, India, Tibet, China and Japan have in common? Like many other cultures of the world, they share a particular form of cultural heritage: ancient handwritten documents. In 2007, scholars from some20 countries around the world gathered...

  16. Culture Stories

    DEFF Research Database (Denmark)

    Jensen, Ole B.

    2007-01-01

    by certain representations and embedded in certain norms and values. The analytical framework is applied on a case of cultural urban branding. The case is the harbour front in Aalborg, Denmark where a number of flagship architecture projects and cultural institutions are being planned. It is shown how...

  17. Cultural Communications.

    Science.gov (United States)

    Armas, Jose

    It is too often taken for granted that the communication process with culturally different children takes place as readily as it might with children from Anglo cultures. Most teachers receive training in verbal and formal communication skills; children come to school with nonverbal and informal communication skills. This initially can create…

  18. Amniotic fluid embolism

    OpenAIRE

    Thongrong, Cattleya; Kasemsiri, Pornthep; Hofmann, James P; Bergese, Sergio D.; Thomas J Papadimos; Gracias, Vicente H.; Adolph, Michael D.; Stawicki, Stanislaw P A

    2013-01-01

    Amniotic fluid embolism (AFE) is an unpredictable and as-of-yet unpreventable complication of maternity. With its low incidence it is unlikely that any given practitioner will be confronted with a case of AFE. However, this rare occurrence carries a high probability of serious sequelae including cardiac arrest, ARDS, coagulopathy with massive hemorrhage, encephalopathy, seizures, and both maternal and infant mortality. In this review the current state of medical knowledge about AFE is outline...

  19. Galilean relativistic fluid mechanics

    OpenAIRE

    Ván, Péter

    2015-01-01

    Single component nonrelativistic dissipative fluids are treated independently of reference frames and flow-frames. First the basic fields and their balances, then the related thermodynamic relations and the entropy production are calculated and the linear constitutive relations are given. The usual basic fields of mass, momentum, energy and their current densities, the heat flux, pressure tensor and diffusion flux are the time- and spacelike components of the third order mass-momentum-energy ...

  20. Physics of Fluids

    OpenAIRE

    Luton, J. A.; Ragab, Saad A.

    1997-01-01

    The interaction of vortices passing near a solid surface has been examined using direct numerical simulation. The configuration studied is a counter-rotating vortex pair approaching a wall in an otherwise quiescent fluid. The focus of these simulations is on the three-dimensional effects, of which little is known. To the authors' knowledge, this is the first three-dimensional simulation that lends support to the short-wavelength instability of the secondary vortex. It has been shown how this ...

  1. Soluble oil cutting fluid

    Energy Technology Data Exchange (ETDEWEB)

    Rawlinson, A.P.; White, J.

    1987-06-23

    A soluble oil, suitable when diluted with water, for use as a cutting fluid comprises an alkali or alkaline-earth metal alkyl benzene sulphonate, a fatty acid diethanolamide, a mixed alkanolamine borate, a polyisobutenesuccinimide and a major proportion of mineral oil. The soluble oil is relatively stable without the need for a conventional coupling agent and some soluble oil emulsions are bio-static even though conventional biocides are not included.

  2. Fluid Dynamics and Viscosity in Strongly Correlated Fluids

    CERN Document Server

    Schaefer, Thomas

    2014-01-01

    We review the modern view of fluid dynamics as an effective low energy, long wavelength theory of many body systems at finite temperature. We introduce the notion of a nearly perfect fluid, defined by a ratio $\\eta/s$ of shear viscosity to entropy density of order $\\hbar/k_B$ or less. Nearly perfect fluids exhibit hydrodynamic behavior at all distances down to the microscopic length scale of the fluid. We summarize arguments that suggest that there is fundamental limit to fluidity, and review the current experimental situation with regard to measurements of $\\eta/s$ in strongly coupled quantum fluids.

  3. Inhibition of oncostatin M in osteoarthritic synovial fluid enhances GAG production in osteoarthritic cartilage repair

    OpenAIRE

    Beekhuizen, M.; GJVM van Osch; AGJ Bot; MCL Hoekstra; DBF Saris; WJA Dhert; LB Creemers

    2013-01-01

    Mediators in the synovial fluid are thought to play a major role in osteoarthritic cartilage turnover. The purpose of the current study was to investigate the role of oncostatin M (OSM) in osteoarthritis (OA) by evaluating the presence of the cytokine and its receptors in the OA joint and interfering with its activity in synovial fluid co-cultured with cartilage explants. OSM levels were increased in the synovial fluid of osteoarthritic patients compared to healthy donors. Immunohistochemistr...

  4. Downhole Fluid Analyzer Development

    Energy Technology Data Exchange (ETDEWEB)

    Bill Turner

    2006-11-28

    A novel fiber optic downhole fluid analyzer has been developed for operation in production wells. This device will allow real-time determination of the oil, gas and water fractions of fluids from different zones in a multizone or multilateral completion environment. The device uses near infrared spectroscopy and induced fluorescence measurement to unambiguously determine the oil, water and gas concentrations at all but the highest water cuts. The only downhole components of the system are the fiber optic cable and windows. All of the active components--light sources, sensors, detection electronics and software--will be located at the surface, and will be able to operate multiple downhole probes. Laboratory testing has demonstrated that the sensor can accurately determine oil, water and gas fractions with a less than 5 percent standard error. Once installed in an intelligent completion, this sensor will give the operating company timely information about the fluids arising from various zones or multilaterals in a complex completion pattern, allowing informed decisions to be made on controlling production. The research and development tasks are discussed along with a market analysis.

  5. Astrophysical fluid dynamics

    Science.gov (United States)

    Ogilvie, Gordon I.

    2016-06-01

    > These lecture notes and example problems are based on a course given at the University of Cambridge in Part III of the Mathematical Tripos. Fluid dynamics is involved in a very wide range of astrophysical phenomena, such as the formation and internal dynamics of stars and giant planets, the workings of jets and accretion discs around stars and black holes and the dynamics of the expanding Universe. Effects that can be important in astrophysical fluids include compressibility, self-gravitation and the dynamical influence of the magnetic field that is `frozen in' to a highly conducting plasma. The basic models introduced and applied in this course are Newtonian gas dynamics and magnetohydrodynamics (MHD) for an ideal compressible fluid. The mathematical structure of the governing equations and the associated conservation laws are explored in some detail because of their importance for both analytical and numerical methods of solution, as well as for physical interpretation. Linear and nonlinear waves, including shocks and other discontinuities, are discussed. The spherical blast wave resulting from a supernova, and involving a strong shock, is a classic problem that can be solved analytically. Steady solutions with spherical or axial symmetry reveal the physics of winds and jets from stars and discs. The linearized equations determine the oscillation modes of astrophysical bodies, as well as their stability and their response to tidal forcing.

