27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

27 CFR 21.94 - Acetaldol.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 5 ml of this solution to a 250 ml glass-stoppered flask containing 25 ml distilled water. Add 25 ml of a freshly prepared 1 percent sodium bisulfite solution. Prepare a blank omitting the acetaldol solution. Place the flasks in a dark place away from excessive heat or cold and allow to stand six hours...

[Studies for analyzing the prohibited ingredients such as acetohexamide in cosmetics].

PubMed

Tokunaga, Hiroshi; Uchino, Tadashi

2005-01-01

Acetohexamide (AH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for AH was investigated by HPLC. The lotion or milky lotion of 0.5g was put into a 10-ml volumetric flask. After adding 1.0ml of AH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol as the testing solution. Creams were procedured as follows; 0.5 g of cream was put into a 10-ml volumetric flask. After adding 1.0ml of tetrahydrofuran into the volumetric flask, the mixture was stirred for several minutes and the ingredients of the creams were dissolved. After adding 1.0ml of AH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol. One milliliter of the mixture including AH at 5 microg/ml was exactly put into a test tube with a cap and then 1 ml of water and 1 ml of hexane were added. After shaking vigorously, stand for several minutes. After centrifuging, the hexane layer was eliminated and the residual mixture was used as the test solution. The testing solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250mm), the mixture of acetonitrile and 50 mmol/l phosphate buffer(pH 5.3)(3:1) and the detection wavelength of 247 nm. The working curve from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of AH and the peak areas. There was no interference of peak of AH with the ingredients such as methylparaben, ethylparaben in the lotions, milky lotion and creams.

[Studies for analyzing the prohibited ingredients such as cyproheptadine hydrochloride in cosmetics].

PubMed

Tokunaga, Hiroshi; Uchino, Tadashi

2005-01-01

Cyproheptadine hydrochloride (CH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for CH was investigated by HPLC. The lotion or milky lotion of 0.5 g was put into a 10-ml volumetric flask. After adding 1.0ml of CH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol as the test solution. Creams were procedured as follows; 0.5 g of cream was put into a 10-ml volumetric flask. After adding 1.0 ml of tetrahydrofuran into the volumetric flask, the mixture was stirred for several minutes and the ingredients of the creams were dissolved. After adding 1.0 ml of CH solution at 50 microg/ml into the volumetric flask, the mixture was made up to 10ml with methanol. This mixture was transferred to a centrifuging tube with a cap and then the tube was centrifuged for 5 minutes at 3000 rpm. The supernatant was used as the test solution. The test solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of 1% acetic acid with 10 mmol/l sodium octanesulfonate and acetonitrile (11:9) and the detection wavelength of 286 nm. The working curve from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of CH and the peak areas. There was no interference of peak of CH with the ingredients such as methylparaben, ethylparaben in the lotions, milky lotion and creams.

Cultivation of recombinant Chinese hamster ovary cells grown as suspended aggregates in stirred vessels.

PubMed

Han, Yi; Liu, Xing-Mao; Liu, Hong; Li, Shi-Chong; Wu, Ben-Chuan; Ye, Ling-Ling; Wang, Qu-Wei; Chen, Zhao-Lie

2006-11-01

Recombinant Chinese hamster ovary (rCHO) cells capable of producing a prourokinase mutant (mPro-uk) grown as suspended aggregates in stirred vessels were described and characterized. The addition of chitosan to a mixture of DMEM and Ham's F12 (D-MEM/F-12) medium promoted cell aggregation and spheroid formation efficiently. Multicellular aggregates formed immediately after the rCHO cells were inoculated into the chitosan-added medium, and the mean diameter of the cell aggregates reflecting the aggregate size increased with culture time, shifting from 65 to 163 mum after 2 and 9 d of culture in spinner flasks. No significant difference in the metabolism performance of the rCHO cells was observed between suspended aggregates and anchored monolayers. However, the cells cultured as suspended aggregates showed a marked decrease in growth rate as evaluated from specific growth rate (mu). Replacing D-MEM/F-12 medium with CD 293 medium caused compact spherical cell aggregates to dissociate into small irregular aggregates and single cells without apparent effects on cell performance in subcultures. The perfusion culture of the rCHO cells grown as suspended aggregates in a 2-l stirred tank bioreactor for 15 d resulted in a maximum viable cell density of 5.6 x 10(6) cells ml(-1) and an mPro-uk concentration of about 2.6 x 10(3) IU ml(-1), and cell viability was remained at roughly 90% during the entire run.

[Studies for analyzing restricted ingredients such as phenylbenzoimidazole sulfonic acid].

PubMed

Tokunaga, Hiroshi; Mori, Kenichiro; Onuki, Nahomi; Nosaka, Tomio; Doi, Kayo; Sakaguchi, Hiroshi; Fujii, Makiko; Takano, Katuhiro; Hayashi, Masato; Yoshizawa, Kenichi; Shimamura, Kimio; Sato, Nobuo

2006-01-01

Phenylbenzoimidazol sulfonic acid (PBS) is a kind of sunscreens in cosmetics and is nominated as the restricted ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for PBS was investigated by HPLC. 1.0 g of the lotions with 1.0% PBS was exactly weighed, put into a 50-mL volumetric flask. Water was added to make exactly 50 mL and this mixture was used as the sample solution. On the other hand, 1.0 g of the creams with 1.0% PBS was exactly weighed, put into a beaker. After adding 1 mL of tetrahydrofuran and dissolving the cream, that mixture was transferred to a 50-mL volumetric flask. And then the beaker was rinsed with 1 mL of tetrahydrofuran and the rinsed solution was put together into the volumetric flask. After adding water to the volumetric flask to make exactly 50 mL, this mixture was used as the sample solution. If necessary, the mixture was filtrated with a membrane filter (0.45 microm). 5.0 mL of the sample solution was pipetted and put into a 200-mL volumetric flask. After adding water to make exactly 200 mL, 20 microL of this solution was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 mm i.d. x 250 mm), the mixture of 40 mmol/L acetic buffer (pH 3.4) and acetonitrile (3:1) with 0.8 mmol/L dodecyltrimethyl ammonium bromide and the detection wavelength of 305 nm. The working curve from 0.5 to 20.0 microg/mL showed a linear line between the concentrations of PBS and the peak areas. There was no interference of peak of PBS from the lotion and cream.

Comparison of a 50 mL pycnometer and a 500 mL flask, EURAMET.M.FF.S8 (EURAMET 1297)

NASA Astrophysics Data System (ADS)

Mićić, Ljiljana; Batista, Elsa

2018-01-01

The purpose of this comparison was to compare the results of the participating laboratories in the calibration of 50 mL pycnometer and 500 mL volumetric flask using the gravimetric method. Laboratories were asked to determined the 'contained' volume of the 50 mL pycnometer and of the 500 mL flask at a reference temperature of 20 °C. The gravimetric method was used for both instruments by all laboratories. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCM, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells.

PubMed

de Soure, António M; Fernandes-Platzgummer, Ana; Moreira, Francisco; Lilaia, Carla; Liu, Shi-Hwei; Ku, Chen-Peng; Huang, Yi-Feng; Milligan, William; Cabral, Joaquim M S; da Silva, Cláudia L

2017-05-01

Umbilical cord matrix (UCM)-derived mesenchymal stem/stromal cells (MSCs) are promising therapeutic candidates for regenerative medicine settings. UCM MSCs have advantages over adult cells as these can be obtained through a non-invasive harvesting procedure and display a higher proliferative capacity. However, the high cell doses required in the clinical setting make large-scale manufacturing of UCM MSCs mandatory. A commercially available human platelet lysate-based culture supplement (UltraGRO TM , AventaCell BioMedical) (5%(v/v)) was tested to effectively isolate UCM MSCs and to expand these cells under (1) static conditions, using planar culture systems and (2) stirred culture using plastic microcarriers in a spinner flask. The MSC-like cells were isolated from UCM explant cultures after 11 ± 2 days. After five passages in static culture, UCM MSCs retained their immunophenotype and multilineage differentiation potential. The UCM MSCs cultured under static conditions using UltraGRO TM -supplemented medium expanded more rapidly compared with UCM MSCs expanded using a previously established protocol. Importantly, UCM MSCs were successfully expanded under dynamic conditions on plastic microcarriers using UltraGRO TM -supplemented medium in spinner flasks. Upon an initial 54% cell adhesion to the beads, UCM MSCs expanded by >13-fold after 5-6 days, maintaining their immunophenotype and multilineage differentiation ability. The present paper reports the establishment of an easily scalable integrated culture platform based on a human platelet lysate supplement for the effective isolation and expansion of UCM MSCs in a xenogeneic-free microcarrier-based system. This platform represents an important advance in obtaining safer and clinically meaningful MSC numbers for clinical translation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Effect of convection on osteoblastic cell growth and function in biodegradable polymer foam scaffolds

NASA Technical Reports Server (NTRS)

Goldstein, A. S.; Juarez, T. M.; Helmke, C. D.; Gustin, M. C.; Mikos, A. G.; McIntire, L. V. (Principal Investigator)

2001-01-01

Culture of seeded osteoblastic cells in three-dimensional osteoconductive scaffolds in vitro is a promising approach to produce an osteoinductive material for repair of bone defects. However, culture of cells in scaffolds sufficiently large to bridge critical-sized defects is a challenge for tissue engineers. Diffusion may not be sufficient to supply nutrients into large scaffolds and consequently cells may grow preferentially at the periphery under static culture conditions. Three alternative culturing schemes that convect media were considered: a spinner flask, a rotary vessel, and a perfusion flow system. Poly(DL-lactic-co-glycolic acid) (PLGA) foam discs (12.7 mm diameter, 6.0 mm thick, 78.8% porous) were seeded with osteoblastic marrow stromal cells and cultured in the presence of dexamethasone and L-ascorbic acid for 7 and 14 days. Cell numbers per foam were found to be similar with all culturing schemes indicating that cell growth could not be enhanced by convection, but histological analysis indicated that the rotary vessel and flow system produced a more uniform distribution of cells throughout the foams. Alkaline phosphatase (ALP) activity per cell was higher with culture in the flow system and spinner flask after 7 days, while no differences in osteocalcin (OC) activity per cell were observed among culturing methods after 14 days in culture. Based on the higher ALP activity and better cell uniformity throughout the cultured foams, the flow system appears to be the superior culturing method, although equally important is the fact that in none of the tests did any of the alternative culturing techniques underperform the static controls. Thus, this study demonstrates that culturing techniques that utilize fluid flow, and in particular the flow perfusion system, improve the properties of the seeded cells over those maintained in static culture.

Design of a flow perfusion bioreactor system for bone tissue-engineering applications.

PubMed

Bancroft, Gregory N; Sikavitsas, Vassilios I; Mikos, Antonios G

2003-06-01

Several different bioreactors have been investigated for tissue-engineering applications. Among these bioreactors are the spinner flask and the rotating wall vessel reactor. In addition, a new type of culture system has been developed and investigated, the flow perfusion culture bioreactor. Flow perfusion culture offers several advantages, notably the ability to mitigate both external and internal diffusional limitations as well as to apply mechanical stress to the cultured cells. For such investigation, a flow perfusion culture system was designed and built. This design is the outgrowth of important design requirements and incorporates features crucial to successful experimentation with such a system.

Laboratory and Field Evaluation of a Waterless Food Service Sanitation System Used by Military Mobile Kitchen Trailer Crews

DTIC Science & Technology

1993-12-01

Stopper the flask and shake vigorously. The mixture was titrated with 0.003 N sodium lauryl sulfate drcpwise. The endpoint was the first definite...100 g sodium sulfate and 1000 mL distilled water, pill0) and three drops of 0.1% bracu 1henol blue indicator to 50 ML of sample in a 250 mL flask...Product QDS (Syncide Plus) (1) was determined by a bromcphenol blue nethod (7). Add 25 mL of chloroform, 25 mL salt buffer solution (7 g sodium carbonate

Simplified Method of the Growth of Human Tumor Infiltrating Lymphocytes (TIL) in Gas-Permeable Flasks to Numbers Needed for Patient Treatment

PubMed Central

Jin, Jianjian; Sabatino, Marianna; Somerville, Robert; Wilson, John R.; Dudley, Mark E.; Stroncek, David F.; Rosenberg, Steven A.

2012-01-01

Adoptive cell therapy (ACT) of metastatic melanoma with autologous tumor infiltrating lymphocytes (TIL) is clinically effective, but TIL production can be challenging. Here we describe a simplified method for initial TIL culture and rapid expansion in gas-permeable flasks. TIL were initially cultured from tumor digests and fragments in 40 mL capacity flasks with a 10 cm2 gas-permeable silicone bottom, G-Rex10. A TIL rapid expansion protocol (REP) was developed using 500 mL capacity flasks with a 100 cm2 gas-permeable silicone bottom, G-Rex100. TIL growth was successfully initiated in G-Rex10 flasks from tumor digests from 13 of 14 patients and from tumor fragments in all 11 tumor samples tested. TIL could then be expanded to 8–10×109 cells in a two-step REP which began by seeding 5 × 106 TIL into a G-Rex100 flask, followed by expansion at day 7 into 3 G-Rex100 flasks. To obtain the 30 to 60 × 109 cells used for patient treatment we seeded 6 G-Rex100 flasks with 5×106 cells and expanded into 18 G-Rex100 flasks. Large scale TIL REP in gas-permeable flasks requires approximately 9 to 10 liters of media, about 3 to 4 times less than other methods. In conclusion, TIL initiation and REP in gas-permeable G-Rex flasks require fewer total vessels, less media, less incubator space and less labor than initiation and REP in 24-well plates, tissue culture flasks and bags. TIL culture in G-Rex flasks will facilitate the production of TIL at the numbers required for patient treatment at most cell processing laboratories. PMID:22421946

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

PubMed Central

2011-01-01

Background Influenza virus is a major health concern that has huge impacts on the human society, and vaccination remains as one of the most effective ways to mitigate this disease. Comparing the two types of commercially available Influenza vaccine, the live attenuated virus vaccine is more cross-reactive and easier to administer than the traditional inactivated vaccines. One promising live attenuated Influenza vaccine that has completed Phase I clinical trial is deltaFLU, a deletion mutant lacking the viral Nonstructural Protein 1 (NS1) gene. As a consequence of this gene deletion, this mutant virus can only propagate effectively in cells with a deficient interferon-mediated antiviral response. To demonstrate the manufacturability of this vaccine candidate, a batch bioreactor production process using adherent Vero cells on microcarriers in commercially available animal-component free, serum-free media is described. Results Five commercially available animal-component free, serum-free media (SFM) were evaluated for growth of Vero cells in agitated Cytodex 1 spinner flask microcarrier cultures. EX-CELL Vero SFM achieved the highest cell concentration of 2.6 × 10^6 cells/ml, whereas other SFM achieved about 1.2 × 10^6 cells/ml. Time points for infection between the late exponential and stationary phases of cell growth had no significant effect in the final virus titres. A virus yield of 7.6 Log10 TCID50/ml was achieved using trypsin concentration of 10 μg/ml and MOI of 0.001. The Influenza vaccine production process was scaled up to a 3 liter controlled stirred tank bioreactor to achieve a cell density of 2.7 × 10^6 cells/ml and virus titre of 8.3 Log10 TCID50/ml. Finally, the bioreactor system was tested for the production of the corresponding wild type H1N1 Influenza virus, which is conventionally used in the production of inactivated vaccine. High virus titres of up to 10 Log10 TCID50/ml were achieved. Conclusions We describe for the first time the production of Influenza viruses using Vero cells in commercially available animal-component free, serum-free medium. This work can be used as a basis for efficient production of attenuated as well as wild type Influenza virus for research and vaccine production. PMID:21835017

Simulated conditions of microgravity suppress progesterone production by luteal cells of the pregnant rat

NASA Technical Reports Server (NTRS)

Bhat, G. K.; Yang, H.; Sridaran, R.

2001-01-01

The purpose of this study was to assess whether simulated conditions of microgravity induce changes in the production of progesterone by luteal cells of the pregnant rat ovary using an in vitro model system. The microgravity environment was simulated using either a high aspect ratio vessel (HARV) bioreactor with free fall or a clinostat without free fall of cells. A mixed population of luteal cells isolated from the corpora lutea of day 8 pregnant rats was attached to cytodex microcarrier beads (cytodex 3). These anchorage dependent cells were placed in equal numbers in the HARV or a spinner flask control vessel in culture conditions. It was found that HARV significantly reduced the daily production of progesterone from day 1 through day 8 compared to controls. Scanning electron microscopy showed that cells attached to the microcarrier beads throughout the duration of the experiment in both types of culture vessels. Cells cultured in chamber slide flasks and placed in a clinostat yielded similar results when compared to those in the HARV. Also, when they were stained by Oil Red-O for lipid droplets, the clinostat flasks showed a larger number of stained cells compared to control flasks at 48 h. Further, the relative amount of Oil Red-O staining per milligram of protein was found to be higher in the clinostat than in the control cells at 48 h. It is speculated that the increase in the level of lipid content in cells subjected to simulated conditions of microgravity may be due to a disruption in cholesterol transport and/or lesions in the steroidogenic pathway leading to a fall in the synthesis of progesterone. Additionally, the fall in progesterone in simulated conditions of microgravity could be due to apoptosis of luteal cells.

Efficient expansion of mesenchymal stromal cells in a disposable fixed bed culture system.

PubMed

Mizukami, Amanda; Orellana, Maristela D; Caruso, Sâmia R; de Lima Prata, Karen; Covas, Dimas T; Swiech, Kamilla

2013-01-01

The need for efficient and reliable technologies for clinical-scale expansion of mesenchymal stromal cells (MSC) has led to the use of disposable bioreactors and culture systems. Here, we evaluate the expansion of cord blood-derived MSC in a disposable fixed bed culture system. Starting from an initial cell density of 6.0 × 10(7) cells, after 7 days of culture, it was possible to produce of 4.2(±0.8) × 10(8) cells, which represents a fold increase of 7.0 (±1.4). After enzymatic retrieval from Fibra-Cell disks, the cells were able to maintain their potential for differentiation into adipocytes and osteocytes and were positive for many markers common to MSC (CD73, CD90, and CD105). The results obtained in this study demonstrate that MSC can be efficiently expanded in the culture system. This novel approach presents several advantages over the current expansion systems, based on culture flasks or microcarrier-based spinner flasks and represents a key element for MSC cellular therapy according to GMP compliant clinical-scale production system. Copyright © 2013 American Institute of Chemical Engineers.

Influence of the mechanical environment on the engineering of mineralised tissues using human dental pulp stem cells and silk fibroin scaffolds.

PubMed

Woloszyk, Anna; Holsten Dircksen, Sabrina; Bostanci, Nagihan; Müller, Ralph; Hofmann, Sandra; Mitsiadis, Thimios A

2014-01-01

Teeth constitute a promising source of stem cells that can be used for tissue engineering and regenerative medicine purposes. Bone loss in the craniofacial complex due to pathological conditions and severe injuries could be treated with new materials combined with human dental pulp stem cells (hDPSCs) that have the same embryonic origin as craniofacial bones. Optimising combinations of scaffolds, cells, growth factors and culture conditions still remains a great challenge. In the present study, we evaluate the mineralisation potential of hDPSCs seeded on porous silk fibroin scaffolds in a mechanically dynamic environment provided by spinner flask bioreactors. Cell-seeded scaffolds were cultured in either standard or osteogenic media in both static and dynamic conditions for 47 days. Histological analysis and micro-computed tomography of the samples showed low levels of mineralisation when samples were cultured in static conditions (0.16±0.1 BV/TV%), while their culture in a dynamic environment with osteogenic medium and weekly µCT scans (4.9±1.6 BV/TV%) significantly increased the formation of homogeneously mineralised structures, which was also confirmed by the elevated calcium levels (4.5±1.0 vs. 8.8±1.7 mg/mL). Molecular analysis of the samples showed that the expression of tooth correlated genes such as Dentin Sialophosphoprotein and Nestin were downregulated by a factor of 6.7 and 7.4, respectively, in hDPSCs when cultured in presence of osteogenic medium. This finding indicates that hDPSCs are able to adopt a non-dental identity by changing the culture conditions only. Also an increased expression of Osteocalcin (1.4x) and Collagen type I (1.7x) was found after culture under mechanically dynamic conditions in control medium. In conclusion, the combination of hDPSCs and silk scaffolds cultured under mechanical loading in spinner flask bioreactors could offer a novel and promising approach for bone tissue engineering where appropriate and rapid bone regeneration in mechanically loaded tissues is required.

Evaluation of an Immobilized Cell Bioreactor for Degradation of Meta- and Para-Nitrobenzoate

DTIC Science & Technology

1994-01-18

AFB IWTP. 4 Shake flask tests and continuous flow, bench-scale bioreactor tests were conducted using EDA or spent CLEPO 204 as the substrate. It was...found that the shake flask cultures completely degraded EDA when it was the sole substrate. However, using spent CLEPO 204 as the substrate caused a...microorganisms isolated, Kelly 4. Erlenmeyer flasks (250 mL) were used in studies to determine the maximal growth rate of Kelly 4 at 30" C in SMSB

Dissolved oxygen levels affect dimorphic growth by the entomopathogenic fungus Isaria fumosorosea

USDA-ARS?s Scientific Manuscript database

The entomopathogenic fungus Isaria fumosorosea is capable of dimorphic growth (hyphal or yeast-like) in submerged culture. In shake flask studies, we evaluated the impact of aeration on the mode of growth of I. fumosorosea. Using 250 mL baffled Erlenmeyer flasks, culture volumes of 50, 100, 150, a...

Algal Cell Response to Pulsed Waved Stimulation and Its Application to Increase Algal Lipid Production

NASA Astrophysics Data System (ADS)

Savchenko, Oleksandra; Xing, Jida; Yang, Xiaoyan; Gu, Quanrong; Shaheen, Mohamed; Huang, Min; Yu, Xiaojian; Burrell, Robert; Patra, Prabir; Chen, Jie

2017-02-01

Generating renewable energy while sequestering CO2 using algae has recently attracted significant research attention, mostly directing towards biological methods such as systems biology, genetic engineering and bio-refining for optimizing algae strains. Other approaches focus on chemical screening to adjust culture conditions or culture media. We report for the first time the physiological changes of algal cells in response to a novel form of mechanical stimulation, or a pulsed wave at the frequency of 1.5 MHz and the duty cycle of 20%. We studied how the pulsed wave can further increase algal lipid production on top of existing biological and chemical methods. Two commonly used algal strains, fresh-water Chlorella vulgaris and seawater Tetraselmis chuii, were selected. We have performed the tests in shake flasks and 1 L spinner-flask bioreactors. Conventional Gravimetric measurements show that up to 20% increase for algal lipid could be achieved after 8 days of stimulation. The total electricity cost needed for the stimulations in a one-liter bioreactor is only one-tenth of a US penny. Gas liquid chromatography shows that the fatty acid composition remains unchanged after pulsed-wave stimulation. Scanning electron microscope results also suggest that pulsed wave stimulation induces shear stress and thus increases algal lipid production.

Clinical-Grade Manufacturing of Therapeutic Human Mesenchymal Stem/Stromal Cells in Microcarrier-Based Culture Systems.

PubMed

Fernandes-Platzgummer, Ana; Carmelo, Joana G; da Silva, Cláudia Lobato; Cabral, Joaquim M S

2016-01-01

The therapeutic potential of mesenchymal stem/stromal cells (MSC) has triggered the need for high cell doses in a vast number of clinical applications. This demand requires the development of good manufacturing practices (GMP)-compliant ex vivo expansion protocols that should be effective to deliver a robust and reproducible supply of clinical-grade cells in a safe and cost-effective manner. Controlled stirred-tank bioreactor systems under xenogeneic (xeno)-free culture conditions offer ideal settings to develop and optimize cell manufacturing to meet the standards and needs of human MSC for cellular therapies. Herein we describe two microcarrier-based stirred culture systems using spinner flasks and controlled stirred-tank bioreactors under xeno-free conditions for the efficient ex vivo expansion of human bone marrow and adipose tissue-derived MSC.

Formation, Structure and Properties of Boron Nitride Fibers from Polymer Precursors

DTIC Science & Technology

1992-02-25

conversion, have been demonstrated in this study. Poly(ethylene oxide) (PEO), poly(N-vinyl-2-pyrrolidinone) ( PVP ), and poly(methyl methacrylate) (PMMA) were...vinylpyrrolidinone ( PVP , Mw = 360,000). A 100 ml reaction flask with teflon boiling chips and a 100 ml receiving flask were used on a high boiling distilling...CH 3 OBO) 3 : B(OH)3 Predicted 2 Experiment Experiment, 1 wt% PVP 2:1 47.8 mole% CH 3 OH 39.6 mole% CH 3OH 35.2 mole% CH3 OH 3:1 47.8 - 35.5 4:1 47.8

Stirred tank bioreactor culture combined with serum-/xenogeneic-free culture medium enables an efficient expansion of umbilical cord-derived mesenchymal stem/stromal cells.