  6. Corrosion in supercritical fluids

    Energy Technology Data Exchange (ETDEWEB)

    Propp, W.A.; Carleson, T.E.; Wai, Chen M.; Taylor, P.R.; Daehling, K.W.; Huang, Shaoping; Abdel-Latif, M.

    1996-05-01

    Integrated studies were carried out in the areas of corrosion, thermodynamic modeling, and electrochemistry under pressure and temperature conditions appropriate for potential applications of supercritical fluid (SCF) extractive metallurgy. Carbon dioxide and water were the primary fluids studied. Modifiers were used in some tests; these consisted of 1 wt% water and 10 wt% methanol for carbon dioxide and of sulfuric acid, sodium sulfate, ammonium sulfate, and ammonium nitrate at concentrations ranging from 0.00517 to 0.010 M for the aqueous fluids. The materials studied were Types 304 and 316 (UNS S30400 and S31600) stainless steel, iron, and AISI-SAE 1080 (UNS G10800) carbon steel. The thermodynamic modeling consisted of development of a personal computer-based program for generating Pourbaix diagrams at supercritical conditions in aqueous systems. As part of the model, a general method for extrapolating entropies and related thermodynamic properties from ambient to SCF conditions was developed. The experimental work was used as a tool to evaluate the predictions of the model for these systems. The model predicted a general loss of passivation in iron-based alloys at SCF conditions that was consistent with experimentally measured corrosion rates and open circuit potentials. For carbon-dioxide-based SCFs, measured corrosion rates were low, indicating that carbon steel would be suitable for use with unmodified carbon dioxide, while Type 304 stainless steel would be suitable for use with water or methanol as modifiers.

  7. Geophysical fluid dynamics

    CERN Document Server

    Pedlosky, Joseph

    1979-01-01

    The content of this book is based, largely, on the core curriculum in geophys­ ical fluid dynamics which I and my colleagues in the Department of Geophysical Sciences at The University of Chicago have taught for the past decade. Our purpose in developing a core curriculum was to provide to advanced undergraduates and entering graduate students a coherent and systematic introduction to the theory of geophysical fluid dynamics. The curriculum and the outline of this book were devised to form a sequence of courses of roughly one and a half academic years (five academic quarters) in length. The goal of the sequence is to help the student rapidly advance to the point where independent study and research are practical expectations. It quickly became apparent that several topics (e. g. , some aspects of potential theory) usually thought of as forming the foundations of a fluid-dynamics curriculum were merely classical rather than essential and could be, however sadly, dispensed with for our purposes. At the same ti...

  8. Geophysical fluid dynamics

    CERN Document Server

    Pedlosky, Joseph

    1982-01-01

    The content of this book is based, largely, on the core curriculum in geophys­ ical fluid dynamics which land my colleagues in the Department of Geophysical Sciences at The University of Chicago have taught for the past decade. Our purpose in developing a core curriculum was to provide to advanced undergraduates and entering graduate students a coherent and systematic introduction to the theory of geophysical fluid dynamics. The curriculum and the outline of this book were devised to form a sequence of courses of roughly one and a half academic years (five academic quarters) in length. The goal of the sequence is to help the student rapidly advance to the point where independent study and research are practical expectations. It quickly became apparent that several topics (e. g. , some aspects of potential theory) usually thought of as forming the foundations of a fluid-dynamics curriculum were merely classical rather than essential and could be, however sadly, dispensed with for our purposes. At the same tim...

  9. Galilean relativistic fluid mechanics

    Science.gov (United States)

    Ván, P.

    2017-01-01

    Single-component nonrelativistic dissipative fluids are treated independently of reference frames and flow-frames. First the basic fields and their balances are derived, then the related thermodynamic relations and the entropy production are calculated and the linear constitutive relations are given. The usual basic fields of mass, momentum, energy and their current densities, the heat flux, pressure tensor and diffusion flux are the time- and spacelike components of the third-order mass-momentum-energy density-flux four-tensor. The corresponding Galilean transformation rules of the physical quantities are derived. It is proved that the non-equilibrium thermodynamic frame theory, including the thermostatic Gibbs relation and extensivity condition and also the entropy production, is independent of the reference frame and also the flow-frame of the fluid. The continuity-Fourier-Navier-Stokes equations are obtained almost in the traditional form if the flow of the fluid is fixed to the temperature. This choice of the flow-frame is the thermo-flow. A simple consequence of the theory is that the relation between the total, kinetic and internal energies is a Galilean transformation rule.

  10. The Viscosity of Polymeric Fluids.

    Science.gov (United States)

    Perrin, J. E.; Martin, G. C.

    1983-01-01

    To illustrate the behavior of polymeric fluids and in what respects they differ from Newtonian liquids, an experiment was developed to account for the shear-rate dependence of non-Newtonian fluids. Background information, procedures, and results are provided for the experiment. Useful in transport processes, fluid mechanics, or physical chemistry…

  11. CT of retrorenal fluid collections

    Energy Technology Data Exchange (ETDEWEB)

    Love, L.; Demos, T.C.; Posniak, H.

    1985-07-01

    Fluid collections dorsal to one or both kidneys are often observed on CT. Most of these collections are located in the posterior pararenal space, but occasionally fluid collections that do not originate in this space also occur. The authors review retrorenal fluid collections with explanations for their occurrence.

  12. Basic concepts of fluid responsiveness

    NARCIS (Netherlands)

    T.G.V. Cherpanath (Thomas); B.F. Geerts (Bart); W.K. Lagrand (Wim); M.J. Schultz (Marcus); A.B.J. Groeneveld (Johan)

    2013-01-01

    textabstractPredicting fluid responsiveness, the response of stroke volume to fluid loading, is a relatively novel concept that aims to optimise circulation, and as such organ perfusion, while avoiding futile and potentially deleterious fluid administrations in critically ill patients. Dynamic param

  13. CULTURAL TOURISM

    Directory of Open Access Journals (Sweden)

    Dana POP

    2016-07-01

    Full Text Available In this paper we will try to analyse the cultural tourism. We will start by referring to the complex concepts of tourism and culture and to the synergies existing between them. We will define cultural tourism and present its appearance and evolution as well as its importance as a modern form of tourism. We will present the various types of cultural tourism with their characteristics and the specific features of cultural tourists according to their interests. We will also mention that there are advantages and disadvantages for any kind of tourism depending on the position – local communities, companies or tourists. For the future we will refer to the new partnership between UNWTO and UNESCO.

  14. Fluid transport due to nonlinear fluid-structure interaction

    DEFF Research Database (Denmark)

    Jensen, Jakob Søndergaard

    1997-01-01

    This work considers nonlinear fluid-structure interaction for a vibrating pipe containing fluid. Transverse pipe vibrations will force the fluid to move relative to the pipe creating unidirectional fluid flow towards the pipe end. The fluid flow induced affects the damping and the stiffness...... of the pipe. The behavior of the system in response to lateral resonant base excitation is analysed numerically and by the use of a perturbation method (multiple scales). Exciting the pipe in the fundamental mode of vibration seems to be most effective for transferring energy from the shaker to the fluid......, whereas higher modes of vibration can be used to transport fluid with pipe vibrations of smaller amplitude. The effect of the nonlinear geometrical terms is analysed and these terms are shown to affect the response for higher modes of vibration. Experimental investigations show good agreement...

  15. Automobile windshield washer fluid: A potential source of transmission for Legionella

    Energy Technology Data Exchange (ETDEWEB)

    Schwake, David Otto [School of Life Sciences, Arizona State University, P.O. Box 875306, Tempe, AZ 85287 (United States); Alum, Absar [School of Sustainable Engineering and the Built Environment, Arizona State University, P.O. Box 875306, Tempe, AZ 85287 (United States); Abbaszadegan, Morteza, E-mail: abbaszadegan@asu.edu [School of Sustainable Engineering and the Built Environment, Arizona State University, P.O. Box 875306, Tempe, AZ 85287 (United States)

    2015-09-01

    Epidemiological evidence suggesting driving cars to be a risk factor for legionellosis has prompted public health studies to investigate vehicle windshield washer fluid as a novel transmission source of this disease. The goal of the current study was to investigate whether or not windshield washer fluid could serve as a potential source of transmission for Legionella. A wide variation in the survival of L. pneumophila was observed when incubated in different washer fluids at 25 and 37 °C, however, one brand tested supported Legionella survival similar to or greater than sterilized deionized water. In addition, 1 L of tap water contained in a washer fluid reservoir was able to support population growth and survival of Legionella for several months. In a field study examining the windshield washer fluid of 12 elementary school buses, Legionella were detected from 84% of samples at a high concentration of 8.1 × 10{sup 4} CFU/mL. Culturable cells were also detected in aerosolized washer fluid during washer fluid spray. By demonstrating survival in certain windshield washer fluids, growth within washer fluid reservoirs, and the presence of viable cells in bus washer fluid spray, we have provided evidence suggesting the potential for a novel route of Legionella exposure. - Highlights: • L. pneumophila survival in one brand of washer fluid and sterilized deionized water were similar. • Legionella population was maintained in tap water for several months. • Culturable Legionella were detected in 10 of 12 school bus washer fluid reservoirs sampled. • Legionella concentrations up to 8.1 × 104 CFU/mL were detected in school bus washer fluid. • L. pneumophila was detected in washer fluid reservoirs and aerosolized washer fluid.

  16. Supercritical fluid reverse micelle separation

    Science.gov (United States)

    Fulton, John L.; Smith, Richard D.

    1993-01-01

    A method of separating solute material from a polar fluid in a first polar fluid phase is provided. The method comprises combining a polar fluid, a second fluid that is a gas at standard temperature and pressure and has a critical density, and a surfactant. The solute material is dissolved in the polar fluid to define the first polar fluid phase. The combined polar and second fluids, surfactant, and solute material dissolved in the polar fluid is maintained under near critical or supercritical temperature and pressure conditions such that the density of the second fluid exceeds the critical density thereof. In this way, a reverse micelle system defining a reverse micelle solvent is formed which comprises a continuous phase in the second fluid and a plurality of reverse micelles dispersed in the continuous phase. The solute material is dissolved in the polar fluid and is in chemical equilibrium with the reverse micelles. The first polar fluid phase and the continuous phase are immiscible. The reverse micelles each comprise a dynamic aggregate of surfactant molecules surrounding a core of the polar fluid. The reverse micelle solvent has a polar fluid-to-surfactant molar ratio W, which can vary over a range having a maximum ratio W.sub.o that determines the maximum size of the reverse micelles. The maximum ratio W.sub.o of the reverse micelle solvent is then varied, and the solute material from the first polar fluid phase is transported into the reverse micelles in the continuous phase at an extraction efficiency determined by the critical or supercritical conditions.

  17. Supercritical fluid reverse micelle separation

    Science.gov (United States)

    Fulton, J.L.; Smith, R.D.

    1993-11-30

    A method of separating solute material from a polar fluid in a first polar fluid phase is provided. The method comprises combining a polar fluid, a second fluid that is a gas at standard temperature and pressure and has a critical density, and a surfactant. The solute material is dissolved in the polar fluid to define the first polar fluid phase. The combined polar and second fluids, surfactant, and solute material dissolved in the polar fluid is maintained under near critical or supercritical temperature and pressure conditions such that the density of the second fluid exceeds the critical density thereof. In this way, a reverse micelle system defining a reverse micelle solvent is formed which comprises a continuous phase in the second fluid and a plurality of reverse micelles dispersed in the continuous phase. The solute material is dissolved in the polar fluid and is in chemical equilibrium with the reverse micelles. The first polar fluid phase and the continuous phase are immiscible. The reverse micelles each comprise a dynamic aggregate of surfactant molecules surrounding a core of the polar fluid. The reverse micelle solvent has a polar fluid-to-surfactant molar ratio W, which can vary over a range having a maximum ratio W[sub o] that determines the maximum size of the reverse micelles. The maximum ratio W[sub o] of the reverse micelle solvent is then varied, and the solute material from the first polar fluid phase is transported into the reverse micelles in the continuous phase at an extraction efficiency determined by the critical or supercritical conditions. 27 figures.