PubMed

Mizukami, Amanda; Fernandes-Platzgummer, Ana; Carmelo, Joana G; Swiech, Kamilla; Covas, Dimas T; Cabral, Joaquim M S; da Silva, Cláudia L

2016-08-01

Mesenchymal stem/stromal cells (MSC) are being widely explored as promising candidates for cell-based therapies. Among the different human MSC origins exploited, umbilical cord represents an attractive and readily available source of MSC that involves a non-invasive collection procedure. In order to achieve relevant cell numbers of human MSC for clinical applications, it is crucial to develop scalable culture systems that allow bioprocess control and monitoring, combined with the use of serum/xenogeneic (xeno)-free culture media. In the present study, we firstly established a spinner flask culture system combining gelatin-based Cultispher(®) S microcarriers and xeno-free culture medium for the expansion of umbilical cord matrix (UCM)-derived MSC. This system enabled the production of 2.4 (±1.1) x10(5) cells/mL (n = 4) after 5 days of culture, corresponding to a 5.3 (±1.6)-fold increase in cell number. The established protocol was then implemented in a stirred-tank bioreactor (800 mL working volume) (n = 3) yielding 115 million cells after 4 days. Upon expansion under stirred conditions, cells retained their differentiation ability and immunomodulatory potential. The development of a scalable microcarrier-based stirred culture system, using xeno-free culture medium that suits the intrinsic features of UCM-derived MSC represents an important step towards a GMP compliant large-scale production platform for these promising cell therapy candidates. Copyright © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Bioreactor Cultivation of Anatomically Shaped Human Bone Grafts

PubMed Central

Temple, Joshua P.; Yeager, Keith; Bhumiratana, Sarindr; Vunjak-Novakovic, Gordana; Grayson, Warren L.

2015-01-01

In this chapter, we describe a method for engineering bone grafts in vitro with the specific geometry of the temporomandibular joint (TMJ) condyle. The anatomical geometry of the bone grafts was segmented from computed tomography (CT) scans, converted to G-code, and used to machine decellularized trabecular bone scaffolds into the identical shape of the condyle. These scaffolds were seeded with human bone marrow-derived mesenchymal stem cells (MSCs) using spinner flasks and cultivated for up to 5 weeks in vitro using a custom-designed perfusion bioreactor system. The flow patterns through the complex geometry were modeled using the FloWorks module of SolidWorks to optimize bioreactor design. The perfused scaffolds exhibited significantly higher cellular content, better matrix production, and increased bone mineral deposition relative to non-perfused (static) controls after 5 weeks of in vitro cultivation. This technology is broadly applicable for creating patient-specific bone grafts of varying shapes and sizes. PMID:24014312

The Fluid Mechanics of a Wavy-Wall Bioreactor

NASA Astrophysics Data System (ADS)

Sucosky, Philippe; Bilgen, Bahar; Aleem, Alexander; Neitzel, Paul; Barabino, Gilda

2004-11-01

Bioreactors are devices used for the production of mammalian tissue in vitro. Although mixing has been shown to stimulate the growth of cartilage constructs, high shear-stress levels can damage the cells. In order to enhance mixing while minimizing shear, a wavy-wall bioreactor (WWB) featuring a sinusoidal internal profile has been designed. The turbulent hydrodynamic environment produced in this device is investigated experimentally using particle-image velocimetry. A model bioreactor made of acrylic and filled with an index-matching solution of zinc iodide is used to compensate for the refraction of light at the walls. The flow observed in different planes is shown to be periodic, spatially dependent, and dominated by mean-shear rather than Reynolds stresses in the vicinity of constructs. Finally, a comparison between the mean-shear stresses obtained in the WWB and in a standard spinner flask reveals similar stress levels near the construct walls.

Experimental study of icing accretion on a rotating conical spinner

NASA Astrophysics Data System (ADS)

Chen, Ningli; Ji, Honghu; Hu, Yaping; Wang, Jian; Cao, Guangzhou

2015-12-01

A reduced scale experiment has been conducted to investigate the icing accretion procedure on a rotating spinner of 60° cone angle. The experiment was carried out in a small scale ice wind tunnel with three different rotating speeds of the spinner. The experimental conditions were determined from the actual icing condition of the spinner of a turbofan engine by using the similarity theory, which considers the rotating effects. The ice thickness on the spinner was got from the image taken by the high speed camera, by image processing. The results of this investigation show that under the experimental condition, ice on the spinner's tip of three different rotating speeds are all glaze ice and about the same thick. However, on the downstream surface of the spinner, ice shape on the rotating spinner is different from that on the stationary spinner. It is uneven glaze ice on the stationary spinner while it is `particle ice' when the rotating speed is 8240 rpm and it is `needle ice' when the rotating speed is 15,200 rpm. The experiment also reveals that when the rotating speed is higher, the ice layer is thicker.

In vitro penetration of swine oocytes by homologous spermatozoa: Distinct systems for gamete's co-incubation and oocyte's cryopreservation.

PubMed

Macedo, M C; Lucia, T; Rambo, G; Ferreira Filho, E B; Rosa, A P; Fabiane, C; Cabral, M; Deschamps, J C

2010-02-01

In vitro penetration (IVP) of swine oocytes by homologous spermatozoa was evaluated in two experiments using four boars as semen donors. In experiment 1, the IVP rate and the number of penetrating spermatozoa (PSP) were compared using three co-incubation systems for vitrified oocytes and fresh sperm: (1) 35mL petri dishes in a CO(2) incubator, (2) 35mL petri dishes in bags (submarine system) and (3) glass flasks partially submerged in water bath with the same gas mixture used for the bag system. Mean PSP was 8.2+/-10.1 and the IVP rate was 90.5%. The PSP differed across all systems (P=0.0006): 15.5+/-0.5 for flasks, 6.3+/-0.4 for CO(2), and 3.9+/-0.4 for bags. The IVP rate for flasks (95.0%) was greater (P=0.01) than for CO(2) and bags (90.8% and 85.0%, respectively), but it did not differ between flasks and CO(2) for three boars (P>0.05). In experiment 2, co-incubation was done as described for glass flasks in experiment 1. The IVP rate and PSP were compared for cryopreserved oocytes: either vitrified in open pulled straws (OPS), or frozen in cryotubes. Mean PSP was 5.4+/-6.5 and IVP rate was 89.6%. Both PSP and IVP rate were greater (P<0.0001) for oocytes frozen in cryotubes (7.0+/-0.3% and 95.8%, respectively) than those frozen in OPS (3.7+/-0.3% and 83.4%, respectively), with no differences found for three boars (P>0.05). In summary, successful IVP of swine oocytes by homologous spermatozoa can be achieved using gametes incubated in glass flasks and oocytes frozen in cryotubes.

[Studies for analyzing prohibited ingredients such as estriol in cosmetics].

PubMed

Tokunaga, Hiroshi; Takeuchi, Orie; Uchino, Tadashi; Ando, Masanori

2004-01-01

Estriol (EO) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method using HPLC for EO was investigated. After placing 1.0 ml of EO solution at 50 microg/ml and 0.5 g of the lotion into a 10-ml volumetric flask, the methanol was added to make until that volume and this solution was used as the testing solution. Milky lotion was procedured as follows: After placing 1.0 ml of EO solution at 50 microg/ml and 0.5 g of the milky lotion into a 10-ml volumetric flask, the methanol was added to make until that volume. The suspending mixture was moved to a centrifuging tube with a cap. After centrifuging at 3000 rpm for 5 minutes, the supernatant was used as the testing solution. The testing solution of 20 microl was determined by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of water and acetonitrile (31:9) and the detection wavelength of 285 nm. The working curve from 1.0 to 6.0 microg/ml showed a linear line between the concentrations of EO and the peak area. There was no interference of peak of EO from the lotion and milky lotion.

Modeling Impact of BRCA1 and BRCA2 Mutations in Mammary Epithelial Cells

DTIC Science & Technology

2012-09-01

remaining tissue was lacerated and minced with opposing scalpels. Minced pieces were placed into a 50mL Erlenmeyer flask, containing 10mL digestion medium...Ham’s F- 12 containing insulin, penicillin, streptomycin, polymyxin B, Fungizone, 10% fetal bovine serum, collagenase, and hyaluronidase) for one

Active turbulence in a gas of self-assembled spinners

PubMed Central

Kokot, Gašper; Das, Shibananda; Winkler, Roland G.; Aranson, Igor S.; Snezhko, Alexey

2017-01-01

Colloidal particles subject to an external periodic forcing exhibit complex collective behavior and self-assembled patterns. A dispersion of magnetic microparticles confined at the air–liquid interface and energized by a uniform uniaxial alternating magnetic field exhibits dynamic arrays of self-assembled spinners rotating in either direction. Here, we report on experimental and simulation studies of active turbulence and transport in a gas of self-assembled spinners. We show that the spinners, emerging as a result of spontaneous symmetry breaking of clock/counterclockwise rotation of self-assembled particle chains, generate vigorous vortical flows at the interface. An ensemble of spinners exhibits chaotic dynamics due to self-generated advection flows. The same-chirality spinners (clockwise or counterclockwise) show a tendency to aggregate and form dynamic clusters. Emergent self-induced interface currents promote active diffusion that could be tuned by the parameters of the external excitation field. Furthermore, the erratic motion of spinners at the interface generates chaotic fluid flow reminiscent of 2D turbulence. Our work provides insight into fundamental aspects of collective transport in active spinner materials and yields rules for particle manipulation at the microscale. PMID:29158382

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2013 CFR

2013-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2012 CFR

2012-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

9 CFR 147.15 - Laboratory procedure recommended for the bacteriological examination of mycoplasma reactors. 12

Code of Federal Regulations, 2014 CFR

2014-01-01

... 10523. (5) Use sterile deionized distilled water to reconstitute penicillin. (6) Sterile serum should be... samples (ml)—5.0 (1:2000) Mycoplasma Broth Base (g)—22.5 Aqueous penicillin (units)—500,000 Sterile serum... Aqueous penicillin (units)—400,000 NAD (ml)—12.5 Cysteine hydrochloride (ml)—12.5 (1) Rotate flask gently...

Active turbulence in a gas of self-assembled spinners

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kokot, Gasper; Das, Shibananda; Winkler, Roland G.

Colloidal particles subject to an external periodic forcing exhibit complex collective behavior and self-assembled patterns. A dispersion of magnetic microparticles confined at the air-liquid interface and energized by a uniform uniaxial alternating magnetic field exhibits dynamic arrays of self-assembled spinners rotating in either direction. Here, we report on experimental and simulation studies of active turbulence and transport in a gas of self-assembled spinners. We show that the spinners, emerging as a result of spontaneous symmetry breaking of clock/counterclockwise rotation of self-assembled particle chains, generate vigorous vortical flows at the interface. An ensemble of spinners exhibits chaotic dynamics due to self-generatedmore » advection flows. The same-chirality spinners (clockwise or counterclock-wise) show a tendency to aggregate and form dynamic clusters. Emergent self-induced interface currents promote active diffusion that could be tuned by the parameters of the external excitation field. Furthermore, the erratic motion of spinners at the interface generates chaotic fluid flow reminiscent of 2D turbulence. As a result, our work provides insight into fundamental aspects of collective transport in active spinner materials and yields rules for particle manipulation at the microscale.« less

Active turbulence in a gas of self-assembled spinners

DOE PAGES

Kokot, Gasper; Das, Shibananda; Winkler, Roland G.; ...

2017-11-20

Colloidal particles subject to an external periodic forcing exhibit complex collective behavior and self-assembled patterns. A dispersion of magnetic microparticles confined at the air-liquid interface and energized by a uniform uniaxial alternating magnetic field exhibits dynamic arrays of self-assembled spinners rotating in either direction. Here, we report on experimental and simulation studies of active turbulence and transport in a gas of self-assembled spinners. We show that the spinners, emerging as a result of spontaneous symmetry breaking of clock/counterclockwise rotation of self-assembled particle chains, generate vigorous vortical flows at the interface. An ensemble of spinners exhibits chaotic dynamics due to self-generatedmore » advection flows. The same-chirality spinners (clockwise or counterclock-wise) show a tendency to aggregate and form dynamic clusters. Emergent self-induced interface currents promote active diffusion that could be tuned by the parameters of the external excitation field. Furthermore, the erratic motion of spinners at the interface generates chaotic fluid flow reminiscent of 2D turbulence. As a result, our work provides insight into fundamental aspects of collective transport in active spinner materials and yields rules for particle manipulation at the microscale.« less

[Cultivated keratinocytes on micro-carriers: in vitro studies of a new carrier system].

PubMed

Hecht, J; Hoefter, E A; Hecht, J; Haraida, S; Nerlich, A; Hartinger, A; Mühlbauer, W; Dimoudis, N

1997-03-01

Epidermal grafts from confluently cultivated keratinocytes have been used since the early eighties for the treatment of severe burns, where the shortage of donor sites for split-thickness skin grafts did not allow for adequate wound coverage. The difficult handling of these grafts as well as the advanced differentiation of their epithelial cells into a multilayer sheet poses a problem for their clinical application. The aim of the study was to characterize cultivated keratinocytes, as well as to observe their migration and proliferation from the MC onto a surface. Keratinocytes were isolated from human foreskin and cultivated in serum-free and serum-containing medium according to a modified method by Rheinwald and Green. Collagen-coated Dextran beads were used as MC. The MC were colonized with keratinocytes using the Spinner culture technique. After seeding the colonized MC into culture flasks, their migration and proliferation was monitored regularly through immunohistochemical studies and measurement of the metabolic cell activity. Immunohistological staining proved that the cells isolated from human foreskin represent keratinocytes of the basal type. Keratinocytes, cultivated with serum-containing and serum free medium, both adhered to the surface of the MC, then migrated onto the surface of the flasks and proliferated to form a multilayer of epithelial cells. In the long-term, a flexible epithelial graft consisting of poorly differentiated keratinocytes should be available, which is simple to produce and easy to handle. This would be an alternative method for treating wounds, where the conventional multilayer epithelial graft (ET) is insufficient.

Effects of Cultivating Conditions on the Water Soluble Polysaccharides Content of Ganoderma lucidum Mycelium in Submerged Flask Culture

NASA Astrophysics Data System (ADS)

Rosyida, V. T.; Hayati, S. N.; Apriyana, W.; Darsih, C.; Hernawan; Poeloengasih, C. D.

2017-12-01

The carcinostatic substance in Ganoderma lucidum (Fr.) Karst (Polyporaceae) is a water soluble polysaccharides (WSP) which might be useful in immunotherapy. Attempt to produce effective substances from cultured mycelia is important to carry out since solid cultivation is a time consuming and quality fluctuating. The effects of cultivating conditions on the water soluble polysaccharides content of G. Lucidum mycelium were investigated in submerged flask cultures. Culture from fruiting bodies was maintained on potato dextrose-agar slope. Slopes were inoculated and incubated at 30°C for 7 days, and stored at 4°C. The flask experiments were performed in 100 ml erlenmeyer flasks containing 20 ml of the sterilized media. Actively growing mycelia (1 piece, 5 mm X 5 mm) from a newly prepared slant culture (about 7 days incubation at 30°C) were inoculated into the flask. The pH was measured and adjusted to the desired value by addition of either 4 M HCl or 2.5 M NaOH. Incubation temperature were 20, 25, and 30°C. At the end of inoculation period (14 days) mycelium consisting of individual pellets was harvested and wash for the analysis. WSP content was analysed using phenol-sulfuric acid method. The optimal initial pH for metabolite production would depend on the culture medium. Generally, high values of pH, such as 9, negatively affect both cell growth and WSP production. The optimum temperature range for the high G. lucidum mycelium and WSP production were found to be 25 - 30 °C at pH values 5 - 7 in both of media.

Investigation of a Systematic Group of NACA 1-Series Cowlings with and Without Spinners

NASA Technical Reports Server (NTRS)

Nichols, Mark R; Keith, Arvid L , Jr

1949-01-01

Report presents the results of an investigation conducted in the Langley propeller research tunnel to study cowling-spinner combinations based on the NACA 1-series nose inlets and to obtain systematic design data for one family of approximately ellipsoidal spinners. In the main part of the investigation, 11 of the related spinners were tested in various combinations with 9 NACA open-nose cowlings, which were also tested without spinners. The effects of location and shape of the spinner, shape of the inner surface of the cowling lip, and operation of a propeller having approximately oval shanks were investigated briefly. In addition, a study was conducted to determine the correct procedure for extrapolating design conditions determined from the low-speed test data to the design conditions at the actual flight Mach number.

Can the Short-Term Toxicity of Water-Soluble Jet Fuel Hydrocarbons Produce Long-Lasting Effects in Lake Plankton Communities?

DTIC Science & Technology

1983-09-01

Richard Starr Algal collection or Ankistrodesmus sp. or Chlorella sp. from the Carolina Biological collection. The cultures were unialgal but not... Chlorella were grown in BBM medium. After five days ot growth, half ot the flasks (3) for each species were spiked with 0.1 ml of toluene. The , .. flasks...quickly recovered and grew at rates equa’ the = toluene-free cells, finally reaching the same final biomass. Ankistrodesmus and Chlorella however

Fidget spinners: Purported benefits, adverse effects and accepted alternatives.

PubMed

Schecter, Rachel A; Shah, Jay; Fruitman, Kate; Milanaik, Ruth Lynn

2017-10-01

In the span of a few months, fidget spinners have caught the eyes of millions of children, parents, educators and paediatricians. Fidget spinners, hand-held toys designed to spin freely in your grasp, have become a source of entertainment for consumers of all ages. Despite a lack of scientific evidence, toy marketers have advertised the benefits of fidget spinners for children with attention-deficit/hyperactivity disorder and other disorders (e.g. autism, anxiety, sensory issues). Parents are incentivized by these purported benefits to purchase fidget spinners to improve their child's concentration and decrease stress. While fidget spinners are a new phenomenon, existing therapy toys (e.g. sensory putty) have been used by occupational therapists for similar reasons, with comparably little research supporting these claims. The purpose of this review is to explore literature regarding sensory toys and examine educator/professional-reported concerns and medical adverse effects of using fidget spinners. Due to a recent surge in popularity, fidget spinners and other self-regulatory occupational therapy toys have yet to be subjected to rigorous scientific research. Thus, their alleged benefits remain scientifically unfounded. Paediatricians should be aware of potential choking hazards with this new fad, and inform parents that peer-reviewed studies do not support the beneficial claims.

Microscale screening systems for 3D cellular microenvironments: platforms, advances, and challenges

PubMed Central

Montanez-Sauri, Sara I.; Beebe, David J.; Sung, Kyung Eun

2015-01-01

The increasing interest in studying cells using more in vivo-like three-dimensional (3D) microenvironments has created a need for advanced 3D screening platforms with enhanced functionalities and increased throughput. 3D screening platforms that better mimic in vivo microenvironments with enhanced throughput would provide more in-depth understanding of the complexity and heterogeneity of microenvironments. The platforms would also better predict the toxicity and efficacy of potential drugs in physiologically relevant conditions. Traditional 3D culture models (e.g. spinner flasks, gyratory rotation devices, non-adhesive surfaces, polymers) were developed to create 3D multicellular structures. However, these traditional systems require large volumes of reagents and cells, and are not compatible with high throughput screening (HTS) systems. Microscale technology offers the miniaturization of 3D cultures and allows efficient screening of various conditions. This review will discuss the development, most influential works, and current advantages and challenges of microscale culture systems for screening cells in 3D microenvironments. PMID:25274061

Effects of different fermentation methods on bacterial cellulose and acid production by Gluconacetobacter xylinus in Cantonese-style rice vinegar.

PubMed

Fu, Liang; Chen, Siqian; Yi, Jiulong; Hou, Zongxia

2014-07-01

A strain of acidogenic bacterium was isolated from the fermentation liquid of Cantonese-style rice vinegar produced by traditional surface fermentation. 16S rDNA identification confirmed the bacterium as Gluconacetobacter xylinus, which synthesizes bacterial cellulose, and the acid productivity of the strain was investigated. In the study, the effects of the membrane integrity and the comparison of the air-liquid interface membrane with immerged membrane on total acidity, cellulose production, alcohol dehydrogenase (ADH) activity and number of bacteria were investigated. The cellulose membrane and the bacteria were observed under SEM for discussing their relationship. The correlations between oxygen consumption and total acid production rate were compared in surface and shake flask fermentation. The results showed the average acid productivity of the strain was 0.02g/(100mL/h), and the integrity of cellulose membrane in surface fermentation had an important effect on total acidity and cellulose production. With a higher membrane integrity, the total acidity after 144 h of fermentation was 3.75 g/100 mL, and the cellulose production was 1.71 g/100 mL after 360 h of fermentation. However, when the membrane was crushed by mechanical force, the total acidity and the cellulose production were as low as 0.36 g/100 mL and 0.14 g/100 mL, respectively. When the cellulose membrane was forced under the surface of fermentation liquid, the total acid production rate was extremely low, but the activity of ADH in the cellulose membrane was basically the same with the one above the liquid surface. The bacteria were mainly distributed in the cellulose membrane during the fermentation. The bacterial counts in surface fermentation were more than in the shake flask fermentation and G. xylinus consumed the substrate faster, in surface fermentation than in shake flask fermentation. The oxygen consumption rate and total acid production rate of surface fermentation were respectively 26.13 times and 2.92 times that of shake flask fermentation.

The Probability Distribution for a Biased Spinner

ERIC Educational Resources Information Center

Foster, Colin

2012-01-01

This article advocates biased spinners as an engaging context for statistics students. Calculating the probability of a biased spinner landing on a particular side makes valuable connections between probability and other areas of mathematics. (Contains 2 figures and 1 table.)

Structure and phylogeography of two tropical predators, spinner (Stenella longirostris) and pantropical spotted (S. attenuata) dolphins, from SNP data

PubMed Central

Morin, Phillip A.

2018-01-01

Little is known about global patterns of genetic connectivity in pelagic dolphins, including how circumtropical pelagic dolphins spread globally following the rapid and recent radiation of the subfamily delphininae. In this study, we tested phylogeographic hypotheses for two circumtropical species, the spinner dolphin (Stenella longirostris) and the pantropical spotted dolphin (Stenella attenuata), using more than 3000 nuclear DNA single nucleotide polymorphisms (SNPs) in each species. Analyses for population structure indicated significant genetic differentiation between almost all subspecies and populations in both species. Bayesian phylogeographic analyses of spinner dolphins showed deep divergence between Indo-Pacific, Atlantic and eastern tropical Pacific Ocean (ETP) lineages. Despite high morphological variation, our results show very close relationships between endemic ETP spinner subspecies in relation to global diversity. The dwarf spinner dolphin is a monophyletic subspecies nested within a major clade of pantropical spinner dolphins from the Indian and western Pacific Ocean populations. Population-level division among the dwarf spinner dolphins was detected—with the northern Australia population being very different from that in Indonesia. In contrast to spinner dolphins, the major boundary for spotted dolphins is between offshore and coastal habitats in the ETP, supporting the current subspecies-level taxonomy. Comparing these species underscores the different scale at which population structure can arise, even in species that are similar in habitat (i.e. pelagic) and distribution. PMID:29765639

Combustion Product Evaluation of Various Charge Sizes and Propellant Formulations. Task 1 Report: Sampling and Analytical Procedures Proposed for Use in Gun Propellant Combustion Product Characterization

DTIC Science & Technology

1988-01-01

ml, 0.01N, NaOH + 1 ml 30% H202 H2S 10 ml. 0.01M CdSO4 + STRactan 10 in H2 Aldehydes 10 ml of 3.1 WM/ml 2.4- dinitrophenylhydrazine (DNPH) soln. in...3 15 IITRI C06673-TR1 I,ý 3. 3.1.2.6 Aldehydes I After sampling, the 11 flasks will be injected with 10 mi of a 3.1 14/mX -2,4- dinitrophenylhydrazine

Thermal Transitions and Reaction Kinetics of Polyhederal Silsesquioxane Containing Phenylethynylphthalimides (Preprint)

DTIC Science & Technology

2010-03-18

Grignard reagent was added drop-wise to a well-stirred THF solution (10 mL) of trichloromethylsilane (5.431 g, 36.3 mmol) in a 250 mL flask. After...chloromethylsilane) The Grignard reagent , (Me3Si)2NC6H4MgBr was prepared exactly as previously described from 37.6 mmol of magnesium and 30 mmol of (N...trimethylsilyl)2-4-bromoaniline in THF (25 mL). This Grignard reagent was added drop-wise to a well-stirred THF solution (10 mL) of trichloromethylsilane

Vitality Stains and Real Time PCR Studies to Delineate the Interactions of Pichia anomala and Aspergillus flavus

USDA-ARS?s Scientific Manuscript database

The objectives of this study were to probe the effect of the yeast, P. anomala against A flavus by using real time RT-PCR technique and vitality fluorescent stains. Yeast and fungi were inoculated into a 250 ml-flask containing 50 ml potato dextrose broth (PDB) at yeast to fungus (Y : F) ratios of ...