  18. Fluid-fluid versus fluid-solid demixing in mixtures of parallel hard hypercubes

    Science.gov (United States)

    Lafuente, Luis; Martínez-Ratón, Yuri

    2011-02-01

    It is well known that increase of the spatial dimensionality enhances the fluid-fluid demixing of a binary mixture of hard hyperspheres, i.e. the demixing occurs for lower mixture size asymmetry as compared to the three-dimensional case. However, according to simulations, in the latter dimension the fluid-fluid demixing is metastable with respect to the fluid-solid transition. According to the results obtained from approximations to the equation of state of hard hyperspheres in higher dimensions, the fluid-fluid demixing might become stable for high enough dimension. However, this conclusion is rather speculative since none of these works have taken into account the stability of the crystalline phase (by a minimization of a given density functional, by spinodal calculations or by MC simulations). Of course, the lack of results is justified by the difficulty of performing density functional calculations or simulations in high dimensions and, in particular, for highly asymmetric binary mixtures. In the present work, we will take advantage of a well tested theoretical tool, namely the fundamental measure density functional theory for parallel hard hypercubes (in the continuum and in the hypercubic lattice). With this, we have calculated the fluid-fluid and fluid-solid spinodals for different spatial dimensions. We have obtained, no matter what the dimensionality, the mixture size asymmetry or the polydispersity (included as a bimodal distribution function centered around the asymmetric edge lengths), that the fluid-fluid critical point is always located above the fluid-solid spinodal. In conclusion, these results point to the existence of demixing between at least one solid phase rich in large particles and one fluid phase rich in small ones, preempting a fluid-fluid demixing, independently of the spatial dimension or the polydispersity.

  19. Structural Transition in Supercritical Fluids

    Directory of Open Access Journals (Sweden)

    Boris I. Sedunov

    2011-01-01

    Full Text Available The extension of the saturation curve ( on the PT diagram in the supercritical region for a number of monocomponent supercritical fluids by peak values for different thermophysical properties, such as heat capacities and and compressibility has been studied. These peaks signal about some sort of fluid structural transition in the supercritical region. Different methods give similar but progressively diverging curves st( for this transition. The zone of temperatures and pressures near these curves can be named as the zone of the fluid structural transition. The outstanding properties of supercritical fluids in this zone help to understand the physical sense of the fluid structural transition.

  20. Undulatory swimming in viscoelastic fluids

    CERN Document Server

    Shen, Xiaoning

    2011-01-01

    The effects of fluid elasticity on the swimming behavior of the nematode \\emph{Caenorhabditis elegans} are experimentally investigated by tracking the nematode's motion and measuring the corresponding velocity fields. We find that fluid elasticity hinders self-propulsion. Compared to Newtonian solutions, fluid elasticity leads to 35% slower propulsion speed. Furthermore, self-propulsion decreases as elastic stresses grow in magnitude in the fluid. This decrease in self-propulsion in viscoelastic fluids is related to the stretching of flexible molecules near hyperbolic points in the flow.

  1. Heat transfer fluids containing nanoparticles

    Science.gov (United States)

    Singh, Dileep; Routbort, Jules; Routbort, A.J.; Yu, Wenhua; Timofeeva, Elena; Smith, David S.; France, David M.

    2016-05-17

    A nanofluid of a base heat transfer fluid and a plurality of ceramic nanoparticles suspended throughout the base heat transfer fluid applicable to commercial and industrial heat transfer applications. The nanofluid is stable, non-reactive and exhibits enhanced heat transfer properties relative to the base heat transfer fluid, with only minimal increases in pumping power required relative to the base heat transfer fluid. In a particular embodiment, the plurality of ceramic nanoparticles comprise silicon carbide and the base heat transfer fluid comprises water and water and ethylene glycol mixtures.

  2. Undulatory swimming in viscoelastic fluids.

    Science.gov (United States)

    Shen, X N; Arratia, P E

    2011-05-20

    The effects of fluid elasticity on the swimming behavior of the nematode Caenorhabditis elegans are experimentally investigated by tracking the nematode's motion and measuring the corresponding velocity fields. We find that fluid elasticity hinders self-propulsion. Compared to Newtonian solutions, fluid elasticity leads to up to 35% slower propulsion. Furthermore, self-propulsion decreases as elastic stresses grow in magnitude in the fluid. This decrease in self-propulsion in viscoelastic fluids is related to the stretching of flexible molecules near hyperbolic points in the flow.

  3. Cultural History and Cultural Materialism.

    Science.gov (United States)

    Berman, Ronald

    1990-01-01

    Historicism critiques cultural history and cultural materialism as a methodology for literary analysis. Questions the finality of interpretation, how original values change, and whether dramatic history implies actual history. Using Shakespearean plays, analyzes the power and politics of a play in relation to its audience; posits that cultural…

  4. Cultural History and Cultural Materialism.

    Science.gov (United States)

    Berman, Ronald

    1990-01-01

    Historicism critiques cultural history and cultural materialism as a methodology for literary analysis. Questions the finality of interpretation, how original values change, and whether dramatic history implies actual history. Using Shakespearean plays, analyzes the power and politics of a play in relation to its audience; posits that cultural…

  5. Digging culture and doing culture

    NARCIS (Netherlands)

    Andries van den Broek; Jos de Haan; Frank Huysmans

    2009-01-01

    Original title: Cultuurbewonderaars en cultuurbeoefenaars. There are people who love art and culture and there are people who practise it; people who enjoy it and people who are themselves creative in their leisure time. Who are these culture-lovers and practitioners? How has participation in

  6. Culture Matters

    Directory of Open Access Journals (Sweden)

    Gillian Warner-Søderholm

    2012-12-01

    Full Text Available Whether managers are concerned with financial issues, marketing, or human resource management (HRM, cultural values and practices do matter. The purpose of this article is to understand Norwegian managers’ cultural values within the cross-cultural landscape of her neighbors in the “Scandinavian cluster.” Clearly, subtle but disturbing differences may surface even when representatives from similar cultures work together. As a follow on from the GLOBE project, data based on the GLOBE instrument were collected on culture and communication values in Norway from 710 Norwegian middle managers for this present study. Although the Scandinavian cultures appear ostensibly similar, the results illustrate that research can reveal subtle but important cultural differences in nations that are similar yet dissimilar. All three Scandinavian societies appear intrinsically egalitarian; they appear to value low Power Distance, directness, and consensus in decision making and to promote Gender Egalitarianism. Nevertheless, there are significant differences in the degrees of commitment to these values by each individual Scandinavian partner. These differences need to be understood and appreciated to avoid misunderstandings.