Type VI Secretion is a Major Virulence Determinant in Burkholderia Mallei

DTIC Science & Technology

2007-06-01

Strain or plasmid Relevant characteristicsb Reference E. coli TOP10 General cloning and blue/white screening Invitrogen Origami (DE3) D3 lysogen, KmR, SmR...was transformed into E. coli Origami (DE3) and the strain was grown in a 125 ml dispos- able Erlenmeyer flask containing 50 ml of LB with Cb, Tc and Km

Scale-up of an alkaline protease from Bacillus pumilus MTCC 7514 utilizing fish meal as a sole source of nutrients.

PubMed

Gupta, Rishikesh Kumar; Prasad, Dinesh; Sathesh, Jaykumar; Naidu, Ramachandra Boopathy; Kamini, Numbi Ramudu; Palanivel, Saravanan; Gowthaman, Marichetti Kuppuswami

2012-09-01

Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits.

A high efficiency stirrer for suspension cell culture with or without microcarriers.

PubMed

de Bruyne, N A

1984-01-01

The stirrer described in this paper employs "Teaspoon stirring" in which a secondary motion is superimposed on the rotation of the liquid. This secondary motion arises from the viscous drag from the wall and bottom of the flask. The culture medium is rotated by a bulb-ended rod suspended from inside the flask cap: the bulb orbits around in a circular trough formed between a central indent in the base and the rounded periphery of the base. There are no bearings and no stagnant areas. The power used to stir 4 flasks each holding 500 ml of medium is less than 2 watts. The speed is electronically controlled by a tachometer and automatically gives a smooth start and stop as well as "Interval stirring" to assist initial attachment of cells.

Predictive modeling of spinner dolphin (Stenella longirostris) resting habitat in the main Hawaiian Islands.

PubMed

Thorne, Lesley H; Johnston, David W; Urban, Dean L; Tyne, Julian; Bejder, Lars; Baird, Robin W; Yin, Suzanne; Rickards, Susan H; Deakos, Mark H; Mobley, Joseph R; Pack, Adam A; Chapla Hill, Marie

2012-01-01

Predictive habitat models can provide critical information that is necessary in many conservation applications. Using Maximum Entropy modeling, we characterized habitat relationships and generated spatial predictions of spinner dolphin (Stenella longirostris) resting habitat in the main Hawaiian Islands. Spinner dolphins in Hawai'i exhibit predictable daily movements, using inshore bays as resting habitat during daylight hours and foraging in offshore waters at night. There are growing concerns regarding the effects of human activities on spinner dolphins resting in coastal areas. However, the environmental factors that define suitable resting habitat remain unclear and must be assessed and quantified in order to properly address interactions between humans and spinner dolphins. We used a series of dolphin sightings from recent surveys in the main Hawaiian Islands and a suite of environmental variables hypothesized as being important to resting habitat to model spinner dolphin resting habitat. The model performed well in predicting resting habitat and indicated that proximity to deep water foraging areas, depth, the proportion of bays with shallow depths, and rugosity were important predictors of spinner dolphin habitat. Predicted locations of suitable spinner dolphin resting habitat provided in this study indicate areas where future survey efforts should be focused and highlight potential areas of conflict with human activities. This study provides an example of a presence-only habitat model used to inform the management of a species for which patterns of habitat availability are poorly understood.

Predictive Modeling of Spinner Dolphin (Stenella longirostris) Resting Habitat in the Main Hawaiian Islands

PubMed Central

Thorne, Lesley H.; Johnston, David W.; Urban, Dean L.; Tyne, Julian; Bejder, Lars; Baird, Robin W.; Yin, Suzanne; Rickards, Susan H.; Deakos, Mark H.; Mobley, Joseph R.; Pack, Adam A.; Chapla Hill, Marie

2012-01-01

Predictive habitat models can provide critical information that is necessary in many conservation applications. Using Maximum Entropy modeling, we characterized habitat relationships and generated spatial predictions of spinner dolphin (Stenella longirostris) resting habitat in the main Hawaiian Islands. Spinner dolphins in Hawai'i exhibit predictable daily movements, using inshore bays as resting habitat during daylight hours and foraging in offshore waters at night. There are growing concerns regarding the effects of human activities on spinner dolphins resting in coastal areas. However, the environmental factors that define suitable resting habitat remain unclear and must be assessed and quantified in order to properly address interactions between humans and spinner dolphins. We used a series of dolphin sightings from recent surveys in the main Hawaiian Islands and a suite of environmental variables hypothesized as being important to resting habitat to model spinner dolphin resting habitat. The model performed well in predicting resting habitat and indicated that proximity to deep water foraging areas, depth, the proportion of bays with shallow depths, and rugosity were important predictors of spinner dolphin habitat. Predicted locations of suitable spinner dolphin resting habitat provided in this study indicate areas where future survey efforts should be focused and highlight potential areas of conflict with human activities. This study provides an example of a presence-only habitat model used to inform the management of a species for which patterns of habitat availability are poorly understood. PMID:22937022

A unique noninvasive approach to monitoring dissolved O2 and CO2 in cell culture.

PubMed

Chatterjee, Madhubanti; Ge, Xudong; Uplekar, Shaunak; Kostov, Yordan; Croucher, Leah; Pilli, Manohar; Rao, Govind

2015-01-01

Although online monitoring of dissolved oxygen (DO) and carbon dioxide (DCO2 ) is highly desirable in bioprocesses, small-scale bioreactors are usually not monitored due to the lack of suitable sensors. Traditional electrochemical sensors are usually not used because they are bulky and invasive. Disposable optical sensors are small and only partially invasive, but there are concerns regarding the toxicity of the patch and the phototoxicity of the illuminating light. Here we present a novel, noninvasive, rate-based technique for monitoring DO and DCO2 in cell cultures. A silicone sampling loop which allowed the diffusion of O2 and CO2 through its wall was inserted inside a bioreactor, and then flushed with N2 until the CO2 and O2 inside the loop were completely removed. The gas inside the loop was then allowed to recirculate through gas impermeable tubing to the O2 and CO2 sensors. We have shown that by measuring the initial diffusion rate we were able to determine the partial pressures of the two gases in the culture. The technique could be readily automated and measurements could be made in minutes. It was tested in demonstration experiments by growing murine hybridoma cells in a T-flask and a spinner-flask at 37°C. The results were comparable to those measured with commercially available fluorescence-based patch sensors. These results show that the rate-based method is an effective way to monitor small-scale cell cultures. This measurement mechanism can be easily built into disposable cell culture vessels for facile use. © 2014 Wiley Periodicals, Inc.

Geographic variations in the whistles of spinner dolphins (Stenella longirostris) of the Main Hawai‵ian Islands

NASA Astrophysics Data System (ADS)

Bazúa-Durán, Carmen; Au, Whitlow W. L.

2004-12-01

Geographic variations in the whistles of Hawai‵ian spinner dolphins are discussed by comparing 27 spinner dolphin pods recorded in waters off the Islands of Kaua‵i, O‵ahu, Lana‵i, and Hawai‵i. Three different behavioral states, the number of dolphins observed in each pod, and ten parameters extracted from each whistle contour were considered by using clustering and discriminant function analyses. The results suggest that spinner dolphin pods in the Main Hawai‵ian Islands share characteristics in approximately 48% of their whistles. Spinner dolphin pods had similar whistle parameters regardless of the island, location, and date when they were sampled and the dolphins' behavioral state and pod size. The term ‵‵whistle-specific subgroup'' (WSS) was used to designate whistle groups with similar whistles parameters (which could have been produced in part by the same dolphins). The emission rate of whistles was higher when spinner dolphins were socializing than when they were traveling or resting, suggesting that whistles are mainly used during close-range interactions. Spinner dolphins also seem to vary whistle duration according to their general behavioral state. Whistle duration and the number of turns and steps of a whistle may be more important in delivering information at the individual level than whistle frequency parameters. .

Low Cost, High-Throughput 3-D Pulmonary Imager Using Hyperpolarized Contrast Agents and Low-Field MRI

DTIC Science & Technology

2017-10-01

capacitive matching element (see Figs. S3 and S4, SI). The solenoid was constructed with 170 windings using 28 AWG gauge magnet wire with a measured...10.1002/9780470034590.emrstm0491. (59) Minard, K. R.; Wind , R. A. Solenoidal microcoil designPart II: Optimizing winding parameters for maximum...TiO2 catalyst to an NMR tube containing 0.5 mL of acetone-d6. The position of the NMR tube inside the NMR spinner turbine was adjusted for the

Fidget Spinner Ingestions in Children-A Problem that Spun Out of Nowhere.

PubMed

Reeves, Patrick T; Nylund, Cade M; Noel, James M; Jones, David S; Chumpitazi, Bruno P; Milczuk, Henry A; Noel, R Adam

2018-06-01

The Consumer Product Safety Risk Management System's injury and potential injury database records 13 cases of fidget spinner ingestion since 2016. In addition to a database query, we report 3 additional cases of fidget spinner ingestion to describe patient presentations and subsequent management strategies. Published by Elsevier Inc.

Development of Synthesis and Large Scale Production Technology for Ultrahigh Energy and Density Fluoro-Organic Compounds

DTIC Science & Technology

2005-12-01

M-I (H2C=CHSF5). To 3.5 g potassium hydroxide in 4 mL of water was added 10.5 mL of isopropanol in a 50 mL three-necked flask equipped with an...S- 5 2006 Ne Tech New Era Technology, Inc. 3720 NW 4 3rd Street, Suite 105, Gainesville, Florida 32606-6190, USA Final Report TITLE: Development of ...Senior Scientist, NeTech Research Institute (Subcontractor): University of Florida Dr. William R. Dolbier, Jr., P1 - University of Florida, Crow

An Investigation of Single- and Dual-Rotation Propellers at Positive and Negative Thrust, and in Combination with an NACA 1-series D-Type Cowling at Mach Numbers up to 0.84

NASA Technical Reports Server (NTRS)

Reynolds, Robert M; Samonds, Robert I; Walker, John H

1957-01-01

An investigation has been made to determine the aerodynamic characteristics of the NACA 4-(5)(05)-041 four-blade, single-relation propeller and the NACA 4-(5)(05)-037 six- and eight-blade, dual-rotation propellers in combination with various spinners and NACA d-type spinner-cowling combinations at Mach numbers up to 0.84. Propeller force characteristics, local velocity distributions in the propeller planes, inlet pressure recoveries, and static-pressure distributions on the cowling surfaces were measured for a wide range of blade angles, advance ratios, and inlet-velocity ratios. Included are data showing: (a) the effect of extended cylindrical spinners on the characteristics of the single-rotation propeller, (b) the effect of variation of the difference in blade angle setting between the front and rear components of the dual-rotation propellers, (c) the negative- and static-thrust characteristics of the propellers with 1 series spinners, and (d) the effects of ideal- and platform-type propeller-spinner junctures on the pressure-recovery characteristics of the single-rotation propeller-spinner-cowling combination.

Production of nattokinase by batch and fed-batch culture of Bacillus subtilis.

PubMed

Cho, Young-Han; Song, Jae Yong; Kim, Kyung Mi; Kim, Mi Kyoung; Lee, In Young; Kim, Sang Bum; Kim, Hyeon Shup; Han, Nam Soo; Lee, Bong Hee; Kim, Beom Soo

2010-09-30

Nattokinase was produced by batch and fed-batch culture of Bacillus subtilis in flask and fermentor. Effect of supplementing complex media (peptone, yeast extract, or tryptone) was investigated on the production of nattokinase. In flask culture, the highest cell growth and nattokinase activity were obtained with 50 g/L of peptone supplementation. In this condition, nattokinase activity was 630 unit/ml at 12 h. In batch culture of B. subtilis in fermentor, the highest nattokinase activity of 3400 unit/ml was obtained at 10h with 50 g/L of peptone supplementation. From the batch kinetics data, it was shown that nattokinase production was growth-associated and culture should be harvested before stationary phase for maximum nattokinase production. In fed-batch culture of B. subtilis using pH-stat feeding strategy, cell growth (optical density monitored at 600 nm) increased to ca. 100 at 22 h, which was 2.5 times higher than that in batch culture. The highest nattokinase activity was 7100 unit/ml at 19 h, which was also 2.1 times higher than that in batch culture. Copyright 2010 Elsevier B.V. All rights reserved.

Manual centrifuge system: Bearing-based hand spinner made with 3-D printer.

PubMed

Sun-Young Yoo; Seung Jae Lee; Jong-Mo Seo

2017-07-01

Compact disk (CD) Microfluidic platforms are being studied for medical applications such as blood tests. However, its size is bulky and electricity is needed to realize centrifuge force. In this paper, bearing-based hand spinner is designed using three-dimensional printer. This spinner does not need electricity and keeps rotating direction unlike paperfuge while it is spinning. The properties of spinner vary depending on bearing type which is positioned at the center. The type of weighting area also affects change in RPM over time. When a separation experiment is implemented, separating mixture into red ink and oil and whole blood into red blood cell and plasma are achieved properly with ceramic ball bearing.

Simple suspension culture system of human iPS cells maintaining their pluripotency for cardiac cell sheet engineering.

PubMed

Haraguchi, Yuji; Matsuura, Katsuhisa; Shimizu, Tatsuya; Yamato, Masayuki; Okano, Teruo

2015-12-01

In this study, a simple three-dimensional (3D) suspension culture method for the expansion and cardiac differentiation of human induced pluripotent stem cells (hiPSCs) is reported. The culture methods were easily adapted from two-dimensional (2D) to 3D culture without any additional manipulations. When hiPSCs were directly applied to 3D culture from 2D in a single-cell suspension, only a few aggregated cells were observed. However, after 3 days, culture of the small hiPSC aggregates in a spinner flask at the optimal agitation rate created aggregates which were capable of cell passages from the single-cell suspension. Cell numbers increased to approximately 10-fold after 12 days of culture. The undifferentiated state of expanded hiPSCs was confirmed by flow cytometry, immunocytochemistry and quantitative RT-PCR, and the hiPSCs differentiated into three germ layers. When the hiPSCs were subsequently cultured in a flask using cardiac differentiation medium, expression of cardiac cell-specific genes and beating cardiomyocytes were observed. Furthermore, the culture of hiPSCs on Matrigel-coated dishes with serum-free medium containing activin A, BMP4 and FGF-2 enabled it to generate robust spontaneous beating cardiomyocytes and these cells expressed several cardiac cell-related genes, including HCN4, MLC-2a and MLC-2v. This suggests that the expanded hiPSCs might maintain the potential to differentiate into several types of cardiomyocytes, including pacemakers. Moreover, when cardiac cell sheets were fabricated using differentiated cardiomyocytes, they beat spontaneously and synchronously, indicating electrically communicative tissue. This simple culture system might enable the generation of sufficient amounts of beating cardiomyocytes for use in cardiac regenerative medicine and tissue engineering. Copyright © 2013 John Wiley & Sons, Ltd.

Smartphone Application Enabling Global Graph Exploitation and Research

DTIC Science & Technology

2013-05-01

employed, blue collar, white collar Religion Mild theology, radical theology Skill Photography, writing, electrical, mechanical, computer, driving...findViewById( R.id.religion_spinner ); 480: String religion = s9.getSelectedItem().toString(); 481: 482: Spinner s10 = (Spinner) findViewById...34Choose Suspect Religion ") || skill.equals("Choose Suspect Skill") || address.equals("Choose Suspect Address")) 504: { 505: Toast msg

[Isolation, identification and characterization of a diethylstilbestrol-degrading bacterial strain Serratia sp].

PubMed

Xu, Ran-Fang; Sun, Min-Xia; Liu, Juan; Wang, Hong; Li, Xin; Zhu, Xue-Zhu; Ling, Wan-Ting

2014-08-01

Utilizing the diethylstilbestrol (DES)-degrading bacteria to biodegrade DES is a most reliable technique for cleanup of DES pollutants from the environment. However, little information is available heretofore on the isolation of DES-degrading bacteria and their DES removal performance in the environment. A novel bacterium capable of degrading DES was isolated from the activated sludge of a wastewater treatment plant. According to its morphology, physiochemical characteristics, and 16S rDNA sequence analysis, this strain was identified as Serratia sp.. The strain was an aerobic bacterium, and it could degrade 68.3% of DES (50 mg x L(-1)) after culturing for 7 days at 30 degrees C, 150 r x min(-1) in shaking flasks. The optimal conditions for DES biodegradation by the obtained strain were 30 degrees C, 40-60 mg x L(-1) DES, pH 7.0, 5% of inoculation volume, 0 g x L(-1) of added NaCl, and 10 mL of liquid medium volume in 100 mL flask.

Long Range Materials Research. Appendix 1. Synthesis and Characterization of Supported Organometallic Rhodium (I) Catalysts

DTIC Science & Technology

1974-06-30

hydrosilates, 130 including irradiation by ultraviolet light, elevated temperatures (up to 3500) in sealed tubes, and free radical initiators (up to 10 mol...to 160 ml dry pyridine and stirred at 00 in a 250 ml Erlenmeyer flask fitted with a septum. After the temperature had equilibrated, p-toluene sulfonyl...chloride (80 g, 0.41 mol) was added slowly, carefully maintaining the temperature below 100. The vessel was tightly capped, the mixture stirred an

Processing Method for Creating Ultra-Thin Lead Zirconate Titanate (PZT) Films Via Chemical Solution Deposition

DTIC Science & Technology

2008-12-01

n-propoxide and titanium isopropoxide , were measured with a graduated auto pipet and combined with 45 mL of 2-MOE in a 125 mL flask. The solution...nitrogen (N2). This anneal procedure was used to remove trapped hydrogen from the thin film. Following the anneal, a bi-layer of titanium (Ti) and...dioxide Ti titanium 10 NO. OF COPIES ORGANIZATION 1 ADMNSTR ELEC DEFNS TECHL INFO CTR ATTN DTIC OCP 8725 JOHN J KINGMAN RD STE

40 CFR Appendix 5 to Subpart A of... - Determination of Crude Oil Contamination in Non-Aqueous Drilling Fluids by Gas Chromatography...

Code of Federal Regulations, 2011 CFR

2011-07-01

.... 6.2.2Analytical balance—Capable of weighing 0.1 mg. 6.2.3Glassware. 6.2.3.1Disposable pipettes... standard at 62.5 mg/mL in methylene chloride. Prepare a medium concentration SPTM standard at 1.25 mg/mL by transferring 1.0 mL of the 62.5 mg/mL solution into a 50 mL volumetric flask and diluting to the mark with...

Towards Improved Optical Limiters

DTIC Science & Technology

2002-05-03

nitrophthalonitrile, 3.37 g of potassium carbonate, and 0.54 g of neopentyl glycol were added to 10 mL of dry DMSO in a round bottom flask. The... glycol oxygens which in turn replace the nitro groups of the phthalonitrile and form a bond. This substitution reaction produces the bridge unit as

USSR and Eastern Europe Scientific Abstracts, Biomedical and Behavioral Sciences, Number 92

DTIC Science & Technology

1978-07-26

EFFECT OF NUTRIENT MEDIUM COMPOSITION ON THE BIOSYNTHESIS OF CELLULOYTIC ENZYMES WITH SUBMERGED CULTIVATION OF FUNGUS TRICHODERMA LONGIBRACHIATUM 7-26...enzymes using submerged cultivation of Trichoderma longibrachiatum 7-26 was sought. The culture was in a rocker (200 rot/min) in 750 ml flasks at 28-30

Synthesis and Characterization of Furanic Compounds

DTIC Science & Technology

2013-09-01

trifluoroacetic acid, 1.6-M solution of BuLi in hexane, dichloromethane, sodium bicarbonate, pyridinium chlorochromate, potassium permanganate , sodium...intermediate, 2,5-diformylfuran (2.29 g, 18.47 mmol), was oxidized in a 100-mL round-bottom flask with potassium permanganate (4.47 g, 153.1 mmol) in a NaOH

Optimization of flask culture medium and conditions for hyaluronic acid production by a Streptococcus equisimilis mutant nc2168.

PubMed

Chen, Yong-Hao; Li, Jun; Liu, Li; Liu, Hong-Zhi; Wang, Qiang

2012-10-01

A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116 by ultraviolet ray combined with(60)Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA). In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33°C, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.

Full-Scale Tests of Several Propellers Equipped with Spinners, Cuffs, Airfoil and Round Shanks, and NACA 16-Series Sections, Special Report

NASA Technical Reports Server (NTRS)

Biermann, David; Hartman, Edwin P.; Pepper, Edward

1940-01-01

Wind-tunnel tests of several propeller, cuff, and spinner combinations were conducted in the 20 foot propeller-research tunnel. Three propellers, which ranged in diameter from 8.4 to 11.25 feet, were tested at the front end of a streamline body incorporating spinners of two diameters. The tests covered a blade angle range from 20 deg to 65 deg. The effect of spinner diameter and propeller cuffs on the characteristics of one propeller was determined. Test were also conducted using a propeller which incorporated aerodynamically good shank sections and using one which incorporated the NACA 16 series sections for the outer 20 percent of the blades. Compressibility effects were not measured, owing to the low testing speeds. The results indicated that a conventional propeller was slightly more efficient when tested in conjunction with a 28 inch diameter spinner than with a 23 inch spinner, and that cuffs increased the efficiency as well as the power absorption characteristics. A propeller having good aerodynamic shanks was found to be definitely superior from the efficiency standpoint to a conventional round-shank propeller with or without cuffs; this propeller would probably be considered structurally impracticable, however. The propeller incorporating the NACA 16 series sections at the tims were found to have a slightly higher efficiency than a conventional propeller; the take-off characteristics appeared to be equally good. The effects noted above probably would be accentuated at helical speeds at which compressibility effects would enter.

Reduction of C Bonds Proceeds with Retention of Configuration: Stereochemical Investigation of the Heterogeneous Reduction by Dideuterium of (Homohypostrophene)Neopentyl(2-Norbornyl)Platinum(II) Complexes on Platinum Black.

DTIC Science & Technology

1991-04-23

ClIH16: C, 89.12; H, 10.88; Found: C, 88.85; H, 10.90. Synthesis of Grignard Reagents . Neopentylmagnesium chloride. Into a 500-mL round-bottomed flask... Grignard reagent several times using the following procedure. We placed 5.0 g (0.206 mol) of magnesium chips and a magnetic stir bar to a 200-mL round...Norbornylmagnesium bromide. This Grignard reagent was synthesized using a variation on established procedures. 58,60 We transferred under argon 50.0 mL (30.0 mmol

Aggregation of Culture Expanded Human Mesenchymal Stem Cells in Microcarrier-based Bioreactor.

PubMed

Yuan, Xuegang; Tsai, Ang-Chen; Farrance, Iain; Rowley, Jon; Ma, Teng

2018-03-15

Three-dimensional aggregation of human mesenchymal stem cells (hMSCs) has been used to enhance their therapeutic properties but current fabrication protocols depend on laboratory methods and are not scalable. In this study, we developed thermal responsive poly(N-isopropylacrylamide) grafted microcarriers (PNIPAM-MCs), which supported expansion and thermal detachment of hMSCs at reduced temperature (23.0 °C). hMSCs were cultured on the PNIPAM-MCs in both spinner flask (SF) and PBS Vertical-Wheel (PBS-VW) bioreactors for expansion. At room temperature, hMSCs were detached as small cell sheets, which subsequently self-assembled into 3D hMSC aggregates in PBS-VW bioreactor and remain as single cells in SF bioreactor owing to different hydrodynamic conditions. hMSC aggregates generated from the bioreactor maintained comparable immunomodulation and cytokine secretion properties compared to the ones made from the AggreWell ® . The results of the current study demonstrate the feasibility of scale-up production of hMSC aggregates in the suspension bioreactor using thermal responsive microcarriers for integrated cell expansion and 3D aggregation in a close bioreactor system and highlight the critical role of hydrodynamics in self-assembly of detached hMSC in suspension.

Potential bioremediation of mercury-contaminated substrate using filamentous fungi isolated from forest soil.

PubMed

Kurniati, Evi; Arfarita, Novi; Imai, Tsuyoshi; Higuchi, Takaya; Kanno, Ariyo; Yamamoto, Koichi; Sekine, Masahiko

2014-06-01

The use of filamentous fungi in bioremediation of heavy metal contamination has been developed recently. This research aims to observe the capability of filamentous fungi isolated from forest soil for bioremediation of mercury contamination in a substrate. Six fungal strains were selected based on their capability to grow in 25 mg/L Hg(2+)-contaminated potato dextrose agar plates. Fungal strain KRP1 showed the highest ratio of growth diameter, 0.831, thus was chosen for further observation. Identification based on colony and cell morphology carried out by 18S rRNA analysis gave a 98% match to Aspergillus flavus strain KRP1. The fungal characteristics in mercury(II) contamination such as range of optimum pH, optimum temperature and tolerance level were 5.5-7 and 25-35°C and 100 mg/L respectively. The concentration of mercury in the media affected fungal growth during lag phases. The capability of the fungal strain to remove the mercury(II) contaminant was evaluated in 100 mL sterile 10 mg/L Hg(2+)-contaminated potato dextrose broth media in 250 mL Erlenmeyer flasks inoculated with 10(8) spore/mL fungal spore suspension and incubation at 30°C for 7 days. The mercury(II) utilization was observed for flasks shaken in a 130 r/min orbital shaker (shaken) and non-shaken flasks (static) treatments. Flasks containing contaminated media with no fungal spores were also provided as control. All treatments were done in triplicate. The strain was able to remove 97.50% and 98.73% mercury from shaken and static systems respectively. A. flavus strain KRP1 seems to have potential use in bioremediation of aqueous substrates containing mercury(II) through a biosorption mechanism. Copyright © 2014 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

Experimental and Theoretical Study of Propeller Spinner/Shank Interference. M.S. Thesis

NASA Technical Reports Server (NTRS)

Cornell, C. C.

1986-01-01

A fundamental experimental and theoretical investigation into the aerodynamic interference associated with propeller spinner and shank regions was conducted. The research program involved a theoretical assessment of solutions previously proposed, followed by a systematic experimental study to supplement the existing data base. As a result, a refined computational procedure was established for prediction of interference effects in terms of interference drag and resolved into propeller thrust and torque components. These quantities were examined with attention to engineering parameters such as two spinner finess ratios, three blade shank forms, and two/three/four/six/eight blades. Consideration of the physics of the phenomena aided in the logical deduction of two individual interference quantities (cascade effects and spinner/shank juncture interference). These interference effects were semi-empirically modeled using existing theories and placed into a compatible form with an existing propeller performance scheme which provided the basis for examples of application.

Space-to-Ground: Space Spinners:11/03/2017

NASA Image and Video Library

2017-11-02

The crew spent this week enabling long term missions and long distance learning...and how long would a fidget spinner spin in space? Space to Ground is your weekly update on what's happening aboard the International Space Station.

Lidar - DTU SpinnerLidar - Reviewed Data

DOE Data Explorer

Herges, Thomas

2017-10-23

Scaled Wind Farm Technology (SWiFT) Facility meteorological tower (MET), turbine, and Technical University of Denmark (DTU) SpinnerLidar data acquired on 20161216 UTC during a neutral atmospheric boundary layer inflow at a single focus distance of 2.5 D (D=27 m).

Discriminating features of echolocation clicks of melon-headed whales (Peponocephala electra), bottlenose dolphins (Tursiops truncatus), and Gray's spinner dolphins (Stenella longirostris longirostris).

PubMed

Baumann-Pickering, Simone; Wiggins, Sean M; Hildebrand, John A; Roch, Marie A; Schnitzler, Hans-Ulrich

2010-10-01

Spectral parameters were used to discriminate between echolocation clicks produced by three dolphin species at Palmyra Atoll: melon-headed whales (Peponocephala electra), bottlenose dolphins (Tursiops truncatus) and Gray's spinner dolphins (Stenella longirostris longirostris). Single species acoustic behavior during daytime observations was recorded with a towed hydrophone array sampling at 192 and 480 kHz. Additionally, an autonomous, bottom moored High-frequency Acoustic Recording Package (HARP) collected acoustic data with a sampling rate of 200 kHz. Melon-headed whale echolocation clicks had the lowest peak and center frequencies, spinner dolphins had the highest frequencies and bottlenose dolphins were nested in between these two species. Frequency differences were significant. Temporal parameters were not well suited for classification. Feature differences were enhanced by reducing variability within a set of single clicks by calculating mean spectra for groups of clicks. Median peak frequencies of averaged clicks (group size 50) of melon-headed whales ranged between 24.4 and 29.7 kHz, of bottlenose dolphins between 26.7 and 36.7 kHz, and of spinner dolphins between 33.8 and 36.0 kHz. Discriminant function analysis showed the ability to correctly discriminate between 93% of melon-headed whales, 75% of spinner dolphins and 54% of bottlenose dolphins.

Simple Laboratory Experiment for Illustrating Soil Respiration.

ERIC Educational Resources Information Center

Hattey, J. A.; Johnson, G. V.

1997-01-01

Describes an experiment to illustrate the effect of food source and added nutrients (N) on microbial activity in the soil. Supplies include air-dried soil, dried plant material, sources of carbon and nitrogen, a trap such as KOH, colored water, and a 500-mL Erlenmeyer flask. Includes a diagram of an incubation chamber to demonstrate microbial…

13. ROOM 40, LOOKING WEST FROM THE RUBENS COIL. SPINNER ...

Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

13. ROOM 40, LOOKING WEST FROM THE RUBENS COIL. SPINNER MAGNETOMETER IS VISIBLE AT FAR LEFT, AND THE RUBENS COIL CONTROL UNIT AT FAR RIGHT. - U.S. Geological Survey, Rock Magnetics Laboratory, 345 Middlefield Road, Menlo Park, San Mateo County, CA

Reflection with a Twist: The Helical Mirror

ERIC Educational Resources Information Center

DeWeerd, Alan J.; Hill, S. Eric

2006-01-01

Over the last few years, the helical mirror or "spinner" has become a popular decoration for gardens and elsewhere. Even casual observation reveals intriguing optical properties, so the spinner is a good teaching tool. To facilitate student exploration, we suggest some questions and provide brief explanations.

Magnetic Spinner

ERIC Educational Resources Information Center

Ouseph, P. J.

2006-01-01

A science toy sometimes called the "magnetic spinner" is an interesting class demonstration to illustrate the principles of magnetic levitation. It can also be used to demonstrate Faraday's law and a horizontally suspended physical pendulum. The levitated part contains two circular magnets encased in a plastic housing. Each magnet stays…

21 CFR 864.5850 - Automated slide spinner.

Code of Federal Regulations, 2010 CFR

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated slide spinner. 864.5850 Section 864.5850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated Hematology Devices § 864...

21 CFR 864.5850 - Automated slide spinner.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Automated slide spinner. 864.5850 Section 864.5850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated Hematology Devices § 864...

21 CFR 864.5850 - Automated slide spinner.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Automated slide spinner. 864.5850 Section 864.5850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated Hematology Devices § 864...

21 CFR 864.5850 - Automated slide spinner.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Automated slide spinner. 864.5850 Section 864.5850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated Hematology Devices § 864...

21 CFR 864.5850 - Automated slide spinner.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Automated slide spinner. 864.5850 Section 864.5850 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Automated and Semi-Automated Hematology Devices § 864...

Production of τ τ jj final states at the LHC and the TauSpinner algorithm: the spin-2 case

NASA Astrophysics Data System (ADS)

Bahmani, M.; Kalinowski, J.; Kotlarski, W.; Richter-Wąs, E.; Wąs, Z.

2018-01-01

The TauSpinner algorithm is a tool that allows one to modify the physics model of the Monte Carlo generated samples due to the changed assumptions of event production dynamics, but without the need of re-generating events. With the help of weights τ -lepton production or decay processes can be modified accordingly to a new physics model. In a recent paper a new version TauSpinner ver.2.0.0 has been presented which includes a provision for introducing non-standard states and couplings and study their effects in the vector-boson-fusion processes by exploiting the spin correlations of τ -lepton pair decay products in processes where final states include also two hard jets. In the present paper we document how this can be achieved taking as an example the non-standard spin-2 state that couples to Standard Model particles and tree-level matrix elements with complete helicity information included for the parton-parton scattering amplitudes into a τ -lepton pair and two outgoing partons. This implementation is prepared as the external (user-provided) routine for the TauSpinner algorithm. It exploits amplitudes generated by MadGraph5 and adapted to the TauSpinner algorithm format. Consistency tests of the implemented matrix elements, re-weighting algorithm and numerical results for observables sensitive to τ polarisation are presented.

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2013 CFR

2013-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2012 CFR

2012-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

40 CFR Appendix A to Part 80 - Test for the Determination of Phosphorus in Gasoline

Code of Federal Regulations, 2014 CFR

2014-07-01

... Specification for Filter Paper for Use in Chemical Analysis. 3. Summary of method. 3.1 Organic matter in the...) during the entire period of sample heating. Note 1: If the temperature of the hot water bath drops below... 100-ml volumetric flasks submerged to the mark in ice water. 4.4 Filter Paper, for quantitative...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2014 CFR

2014-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2012 CFR

2012-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

46 CFR Appendix III to Part 150 - Testing Procedures for Determining Exceptions to the Chart

Code of Federal Regulations, 2013 CFR

2013-10-01

... provided with shields. Testing chemicals other than liquids—The procedure outlined below was developed for... test tube to a stand behind a safety shield (in a hood). Carefully add from a dropper 0.5ml of the... mixture. The Dewar flask is equipped with a magnetic stirrer having a stirring bar coated with an inert...

Efficient Manufacturing of Therapeutic Mesenchymal Stromal Cells Using the Quantum Cell Expansion System

PubMed Central

Hanley, Patrick J.; Mei, Zhuyong; Durett, April G.; Cabreira-Harrison, Marie da Graca; Klis, Mariola; Li, Wei; Zhao, Yali; Yang, Bing; Parsha, Kaushik; Mir, Osman; Vahidy, Farhaan; Bloom, Debra; Rice, R. Brent; Hematti, Peiman; Savitz, Sean I; Gee, Adrian P.

2014-01-01

Background The use of bone marrow-derived mesenchymal stromal cells (MSCs) as a cellular therapy for various diseases, such as graft-versus-host-disease, diabetes, ischemic cardiomyopathy, and Crohn's disease has produced promising results in early-phase clinical trials. However, for widespread application and use in later phase studies, manufacture of these cells needs to be cost effective, safe, and reproducible. Current methods of manufacturing in flasks or cell factories are labor-intensive, involve a large number of open procedures, and require prolonged culture times. Methods We evaluated the Quantum Cell Expansion system for the expansion of large numbers of MSCs from unprocessed bone marrow in a functionally closed system and compared the results to a flask-based method currently in clinical trials. Results After only two passages, we were able to expand a mean of 6.6×108 MSCs from 25 mL of bone marrow reproducibly. The mean expansion time was 21 days, and cells obtained were able to differentiate into all three lineages: chondrocytes, osteoblasts, and adipocytes. The Quantum was able to generate the target cell number of 2.0×108 cells in an average of 9-fewer days and in half the number of passages required during flask-based expansion. We estimated the Quantum would involve 133 open procedures versus 54,400 in flasks when manufacturing for a clinical trial. Quantum-expanded MSCs infused into an ischemic stroke rat model were therapeutically active. Discussion The Quantum is a novel method of generating high numbers of MSCs in less time and at lower passages when compared to flasks. In the Quantum, the risk of contamination is substantially reduced due to the substantial decrease in open procedures. PMID:24726657

Stable expression and purification of a functional processed Fab' fragment from a single nascent polypeptide in CHO cells expressing the mCAT-1 retroviral receptor.

PubMed

Camper, Nicolas; Byrne, Teresa; Burden, Roberta E; Lowry, Jenny; Gray, Breena; Johnston, James A; Migaud, Marie E; Olwill, Shane A; Buick, Richard J; Scott, Christopher J

2011-09-30

Monoclonal antibodies and derivative formats such as Fab' fragments are used in a broad range of therapeutic, diagnostic and research applications. New systems and methodologies that can improve the production of these proteins are consequently of much interest. Here we present a novel approach for the rapid production of processed Fab' fragments in a CHO cell line that has been engineered to express the mouse cationic amino acid transporter receptor 1 (mCAT-1). This facilitated the introduction of the target antibody gene through retroviral transfection, rapidly producing stable expression. Using this system, we designed a single retroviral vector construct for the expression of a target Fab' fragment as a single polypeptide with a furin cleavage site and a FMDV 2A self-cleaving peptide introduced to bridge the light and truncated heavy chain regions. The introduction of these cleavage motifs ensured equimolar expression and processing of the heavy and light domains as exemplified by the production of an active chimeric Fab' fragment against the Fas receptor, routinely expressed in 1-2mg/L yield in spinner-flask cell cultures. These results demonstrate that this method could have application in the facile production of bioactive Fab' fragments. Copyright © 2011 Elsevier B.V. All rights reserved.

Temporally and spatially partitioned behaviours of spinner dolphins: implications for resilience to human disturbance

PubMed Central

Johnston, David W.; Christiansen, Fredrik

2017-01-01

Selective forces shape the evolution of wildlife behavioural strategies and influence the spatial and temporal partitioning of behavioural activities to maximize individual fitness. Globally, wildlife is increasingly exposed to human activities which may affect their behavioural activities. The ability of wildlife to compensate for the effects of human activities may have implications for their resilience to disturbance. Resilience theory suggests that behavioural systems which are constrained in their repertoires are less resilient to disturbance than flexible systems. Using behavioural time-series data, we show that spinner dolphins (Stenella longirostris) spatially and temporally partition their behavioural activities on a daily basis. Specifically, spinner dolphins were never observed foraging during daytime, where resting was the predominant activity. Travelling and socializing probabilities were higher in early mornings and late afternoons when dolphins were returning from or preparing for nocturnal feeding trips, respectively. The constrained nature of spinner dolphin behaviours suggests they are less resilient to human disturbance than other cetaceans. These dolphins experience the highest exposure rates to human activities ever reported for any cetaceans. Over the last 30 years human activities have increased significantly in Hawaii, but the spinner dolphins still inhabit these bays. Recent abundance estimates (2011 and 2012) however, are lower than all previous estimates (1979–1981, 1989–1992 and 2003), indicating a possible long-term impact. Quantification of the spatial and temporal partitioning of wildlife behavioural schedules provides critical insight for conservation measures that aim to mitigate the effects of human disturbance. PMID:28280561

Temporally and spatially partitioned behaviours of spinner dolphins: implications for resilience to human disturbance.

PubMed

Tyne, Julian A; Johnston, David W; Christiansen, Fredrik; Bejder, Lars

2017-01-01

Selective forces shape the evolution of wildlife behavioural strategies and influence the spatial and temporal partitioning of behavioural activities to maximize individual fitness. Globally, wildlife is increasingly exposed to human activities which may affect their behavioural activities. The ability of wildlife to compensate for the effects of human activities may have implications for their resilience to disturbance. Resilience theory suggests that behavioural systems which are constrained in their repertoires are less resilient to disturbance than flexible systems. Using behavioural time-series data, we show that spinner dolphins ( Stenella longirostris ) spatially and temporally partition their behavioural activities on a daily basis. Specifically, spinner dolphins were never observed foraging during daytime, where resting was the predominant activity. Travelling and socializing probabilities were higher in early mornings and late afternoons when dolphins were returning from or preparing for nocturnal feeding trips, respectively. The constrained nature of spinner dolphin behaviours suggests they are less resilient to human disturbance than other cetaceans. These dolphins experience the highest exposure rates to human activities ever reported for any cetaceans. Over the last 30 years human activities have increased significantly in Hawaii, but the spinner dolphins still inhabit these bays. Recent abundance estimates (2011 and 2012) however, are lower than all previous estimates (1979-1981, 1989-1992 and 2003), indicating a possible long-term impact. Quantification of the spatial and temporal partitioning of wildlife behavioural schedules provides critical insight for conservation measures that aim to mitigate the effects of human disturbance.

Abundance and Survival Rates of the Hawai’i Island Associated Spinner Dolphin (Stenella longirostris) Stock

PubMed Central

Tyne, Julian A.; Pollock, Kenneth H.; Johnston, David W.; Bejder, Lars

2014-01-01

Reliable population estimates are critical to implement effective management strategies. The Hawai’i Island spinner dolphin (Stenella longirostris) is a genetically distinct stock that displays a rigid daily behavioural pattern, foraging offshore at night and resting in sheltered bays during the day. Consequently, they are exposed to frequent human interactions and disturbance. We estimated population parameters of this spinner dolphin stock using a systematic sampling design and capture–recapture models. From September 2010 to August 2011, boat-based photo-identification surveys were undertaken monthly over 132 days (>1,150 hours of effort; >100,000 dorsal fin images) in the four main resting bays along the Kona Coast, Hawai’i Island. All images were graded according to photographic quality and distinctiveness. Over 32,000 images were included in the analyses, from which 607 distinctive individuals were catalogued and 214 were highly distinctive. Two independent estimates of the proportion of highly distinctive individuals in the population were not significantly different (p = 0.68). Individual heterogeneity and time variation in capture probabilities were strongly indicated for these data; therefore capture–recapture models allowing for these variations were used. The estimated annual apparent survival rate (product of true survival and permanent emigration) was 0.97 SE±0.05. Open and closed capture–recapture models for the highly distinctive individuals photographed at least once each month produced similar abundance estimates. An estimate of 221±4.3 SE highly distinctive spinner dolphins, resulted in a total abundance of 631±60.1 SE, (95% CI 524–761) spinner dolphins in the Hawai’i Island stock, which is lower than previous estimates. When this abundance estimate is considered alongside the rigid daily behavioural pattern, genetic distinctiveness, and the ease of human access to spinner dolphins in their preferred resting habitats, this Hawai’i Island stock is likely more vulnerable to negative impacts from human disturbance than previously believed. PMID:24465917

Abundance and survival rates of the Hawai'i Island associated spinner dolphin (Stenella longirostris) stock.

PubMed

Tyne, Julian A; Pollock, Kenneth H; Johnston, David W; Bejder, Lars

2014-01-01

Reliable population estimates are critical to implement effective management strategies. The Hawai'i Island spinner dolphin (Stenella longirostris) is a genetically distinct stock that displays a rigid daily behavioural pattern, foraging offshore at night and resting in sheltered bays during the day. Consequently, they are exposed to frequent human interactions and disturbance. We estimated population parameters of this spinner dolphin stock using a systematic sampling design and capture-recapture models. From September 2010 to August 2011, boat-based photo-identification surveys were undertaken monthly over 132 days (>1,150 hours of effort; >100,000 dorsal fin images) in the four main resting bays along the Kona Coast, Hawai'i Island. All images were graded according to photographic quality and distinctiveness. Over 32,000 images were included in the analyses, from which 607 distinctive individuals were catalogued and 214 were highly distinctive. Two independent estimates of the proportion of highly distinctive individuals in the population were not significantly different (p = 0.68). Individual heterogeneity and time variation in capture probabilities were strongly indicated for these data; therefore capture-recapture models allowing for these variations were used. The estimated annual apparent survival rate (product of true survival and permanent emigration) was 0.97 SE ± 0.05. Open and closed capture-recapture models for the highly distinctive individuals photographed at least once each month produced similar abundance estimates. An estimate of 221 ± 4.3 SE highly distinctive spinner dolphins, resulted in a total abundance of 631 ± 60.1 SE, (95% CI 524-761) spinner dolphins in the Hawai'i Island stock, which is lower than previous estimates. When this abundance estimate is considered alongside the rigid daily behavioural pattern, genetic distinctiveness, and the ease of human access to spinner dolphins in their preferred resting habitats, this Hawai'i Island stock is likely more vulnerable to negative impacts from human disturbance than previously believed.

Extending hepatocyte functionality for drug-testing applications using high-viscosity alginate-encapsulated three-dimensional cultures in bioreactors.

PubMed

Miranda, Joana P; Rodrigues, Armanda; Tostões, Rui M; Leite, Sofia; Zimmerman, Heiko; Carrondo, Manuel J T; Alves, Paula M

2010-12-01

The maintenance of differentiated hepatocyte phenotype in vitro depends on several factors-in particular, on extracellular matrix interactions, for example, with three-dimensional (3D) matrices. Alginate hydrogel provides the cells with a good extracellular matrix due to the formation of a massive capsule with semi-permeable properties that allows for diffusion of the medium components into the cells as well as efficient waste product elimination. Simultaneously, alginate protects the cells from shear stress caused by the hydrodynamics when cultured in stirred systems such as bioreactors. We have previously developed a hepatocyte aggregate 3D culture system in a bioreactor where improved hepatocyte functionality could be maintained over longer periods (21 days). In this work, ultra-high-viscosity alginate was used for hepatocyte aggregates entrapment. Hepatocyte biotransformation (phase I and II enzymes), CYP450 inducibility, and secretory capacity (albumin and urea production) were monitored. The analyses were performed in both spinner vessels and bioreactors to test the effect of the pO(2) control, unavailable in the spinners. Performance of alginate-encapsulated hepatocyte aggregates in culture was compared with nonencapsulated aggregate cultures in both bioreactor (controlled environment) and spinner vessels. For both culture systems, hepatocytes' metabolic and biotransformation capacities were maintained for up to 1 month, and encapsulated cells in bioreactors showed the best performance. In particular, albumin production rate increased 2- and 1.5-fold in encapsulated aggregates compared with nonencapsulated aggregates in bioreactor and spinner vessels, respectively. Urea production rate increased twofold in encapsulated cultures compared with nonencapsulated cells, in both bioreactor and spinner vessels. Similarly, in both the bioreactor and the spinner system, cell encapsulation resulted in a 1.5- and 2.8-fold improvement of hepatocyte 7-ethoxycoumarin and uridine diphosphate glucuronosyltransferases (UGT) activities, respectively. For all parameters, but for UGT activity, the bioreactor system resulted better than the spinner vessels; for UGT activity no difference was observed between the two. Furthermore, both encapsulated and nonencapsulated 3D culture systems were inducible by 3-methylcholanthrene and dexamethasone. The encapsulated systems consistently showed improved performance over the nonencapsulated cells, indicating that the protection conferred by the alginate matrix plays a relevant role in maintaining the hepatocyte functionalities in vitro.

Influence of simulated microgravity on the longevity of insect-cell culture

NASA Technical Reports Server (NTRS)

Cowger, N. L.; O'Connor, K. C.; Bivins, J. E.

1997-01-01

Simulated microgravity within the NASA High Aspect Rotating-Wall Vessel (HARV) provides a quiescent environment to culture fragile insect cells. In this vessel, the duration of stationary and death phase for cultures of Spodoptera frugiperda cells was greatly extended over that achieved in shaker-flask controls. For both HARV and control cultures, S. frugiperda cells grew to concentrations in excess of 1 x 10(7) viable cells ml-1 with viabilities greater than 90%. In the HARV, stationary phase was maintained 9-15 days in contrast to 4-5 days in the shaker flask. Furthermore, the rate of cell death was reduced in the HARV by a factor of 20-90 relative to the control culture and was characterized with a death rate constant of 0.01-0.02 day-1. Beginning in the stationary phase and continuing in the death phase, there was a significant decrease in population size in the HARV versus an increase in the shaker flask. This phenomenon could represent cell adaptation to simulated microgravity and/or a change in the ratio of apoptotic to necrotic cells. Differences observed in this research between the HARV and its control were attributed to a reduction in hydrodynamic forces in the microgravity vessel.

Highly Efficient Large-Scale Lentiviral Vector Concentration by Tandem Tangential Flow Filtration

PubMed Central

Cooper, Aaron R.; Patel, Sanjeet; Senadheera, Shantha; Plath, Kathrin; Kohn, Donald B.; Hollis, Roger P.

2014-01-01

Large-scale lentiviral vector (LV) concentration can be inefficient and time consuming, often involving multiple rounds of filtration and centrifugation. This report describes a simpler method using two tangential flow filtration (TFF) steps to concentrate liter-scale volumes of LV supernatant, achieving in excess of 2000-fold concentration in less than 3 hours with very high recovery (>97%). Large volumes of LV supernatant can be produced easily through the use of multi-layer flasks, each having 1720 cm2 surface area and producing ~560 mL of supernatant per flask. Combining the use of such flasks and TFF greatly simplifies large-scale production of LV. As a demonstration, the method is used to produce a very high titer LV (>1010 TU/mL) and transduce primary human CD34+ hematopoietic stem/progenitor cells at high final vector concentrations with no overt toxicity. A complex LV (STEMCCA) for induced pluripotent stem cell generation is also concentrated from low initial titer and used to transduce and reprogram primary human fibroblasts with no overt toxicity. Additionally, a generalized and simple multiplexed real- time PCR assay is described for lentiviral vector titer and copy number determination. PMID:21784103

Using a Fidget Spinner to Teach Physics

ERIC Educational Resources Information Center

Mesquita, Lucas; Brockington, Guilherme; de Almeida, Pamella Aline; Truyol, Maria Elena; Testoni, André; Sousa, Paula, F. F.

2018-01-01

In 2017, almost everywhere in the world, there was a boom in sales of fidget spinners. A toy that has 'multi-lobes' that can rotate freely from a thrust performed with the fingers. Thereby, we analysed the physics concepts that involved in this toy, such as angular and linear velocity. We did the analysis using the software Tracker, a laboratory…

Chutes and Ladders for the Impatient

ERIC Educational Resources Information Center

Cheteyan, Leslie A.; Hengeveld, Stewart; Jones, Michael A.