  7. Fluid viscosity under confined conditions

    Science.gov (United States)

    Rudyak, V. Ya.; Belkin, A. A.

    2014-12-01

    Closed equations of fluid transfer in confined conditions are constructed in this study using ab initio methods of nonequilibrium statistical mechanics. It is shown that the fluid viscosity is not determined by the fluid properties alone, but becomes a property of the "fluid-nanochannel walls" system as a whole. Relations for the tensor of stresses and the interphase force, which specifies the exchange by momentum of fluid molecules with the channel-wall molecules, are derived. It is shown that the coefficient of viscosity is now determined by the sum of three contributions. The first contribution coincides with the expression for the coefficient of the viscosity of fluid in the bulk being specified by the interaction of fluid molecules with each other. The second contribution has the same structure as the first one but is determined by the interaction of fluid molecules with the channel-wall molecules. Finally, the third contribution has no analog in the usual statistical mechanics of transport processes of a simple fluid. It is associated with the correlation of intermolecular forces of the fluid and the channel walls. Thus, it is established that the coefficient of viscosity of fluid in sufficiently small channels will substantially differ from its bulk value.

  8. Noncommutative Fluid and Cosmological Perturbations

    CERN Document Server

    Das, Praloy

    2016-01-01

    In the present paper we have developed a Non-Commutative (NC) generalization of perfect fluid model from first principles, in a Hamiltonian framework. The noncommutativity is introduced at the Lagrangian (particle) coordinate space brackets and the induced NC fluid bracket algebra for the Eulerian (fluid) field variables is derived. Together with a Hamiltonian this NC algebra generates the generalized fluid dynamics that satisfies exact local conservation laws for mass and energy thereby maintaining mass and energy conservation. However, nontrivial NC correction terms appear in charge and energy fluxes. Other non-relativistic spacetime symmetries of the NC fluid are also discussed in detail. This constitutes the NC fluid dynamics and kinematics. In the second part we construct an extension of Friedmann-Robertson-Walker (FRW) cosmological model based on the NC fluid dynamics presented here. We outline the way in which NC effects generate cosmological perturbations bringing in anisotropy and inhomogeneity in th...

  9. Active colloids in complex fluids

    CERN Document Server

    Patteson, Alison E; Arratia, Paulo E

    2016-01-01

    We review recent work on active colloids or swimmers, such as self-propelled microorganisms, phoretic colloidal particles, and artificial micro-robotic systems, moving in fluid-like environments. These environments can be water-like and Newtonian but can frequently contain macromolecules, flexible polymers, soft cells, or hard particles, which impart complex, nonlinear rheological features to the fluid. While significant progress has been made on understanding how active colloids move and interact in Newtonian fluids, little is known on how active colloids behave in complex and non-Newtonian fluids. An emerging literature is starting to show how fluid rheology can dramatically change the gaits and speeds of individual swimmers. Simultaneously, a moving swimmer induces time dependent, three dimensional fluid flows, that can modify the medium (fluid) rheological properties. This two-way, non-linear coupling at microscopic scales has profound implications at meso- and macro-scales: steady state suspension proper...

  10. Fluid dynamics of heart development.

    Science.gov (United States)

    Santhanakrishnan, Arvind; Miller, Laura A

    2011-09-01

    The morphology, muscle mechanics, fluid dynamics, conduction properties, and molecular biology of the developing embryonic heart have received much attention in recent years due to the importance of both fluid and elastic forces in shaping the heart as well as the striking relationship between the heart's evolution and development. Although few studies have directly addressed the connection between fluid dynamics and heart development, a number of studies suggest that fluids may play a key role in morphogenic signaling. For example, fluid shear stress may trigger biochemical cascades within the endothelial cells of the developing heart that regulate chamber and valve morphogenesis. Myocardial activity generates forces on the intracardiac blood, creating pressure gradients across the cardiac wall. These pressures may also serve as epigenetic signals. In this article, the fluid dynamics of the early stages of heart development is reviewed. The relevant work in cardiac morphology, muscle mechanics, regulatory networks, and electrophysiology is also reviewed in the context of intracardial fluid dynamics.

  11. Fluid and Electrolyte Nutrition

    Science.gov (United States)

    Lane, Helen W.; Smith, Scott M.; Leach, Carolyn S.; Rice, Barbara L.

    1999-01-01

    Studies of fluid and electrolyte homeostasis have been completed since the early human space flight programs, with comprehensive research completed on the Spacelab Life Sciences missions SLS-1 and SLS-2 flights, and more recently on the Mir 18 mission. This work documented the known shifts in fluids, the decrease in total blood volume, and indications of reduced thirst. Data from these flights was used to evaluate the nutritional needs for water, sodium, and potassium. Interpretations of the data are confounded by the inadequate energy intakes routinely observed during space flight. This in turn results in reduced fluid intake, as food provides approximately 70% water intake. Subsequently, body weight, lean body mass, total body water, and total body potassium may decrease. Given these issues, there is evidence to support a minimum required water intake of 2 L per day. Data from previous Shuttle flights indicated that water intake is 2285 +/- 715 ml/day (mean +/- SD, n=26). There are no indications that sodium intake or homeostasis is compromised during space flight. The normal or low aldosterone and urinary sodium levels suggest adequate sodium intake (4047 +/- 902 mg/day, n=26). Because excessive sodium intake is associated with hypercalciuria, the recommended maximum amount of sodium intake during flight is 3500 mg/day (i.e., similar to the Recommended Dietary Allowance, RDA). Potassium metabolism appears to be more complex. Data indicate loss of body potassium related to muscle atrophy and low dietary intake (2407 +/- 548 mg/day, n=26). Although possibly related to measurement error, the elevations in blood potassium suggest alterations in potassium homeostasis. The space RDA for minimum potassium intake is 3500 mg/day. With the documented inadequate intakes, efforts are being made to increase dietary consumption of potassium.

  12. In Vitro binding capacity of zeolite A to calcium, phosphorus and magnesium in rumen fluid as influenced by changes in pH

    DEFF Research Database (Denmark)

    Thilsing, Trine; Jørgensen, Rolf Jess; Poulsen, H.D.