2011-01-01

In this paper, we review the rules and game board for "Chutes and Ladders", define a Markov chain to model the game regardless of the spinner range, and describe how properties of Markov chains are used to determine that an optimal spinner range of 15 minimizes the expected number of turns for a player to complete the game. Because the Markov…

A simple eccentric stirred tank mini-bioreactor: mixing characterization and mammalian cell culture experiments.

PubMed

Bulnes-Abundis, David; Carrillo-Cocom, Leydi M; Aráiz-Hernández, Diana; García-Ulloa, Alfonso; Granados-Pastor, Marisa; Sánchez-Arreola, Pamela B; Murugappan, Gayathree; Alvarez, Mario M

2013-04-01

In industrial practice, stirred tank bioreactors are the most common mammalian cell culture platform. However, research and screening protocols at the laboratory scale (i.e., 5-100 mL) rely primarily on Petri dishes, culture bottles, or Erlenmeyer flasks. There is a clear need for simple-easy to assemble, easy to use, easy to clean-cell culture mini-bioreactors for lab-scale and/or screening applications. Here, we study the mixing performance and culture adequacy of a 30 mL eccentric stirred tank mini-bioreactor. A detailed mixing characterization of the proposed bioreactor is presented. Laser induced fluorescence (LIF) experiments and computational fluid dynamics (CFD) computations are used to identify the operational conditions required for adequate mixing. Mammalian cell culture experiments were conducted with two different cell models. The specific growth rate and the maximum cell density of Chinese hamster ovary (CHO) cell cultures grown in the mini-bioreactor were comparable to those observed for 6-well culture plates, Erlenmeyer flasks, and 1 L fully instrumented bioreactors. Human hematopoietic stem cells were successfully expanded tenfold in suspension conditions using the eccentric mini-bioreactor system. Our results demonstrate good mixing performance and suggest the practicality and adequacy of the proposed mini-bioreactor. Copyright © 2012 Wiley Periodicals, Inc.

Automatic dispensing of liquid nitrogen in submilliliter doses

NASA Astrophysics Data System (ADS)

Milner, C. J.

1984-10-01

Well-controlled doses of 0.2 to 0.5 ml of liquid nitrogen are delivered, on electrical signal (not more than once per 5 s), as fills of a miniature bucket raised by an automatic hoist. The bucket is lifted, brimming, from the storage flask and then moved sideways until over the receiver. At this point, a steel ball, which has been resting in and sealing a drain hole in the bucket, is lifted from its seat by a magnet fixed alongside the (now descending) bucket. Design features are outlined: some alternative designs, valving liquid through a short drain tube fixed in the storage flask, are briefly reviewed. In tests the device delivered 74 g (approx. 260 doses) during 63 min, the loss by evaporation meanwhile being 11 g from the bucket, implying a transfer efficiency of 87%. An indirect measure indicated the dose sizes as 354±10 μl approximately.

Blood transport method for chromosome analysis of residents living near Semipalatinsk nuclear test site.

PubMed

Rodzi, Mohd; Ihda, Shozo; Yokozeki, Masako; Takeichi, Nobuo; Tanaka, Kimio; Hoshi, Masaharu

2009-12-01

A study was conducted to compare the storage conditions and transportation period for blood samples collected from residents living in areas near the Semipalatinsk nuclear test site (SNTS). Experiments were performed to simulate storage and shipping environments. Phytohaemagglutinin (PHA)-stimulated blood was stored in 15-ml tubes (condition A: current transport method) in the absence or in 50-ml flasks (condition B: previous transport method) in the presence of RPMI-1640 and 20% fetal bovine serum (FBS). Samples were kept refrigerated at 4 degrees C and cell viability was assessed after 3, 8, 12 and 14 days of storage. RPMI-1640, 20% FBS and further PHA were added to blood samples under condition A in 50-ml flasks for culture. Whole-blood samples under condition B were directly incubated without further sub-culturing process, neither media nor PHA were added, to adopt a similar protocol to that employed in the previous transport method. Samples in condition A and condition B were incubated for 48 hr at 37 degrees C and their mitotic index was determined. The results showed that viable lymphocytes were consistent in both storage conditions but the mitotic index was higher in condition A than in condition B. Although further confirmation studies have to be carried out, previous chromosomal studies and the present experiment have shown that PHA-stimulated blood could be stored without culture medium for up to 8 days under condition A. The present results will be useful for cytogenetic analysis of blood samples that have been transported long distances wherever a radiation accident has occurred.

Effect of oxygen supply on Monascus pigments and citrinin production in submerged fermentation.

PubMed

Yang, Jian; Chen, Qi; Wang, Weiping; Hu, Jiajun; Hu, Chuan

2015-05-01

The influence of oxygen supply on Monascus pigments and citrinin production by Monascus ruber HS.4000 in submerged fermentation was studied. For Monascus cultivation with high pigments and low citrinin production, the initial growth phase, mid-stage phase, and later-stage production phase were separated by shifting oxygen supply. The optimal condition for the fermentation process in shake-flask fermentation was a three-stage rotating rate controlled strategy (0-48 h at 150 rpm, 48-108 h at 250 rpm, 108-120 h at 200 rpm) with medium volume of 100 mL added to 250 mL Erlenmeyer flasks at 30°C for 120 h cultivation. Compared to constant one-stage cultivation (medium volume of 100 mL, rotating rate of 250 rpm), the pigments were reduced by 40.4%, but citrinin was reduced by 64.2%. The most appropriate condition for the fermentation process in a 10 L fermentor is also a three-stage aeration process (0-48 h at 300 L/h, 48-96 h at 500 L/h, 96-120 h at 200 L/h) with agitation of 300 rpm at 30°C for 120 h cultivation, and 237.3 ± 5.7 U/mL pigments were produced in 120 h with 6.05 ± 0.19 mg/L citrinin in a 10 L fermentor. Compared to aeration-constant (500 L/h) cultivation, pigment production was increased by 29.6% and citrinin concentration was reduced by 79.5%. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Radiochemical Separation and Quantification of Tritium in Metallic Radwastes Generated from CANDU Type NPP - 13279

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ahn, H.J.; Choi, K.C.; Choi, K.S.

2013-07-01

As a destructive quantification method of {sup 3}H in low and intermediate level radwastes, bomb oxidation, sample oxidation, and wet oxidation methods have been introduced. These methods have some merits and demerits in the radiochemical separation of {sup 3}H radionuclides. That is, since the bomb oxidation and sample oxidation methods are techniques using heating at high temperature, the separation methods of the radionuclides are relatively simple. However, since {sup 3}H radionuclide has a property of being diffused deeply into the inside of metals, {sup 3}H which is distributed on the surface of the metals can only be extracted if themore » methods are applied. As an another separation method, the wet oxidation method makes {sup 3}H oxidized with an acidic solution, and extracted completely to an oxidized HTO compound. However, incomplete oxidized {sup 3}H compounds, which are produced by reactions of acidic solutions and metallic radwastes, can be released into the air. Thus, in this study, a wet oxidation method to extract and quantify the {sup 3}H radionuclide from metallic radwastes was established. In particular, a complete extraction method and complete oxidation method of incomplete chemical compounds of {sup 3}H using a Pt catalyst were studied. The radioactivity of {sup 3}H in metallic radwastes is extracted and measured using a wet oxidation method and liquid scintillation counter. Considering the surface dose rate of the sample, the appropriate size of the sample was determined and weighed, and a mixture of oxidants was added to a 200 ml round flask with 3 tubes. The flask was quickly connected to the distilling apparatus. 20 mL of 16 wt% H{sub 2}SO{sub 4} was given into the 200-ml round flask through a dropping funnel while under stirring and refluxing. After dropping, the temperature of the mixture was raised to 96 deg. C and the sample was leached and oxidized by refluxing for 3 hours. At that time, the incomplete oxidized {sup 3}H compounds were completely oxidized using the Pt catalysts and produced a stable HTO compound. After that, about a 20 ml solution was distilled in the separation apparatus, and the distillate was mixed with an ultimagold LLT as a cocktail solution. The solution in the vial was left standing for at least 24 hours. The radioactivity of {sup 3}H was counted directly using a liquid scintillation analyzer (Packard, 2500 TR/AB, Alpha and Beta Liquid Scintillation Analyzer). (authors)« less

High-sensitivity multifunctional spinner magnetometer using a magneto-impedance sensor

NASA Astrophysics Data System (ADS)

Kodama, Kazuto

2017-01-01

A novel spinner magnetometer was developed with a wide dynamic range from 10-10 to 10-4 Am2 and a resolution of 10-11 Am2. High sensitivity was achieved with the use of a magneto-impedance (MI) sensor, which is a compact, sensitive magnetic sensor used industrially. Its slow-spinning rate (5 Hz) and the incorporation of a unique mechanism for adjusting the spacing between the sensing unit and the spinning axis allows the measurement of fragile samples sized 10-50 mm. The sensor configuration, in which a pair of MI sensors is connected in opposite serial, along with an amplification circuit with a programmable low-pass filter, reduces the problems of external noise and sensor drift. The signal, with reference to the spinning frequency, is detected with a lock-in amplifier. The MI spinner has two selectable measurement modes: the fundamental mode (F mode) and the harmonic mode (H mode). Measurements in the F mode detect signals of the fundamental frequency (5 Hz), in the same way as conventional spinner magnetometers. In the H mode, the second (10 Hz) and the third (15 Hz) harmonic components are measured, in addition to the fundamental component. Tests in the H mode were performed using a small coil and a natural sample to simulate dipoles with various degrees of offset. The results revealed that the magnitude of the fundamental component of the offset dipole was systematically larger (by several percent) than that of the nonoffset dipole. These findings suggest that this novel MI spinner will be useful in estimating the inhomogeneity of the magnetization of a sample that can equivalently be described by an offset dipole.

Emergence of reconfigurable wires and spinners via dynamic self-assembly

DOE PAGES

Kokot, Gasper; Piet, David; Whitesides, George M.; ...

2015-03-26

Dissipative colloidal materials use energy to generate and maintain structural complexity. The energy injection rate, and properties of the environment are important control parameters that influence the outcome of dynamic self-assembly. Here we demonstrate that dispersions of magnetic microparticles confined at the air-liquid interface, and energized by a uniaxial in-plane alternating magnetic field, self-assemble into a variety of structures that range from pulsating clusters and single-particle-thick wires to dynamic arrays of spinners (self-assembled short chains) rotating in either direction. The spinners emerge via spontaneous breaking of the uniaxial symmetry of the energizing magnetic field. Demonstration of the formation and disaggregationmore » of particle assemblies suggests strategies to form new meso-scale structures with the potential to perform functions such as mixing and sensing.« less

[Comparative evaluation of the functional state of the respiratory system in workers exposed to asbestos and cotton dust].

PubMed

Izycki, J; Gielec, L

1979-01-01

63 spinners exposed to cotton dust and 75 spinners exposed to asbestos dust were examined. In the women working in asbestos spinning room chronic nonspecific respiratory tract diseases prevalence was found to amount to 30% and in the cotton exposed group it was 15%. Mean values of lungs' vital capacity remained within the due values, although they were statistically significantly lower in asbestos exposed spinners (p less than 0,001). On the other hand, the percentage ratio of the forced expiratory capacity of 1 second was statistically significantly lower (0,001 less than p less than 0,01) in those exposed to cotton dust. The authors point to the occurrence of functional disturbances of the respiratory tract prior to radiological symptoms of lung asbestosis and their correlation with clinical symptoms.

DNA integrity determination in marine invertebrates by Fast Micromethod.

PubMed

Jaksić, Zeljko; Batel, Renato

2003-12-10

This study was focused toward the adaptation of the previously developed Fast Micromethod for DNA damage determination to marine invertebrates for the establishment of biomonitoring assessment. The Fast Micromethod detects DNA damage (strand breaks, alkali-labile sites and incomplete excision repair) and determines DNA integrity in cell suspensions or tissue homogenates in single microplates. The procedure is based on the ability of the specific fluorochrome dye PicoGreen to preferentially interact with high integrity DNA molecules, dsDNA, in the presence of ssDNA and proteins in high alkaline medium, thereby allowing direct fluorometric measurements of dsDNA denaturation without sample handling and stepwise DNA separations. The results presented herein describe the influence of the DNA amount and the pH of the denaturation media on slopes of the kinetic denaturation curves and calculated strand scission factors (SSFs). The optimal amount of DNA in Mytilus galloprovincialis gills homogenate was found to be 100 ng ml(-1) and the greatest differences in DNA unwinding kinetics (slopes and SSF values) were reached at pH 11.5. The induction of DNA damage and loss of DNA integrity was measured in native DNA isolated from cotton-spinner Holothuria tubulosa, marine sponge Suberites domuncula cells and mussel M. galloprovincialis gills homogenate. DNA damage and loss of DNA integrity were detected after induction by different doses of (gamma-rays, generated by 137Cs 1800 Ci; 0-500 rad in marine sponge S. domuncula cells up to SSFx(-1) values 0.082 +/- 0.012 for the highest radiation dose). Analysis by chemical xenobiotics based on the in vitro action of bleomycin (bleomycin-Fe(II) complex 0-50 or 0-83 microg ml(-1) (microM)) with native DNA from cotton-spinner H. tubulosa and mussel M. galloprovincialis gills homogenate yielded values of 0.537 +/- 0.072 and 0.130 +/- 0.018, respectively. In vivo experiments with mussel M. galloprovincialis gills homogenate by 4-nitroquinoline-N-oxide (NQO; 0-1 microg g(-1) NQO mussel) and benzo[a]pyrene (B[a]P; 0-20 microg g(-1) B[a]P mussel) indicated SSFx(-1) values of 0.121 +/- 0.016 and 0.090 +/- 0.007, respectively, for the highest applied doses of chemical xenobiotics. The analytical technique described here allows simple and fast analysis of DNA integrity, requires very short time for multiple analyses (less than 3 h) and even less than 100 ng DNA per single well (50 ng DNA isolated from cotton-spinner, 12,500 sponge cells or about 10 mg of mussel gills homogenate) in a microplate. This makes the Fast Micromethod applicable for the measurement of DNA integrity of small samples for genotoxicity assessment (biomonitoring), the effects of genotoxins on lower marine taxa or sessile invertebrates in marine environment (e.g. sponges, mussels) and the estimation of directional changes and harmful effects in the ecosystem.

Production of coagulation factor VII in human cell lines Sk-Hep-1 and HKB-11.

PubMed

Corrêa de Freitas, Marcela Cristina; Bomfim, Aline de Sousa; Mizukami, Amanda; Picanço-Castro, Virgínia; Swiech, Kamilla; Covas, Dimas Tadeu

2017-09-01

Recombinant factor VII (rFVII) is the main therapeutic choice for hemophilia patients who have developed inhibitory antibodies against conventional treatments (FVIII and FIX). Because of the post-translational modifications, rFVII needs to be produced in mammalian cell lines. In this study, for the first time, we have shown efficient rFVII production in HepG2, Sk-Hep-1, and HKB-11 cell lines. Experiments in static conditions for a period of 96 h showed that HepG2-FVII produced the highest amounts of rhFVII, with an average of 1843 ng/mL. Sk-hep-1-FVII cells reached a maximum protein production of 1432 ng/mL and HKB-11-FVII cells reached 1468 ng/mL. Sk-Hep-1-rFVII and HKB-11-rFVII were selected for the first step of scale-up. Over 10 days of spinner flask culture, HKB-11 and SK-Hep-1 cells showed a cumulative production of rFVII of 152 μg and 202.6 μg in 50 mL, respectively. Thus, these human cell lines can be used for an efficient production of recombinant FVII. With more investment in basic research, human cell lines can be optimized for the commercial production of different bio therapeutic proteins. Copyright © 2017 Elsevier Inc. All rights reserved.

Novel strategy to engineer trachea cartilage graft with marrow mesenchymal stem cell macroaggregate and hydrolyzable scaffold.

PubMed

Liu, Liangqi; Wu, Wei; Tuo, Xiaoye; Geng, Wenxin; Zhao, Jie; Wei, Jing; Yan, Xingrong; Yang, Wei; Li, Liwen; Chen, Fulin

2010-05-01

Limited donor sites of cartilage and dedifferentiation of chondrocytes during expansion, low tissue reconstruction efficiency, and uncontrollable immune reactions to foreign materials are the main obstacles to overcome before cartilage tissue engineering can be widely used in the clinic. In the current study, we developed a novel strategy to fabricate tissue-engineered trachea cartilage grafts using marrow mesenchymal stem cell (MSC) macroaggregates and hydrolyzable scaffold of polylactic acid-polyglycolic acid copolymer (PLGA). Rabbit MSCs were continuously cultured to prepare macroaggregates in sheet form. The macroaggregates were studied for their potential for chondrogenesis. The macroaggregates were wrapped against the PLGA scaffold to make a tubular composite. The composites were incubated in spinner flasks for 4 weeks to fabricate trachea cartilage grafts. Histological observation and polymerase chain reaction array showed that MSC macroaggregates could obtain the optimal chondrogenic capacity under the induction of transforming growth factor-beta. Engineered trachea cartilage consisted of evenly spaced lacunae embedded in a matrix rich in proteoglycans. PLGA scaffold degraded totally during in vitro incubation and the engineered cartilage graft was composed of autologous tissue. Based on this novel, MSC macroaggregate and hydrolyzable scaffold composite strategy, ready-to-implant autologous trachea cartilage grafts could be successfully fabricated. The strategy also had the advantages of high efficiency in cell seeding and tissue regeneration, and could possibly be used in future in vivo experiments.

Biofabrication of tissue constructs by 3D bioprinting of cell-laden microcarriers.

PubMed

Levato, Riccardo; Visser, Jetze; Planell, Josep A; Engel, Elisabeth; Malda, Jos; Mateos-Timoneda, Miguel A

2014-09-01

Bioprinting allows the fabrication of living constructs with custom-made architectures by spatially controlled deposition of multiple bioinks. This is important for the generation of tissue, such as osteochondral tissue, which displays a zonal composition in the cartilage domain supported by the underlying subchondral bone. Challenges in fabricating functional grafts of clinically relevant size include the incorporation of cues to guide specific cell differentiation and the generation of sufficient cells, which is hard to obtain with conventional cell culture techniques. A novel strategy to address these demands is to combine bioprinting with microcarrier technology. This technology allows for the extensive expansion of cells, while they form multi-cellular aggregates, and their phenotype can be controlled. In this work, living constructs were fabricated via bioprinting of cell-laden microcarriers. Mesenchymal stromal cell (MSC)-laden polylactic acid microcarriers, obtained via static culture or spinner flask expansion, were encapsulated in gelatin methacrylamide-gellan gum bioinks, and the printability of the composite material was studied. This bioprinting approach allowed for the fabrication of constructs with high cell concentration and viability. Microcarrier encapsulation improved the compressive modulus of the hydrogel constructs, facilitated cell adhesion, and supported osteogenic differentiation and bone matrix deposition by MSCs. Bilayered osteochondral models were fabricated using microcarrier-laden bioink for the bone compartment. These findings underscore the potential of this new microcarrier-based biofabrication approach for bone and osteochondral constructs.

Evaluation of secretome of highly efficient lignocellulolytic Penicillium sp. Dal 5 isolated from rhizosphere of conifers.

PubMed

Rai, Rohit; Kaur, Baljit; Singh, Surender; Di Falco, Macros; Tsang, Adrian; Chadha, B S

2016-09-01

Penicillium sp. (Dal 5) isolated from rhizosphere of conifers from Dalhousie (Himachal Pradesh, India) was found to be an efficient cellulolytic strain. The culture under shake flask on CWR (cellulose, wheat bran and rice straw) medium produced appreciably higher levels of endoglucanase (35.69U/ml), β-glucosidase (4.20U/ml), cellobiohydrolase (2.86U/ml), FPase (1.2U/ml) and xylanase (115U/ml) compared to other Penicillium strains reported in literature. The mass spectroscopy analysis of Penicillium sp. Dal 5 secretome identified 108 proteins constituting an array of CAZymes including glycosyl hydrolases (GH) belonging to 24 different families, polysaccharide lyases (PL), carbohydrate esterases (CE), lytic polysaccharide mono-oxygenases (LPMO) in addition to swollenin and a variety of carbohydrate binding modules (CBM) indicating an elaborate genetic potential of this strain for hydrolysis of lignocellulosics. Further, the culture extract was evaluated for hydrolysis of alkali treated rice straw, wheat straw, bagasse and corn cob at 10% substrate loading rate. Copyright © 2016 Elsevier Ltd. All rights reserved.

Methodology for enabling high-throughput simultaneous saccharification and fermentation screening of yeast using solid biomass as a substrate.

PubMed

Elliston, Adam; Wood, Ian P; Soucouri, Marie J; Tantale, Rachelle J; Dicks, Jo; Roberts, Ian N; Waldron, Keith W

2015-01-01

High-throughput (HTP) screening is becoming an increasingly useful tool for collating biological data which would otherwise require the employment of excessive resources. Second generation biofuel production is one such process. HTP screening allows the investigation of large sample sets to be undertaken with increased speed and cost effectiveness. This paper outlines a methodology that will enable solid lignocellulosic substrates to be hydrolyzed and fermented at a 96-well plate scale, facilitating HTP screening of ethanol production, whilst maintaining repeatability similar to that achieved at a larger scale. The results showed that utilizing sheets of biomass of consistent density (handbills), for paper, and slurries of pretreated biomass that could be pipetted allowed standardized and accurate transfers to 96-well plates to be achieved (±3.1 and 1.7%, respectively). Processing these substrates by simultaneous saccharification and fermentation (SSF) at various volumes showed no significant difference on final ethanol yields, either at standard shake flask (200 mL), universal bottle (10 mL) or 96-well plate (1 mL) scales. Substrate concentrations of up to 10% (w/v) were trialed successfully for SSFs at 1 mL volume. The methodology was successfully tested by showing the effects of steam explosion pretreatment on both oilseed rape and wheat straws. This methodology could be used to replace large shake flask reactions with comparatively fast 96-well plate SSF assays allowing for HTP experimentation. Additionally this method is compatible with a number of standardized assay techniques such as simple colorimetric, High-performance liquid chromatography (HPLC) and Nuclear magnetic resonance (NMR) spectroscopy. Furthermore this research has practical uses in the biorefining of biomass substrates for second generation biofuels and novel biobased chemicals by allowing HTP SSF screening, which should allow selected samples to be scaled up or studied in more detail.

Nanoscale Imaging with a Single Quantum Dot

DTIC Science & Technology

2011-12-19

mL round-bottom flask equipped with a condenser , a thermometer and a magnetic stirring bar. After the EG was heated to 160 °C in an oil bath, 0.5 mL...radiatively transferred to the wire’s SPP mode through an electric dipole interaction19. The efficiency of this process scales as the spontaneous emission...Liu, J., Gao, D., Li, H.-F. & Lin, J.-M. Controlled photopolymerization of hydrogel microstructures inside microchannels for bioassays. Lab. Chip 9

Adsorption of Crystal Violet Dye Using Zeolite A Synthesized From Coal Fly Ash

NASA Astrophysics Data System (ADS)

Jumaeri; Kusumastuti, E.; Santosa, S. J.; Sutarno

2017-02-01

Adsorption of Crystal Violet (CV) dye using zeolite A synthesized from coal fly ash (ZA) has been done. Effect of pH, contact time, and the initial concentration of dye adsorption was studied in this adsorption. Model experimental of adsorption isotherms and adsorption kinetics were also studied. The adsorption is done in a batch reactor at room temperature. A total of 0.01 g of zeolite A was added to the Erlenmeyer flask 50 mL containing 20 mL of the dye solution of Crystal Violet in a variety of conditions of pH, contact time and initial concentration. Furthermore, Erlenmeyer flask and its contents were shaken using an orbital shaker at a speed of 200 rpm. After a specified period of adsorption, the solution was centrifuged for 2 minutes so that the solids separated from the solution. The concentration of the dye after adsorption determined using Genesis-20 Spectrophotometer. The results showed that the Zeolite A synthesized from coal fly ash could be used as an effective adsorbent for Crystal Violet dye. The optimum adsorption occurs at pH 6, and contact time 45 minutes. At the initial concentration of 2 to 6 mg/L, adsorption is reduced from 79 to 62.8%. Crystal Violet dye adsorption in zeolite A fulfilled kinetic model of pseudo-order 2 and model of Freundlich adsorption isotherm.

Comparison of occupational exposure to carbon disulphide in a viscose rayon factory before and after technical adjustments.

PubMed

Bulat, Petar; Daemen, Edgard; Van Risseghem, Marleen; De Bacquer, Dirk; Tan, Xiaodong; Braeckman, Lutgart; Vanhoorne, Michel

2002-01-01

The objective of this follow-up study was to verify the efficacy of the technical adjustments gradually introduced in departments of a viscose rayon factory from 1989 onward. Personal exposure to carbon disulphide was assessed by means of personal monitoring through active sampling. Six job titles in three departments of the factory were sampled. Geometric means were calculated and used as estimates of time-weighted average (TWA) concentrations. The results from the present study were compared with similar measurements from a previous study in the same factory. Due to organizational changes, only three job titles (spinner, first spinner, and viscose preparator) could be compared directly. Two new job titles were identified, although tasks performed in these two job titles already existed. The measurements from one job title could not be compared, due to a substantial reorganization and automation of the tasks carried out in the department. The comparison before and after technical improvements shows that personal exposure of spinner and first spinner has been substantially reduced. Even the geometric means of measurements outside the fresh air mask are below the TWA-TLV (Threshold Limit Value). Despite the difficulties in comparing the results from the two studies, it is concluded that the technical measures reduced up to tenfold personal exposure to carbon disulphide and personal protection reduced it further by a factor two.

Tests of Five Full-Scale Propellers in the Presence of a Radial and a Liquid-Cooled Engine Nacelle, Including Tests of Two Spinners

NASA Technical Reports Server (NTRS)

Biermann, David; Hartman, Edwin P

1938-01-01

Wind-tunnel tests are reported of five 3-blade 10-foot propellers operating in front of a radial and a liquid-cooled engine nacelle. The range of blade angles investigated extended from 15 degrees to 45 degrees. Two spinners were tested in conjunction with the liquid-cooled engine nacelle. Comparisons are made between propellers having different blade-shank shapes, blades of different thickness, and different airfoil sections. The results show that propellers operating in front of the liquid-cooled engine nacelle had higher take-off efficiencies than when operating in front of the radial engine nacelle; the peak efficiency was higher only when spinners were employed. One spinner increased the propulsive efficiency of the liquid-cooled unit 6 percent for the highest blade-angle setting investigated and less for lower blade angles. The propeller having airfoil sections extending into the hub was superior to one having round blade shanks. The thick propeller having a Clark y section had a higher take-off efficiency than the thinner one, but its maximum efficiency was possibly lower. Of the three blade sections tested, Clark y, R.A.F. 6, and NACA 2400-34, the Clark y was superior for the high-speed condition, but the R.A.F. 6 excelled for the take-off condition.

Pigeon pea waste as a novel, inexpensive, substrate for production of a thermostable alkaline protease from thermoalkalophilic Bacillus sp. JB-99.

PubMed

Johnvesly, B; Manjunath, B R; Naik, G R

2002-03-01

Thermoalkaliphilic Bacillus sp. JB-99 was grown in a 250 ml Erlenmeyer flask containing 50 ml medium containing (g/l) Pigeon pea waste 10; NaNO3, 5.0; K2HPO4, 5.0; MgSO4 x 2H2O, 0.2 and Na2CO3, 10.0. Incubations were carried out at 50 degrees C on a rotary incubator shaker for 15 h. A high level of extra cellular thermostable protease activity was observed after 24 h incubation. The optimum temperature and pH for activity were 70 degrees C and 11, respectively, so this enzyme showed stable activity at high temperature and under alkaline conditions.

Culture medium optimization for acetic acid production by a persimmon vinegar-derived bacterium.

PubMed

Kim, Jin-Nam; Choo, Jong-Sok; Wee, Young-Jung; Yun, Jong-Sun; Ryu, Hwa-Won

2005-01-01

A new acetic acid-producing microorganism, Acetobacter sp. RKY4, was isolated from Korean traditional persimmon vinegar, and we optimized the culture medium for acetic acid production from ethanol using the newly isolated Acetobacter sp. RKY4. The optimized culture medium for acetic acid production using this microorganism was found to be 40 g/L ethanol, 10 g/L glycerol, 10 g/L corn steep liquor, 0.5 g/L MgSO4.7H2O, and 1.0 g/L (NH4)H2PO4. Acetobacter sp. RKY4 produced 47.1 g/L of acetic acid after 48 h of fermentation in a 250 mL Erlenmeyer flask containing 50 mL of the optimized medium.

An effective system to produce smoke solutions from dried plant tissue for seed germination studies1

PubMed Central

Coons, Janice; Coutant, Nancy; Lawrence, Barbara; Finn, Daniel; Finn, Stephanie

2014-01-01

• Premise of the study: An efficient and inexpensive system was developed to produce smoke solutions from plant material to research the influence of water-soluble compounds from smoke on seed germination. • Methods and Results: Smoke solutions (300 mL per batch) were produced by burning small quantities (100–200 g) of dried plant material from a range of species in a bee smoker attached by a heater hose to a side-arm flask. The flask was attached to a vacuum water aspirator, to pull the smoke through the water. The entire apparatus was operated in a laboratory fume hood. • Conclusions: Compared with other smoke solution preparation systems, the system described is easy to assemble and operate, inexpensive to build, and effective at producing smoke solutions from desired species in a small indoor space. Quantitative measurements can be made when using this system, allowing for replication of the process. PMID:25202613

Effect of simulated microgravity and shear stress on microcin B17 production by Escherichia coli and on its excretion into the medium

NASA Technical Reports Server (NTRS)

Fang, A.; Pierson, D. L.; Koenig, D. W.; Mishra, S. K.; Demain, A. L.

1997-01-01

Production of the antibacterial polypeptide microcin B17 (MccB17) by Escherichia coli ZK650 was inhibited by simulated microgravity. The site of MccB17 accumulation was found to be different, depending on whether the organism was grown in shaking flasks or in rotating bioreactors designed to establish a simulated microgravity environment. In flasks, the accumulation was cellular, but in the reactors, virtually all the microcin was found in the medium. The change from a cellular site to an extracellular one was apparently not a function of gravity, since extracellular production occurred in these bioreactors, irrespective of whether they were operated in the simulated microgravity or normal gravity mode. More probably, excretion is due to the much lower degree of shear stress in the bioreactors. Addition of even a single glass bead to the 50-ml medium volume in the bioreactor created enough shear to change the site of MccB17 accumulation from the medium to the cells.

Impact of carbon and nitrogen feeding strategy on high production of biomass and docosahexaenoic acid (DHA) by Schizochytrium sp. LU310.

PubMed

Ling, Xueping; Guo, Jing; Liu, Xiaoting; Zhang, Xia; Wang, Nan; Lu, Yinghua; Ng, I-Son

2015-05-01

A new isolated Schizochytrium sp. LU310 from the mangrove forest of Wenzhou, China, was found as a high producing microalga of docosahexaenoic acid (DHA). In this study, the significant improvements for DHA fermentation by the batch mode in the baffled flasks (i.e. higher oxygen supply) were achieved. By applied the nitrogen-feeding strategy in 1000 mL baffled flasks, the biomass, DHA concentration and DHA productivity were increased by 110.4%, 117.9% and 110.4%, respectively. Moreover, DHA concentration of 21.06 g/L was obtained by feeding 15 g/L of glucose intermittently, which was an increase of 41.25% over that of the batch mode. Finally, an innovative strategy was carried out by intermittent feeding carbon and simultaneously feeding nitrogen. The maximum DHA concentration and DHA productivity in the fed-batch cultivation reached to 24.74 g/L and 241.5 mg/L/h, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

A new spinner magnetometer using high sensitivity magneto-impedance sensor

NASA Astrophysics Data System (ADS)

Kodama, Kazuto

2016-04-01

A sensitive spinner magnetometer was developed using a pair of high-resolution Magneto-Impedance sensors. The MI sensor generally utilizes the MI effect of amorphous wire whose impedance changes by the application of a small magnetic field. Various kinds of MI sensors are currently used in many electric devices, for example, a magnetic compass chip built-in smart phones and car navigations. The MI sensor employed in this study is a pico-Tesla MI sensor, an especially sensitive MI sensor originally manufactured for industrial use to detect contamination of small magnetic particles in industrial materials such as fabrics. To detect weak magnetic signals from natural samples and avoid DC drift, a gradiometer system was employed that consists of a pair of the MI sensors and the electronics with analog filter and pre-amplification circuit. This MI gradiometer system was equipped to a commercial spinner magnetometer (SMD-88, Natsuhara Giken, Osaka) with the spinning rate of 5 Hz. It is demonstrated that this new spinner magnetometer is capable of measuring weak magnetic samples of 10-6 mAm2, with the highest resolution being 10-8 mAm2, approximately two orders of magnitude better than the previous one using a ring-core flux-gate sensor. One of the advantages of the MI spinner magnetometer is that it can be easily modified to accommodate samples of any shape and size. Moreover the slow-rotating speed (5 Hz) allows to measure samples for archeomagnetic studies that are usually irregular and fragile. Because the irregularity of shape increases errors in measuring the dipole component of the total magnetization, it is necessary to increase the distance between the sample and sensor, resulting in poorer sensitivity. The high-sensitivity MI sensor enables to measure the NRM of such irregular-shaped samples from an appropriate distance to the sample, with no significant loss of sensitivity.

Orally Administered Bioadherent Sustained Release Microencapsulated Vaccines

DTIC Science & Technology

1996-09-01

ml flask culture (MMY media) was inoculated with a recombinant Pichia yeast clone containing the vpB gene integrated at the alcohol oxidase (AOX...measured. 6 DAMD17-95-C-5099 2. Materials and Methods 2.1. Materials vpB is expressed in Pichia yeast and isolated and purified as described below. SEA...locus in the Pichia genome. The culture was grown at 30"C for approximately 48 hours until an optical density ( OD 650) of 3.0 was reached. The culture

Feasibility of Biomonitoring of Exposure to Permethrin Through Analysis of Long-Lived (Metabolite) Adducts to Proteins

DTIC Science & Technology

2007-09-01

Preparation of the Grignard reagent To flask E, containing magnesium (52 mg, 2 mmol) and a few crystals of iodine was added a solution of 3...bromodiphenylether (750 mg, 3 mmol) in diethyl ether (3 mL). The entire system was kept under nitrogen. The preparation of the Grignard reagent was initiated by...device represented below was used for the multi-step synthesis of 3-PBA; all reagents were present in the manifold prior to the beginning of the

Self contained, independent, in-vacuum spinner motor

DOEpatents

Ayers, Marion J.

2002-01-01

An independent, self contained apparatus for operation within a vacuum chamber. A sealed enclosure is located in the chamber. The enclosure contains its own atmosphere independent of the vacuum in the chamber. A motor, power unit, and controls are located entirely within the enclosure. They do not have a direct structural connection outside of the enclosure in any way that would effect the atmosphere within the enclosure. The motor, power unit, and controls drive a spinner plate located outside the enclosure but within the vacuum chamber.

[Eyeball perforation of a 7-year-old girl caused by the "Must-Have Office Toy 2017" : The potential risk of injury by a spinning toy (fidget spinner)].

PubMed

Nessmann, A; Chaloupka, K; Böni, C

2018-03-01

A 7-year-old girl presented to the emergency department of the eye clinic with an eyeball perforation with iris prolapse and corneal as well as scleral wounds of the left eye caused by a spinning toy (fidget spinner). A surgical wound inspection with iris reposition and corneal and scleral suture were performed without delay. This case report demonstrates the potential risk of this popular toy.

The broadband social acoustic signaling behavior of spinner and spotted dolphins

NASA Astrophysics Data System (ADS)

Lammers, Marc O.; Au, Whitlow W. L.; Herzing, Denise L.

2003-09-01

Efforts to study the social acoustic signaling behavior of delphinids have traditionally been restricted to audio-range (<20 kHz) analyses. To explore the occurrence of communication signals at ultrasonic frequencies, broadband recordings of whistles and burst pulses were obtained from two commonly studied species of delphinids, the Hawaiian spinner dolphin (Stenella longirostris) and the Atlantic spotted dolphin (Stenella frontalis). Signals were quantitatively analyzed to establish their full bandwidth, to identify distinguishing characteristics between each species, and to determine how often they occur beyond the range of human hearing. Fundamental whistle contours were found to extend beyond 20 kHz only rarely among spotted dolphins, but with some regularity in spinner dolphins. Harmonics were present in the majority of whistles and varied considerably in their number, occurrence, and amplitude. Many whistles had harmonics that extended past 50 kHz and some reached as high as 100 kHz. The relative amplitude of harmonics and the high hearing sensitivity of dolphins to equivalent frequencies suggest that harmonics are biologically relevant spectral features. The burst pulses of both species were found to be predominantly ultrasonic, often with little or no energy below 20 kHz. The findings presented reveal that the social signals produced by spinner and spotted dolphins span the full range of their hearing sensitivity, are spectrally quite varied, and in the case of burst pulses are probably produced more frequently than reported by audio-range analyses.

The broadband social acoustic signaling behavior of spinner and spotted dolphins.

PubMed

Lammers, Marc O; Au, Whitlow W L; Herzing, Denise L

2003-09-01

Efforts to study the social acoustic signaling behavior of delphinids have traditionally been restricted to audio-range (<20 kHz) analyses. To explore the occurrence of communication signals at ultrasonic frequencies, broadband recordings of whistles and burst pulses were obtained from two commonly studied species of delphinids, the Hawaiian spinner dolphin (Stenella longirostris) and the Atlantic spotted dolphin (Stenella frontalis). Signals were quantitatively analyzed to establish their full bandwidth, to identify distinguishing characteristics between each species, and to determine how often they occur beyond the range of human hearing. Fundamental whistle contours were found to extend beyond 20 kHz only rarely among spotted dolphins, but with some regularity in spinner dolphins. Harmonics were present in the majority of whistles and varied considerably in their number, occurrence, and amplitude. Many whistles had harmonics that extended past 50 kHz and some reached as high as 100 kHz. The relative amplitude of harmonics and the high hearing sensitivity of dolphins to equivalent frequencies suggest that harmonics are biologically relevant spectral features. The burst pulses of both species were found to be predominantly ultrasonic, often with little or no energy below 20 kHz. The findings presented reveal that the social signals produced by spinner and spotted dolphins span the full range of their hearing sensitivity, are spectrally quite varied, and in the case of burst pulses are probably produced more frequently than reported by audio-range analyses.

A comparison of bioreactors for culture of fetal mesenchymal stem cells for bone tissue engineering.

PubMed

Zhang, Zhi-Yong; Teoh, Swee Hin; Teo, Erin Yiling; Khoon Chong, Mark Seow; Shin, Chong Woon; Tien, Foo Toon; Choolani, Mahesh A; Chan, Jerry K Y

2010-11-01

Bioreactors provide a dynamic culture system for efficient exchange of nutrients and mechanical stimulus necessary for the generation of effective tissue engineered bone grafts (TEBG). We have shown that biaxial rotating (BXR) bioreactor-matured human fetal mesenchymal stem cell (hfMSC) mediated-TEBG can heal a rat critical sized femoral defect. However, it is not known whether optimal bioreactors exist for bone TE (BTE) applications. We systematically compared this BXR bioreactor with three most commonly used systems: Spinner Flask (SF), Perfusion and Rotating Wall Vessel (RWV) bioreactors, for their application in BTE. The BXR bioreactor achieved higher levels of cellularity and confluence (1.4-2.5x, p < 0.05) in large 785 mm(3) macroporous scaffolds not achieved in the other bioreactors operating in optimal settings. BXR bioreactor-treated scaffolds experienced earlier and more robust osteogenic differentiation on von Kossa staining, ALP induction (1.2-1.6×, p < 0.01) and calcium deposition (1.3-2.3×, p < 0.01). We developed a Micro CT quantification method which demonstrated homogenous distribution of hfMSC in BXR bioreactor-treated grafts, but not with the other three. BXR bioreactor enabled superior cellular proliferation, spatial distribution and osteogenic induction of hfMSC over other commonly used bioreactors. In addition, we developed and validated a non-invasive quantitative micro CT-based technique for analyzing neo-tissue formation and its spatial distribution within scaffolds. Copyright © 2010 Elsevier Ltd. All rights reserved.

Optimization of trehalose production by a novel strain Brevibacterium sp. SY361.

PubMed

Wang, Lei; Huang, Rui; Gu, Guanbin; Fang, Hongying

2008-10-01

Trehalose production by a novel strain of Brevibacterium sp. SY361 was optimized in submerged fermentation. Different chemical and physical parameters such as carbon and nitrogen sources, inoculum level, initial pH, incubation temperature, aeration and time-course of fermentation, were studied in order to increase trehalose productivity. An optimal production medium containing 3% (w/v) glucose, 0.9% (v/v) corn steep liquor, 0.5% (w/v) KH(2)PO(4) and 0.4% (w/v) MgSO(4).7 H(2)O was found suitable for trehalose production. An optimal volume of medium in a 500 ml flask was 80 ml. The optimal levels of other parameters were 4.0% (v/v) of inoculum, initial pH of 6.0, incubation temperature of 28-32 degrees C and time-course of 60 h. Optimized parameters gave a maximum trehalose of 12.2 mg/ml with a conversion rate of 58.4%. (c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Enhanced production of alkaline thermostable keratinolytic protease from calcium alginate immobilized cells of thermoalkalophilic Bacillus halodurans JB 99 exhibiting dehairing activity.

PubMed

Shrinivas, Dengeti; Kumar, Raghwendra; Naik, G R

2012-01-01

The thermoalkalophilic Bacillus halodurans JB 99 cells known for production of novel thermostable alkaline keratinolytic protease were immobilized in calcium alginate matrix. Batch and repeated batch cultivation using calcium alginate immobilized cells were studied for alkaline protease production in submerged fermentation. Immobilized cells with 2.5% alginate and 350 beads/flask of initial cell loading showed enhanced production of alkaline protease by 23.2% (5,275 ± 39.4 U/ml) as compared to free cells (4,280 ± 35.4 U/ml) after 24 h. In the semicontinuous mode of cultivation, immobilized cells under optimized conditions produced an appreciable level of alkaline protease in up to nine cycles and reached a maximal value of 5,975 U/ml after the seventh cycle. The enzyme produced from immobilized cells efficiently degraded chicken feathers in the presence of a reducing agent which can help the poultry industry in the management of keratin-rich waste and obtaining value-added products.

Soundscape Ecology of Hawaiian Spinner Dolphin Resting Bays

NASA Astrophysics Data System (ADS)

Heenehan, Heather Leigh

Sound is a key sensory modality for Hawaiian spinner dolphins. Like many other marine animals, these dolphins rely on sound and their acoustic environment for many aspects of their daily lives, making it is essential to understand soundscape in areas that are critical to their survival. Hawaiian spinner dolphins rest during the day in shallow coastal areas and forage offshore at night. In my dissertation I focus on the soundscape of the bays where Hawaiian spinner dolphins rest taking a soundscape ecology approach. I primarily relied on passive acoustic monitoring using four DSG-Ocean acoustic loggers in four Hawaiian spinner dolphin resting bays on the Kona Coast of Hawai'i Island. 30-second recordings were made every four minutes in each of the bays for 20 to 27 months between January 8, 2011 and March 30, 2013. I also utilized concomitant vessel-based visual surveys in the four bays to provide context for these recordings. In my first chapter I used the contributions of the dolphins to the soundscape to monitor presence in the bays and found the degree of presence varied greatly from less than 40% to nearly 90% of days monitored with dolphins present. Having established these bays as important to the animals, in my second chapter I explored the many components of their resting bay soundscape and evaluated the influence of natural and human events on the soundscape. I characterized the overall soundscape in each of the four bays, used the tsunami event of March 2011 to approximate a natural soundscape and identified all loud daytime outliers. Overall, sound levels were consistently louder at night and quieter during the daytime due to the sounds from snapping shrimp. In fact, peak Hawaiian spinner dolphin resting time co-occurs with the quietest part of the day. However, I also found that humans drastically alter this daytime soundscape with sound from offshore aquaculture, vessel sound and military mid-frequency active sonar. During one recorded mid-frequency active sonar event in August 2011, sound pressure levels in the 3.15 kHz 1/3 rd-octave band were as high as 45.8 dB above median ambient noise levels. Human activity both inside (vessels) and outside (sonar and aquaculture) the bays significantly altered the resting bay soundscape. Inside the bays there are high levels of human activity including vessel-based tourism directly targeting the dolphins. The interactions between humans and dolphins in their resting bays are of concern; therefore, my third chapter aimed to assess the acoustic response of the dolphins to human activity. Using days where acoustic recordings overlapped with visual surveys I found the greatest response in a bay with dolphin-centric activities, not in the bay with the most vessel activity, indicating that it is not the magnitude that elicits a response but the focus of the activity. In my fourth chapter I summarize the key results from my first three chapters to illustrate the power of multiple site design to prioritize action to protect Hawaiian spinner dolphins in their resting bays, a chapter I hope will be useful for managers should they take further action to protect the dolphins.

Organo-Metallic Elements for Associative Information Processing

DTIC Science & Technology

1989-01-15

7844 400 MHz oscilloscope Hamamatsu Model B2297-3 Ge photodiodes Scientech Model 365 power meter Miscellaneous optical components (i.e., harmonic...dried in vacuo at 80 °C for 3 hours to give a tan colored powder 234.2 g, yield I 84%, mp > 400 oC. IR(KBr) 1787, 1733, 1683, 1621, 1557, 1467, 1375...flask was cooled in an ice-water bath for 3 hours and the solid was collected on a filter and washed thoroughly with water 2 x 400 mL and ethanol 3 x

The conversion of BTEX compounds by single and defined mixed cultures to medium-chain-length polyhydroxyalkanoate.

PubMed

Nikodinovic, Jasmina; Kenny, Shane T; Babu, Ramesh P; Woods, Trevor; Blau, Werner J; O'Connor, Kevin E

2008-09-01

Here, we report the use of petrochemical aromatic hydrocarbons as a feedstock for the biotechnological conversion into valuable biodegradable plastic polymers--polyhydroxyalkanoates (PHAs). We assessed the ability of the known Pseudomonas putida species that are able to utilize benzene, toluene, ethylbenzene, p-xylene (BTEX) compounds as a sole carbon and energy source for their ability to produce PHA from the single substrates. P. putida F1 is able to accumulate medium-chain-length (mcl) PHA when supplied with toluene, benzene, or ethylbenzene. P. putida mt-2 accumulates mcl-PHA when supplied with toluene or p-xylene. The highest level of PHA accumulated by cultures in shake flask was 26% cell dry weight for P. putida mt-2 supplied with p-xylene. A synthetic mixture of benzene, toluene, ethylbenzene, p-xylene, and styrene (BTEXS) which mimics the aromatic fraction of mixed plastic pyrolysis oil was supplied to a defined mixed culture of P. putida F1, mt-2, and CA-3 in the shake flasks and fermentation experiments. PHA was accumulated to 24% and to 36% of the cell dry weight of the shake flask and fermentation grown cultures respectively. In addition a three-fold higher cell density was achieved with the mixed culture grown in the bioreactor compared to shake flask experiments. A run in the 5-l fermentor resulted in the utilization of 59.6 g (67.5 ml) of the BTEXS mixture and the production of 6 g of mcl-PHA. The monomer composition of PHA accumulated by the mixed culture was the same as that accumulated by single strains supplied with single substrates with 3-hydroxydecanoic acid occurring as the predominant monomer. The purified polymer was partially crystalline with an average molecular weight of 86.9 kDa. It has a thermal degradation temperature of 350 degrees C and a glass transition temperature of -48.5 degrees C.

Expansion in microcarrier-spinner cultures improves the chondrogenic potential of human early mesenchymal stromal cells.

PubMed

Lin, Youshan Melissa; Lim, Jessica Fang Yan; Lee, Jialing; Choolani, Mahesh; Chan, Jerry Kok Yen; Reuveny, Shaul; Oh, Steve Kah Weng

2016-06-01

Cartilage tissue engineering with human mesenchymal stromal cells (hMSC) is promising for allogeneic cell therapy. To achieve large-scale hMSC propagation, scalable microcarrier-based cultures are preferred over conventional static cultures on tissue culture plastic. Yet it remains unclear how microcarrier cultures affect hMSC chondrogenic potential, and how this potential is distinguished from that of tissue culture plastic. Hence, our study aims to compare the chondrogenic potential of human early MSC (heMSC) between microcarrier-spinner and tissue culture plastic cultures. heMSC expanded on either collagen-coated Cytodex 3 microcarriers in spinner cultures or tissue culture plastic were harvested for chondrogenic pellet differentiation with empirically determined chondrogenic inducer bone morphogenetic protein 2 (BMP2). Pellet diameter, DNA content, glycosaminoglycan (GAG) and collagen II production, histological staining and gene expression of chondrogenic markers including SOX9, S100β, MMP13 and ALPL, were investigated and compared in both conditions. BMP2 was the most effective chondrogenic inducer for heMSC. Chondrogenic pellets generated from microcarrier cultures developed larger pellet diameters, and produced more DNA, GAG and collagen II per pellet with greater GAG/DNA and collagen II/DNA ratios compared with that of tissue culture plastic. Moreover, they induced higher expression of chondrogenic genes (e.g., S100β) but not of hypertrophic genes (e.g., MMP13 and ALPL). A similar trend showing enhanced chondrogenic potential was achieved with another microcarrier type, suggesting that the mechanism is due to the agitated nature of microcarrier cultures. This is the first study demonstrating that scalable microcarrier-spinner cultures enhance the chondrogenic potential of heMSC, supporting their use for large-scale cell expansion in cartilage cell therapy. Copyright © 2016 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Muscle biochemistry of a pelagic delphinid (Stenella longirostris longirostris): insight into fishery-induced separation of mothers and calves.