    2006-01-01

    An in vitro experiment was designed to mimic the transport of ingested zeolite A in the forestomachs and proximal part of the small intestine so as to evaluate the binding capacity of zeolite A to Ca, P and Mg as influenced by changes in pH. This was done by incubation of rumen fluid solutions...... with and without zeolite, as well as varying the content of Ca and/or P. The pH was lowered by addition of HCl so as to mimic abomasal conditions, followed by subsequent HCO3- addition to mimic small intestinal pH. Rumen fluid samples were taken at strategic time points in the experiment. All samples were...... centrifuged and the supernatant analysed for Ca, P and Mg as indicators of the amount of unbound mineral. The addition of zeolite to rumen fluid solutions reduced the amount of supernatant Ca and Mg at rumen pH, whereas the level of P was not reduced. After adding HCl, a large proportion of the zeolite...

  13. Cultural Spectrum

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Exorbitant Performance Fees ProhibitedThe Ministry of Culture has taken measures todiscourage performance artists from charging excessively for their stage appearances. According to the ministry, payment for performers and ticket prices must be in line with average con-

  14. Blood culture

    Science.gov (United States)

    Culture - blood ... A blood sample is needed . The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

  15. Rectal culture

    Science.gov (United States)

    ... 2016:chap 283. Haines CF, Sears CL. Infectious enteritis and proctocolitis. In: Feldman M, Friedman LS, Brandt ... PA: Elsevier; 2017:chap 22. Read More Cryptosporidium enteritis Fecal culture Proctitis Review Date 5/11/2016 ...

  16. callus culture

    African Journals Online (AJOL)

    steve

    and KH2PO4 -NH4NO3 depletion was fatal to grape cells and resulted in formation ... Individual phenolic compounds found in the grape cell cultures were ...... Characterization of in vitro anthocyanin-producing sour cherry ... Process Biochem.

  17. Culture School

    Institute of Scientific and Technical Information of China (English)

    MATTHEW LIM

    2008-01-01

    @@ Until recently, employees posted abroad would, if they were lucky, receive some limited language training before they relocated. How they would cope with living in a completely new culture when they arrived was left up to them to figure out.

  18. Yangshao Culture

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Yangshao culture was born in the Neolithic Age. In 1921, archaeologists unearthed a number of chipped stone implements such as knives and axes; bone objects and everyday ceramic items. Thus Yangshao Village and its matriarchal society have

  19. Acoustic concentration of particles in fluid flow

    Science.gov (United States)

    Ward, Michael D.; Kaduchak, Gregory

    2010-11-23

    An apparatus for acoustic concentration of particles in a fluid flow includes a substantially acoustically transparent membrane and a vibration generator that define a fluid flow path therebetween. The fluid flow path is in fluid communication with a fluid source and a fluid outlet and the vibration generator is disposed adjacent the fluid flow path and is capable of producing an acoustic field in the fluid flow path. The acoustic field produces at least one pressure minima in the fluid flow path at a predetermined location within the fluid flow path and forces predetermined particles in the fluid flow path to the at least one pressure minima.

  20. Acoustic concentration of particles in fluid flow

    Energy Technology Data Exchange (ETDEWEB)

    Ward, Michael W.; Kaduchak, Gregory

    2017-08-15

    Disclosed herein is a acoustic concentration of particles in a fluid flow that includes a substantially acoustically transparent membrane and a vibration generator that define a fluid flow path therebetween. The fluid flow path is in fluid communication with a fluid source and a fluid outlet and the vibration generator is disposed adjacent the fluid flow path and is capable of producing an acoustic field in the fluid flow path. The acoustic field produces at least one pressure minima in the fluid flow path at a predetermined location within the fluid flow path and forces predetermined particles in the fluid flow path to the at least one pressure minima.

  1. Transport Coefficients of Fluids

    CERN Document Server

    Eu, Byung Chan

    2006-01-01

    Until recently the formal statistical mechanical approach offered no practicable method for computing the transport coefficients of liquids, and so most practitioners had to resort to empirical fitting formulas. This has now changed, as demonstrated in this innovative monograph. The author presents and applies new methods based on statistical mechanics for calculating the transport coefficients of simple and complex liquids over wide ranges of density and temperature. These molecular theories enable the transport coefficients to be calculated in terms of equilibrium thermodynamic properties, and the results are shown to account satisfactorily for experimental observations, including even the non-Newtonian behavior of fluids far from equilibrium.

  2. Essential Computational Fluid Dynamics

    CERN Document Server

    Zikanov, Oleg

    2011-01-01

    This book serves as a complete and self-contained introduction to the principles of Computational Fluid Dynamic (CFD) analysis. It is deliberately short (at approximately 300 pages) and can be used as a text for the first part of the course of applied CFD followed by a software tutorial. The main objectives of this non-traditional format are: 1) To introduce and explain, using simple examples where possible, the principles and methods of CFD analysis and to demystify the `black box’ of a CFD software tool, and 2) To provide a basic understanding of how CFD problems are set and

  3. Electrochemistry in supercritical fluids

    Science.gov (United States)

    Branch, Jack A.; Bartlett, Philip N.

    2015-01-01

    A wide range of supercritical fluids (SCFs) have been studied as solvents for electrochemistry with carbon dioxide and hydrofluorocarbons (HFCs) being the most extensively studied. Recent advances have shown that it is possible to get well-resolved voltammetry in SCFs by suitable choice of the conditions and the electrolyte. In this review, we discuss the voltammetry obtained in these systems, studies of the double-layer capacitance, work on the electrodeposition of metals into high aspect ratio nanopores and the use of metallocenes as redox probes and standards in both supercritical carbon dioxide–acetonitrile and supercritical HFCs. PMID:26574527

  4. Electrochemistry in supercritical fluids.

    Science.gov (United States)

    Branch, Jack A; Bartlett, Philip N

    2015-12-28

    A wide range of supercritical fluids (SCFs) have been studied as solvents for electrochemistry with carbon dioxide and hydrofluorocarbons (HFCs) being the most extensively studied. Recent advances have shown that it is possible to get well-resolved voltammetry in SCFs by suitable choice of the conditions and the electrolyte. In this review, we discuss the voltammetry obtained in these systems, studies of the double-layer capacitance, work on the electrodeposition of metals into high aspect ratio nanopores and the use of metallocenes as redox probes and standards in both supercritical carbon dioxide-acetonitrile and supercritical HFCs.

  5. Mixture of Anisotropic Fluids

    Science.gov (United States)

    Florkowski, W.; Maj, R.