PubMed

Noren, Shawn R; West, Kristi

2017-04-15

The length of time required for postnatal maturation of the locomotor muscle (longissimus dorsi) biochemistry [myoglobin (Mb) content and buffering capacity] in marine mammals typically varies with nursing duration, but it can be accelerated by species-specific behavioral demands, such as deep-diving and sub-ice transit. We examined how the swimming demands of a pelagic lifestyle influence postnatal maturation of Mb and buffering capacity in spinner dolphins ( Stenella longirostris longirostris ). Mb content of newborn (1.16±0.07 g Mb per 100 g wet muscle mass, n =6) and juvenile (2.77±0.22 g per 100 g, n =4) spinner dolphins were only 19% and 46% of adult levels (6.00±0.74 g per 100 g, n =6), respectively. At birth, buffering capacity was 52.70±4.48 slykes ( n =6) and increased to 78.53±1.91 slykes ( n =6) once a body length of 141 cm was achieved, representing 1.6- to 2.0-year-old dolphins. Based on the age of weaning (1.3-1.6 years post-partum), muscle maturation occurred just after weaning as described for coastal bottlenose dolphins ( Tursiops truncatus ). Thus, a pelagic lifestyle does not promote rapid maturation of muscle biochemistry. Rather, it promotes enhanced muscle biochemistry: newborn and adult spinner dolphins had four- and two-times greater Mb contents than newborn and adult bottlenose dolphins, respectively. Indeed, adult levels rivaled those of deep-diving cetaceans. Nonetheless, the relatively underdeveloped muscle biochemistry of calves likely contributes to documented mother-calf separations for spinner dolphins chased by the tuna purse-seine fishery. © 2017. Published by The Company of Biologists Ltd.

Using a fidget spinner to teach physics

NASA Astrophysics Data System (ADS)

Mesquita, Lucas; Brockington, Guilherme; Aline de Almeida, Pamella; Truyol, Maria Elena; André Testoni, Leonardo; Sousa, Paula F. F.

2018-07-01

In 2017, almost everywhere in the world, there was a boom in sales of fidget spinners. A toy that has ‘multi-lobes’ that can rotate freely from a thrust performed with the fingers. Thereby, we analysed the physics concepts that involved in this toy, such as angular and linear velocity. We did the analysis using the software Tracker, a laboratory light sensor, a free mobile application that turns the device flashlight into a strobe light source. Thus, we aim to show that it is possible to transform a well-known toy into an opportunity to work on physics concepts and to construct experiments in the classroom.

[Continuously perfused cultivation of genetically-engineered CHO cells producing prothrombin in a modified Super-Spinner].

PubMed

Chen, Z L; Iding, K; Lütkemeyer, D; Lehmann, J

2001-01-01

A Super-Spinner was Modified by mounting a stainless steel filter(pore size 75 microns) to the impeller shaft to retain cells while fresh nutrient is perfused. Using Macroporous microcarrier Cytopore 1, continuously perfused cultivation of a recombinant CHO cell line, CHO2DS producing prothrombin was performed with the perfusion of a protein-free medium DF6S. The cell retention rate was more than 90% during the 24 days continuously perfused cultivation. The viable cell density of CHO2DS and prothrombin concentration reached 4.62 x 10(6)(cells.m/L) and 11.3(mg/L) respectively after 9 days culture.

Echolocation click rates and behavior of foraging Hawaiian spinner dolphins

NASA Astrophysics Data System (ADS)

Benoit-Bird, Kelly J.; Au, Whitlow W. L.

2004-05-01

Groups of spinner dolphins work together to actively aggregate small animals in the deep-scattering layer that serve as their prey. Detailed information on dolphin foraging behavior, obtained with a 200-kHz multibeam sonar (Simrad MS2000), made it possible to correlate echolocation and foraging. Fifty-six groups of spinner dolphins foraging at night within a midwater micronekton sound-scattering layer were observed with the sonar. During sonar surveys, the rates of whistles and echolocation clicks were measured using four hydrophones at 6-m depth intervals. Significant differences in click rates were found between depths and between the different stages of foraging. Groups of foraging dolphins ranged in size from 16 to 28 dolphins. Click rates were not significantly affected by the number of dolphins in a foraging group. Contrary to initial predictions, click rates were relatively low when sonar data indicated that pairs of dolphins were actively feeding. Highest echolocation rates occurred within the scattering layer, during transitions between foraging states. Whistles were only detected when dolphins were not in a foraging formation and when animals were surfacing. This suggests clicks may be used directly or indirectly to cue group movement during foraging.

Surface-pressure Distributions on a Systematic Group of NACA 1-series Cowlings with and Without Spinners

NASA Technical Reports Server (NTRS)

Boswinkle, Robert W JR; Keith, Arvid L JR

1948-01-01

A method for calculating the flow fields of axially symmetric bodies from their pressure distributions is reported in NACA RM No. L8I17. In order to facilitate application of this method to the important case of the cowling-spinner combination, for use in the design of propellers, the present paper presents static-pressure distributions on the tops of 79 high-critical-speed NACA 1-series cowling-spinner combinations over wide ranges of inlet-velocity ratio at angles of attack of 0 degrees, 2 degrees, 4 degrees, and 6 degrees. Static-pressure distributions around the nose sections of several cowlings are given in greater detail to aid in estimating the pressures near the stagnation points and to show the effect of changes in the internal lip shape. The effects of the operation of a typical propeller on the surface pressures on the cowling are shown for one configuration. The pressure distributions over the nine NACA 1-series nose inlets used as the basic components of these combinations are also presented ro supplement the existing open-nose-cowling data of NACA ACR No. L5F30a which are applicable to the case of the rotating cowling.

Microbiological Production of Carotenoids

PubMed Central

Ciegler, Alex; Nelson, George E. N.; Hall, Harlow H.

1962-01-01

Synthesis of β-carotene by mated strains of Blakeslea trispora in shaken-flask culture was considerably enhanced by adding either 5% kerosene after 2 days of fermentation or acid-refined kerosene at the start of fermentation to a grain-based medium that also contained a natural lipid, nonionic detergent, and β-ionone; average yields of 17,500 μg per g of dry fermentation solids (86,000 μg per 100 ml of medium) were attained when refined kerosene was used. Almost all of the carotene was retained within the mycelium. Peak yields were achieved in 5 days. PMID:13879500

Biodegradable poly-ε-caprolactone microcarriers for efficient production of human mesenchymal stromal cells and secreted cytokines in batch and fed-batch bioreactors.

PubMed

Lam, Alan Tin-Lun; Li, Jian; Toh, Jessica Pei-Wen; Sim, Eileen Jia-Hui; Chen, Allen Kuan-Liang; Chan, Jerry Kok-Yen; Choolani, Mahesh; Reuveny, Shaul; Birch, William R; Oh, Steve Kah-Weng

2017-03-01

Large numbers of human mesenchymal stromal cells (MSCs) used for a variety of applications in tissue engineering and cell therapy can be generated by scalable expansion in a bioreactor using microcarriers (MCs) systems. However, the enzymatic digestion process needed to detach cells from the growth surface can affect cell viability and potentially the potency and differentiation efficiency. Thus, the main aim of our study was to develop biocompatible and biodegradable MCs that can support high MSC yields while maintaining their differentiation capability and potency. After cell expansion, the cells that covered MCs can be directly implanted in vivo without the need for cell harvesting or use of scaffold. Poly-ε-caprolactone (PCL) is known as a biocompatible and biodegradable material. However, it cannot be used for generation of MCs because its high density (1.14 g/cm 3 ) would exclude its applicability for suspension MCs in stirred reactors. In this article, we describe expansion and potency of MSCs propagated on low-density (1.06 g/cm 3 ) porous PCL MCs coated with extracellular matrices (LPCLs) in suspended stirred reactors. Using these LPCLs, cell yields of about 4 × 10 4 cells/cm 2 and 7- to 10-fold increases were obtained using four different MSC lines (bone marrow, cord blood, fetal and Wharton's jelly). These yields were comparable with those obtained using non-degradable MCs (Cytodex 3) and higher than two-dimensional monolayer (MNL) cultures. A fed-batch process, which demonstrated faster cell expansion (4.5 × 10 4 cells/cm 2 in 5 days as compared with 7 days in batch culture) and about 70% reduction in growth media usage, was developed and scaled up from 100-mL spinner flask to 1-L controlled bioreactor. Surface marker expression, trilineage differentiation and clonogenic potential of the MSCs expanded on LPCL were not affected. Cytokine secretion kinetics, which occurred mostly during late logarithmic phase, was usually comparable with that obtained in Cytodex 3 cultures and higher than MNL cultures. In conclusion, biodegradable LPCL can be used to efficiently expand a variety of MSC lines in stirred scalable reactors in a cost-effective manner while maintaining surface markers expression, differentiation capability and high levels of cytokine secretion. This study is the first step in testing these cell-biodegradable porous MC aggregates for tissue engineering and cell therapy, such as bone and cartilage regeneration, or wound healing. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Automated simultaneous measurement of the δ(13) C and δ(2) H values of methane and the δ(13) C and δ(18) O values of carbon dioxide in flask air samples using a new multi cryo-trap/gas chromatography/isotope ratio mass spectrometry system.

PubMed

Brand, Willi A; Rothe, Michael; Sperlich, Peter; Strube, Martin; Wendeberg, Magnus

2016-07-15

The isotopic composition of greenhouse gases helps to constrain global budgets and to study sink and source processes. We present a new system for high-precision stable isotope measurements of carbon, hydrogen and oxygen in atmospheric methane and carbon dioxide. The design is intended for analyzing flask air samples from existing sampling programs without the need for extra sample air for methane analysis. CO2 and CH4 isotopes are measured simultaneously using two isotope ratio mass spectrometers, one for the analysis of δ(13) C and δ(18) O values and the second one for δ(2) H values. The inlet carousel delivers air from 16 sample positions (glass flasks 1-5 L and high-pressure cylinders). Three 10-port valves take aliquots from the sample stream. CH4 from 100-mL air aliquots is preconcentrated in 0.8-mL sample loops using a new cryo-trap system. A precisely calibrated working reference air is used in parallel with the sample according to the Principle of Identical Treatment. It takes about 36 hours for a fully calibrated analysis of a complete carousel including extractions of four working reference and one quality control reference air. Long-term precision values, as obtained from the quality control reference gas since 2012, account for 0.04 ‰ (δ(13) C values of CO2 ), 0.07 ‰ (δ(18) O values of CO2 ), 0.11 ‰ (δ(13) C values of CH4 ) and 1.0 ‰ (δ(2) H values of CH4 ). Within a single day, the system exhibits a typical methane δ(13) C standard deviation (1σ) of 0.06 ‰ for 10 repeated measurements. The system has been in routine operation at the MPI-BGC since 2012. Consistency of the data and compatibility with results from other laboratories at a high precision level are of utmost importance. A high sample throughput and reliability of operation are important achievements of the presented system to cope with the large number of air samples to be analyzed. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Osteoarthritic human chondrocytes proliferate in 3D co-culture with mesenchymal stem cells in suspension bioreactors.

PubMed

Khurshid, Madiha; Mulet-Sierra, Aillette; Adesida, Adetola; Sen, Arindom

2018-03-01

Osteoarthritis (OA) is a painful disease, characterized by progressive surface erosion of articular cartilage. The use of human articular chondrocytes (hACs) sourced from OA patients has been proposed as a potential therapy for cartilage repair, but this approach is limited by the lack of scalable methods to produce clinically relevant quantities of cartilage-generating cells. Previous studies in static culture have shown that hACs co-cultured with human mesenchymal stem cells (hMSCs) as 3D pellets can upregulate proliferation and generate neocartilage with enhanced functional matrix formation relative to that produced from either cell type alone. However, because static culture flasks are not readily amenable to scale up, scalable suspension bioreactors were investigated to determine if they could support the co-culture of hMSCs and OA hACs under serum-free conditions to facilitate clinical translation of this approach. When hACs and hMSCs (1:3 ratio) were inoculated at 20,000 cells/ml into 125-ml suspension bioreactors and fed weekly, they spontaneously formed 3D aggregates and proliferated, resulting in a 4.75-fold increase over 16 days. Whereas the apparent growth rate was lower than that achieved during co-culture as a 2D monolayer in static culture flasks, bioreactor co-culture as 3D aggregates resulted in a significantly lower collagen I to II mRNA expression ratio and more than double the glycosaminoglycan/DNA content (5.8 vs. 2.5 μg/μg). The proliferation of hMSCs and hACs as 3D aggregates in serum-free suspension culture demonstrates that scalable bioreactors represent an accessible platform capable of supporting the generation of clinical quantities of cells for use in cell-based cartilage repair. Copyright © 2017 John Wiley & Sons, Ltd.

pH-metric solubility. 2: correlation between the acid-base titration and the saturation shake-flask solubility-pH methods.

PubMed

Avdeef, A; Berger, C M; Brownell, C

2000-01-01

The objective of this study was to compare the results of a normal saturation shake-flask method to a new potentiometric acid-base titration method for determining the intrinsic solubility and the solubility-pH profiles of ionizable molecules, and to report the solubility constants determined by the latter technique. The solubility-pH profiles of twelve generic drugs (atenolol, diclofenac.Na, famotidine, flurbiprofen, furosemide, hydrochlorothiazide, ibuprofen, ketoprofen, labetolol.HCl, naproxen, phenytoin, and propranolol.HCl), with solubilities spanning over six orders of magnitude, were determined both by the new pH-metric method and by a traditional approach (24 hr shaking of saturated solutions, followed by filtration, then HPLC assaying with UV detection). The 212 separate saturation shake-flask solubility measurements and those derived from 65 potentiometric titrations agreed well. The analysis produced the correlation equation: log(1/S)titration = -0.063(+/- 0.032) + 1.025(+/- 0.011) log(1/S)shake-flask, s = 0.20, r2 = 0.978. The potentiometrically-derived intrinsic solubilities of the drugs were: atenolol 13.5 mg/mL, diclofenac.Na 0.82 microg/mL, famotidine 1.1 mg/ mL, flurbiprofen 10.6 microg/mL, furosemide 5.9 microg/mL, hydrochlorothiazide 0.70 mg/mL, ibuprofen 49 microg/mL, ketoprofen 118 microg/mL, labetolol.HCl 128 microg/mL, naproxen 14 microg/mL, phenytoin 19 microg/mL, and propranolol.HCl 70 microg/mL. The new potentiometric method was shown to be reliable for determining the solubility-pH profiles of uncharged ionizable drug substances. Its speed compared to conventional equilibrium measurements, its sound theoretical basis, its ability to generate the full solubility-pH profile from a single titration, and its dynamic range (currently estimated to be seven orders of magnitude) make the new pH-metric method an attractive addition to traditional approaches used by preformulation and development scientists. It may be useful even to discovery scientists in critical decision situations (such as calibrating computational prediction methods).

Flow Dynamics In Eccentrically Rotating Flasks Used For Dispersant Effectiveness Testing

EPA Science Inventory

The evaluation of dispersant effectiveness used for oil spills is commonly done using tests conducted in laboratory flasks. We used a Hot Wire Anemometer (HWA) to characterize mixing dynamics in the Swirling Flask (SF) and the Baffled Flask (BF), the latter is being considered b...

Passive acoustic monitoring of coastally associated Hawaiian spinner dolphins, Stenella longirostris, ground-truthed through visual surveys.

PubMed

Heenehan, Heather L; Tyne, Julian A; Bejder, Lars; Van Parijs, Sofie M; Johnston, David W

2016-07-01

Effective decision making to protect coastally associated dolphins relies on monitoring the presence of animals in areas that are critical to their survival. Hawaiian spinner dolphins forage at night and rest during the day in shallow bays. Due to their predictable presence, they are targeted by dolphin-tourism. In this study, comparisons of presence were made between passive acoustic monitoring (PAM) and vessel-based visual surveys in Hawaiian spinner dolphin resting bays. DSG-Ocean passive acoustic recording devices were deployed in four bays along the Kona Coast of Hawai'i Island between January 8, 2011 and August 30, 2012. The devices sampled at 80 kHz, making 30-s recordings every four minutes. Overall, dolphins were acoustically detected on 37.1% to 89.6% of recording days depending on the bay. Vessel-based visual surveys overlapped with the PAM surveys on 202 days across the four bays. No significant differences were found between visual and acoustic detections suggesting acoustic surveys can be used as a proxy for visual surveys. Given the need to monitor dolphin presence across sites, PAM is the most suitable and efficient tool for monitoring long-term presence/absence. Concomitant photo-identification surveys are necessary to address changes in abundance over time.

Electrically conductive connection for an electrode

DOEpatents

Hornack, Thomas R.; Chilko, Robert J.

1986-01-01

An electrically conductive connection for an electrode assembly of an electrolyte cell in which aluminum is produced by electrolysis in a molten salt is described. The electrode assembly comprises an electrode flask and a conductor rod. The flask has a collar above an area of minimum flask diameter. The electrically conductive connection comprises the electrode flask, the conductor rod and a structure bearing against the collar and the conductor rod for pulling the conductor rod into compressive and electrical contact with the flask.

Electrically conductive connection for an electrode

DOEpatents

Hornack, T.R.; Chilko, R.J.

1986-09-02

An electrically conductive connection for an electrode assembly of an electrolyte cell in which aluminum is produced by electrolysis in a molten salt is described. The electrode assembly comprises an electrode flask and a conductor rod. The flask has a collar above an area of minimum flask diameter. The electrically conductive connection comprises the electrode flask, the conductor rod and a structure bearing against the collar and the conductor rod for pulling the conductor rod into compressive and electrical contact with the flask. 2 figs.

Anti-Inflammatory Effect of Carprofen Is Enhanced by Avocado/Soybean Unsaponifiables, Glucosamine and Chondroitin Sulfate Combination in Chondrocyte Microcarrier Spinner Culture.

PubMed

Grzanna, Mark W; Secor, Erica J; Fortuno, Lowella V; Au, Angela Y; Frondoza, Carmelita G

2018-06-01

Objective Osteoarthritis is a painful, chronic joint disease affecting man and animals with no known curative therapies. Palliative nonsteroidal anti-inflammatory drugs (NSAIDs) are commonly used but they cause adverse side effects prompting the search for safer alternatives. To address this need, we evaluated the anti-inflammatory activity of avocado/soybean unsaponifiables (ASU), glucosamine (GLU), and chondroitin sulfate (CS) with or without the NSAID carprofen. Design Canine chondrocytes were propagated in microcarrier spinner culture and incubated with (1) control medium, (2) ASU (8.3 µg/mL) + GLU (11 µg/mL) + CS (20 µg/mL) combination for 24 hours; and/or carprofen (40 ng/mL). Cultures were next incubated with control medium alone or IL-1β (10 ng/mL) for another 24 hours. Production of PGE 2 , IL-6, IL-8, and MCP-1 (also known as CCL-2) were measured by ELISA. Results Chondrocytes proliferated in microcarrier spinner culture and produced type II collagen and aggrecan. Stimulation with IL-1β induced significant increases in PGE 2 , IL-6, IL-8, and MCP-1 production. The increases in production were suppressed by carprofen as well as [ASU+GLU+CS]. The combination of carprofen and [ASU+GLU+CS] reduced PGE 2 production significantly more than either preparation alone. The inhibitory effect of carprofen on IL-6, IL-8, and MCP-1 production was significantly less than that of [ASU+GLU+CS], whereas the combination did not reduce the production of these molecules significantly more than [ASU+GLU+CS] alone. Conclusions The potentiating effect of [ASU+GLU+CS] on low-dose carprofen was identified in chondrocyte microcarrier spinner cultures. Our results suggest that the combination of low-dose NSAIDs like carprofen with [ASU+GLU+CS] could offer a safe, effective management for joint pain.

A Sensitive, Multifunctional Spinner Magnetometer Using Magneto-impedance Sensor: a Rapid and Convenient Tool for the Quantification of Inhomogeneity of Magnetization

NASA Astrophysics Data System (ADS)

Kodama, K.

2016-12-01

A new type of spinner magnetometer with wide dynamic range from 10-7 mAm2 to 10-1 mAm2 and the resolution of 10-8 mAm2 was developed. The high sensitivity was achieved by using magneto-impedance (MI) sensor, a compact, high-performance magnetic sensor used in industrial fields. The slow spinning speed (5 Hz) and the unique mechanism enabling the adjustment of the sample-sensor distance allow measurements of fragile samples in any shape and size. A differential arrangement connecting a pair of the MI sensors in opposite serial reduces external noise and temperature drift. The differential sensor output is transferred to an amplification circuit associated with a programmable low-pass filter. The signal with reference to the spinning frequency is detected with a digital lock-in amplifier. The spinner magnetometer has two selectable measurement modes, the fundamental-mode (F-mode) and the harmonic-mode (H-mode). Measurements in the F-mode detect signals oscillating at the fundamental frequency (5 Hz) as conventional spinner magnetometers do. In the H-mode, additionally, the second (10 Hz) and the third (15 Hz) harmonic components can be measured. Tests in the H-mode were performed using a small coil and changing its position to simulate an offset-dipole. The results demonstrate that the dipole moment of the fundamental component is systematically biased by both quadrupole and octupole components arising in practice from inhomogeneity of magnetization or irregularity of sample shape. This study proposes, combined with theoretical and numerical analyses, quantification of such non-dipole effects and associated errors in the determination of dipole moment of a sample, as well as their correction that may be necessary, for example, when measuring irregular-shaped samples in the proximity of the sensor.

Monoclonal antibody production using a new supermacroporous cryogel bioreactor.

PubMed

Nilsang, Suthasinee; Nandakumar, Kutty Selva; Galaev, Igor Yu; Rakshit, Sudip Kumar; Holmdahl, Rikard; Mattiasson, Bo; Kumar, Ashok

2007-01-01

A supermacroporous cryogel bioreactor has been developed to culture hybridoma cells for long-term continuous production of monoclonal antibodies (mAb). Hybridoma clone M2139, secreting antibodies against J1 epitope (GERGAAGIAGPK; amino acids, 551-564) of collagen type II, are immobilized in the porous bed matrix of a cryogel column (10 mL bed volume). The cells got attached to the matrix within 48 h after inoculation and grew as a confluent sheet inside the cryogel matrix. Cells were in the lag phase for 15 days and secreted mAb into the circulation medium. Glucose consumption and lactic acid production were also monitored, and during the exponential phase (approximately 20 days), the hybridoma cell line consumed 0.75 mM day-1 glucose, produced 2.48 mM day-1 lactic acid, and produced 6.5 microg mL-1 day-1 mAb during the exponential phase. The mAb concentration reached 130 microg mL-1 after continuous run of the cryogel column for 36 days. The yield of the mAb after purification was 67.5 mg L-1, which was three times greater than the mAb yield obtained from T-flask batch cultivation. Even after the exchange of medium reservoir, cells in the cryogel column were still active and had relatively stable mAb production for an extended period of time. The bioreactor was operated continuously for 55 days without any contamination. The results from ELISA as well as arthritis experiments demonstrate that the antibodies secreted by cells grown on the cryogel column did not differ from antibodies purified from the cells grown in commercial CL-1000 culture flasks. Thus, supermacroporous cryogels can be useful as a supporting material for productive hybridoma cell culture. Cells were found to be viable inside the porous matrix of the cryogel during the study period and secreted antibodies continuously. The antibodies thus obtained from the cryogel reactor were found to be functionally active in vivo, as demonstrated by their capacity to induce arthritis in mice.

Flask sample measurements for CO2, CH4 and CO using cavity ring-down spectrometry

NASA Astrophysics Data System (ADS)

Wang, J.-L.; Jacobson, G.; Rella, C. W.; Chang, C.-Y.; Liu, I.; Liu, W.-T.; Chew, C.; Ou-Yang, C.-F.; Liao, W.-C.; Chang, C.-C.

2013-08-01

In recent years, cavity ring-down spectrometry (CRDS) has been demonstrated to be a highly sensitive, stable and fast analytical technique for real-time in situ measurements of greenhouse gases. In this study, we propose the technique (which we call flask-CRDS) of analyzing whole air flask samples for CO2, CH4 and CO using a custom gas manifold designed to connect to a CRDS analyzer. Extremely stable measurements of these gases can be achieved over a large pressure range in the flask, from 175 to 760 Torr. The wide pressure range is conducive to flask sample measurement in three ways: (1) flask samples can be collected in low-pressure environments (e.g. high-altitude locations); (2) flask samples can be first analyzed for other trace gases with the remaining low-pressure sample for CRDS analysis of CO2, CH4 and CO; and (3) flask samples can be archived and re-analyzed for validation. The repeatability of this method (1σ of 0.07 ppm for CO2, 0.4 ppb for CH4, and 0.5 ppb for CO) was assessed by analyzing five canisters filled with the same air sample to a pressure of 200 Torr. An inter-comparison of the flask-CRDS data with in-situ CRDS measurements at a high-altitude mountain baseline station revealed excellent agreement, with differences of 0.10 ± 0.09 ppm (1σ) for CO2 and 0.9 ± 1.0 ppb for CH4. This study demonstrated that the flask-CRDS method was not only simple to build and operate but could also perform highly accurate and precise measurements of atmospheric CO2, CH4 and CO in flask samples.