    The recently introduced approach describing coupled quark and gluon anisotropic fluids is generalized to include explicitly the transitions between quarks and gluons. We study the effects of such processes on the thermalization rate of anisotropic systems. We find that the quark-gluon transitions may enhance the overall thermalization rate in the cases where the initial momentum anisotropies correspond to mixed oblate-prolate or prolate configurations. On the other hand, no effect on the thermalization rate is found in the case of oblate configurations. The observed regularities are connected with the late-time behavior of the analyzed systems which is described either by the exponential decay or the power law.

  6. Mixture of anisotropic fluids

    CERN Document Server

    Florkowski, Wojciech

    2013-01-01

    The recently introduced approach describing coupled quark and gluon anisotropic fluids is generalized to include explicitly the transitions between quarks and gluons. We study the effects of such processes on the thermalization rate of anisotropic systems. We find that the quark-gluon transitions may enhance the overall thermalization rate in the cases where the initial momentum anisotropies correspond to mixed oblate-prolate or prolate configurations. On the other hand, no effect on the thermalization rate is found in the case of oblate configurations. The observed regularities are connected with the late-time behavior of the analyzed systems which is described either by the exponential decay or the power law.

  7. Conformal fluid dynamics

    CERN Document Server

    Jarvis, P D

    2006-01-01

    We present a conformal theory of a dissipationless relativistic fluid in 2 space-time dimensions. The theory carries with it a representation of the algebra of 2-$D$ area-preserving diffeomorphisms in the target space of the complex scalar potentials. A complete canonical description is given, and the central charge of the current algebra is calculated. The passage to the quantum theory is discussed in some detail; as a result of operator ordering problems, full quantization at the level of the fields is as yet an open problem.

  8. Reliability of fluid systems

    Directory of Open Access Journals (Sweden)

    Kopáček Jaroslav

    2016-01-01

    Full Text Available This paper focuses on the importance of detection reliability, especially in complex fluid systems for demanding production technology. The initial criterion for assessing the reliability is the failure of object (element, which is seen as a random variable and their data (values can be processed using by the mathematical methods of theory probability and statistics. They are defined the basic indicators of reliability and their applications in calculations of serial, parallel and backed-up systems. For illustration, there are calculation examples of indicators of reliability for various elements of the system and for the selected pneumatic circuit.

  9. Culture perspectives.

    Science.gov (United States)

    Locsin, Rozzano C

    2002-10-01

    All cultures have had means and techniques that express their immediate aims. The thing that interests me is that today, painters do not have to go to a subject matter outside of themselves. They work from a different source. They work from within. It seems to me that the modern artist cannot express this age, the airplane, the atom bomb, the radio, in the old form of the Renaissance or of any of the old cultures.

  10. Cultural Imports

    Institute of Scientific and Technical Information of China (English)

    1996-01-01

    FOREIGN commodities have flowed into China since the country opened its doors to the outside world. China is an expansive territory with a huge population offering a vast potential consumer market. There are absolutely no limits to the world of culture in China, with Chinese people having access to foreign films, dramas, music and books, all of which have helped to strengthen exchanges between Chinese and Western cultures.

  11. Cell culture plastics with immobilized interleukin-4 for monocyte differentiation

    DEFF Research Database (Denmark)

    Hansen, Morten; Hjortø, Gertrud Malene; Met, Özcan;

    2011-01-01

    Standard cell culture plastic was surface modified by passive adsorption or covalent attachment of interleukin (IL)-4 and investigated for its ability to induce differentiation of human monocytes into mature dendritic cells, a process dose-dependently regulated by IL-4. Covalent attachment of IL-4...... proceeded via anthraquinone photochemistry to introduce amine functionalities at the surface followed by coupling of IL-4 through a bifunctional amine-reactive linker. X-ray photoelectron spectroscopy showed that undesirable multilayer formation of the photoactive compound could be avoided by reaction...... in water instead of phosphate-buffered saline. Passively adsorbed IL-4 was observed to induce differentiation to dendritic cells, but analysis of cell culture supernatants revealed that leakage of IL-4 into solution could account for the differentiation observed. Covalent attachment resulted in bound IL-4...

  12. Null Fluids - A New Viewpoint of Galilean Fluids

    CERN Document Server

    Banerjee, Nabamita; Jain, Akash

    2015-01-01

    This article is a detailed version of our short letter `On equilibrium partition function for non-relativistic fluid' [arXiv:1505.05677] extended to include an anomalous $U(1)$ symmetry. We construct a relativistic system, which we call null fluid and show that it is in one-to-one correspondence with a Galilean fluid living in one lower dimension. The correspondence is based on light cone reduction, which is known to reduce the Poincare symmetry of a theory to Galilean in one lower dimension. We show that the proposed null fluid and the corresponding Galilean fluid have exactly same symmetries, thermodynamics, constitutive relations, and equilibrium partition to all orders in derivative expansion. We also devise a mechanism to introduce $U(1)$ anomaly in even dimensional Galilean theories using light cone reduction, and study its effect on the constitutive relations of a Galilean Fluid.

  13. Null fluids: A new viewpoint of Galilean fluids

    Science.gov (United States)

    Banerjee, Nabamita; Dutta, Suvankar; Jain, Akash

    2016-05-01

    In this article, we study a Galilean fluid with a conserved U (1 ) current up to anomalies. We construct a relativistic system, which we call a null fluid and show that it is in one-to-one correspondence with a Galilean fluid living in one lower dimension. The correspondence is based on light cone reduction, which is known to reduce the Poincaré symmetry of a theory to Galilean in one lower dimension. We show that the proposed null fluid and the corresponding Galilean fluid have exactly same symmetries, thermodynamics, constitutive relations, and equilibrium partition to all orders in the derivative expansion. We also devise a mechanism to introduce U (1 ) anomaly in even dimensional Galilean theories using light cone reduction, and study its effect on the constitutive relations of a Galilean fluid.

  14. Amniotic fluid embolism

    Directory of Open Access Journals (Sweden)

    Rudra A

    2009-01-01

    Full Text Available The disastrous entry of amniotic fluid into the maternal circulation leads to dramatic sequelae of clinical events, characteristically referred to as Amniotic fluid embolism (AFE. The underlying mechanism for AFE is still poorly understood. Unfortunately, this situation has very grave maternal and fetal consequences. AFE can occur during labor, caesarean section, dilatation and evacuation or in the immediate postpartum period. The pathophysiology is believed to be immune mediated which affects the respiratory, cardiovascular, neurological and hematological systems. Undetected and untreated it culminates into fulminant pulmonary edema, intractable convulsions, disseminated intravascular coagulation (DIC, malignant arrhythmias and cardiac arrest. Definite diagnosis can be confirmed by identification of lanugo, fetal hair and fetal squamous cells (squames in blood aspirated from the right ventricle. Usually the diagnosis is made clinically and by exclusion of other causes. The cornerstone of management is a multidisciplinary approach with supportive treatment of failing organs systems. Despite improved modalities for diagnosing AFE, and better intensive care support facilities, the mortality is still high.

  15. Urine culture - catheterized specimen

    Science.gov (United States)

    Culture - urine - catheterized specimen; Urine culture - catheterization; Catheterized urine specimen culture ... urinary tract infections may be found in the culture. This is called a contaminant. You may not ...

  16. Suction blister fluid as potential body fluid for biomarker proteins.

    Science.gov (United States)

    Kool, Jeroen; Reubsaet, Léon; Wesseldijk, Feikje; Maravilha, Raquel T; Pinkse, Martijn W; D'Santos, Clive S; van Hilten, Jacobus J; Zijlstra, Freek J; Heck, Albert J R

    2007-10-01

    Early diagnosis is important for effective disease management. Measurement of biomarkers present at the local level of the skin could be advantageous in facilitating the diagnostic process. The analysis of the proteome of suction blister fluid, representative for the interstitial fluid of the skin, is therefore a desirable first step in the search for potential biomarkers involved in biological pathways of particular diseases. Here, we describe a global analysis of the suction blister fluid proteome as potential body fluid for biomarker proteins. The suction blister fluid proteome was compared with a serum proteome analyzed using identical protocols. By using stringent criteria allowing less than 1% false positive identifications, we were able to detect, using identical experimental conditions and amount of starting material, 401 proteins in suction blister fluid and 240 proteins in serum. As a major result of our analysis we construct a prejudiced list of 34 proteins, relatively highly and uniquely detected in suction blister fluid as compared to serum, with established and putative characteristics as biomarkers. We conclude that suction blister fluid might potentially serve as a good alternative biomarker body fluid for diseases that involve the skin.

  17. Development of myiasis vaccine: In vitro detection of immunoprotective responses of peritrophic membrane protein, first instar larva Ll supernatant and pellet antigen of fly Chrysomyia bezziana in sheep

    Directory of Open Access Journals (Sweden)

    Sukarsih

    1999-10-01

    Full Text Available Myiasis control by means of individual treatment of animals which are mainly rised extensively is time consumed and expensive. The alternative way to control this disease by vaccination is considered effective and economically accepted. However the expected vaccine is now still being developed under a collaborative project between CSIRO, Inter-University Centre on Biotechnology-ITB and Research Institute for Veterinary Science and funded by ACIAR. There are several antigens have been identified as vaccine candidates and an in vitro bioassay technique has been developed for assessing the immunoresponses of vaccine in sheep. Three antigens were used for vaccines in this study, these included protein peritrophic membrane (PM, soluble extract (SE and pellet extract (PE of 1st instar larvae of Chrysomya bezziana. Twenty four experimental sheep were divided into 4 groups of 6 animals, 3 groups of animals were injected with PM, SE and PE vaccines with the dose rate of 0.5 g PM/head, 0.8 g PE/head and 4.2 ml LE/head respectively, and the other one group was injected with 4 ml PBS/head as a control group. Vaccination with the same dose was repeated 4 weeks after the 1st vaccination as a booster, and 2 weeks after the booster the sheep were challenged with live larvae, 3 days after challenge animals were killed. Sera were collected at the day of vaccination, 4 weeks after vaccination, 2 weeks after booster, and 3 days after challenge. An in vitro bioassay technique was conducted by culturing 1st instar larvae on five media containing sera collected from each experimental animal. The effects of sera on cultivated larvae were assessed by means of larval weight and larval mortality rate. The results indicated that the growth rate and survival of cultivated larvae in media containing anti-PM sera were significantly lower (P<0.01 compared to the larvae cultivated on media with sera on the day of vaccination. The larval weight depression by anti- PM sera

  18. Cultural tourism and tourism cultures

    DEFF Research Database (Denmark)

    Ooi, Can-Seng

    Presenting a comprehensive and dynamic understanding of cultural tourism, this volume examines cultural mediators and how they help tourists appreciate foreign cultures. It also shows how tourism experiences are strategically crafted by mediators, the complexity of the mediation process, and how...... various products are mediated differently. A number of different products are investigated, including destination brand identities, "living" cultures and everyday life, art and history. The author illustrates his arguments by comparing the tourism strategies of Copenhagen and Singapore, and demonstrates...... how tourism is an agent for social change. The author also offers an original and refreshing way of understanding tourist behaviour through the concept of the "versatile tourist". The book's empirical cases and dialogic framework provide new and deep insights into tourism activities. In his...

  19. Performance Testing of Cutting Fluids

    DEFF Research Database (Denmark)

    Belluco, Walter

    within the whole range of operations, materials, cutting fluids, operating conditions, etc. Cutting fluid performance was evaluated in turning, drilling, reaming and tapping, and with respect to tool life, cutting forces, chip formation and product quality (dimensional accuracy and surface integrity......). A number of different work materials were considered, with emphasis on austenitic stainless steel. Cutting fluids from two main groups were investigated, water miscible (reviewed from previous work) and straight oils. Results show that correlation of cutting fluid performance in different operations exists...... within the same group of cutting fluids, for stainless steel. A possible rationalisation of cutting fluid performance tests is suggested. In order to select a set of basic tests and optimise them for use as general and standardised testing methods, an original approach to the evaluation of cutting force...

  20. Fluid mechanics fundamentals and applications

    CERN Document Server

    Cengel, Yunus

    2013-01-01

    Cengel and Cimbala's Fluid Mechanics Fundamentals and Applications, communicates directly with tomorrow's engineers in a simple yet precise manner. The text covers the basic principles and equations of fluid mechanics in the context of numerous and diverse real-world engineering examples. The text helps students develop an intuitive understanding of fluid mechanics by emphasizing the physics, using figures, numerous photographs and visual aids to reinforce the physics. The highly visual approach enhances the learning of Fluid mechanics by students. This text distinguishes itself from others by the way the material is presented - in a progressive order from simple to more difficult, building each chapter upon foundations laid down in previous chapters. In this way, even the traditionally challenging aspects of fluid mechanics can be learned effectively. McGraw-Hill is also proud to offer ConnectPlus powered by Maple with the third edition of Cengel/Cimbabla, Fluid Mechanics. This innovative and powerful new sy...