[Studies for analyzing prohibited ingredients such as tetracaine hydrochloride in cosmetics].

PubMed

Tokunaga, Hiroshi; Takeuchi, Orie; Uchino, Tadashi; Ando, Masanori

2004-01-01

Tetracaine hydrochloride (TH) is nominated as the prohibited ingredients in cosmetics in Japanese Pharmaceutical Affairs Act. So the analytical method for TH was investigated by HPLC. After adding 5 ml of TH solution at 10 microg/ml and 2 ml of salicylic acid solution at 75 microg/ml as the internal standard to 0.5 g of the lotion, the mixture was made up to 10 ml with a mixture of water and methanol (1:1) as the testing solution. Milky lotion was procedured as follows: After adding 5 ml of TH solution at 10 microg/ml and 2 ml of internal standard solution to 0.5 g of the milky lotion, the mixture was made up to 10 ml with a mixture of water and methanol (1:1). Two milliliter of this mixture was placed into a centrifuging tube with a cap and 2 ml of hexane was added. After shaking vigorously and centrifuging, the lower layer was used as the testing solution. In the case of the cream, the other procedures were used: 0.5 g of cream was placed into a 10-ml volumetric flask and 1 ml of tetrahydrofuran was added. After dissolving, the mixture of methanol and water (1:1) was added to make up 10.0 ml. Two milliliter of this mixture was placed into a centrifuging tube with a cap and 2.0 ml of hexane was added. After shaking vigorously and centrifuging, the lower layer was used as the testing solution. The testing solution of 20 microl was analyzed by HPLC using the ODS column (CAPCELL PAK C18 column, 4.6 x 250 mm), the mixture of acetonitrile and 50 mmol/l phosphate buffer(pH 2.0)(7:3) and the detection wavelength of 303 nm. The working curves from 0.5 to 6.0 microg/ml showed a linear line between the concentrations of TH and the peak area ratio. There was no interference of peak of TH from the lotion, milky lotion and cream.

Continuous production of monoclonal antibody in a packed-bed bioreactor.

PubMed

Golmakany, Naghmeh; Rasaee, Mohammad Javad; Furouzandeh, Mehdi; Shojaosadati, Seyed Abbas; Kashanian, Soheila; Omidfar, Kobra

2005-06-01

In the present study the growth and MAb (monoclonal antibody) production of a mouse x mouse hybridoma cell producing anti-digoxin MAb was evaluated. The hybridoma cells entrapped within the support matrix Fibra-Cel were cultured in batch and continuous mode following special protocols. Cell-culture studies were performed in a 1-litre spinner basket containing 3 g.litre-1 support matrix. Batch culture was operated with the cell density of 42x10(6) cells. During the 7 days of culture, the medium was sampled daily in order to assess glucose and MAb concentrations and the lactate dehydrogenase released into the culture medium. After a culture period of 72 h, the cell density and MAb concentration were found to be 10.4x10(7) cells/3 g of NWPF (non-woven polyester fibre) discs and 250 microg/ml respectively. This yield gradually decreased to 0.55x10(6) cells/3 g of packaging material and 60 microg/ml respectively at the end of the batch culture. In the continuous-culture studies, the batch culture was initially operated for 64.5 h and then continuous flow was started at the dilution rates of 0.15, 0.2, 0.25 and 0.3 day-1 and finally stabilized at 0.25 day-1 within 288 h (12 days). The MAb concentration at steady state was found to be 116-120 microg/day per ml, and the yield of operation was 62.5 mg/day per ml, which was 3.5 times higher than that of batch culture. In conclusion, a packed-bed bioreactor with the support matrix Fibra-Cel, operated in continuous-feeding mode, is more efficient for large-scale MAb production than a batch culture. On the other hand, by using a continuous-culture system, a better supply of nutrients and removal of inhibitory metabolites and proteolytic enzymes was obtained.

Comparative biodegradation of HDPE and LDPE using an indigenously developed microbial consortium.

PubMed

Satlewal, Alok; Soni, Ravindra; Zaidi, Mgh; Shouche, Yogesh; Goel, Reeta

2008-03-01

A variety of bacterial strains were isolated from waste disposal sites of Uttaranchal, India, and some from artificially developed soil beds containing maleic anhydride, glucose, and small pieces of polyethylene. Primary screening of isolates was done based on their ability to utilize high- and low-density polyethylenes (HDPE/LDPE) as a primary carbon source. Thereafter, a consortium was developed using potential strains. Furthermore, a biodegradation assay was carried out in 500-ml flasks containing minimal broth (250 ml) and HDPE/ LDPE at 5 mg/ml concentration. After incubation for two weeks, degraded samples were recovered through filtration and subsequent evaporation. Fourier transform infrared spectroscopy (FTIR) and simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis TG-DTG-DTA) were used to analyze these samples. Results showed that consortium-treated HDPE (considered to be more inert relative to LDPE) was degraded to a greater extent 22.41% weight loss) in comparison with LDPE (21.70% weight loss), whereas, in the case of untreated samples, weight loss was more for LDPE than HDPE (4.5% and 2.5%, respectively) at 400 degrees . Therefore, this study suggests that polyethylene could be degraded by utilizing microbial consortia in an eco-friendly manner.

Pre-treatment of synthetic elastomeric scaffolds by cardiac fibroblasts improves engineered heart tissue

PubMed Central

Radisic, Milica; Park, Hyoungshin; Martens, Timothy P.; Salazar-Lazaro, Johanna E.; Geng, Wenliang; Wang, Yadong; Langer, Robert; Freed, Lisa E.; Vunjak-Novakovic, Gordana

2009-01-01

Native myocardium consists of several cell types, of which approximately one-third are myocytes and most of the nonmyocytes are fibroblasts. By analogy with monolayer culture in which fibroblasts were removed to prevent overgrowth, early attempts to engineer myocardium utilized cell populations enriched for cardiac myocytes (CMs; ~80–90% of total cells). We hypothesized that the pre-treatment of synthetic elastomeric scaffolds with cardiac fibroblasts (CFs) will enhance the functional assembly of the engineered cardiac constructs by creating an environment supportive of cardiomyocyte attachment and function. Cells isolated from neonatal rat ventricles were prepared to form three distinct populations: rapidly plating cells identified as CFs, slowly plating cells identified as CMs, and unseparated initial population of cells (US). The cell fractions (3 × 106 cells total) were seeded into poly(glycerol sebacate) scaffolds (highly porous discs, 5 mm in diameter × 2-mm thick) using Matrigel™, either separately (CM or CF), concurrently (US), or sequentially (CF pre-treatment followed by CM culture, CF + CM), and cultured in spinner flasks. The CF + CM group had the highest amplitude of contraction and the lowest excitation threshold, superior DNA content, and higher glucose consumption rate. The CF + CM group exhibited compact 100- to 200-μm thick layers of elongated myocytes aligned in parallel over layers of collagen-producing fibroblasts, while US and CM groups exhibited scattered and poorly elongated myocytes. The sequential co-culture of CF and CM on a synthetic elastomer scaffold thus created an environment supportive of cardiomyocyte attachment, differentiation, and contractile function, presumably due to scaffold conditioning by cultured fibroblasts. When implanted over the infarcted myocardium in a nude rat model, cell-free poly(glycerol sebacate) remained at the ventricular wall after 2 weeks of in vivo, and was vascularized. PMID:18041719

Bioprocess integration for human mesenchymal stem cells: From up to downstream processing scale-up to cell proteome characterization.

PubMed

Cunha, Bárbara; Aguiar, Tiago; Carvalho, Sofia B; Silva, Marta M; Gomes, Ricardo A; Carrondo, Manuel J T; Gomes-Alves, Patrícia; Peixoto, Cristina; Serra, Margarida; Alves, Paula M

2017-04-20

To deliver the required cell numbers and doses to therapy, scaling-up production and purification processes (at least to the liter-scale) while maintaining cells' characteristics is compulsory. Therefore, the aim of this work was to prove scalability of an integrated streamlined bioprocess compatible with current good manufacturing practices (cGMP) comprised by cell expansion, harvesting and volume reduction unit operations using human mesenchymal stem cells (hMSC) isolated from bone marrow (BM-MSC) and adipose tissue (AT-MSC). BM-MSC and AT-MSC expansion and harvesting steps were scaled-up from spinner flasks to 2L scale stirred tank single-use bioreactor using synthetic microcarriers and xeno-free medium, ensuring high cellular volumetric productivities (50×10 6 cellL -1 day -1 ), expansion factors (14-16 fold) and cell recovery yields (80%). For the concentration step, flat sheet cassettes (FSC) and hollow fiber cartridges (HF) were compared showing a fairly linear scale-up, with a need to slightly decrease the permeate flux (30-50 LMH, respectively) to maximize cell recovery yield. Nonetheless, FSC allowed to recover 18% more cells after a volume reduction factor of 50. Overall, at the end of the entire bioprocess more than 65% of viable (>95%) hMSC could be recovered without compromising cell's critical quality attributes (CQA) of viability, identity and differentiation potential. Alongside the standard quality assays, a proteomics workflow based on mass spectrometry tools was established to characterize the impact of processing on hMSC's CQA; These analytical tools constitute a powerful tool to be used in process design and development. Copyright © 2017 Elsevier B.V. All rights reserved.

Engineering tubular bone using mesenchymal stem cell sheets and coral particles

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geng, Wenxin; Ma, Dongyang; Yan, Xingrong

Highlights: • We developed a novel engineering strategy to solve the limitations of bone grafts. • We fabricated tubular constructs using cell sheets and coral particles. • The composite constructs showed high radiological density and compressive strength. • These characteristics were similar to those of native bone. -- Abstract: The development of bone tissue engineering has provided new solutions for bone defects. However, the cell-scaffold-based approaches currently in use have several limitations, including low cell seeding rates and poor bone formation capacity. In the present study, we developed a novel strategy to engineer bone grafts using mesenchymal stem cell sheetsmore » and coral particles. Rabbit bone marrow mesenchymal stem cells were continuously cultured to form a cell sheet with osteogenic potential and coral particles were integrated into the sheet. The composite sheet was then wrapped around a cylindrical mandrel to fabricate a tubular construct. The resultant tubular construct was cultured in a spinner-flask bioreactor and subsequently implanted into a subcutaneous pocket in a nude mouse for assessment of its histological characteristics, radiological density and mechanical property. A similar construct assembled from a cell sheet alone acted as a control. In vitro observations demonstrated that the composite construct maintained its tubular shape, and exhibited higher radiological density, compressive strength and greater extracellular matrix deposition than did the control construct. In vivo experiments further revealed that new bone formed ectopically on the composite constructs, so that the 8-week explants of the composite sheets displayed radiological density similar to that of native bone. These results indicate that the strategy of using a combination of a cell sheet and coral particles has great potential for bone tissue engineering and repairing bone defects.« less

Biodegradation of trichloroethylene and toluene by indigenous microbial populations in soil.

PubMed Central

Fan, S; Scow, K M

1993-01-01

The biodegradation of trichloroethylene (TCE) and toluene, incubated separately and in combination, by indigenous microbial populations was measured in three unsaturated soils incubated under aerobic conditions. Sorption and desorption of TCE (0.1 to 10 micrograms ml-1) and toluene (1.0 to 20 micrograms ml-1) were measured in two soils and followed a reversible linear isotherm. At a concentration of 1 micrograms ml-1, TCE was not degraded in the absence of toluene in any of the soils. In combination, both 1 microgram of TCE ml-1 and 20 micrograms of toluene ml-1 were degraded simultaneously after a lag period of approximately 60 to 80 h, and the period of degradation lasted from 70 to 90 h. Usually 60 to 75% of the initial 1 microgram of TCE ml-1 was degraded, whereas 100% of the toluene disappeared. A second addition of 20 micrograms of toluene ml-1 to a flask with residual TCE resulted in another 10 to 20% removal of the chemical. Initial rates of degradation of toluene and TCE were similar at 32, 25, and 18 degrees C; however, the lag period increased with decreasing temperature. There was little difference in degradation of toluene and TCE at soil moisture contents of 16, 25, and 30%, whereas there was no detectable degradation at 5 and 2.5% moisture. The addition of phenol, but not benzoate, stimulated the degradation of TCE in Rindge and Yolo silt loam soils, methanol and ethylene slightly stimulated TCE degradation in Rindge soil, glucose had no effect in either soil, and dissolved organic carbon extracted from soil strongly sorbed TCE but did not affect its rate of biodegradation. PMID:8328806

Effects of soy peptone on the inoculum preparation of Streptococcus zooepidemicus for production of hyaluronic acid.

PubMed

Benedini, Leandro Junqueira; Santana, Maria Helena Andrade

2013-02-01

Soy peptone (SP) was studied as nutrient source in replacement of the conventional media as Brain-Heart Infusion (BHI) and sheep blood in the first seed culture medium in Petri plates of Streptococcus zooepidemicus. This substitution, aimed at meeting the claim of the pharmaceutical and cosmetics industries, for the removal of animal sources of the culture media used in obtaining their products for safety reasons. The animal sources were used as a control. The effects of this substitution were studied in fermentations carried out at 37°C and 150rpm in 250mL Erlenmeyer flasks containing 100mL culture medium containing glucose and SP only. The replacement of animal nutrient sources by SP to about twice the BHI concentration did not alter the amount of the produced HA, or caused deviations in the metabolism of the microorganism in favor of HA to the detriment of cell growth. Copyright © 2013 Elsevier Ltd. All rights reserved.

Enhanced production of ATP-binding cassette protein exporter-dependent lipase by modifying the growth medium components of Pseudomonas fluorescens.

PubMed

Eom, Gyeong Tae; Song, Jae Kwang

2014-08-01

The industrially-important thermostable lipase, TliA, was extracellularly produced in the recombinant Pseudomonas fluorescens by the homologous expression of TliA and its cognate ABC protein exporter, TliDEF. To increase the secretory production of TliA, we optimized the growth temperature and the culture medium of P. fluorescens. The total amount and the specific productivity of lipase was highest at 25 °C of cell growth temperature, although maximal cell growth was observed at 30 °C. Using the culture medium composed of 20 g dextrin l(-1), 40 g Tween 80 l(-1) and 30 g peptone l(-1), TliA was produced at a level of 2,200 U ml(-1) in a flask culture. The TliA production increased about 3.8-fold (8,450 U ml(-1)) in batch fermentation using a 2.5 l fermentor, which was about 7.7-fold higher than that of previously reported TliA production.

Struvite Crystallization of Anaerobic Digestive Fluid of Swine Manure Containing Highly Concentrated Nitrogen

PubMed Central

Lee, Eun Young; Oh, Min Hwan; Yang, Seung-Hak; Yoon, Tae Han

2015-01-01

In this study, the optimal operation factors for struvite crystallization for removing and recovering nitrogen and phosphorus from anaerobic digestive fluid of swine manure containing highly concentrated nitrogen was determined. Every experiment for the struvite crystallization reaction was conducted by placing 1,000 mL of digestion fluid in a 2,000 mL Erlenmeyer flask at various temperatures, pH, and mixing speed. Except for special circumstances, the digestion fluid was centrifuged (10,000 rpm, 10 min) and then the supernatant was used for the experiment at room temperature and 100 rpm. The optimal mole ratio of PO43−:Mg2+ was 1:1.5, and the pH effect ranging from 9 to 11 was similar, when mixed for 1 hour. Under this condition, the removal efficiency of NH4+-N and PO43−-P was 40% and 88.6%, respectively. X-shaped crystal was observed by light and scanning electron microscopy. In addition, struvite crystal structure was confirmed through X-ray diffraction analysis. PMID:26104412

Breathing gas perfluorocarbon measurements using an absorber filled with zeolites.

PubMed

Proquitté, H; Rüdiger, M; Wauer, R R; Schmalisch, G

2003-11-01

Perfluorocarbon (PFC) has been widely used in the treatment of respiratory diseases; however, PFC content of the breathing gases remains unknown. Therefore, we developed an absorber using PFC selective zeolites for PFC measurement in gases and investigated its accuracy. To generate a breathing gas with different PFC contents a heated flask was rinsed with a constant air flow of 4 litre x min(-1) and 1, 5, 10, and 20 ml of PFC were infused over 20 min using an infusor. The absorber was placed on an electronic scale and the total PFC volume was calculated from the weight gain. Steady-state increase in weight was achieved 3.5 min after stopping the infusion. The calculated PFC volume was slightly underestimated but the measuring error did not exceed -1% for PFC less than 1 ml. The measurement error decreased with increasing PFC volume. This zeolite absorber is an accurate method to quantitatively determine PFC in breathing gases and can be used as a reference method to validate other PFC sensors.

Preparation of gentiooligosaccharides using Trichoderma viride β-glucosidase.

PubMed

Wang, Fei; Wu, Jing; Chen, Sheng

2018-05-15

The recombinant plasmid pPIC9K-bgl1 containing β-glucosidase bgl1 from Trichoderma viride was constructed by overlapping PCR and integrated into Pichia pastoris KM71. In order to assist the formation of disulfide bonds and thus improve protein folding efficiency, protein disulfide isomerase pdi was co-expressed in the P. pastoris KM71/pPIC9K-bgl1/pPICZ-A-pdi strain, and fermentation in flasks resulted in enzyme activity of 143 U/ml. The enzyme activity of β-glucosidase reached 1402 U/ml following optimisation of fermentation conditions in a 3.6 l bioreactor. With 80% glucose as substrate, gentiooligosaccharides were synthesised by β-glucosidase-based reverse hydrolysis. A yield of 130 g/l was achieved with a conversion rate of 16.25%. With 20% glucose and 40% cellobiose as substrates, gentiooligosaccharides were synthesised by transglycosylation with a yield of 116 g/l and a conversion rate of 19.4%. Copyright © 2017 Elsevier Ltd. All rights reserved.

Identification of significant medium components that affect docosahexaenoic acid production by Schizochytrium sp. SW1

NASA Astrophysics Data System (ADS)

Manikan, Vidyah; Hamid, Aidil A.

2013-11-01

Central composite design (CCD) was employed to investigate the significance of glucose, yeast extract, MSG and sea salt in affecting the amount of docosahexaenoic acid (DHA) accumulated by a locally isolated strain of Schizochytrium. Design Expert software was used to construct a set of experiments where each medium component mentioned above was varied over three levels. Cultivation was carried out in 250mL flasks containing 50mL of medium, incubated at 30°C with 200 rpm agitation for 96 hours. ANOVA was conducted to identify the influential factors and the level of their significance where factors that scored a probability value of less than 0.05 were considered significant. The level of influence for each independent variable was also interpreted using perturbation whereas pattern of interaction between the factors were interpreted using interaction plots. This experiment revealed that yeast extract and monosodium glutamate have significant influence on DHA accumulation process by Schizochytrium sp. SW1.

Enhancement of Rhamnolipid Production in Residual Soybean Oil by an Isolated Strain of Pseudomonas aeruginosa

NASA Astrophysics Data System (ADS)

de Lima, C. J. B.; França, F. P.; Sérvulo, E. F. C.; Resende, M. M.; Cardoso, V. L.

In the present work, the production of rhamnolipid from residual soybean oil (RSO) from food frying facilities was studied using a strain of Pseudomonas aeruginosa of contaminated lagoon, isolated from a hydrocarbon contaminated soil. The optimization of RSO, amonium nitrate, and brewery residual yeast concentrations was accomplished by a central composite experimental design and surface response analysis. The experiments were performed in 500-mL Erlenmeyer flasks containing 50mL of mineral medium, at 170 rpm and 30±1°C, for a 48-h fermentation period. Rhamnolipid production has been monitored by measurements of surface tension, rhamnose concentration, and emulsifying activity. The best-planned results, located on the central point, have corresponded to 22g/L of RSO, 5.625 g/ L of NH4NO3' and 11.5 g/L of brewery yeast. At the maximum point the values for rhamnose and emulsifying index were 2.2g/L and 100%, respectively.

Cooling of Water in a Flask: Convection Currents in a Fluid with a Density Maximum

ERIC Educational Resources Information Center

Velasco, S.; White, J. A.; Roman, F. L.

2010-01-01

The effect of density inversion on the convective flow of water in a spherical glass flask cooled with the help of an ice-water bath is shown. The experiment was carried out by temperature measurements (cooling curves) taken at three different heights along the vertical diameter of the flask. Flows inside the flask are visualized by seeding the…

A new wireless system for decentralised measurement of physiological parameters from shake flasks

PubMed Central

Vasala, Antti; Panula, Johanna; Bollók, Monika; Illmann, Lutz; Hälsig, Christian; Neubauer, Peter

2006-01-01

Background Shake flasks are widely used because of their low price and simple handling. Many researcher are, however, not aware of the physiological consequences of oxygen limitation and substrate overflow metabolism that occur in shake flasks. Availability of a wireless measuring system brings the possibilities for quality control and design of cultivation conditions. Results Here we present a new wireless solution for the measurement of pH and oxygen from shake flasks with standard sensors, which allows data transmission over a distance of more than 100 metres in laboratory environments. This new system was applied to monitoring of cultivation conditions in shake flasks. The at-time monitoring of the growth conditions became possible by simple means. Here we demonstrate that with typical protocols E. coli shake flask cultures run into severe oxygen limitation and the medium is strongly acidified. Additionally the strength of the new system is demonstrated by continuous monitoring of the oxygen level in methanol-fed Pichia pastoris shake flask cultures, which allows the optimisation of substrate feeding for preventing starvation or methanol overfeed. 40 % higher cell density was obtained by preventing starvation phases which occur in standard shake flask protocols by adding methanol when the respiration activity decreased in the cultures. Conclusion The here introduced wireless system can read parallel sensor data over long distances from shake flasks that are under vigorous shaking in cultivation rooms or closed incubators. The presented technology allows centralised monitoring of decentralised targets. It is useful for the monitoring of pH and dissolved oxygen in shake flask cultures. It is not limited to standard sensors, but can be easily adopted to new types of sensors and measurement places (e.g., new sensor points in large-scale bioreactors). PMID:16504107

Practices of shake-flask culture and advances in monitoring CO2 and O2.

PubMed

Takahashi, Masato; Aoyagi, Hideki

2018-05-01

About 85 years have passed since the shaking culture was devised. Since then, various monitoring devices have been developed to measure culture parameters. O 2 consumed and CO 2 produced by the respiration of cells in shaking cultures are of paramount importance due to their presence in both the culture broth and headspace of shake flask. Monitoring in situ conditions during shake-flask culture is useful for analysing the behaviour of O 2 and CO 2 , which interact according to Henry's law, and is more convenient than conventional sampling that requires interruption of shaking. In situ monitoring devices for shake-flask cultures are classified as direct or the recently developed bypass type. It is important to understand the characteristics of each type along with their unintended effect on shake-flask cultures, in order to improve the existing devices and culture conditions. Technical developments in the bypass monitoring devices are strongly desired in the future. It is also necessary to understand the mechanism underlying conventional shake-flask culture. The existing shaking culture methodology can be expanded into next-generation shake-flask cultures constituting a novel culture environment through a judicious selection of monitoring devices depending on the intended purpose of shake-flask culture. Construction and sharing the databases compatible with the various types of the monitoring devices and measurement instruments adapted for shaking culture can provide a valuable resource for broadening the application of cells with shake-flask culture.

Production of a biodegradable plastic-degrading enzyme from cheese whey by the phyllosphere yeast Pseudozyma antarctica GB-4(1)W.

PubMed

Watanabe, Takashi; Shinozaki, Yukiko; Suzuki, Ken; Koitabashi, Motoo; Yoshida, Shigenobu; Sameshima-Yamashita, Yuka; Kuze Kitamoto, Hiroko

2014-08-01

Cheese whey is a by-product of cheese production and has high concentrations of lactose (about 5%) and other nutrients. Pseudozyma antarctica produces a unique cutinase-like enzyme, named PaE, that efficiently degrades biodegradable plastics. A previous study showed that a combination of 1% oil and 0.5% lactose increased cutinase-like enzyme production by another species of yeast. In this study, to produce PaE from cheese whey, we investigated the effects of soybean oil on PaE production (expressed as biodegradable plastic-degrading activity) by P. antarctica growing on lactose or cheese whey. In flask cultures, the final PaE activity was only 0.03 U/ml when soybean oil was used as the sole carbon source, but increased to 1.79 U/ml when a limited amount of soybean oil (under 0.5%) was combined with a relatively high concentration of lactose (6%). Using a 5-L jar fermentor with lactose fed-batch cultivation and periodic soybean oil addition, about 14.6 U/ml of PaE was obtained after 5 days of cultivation. When the lactose was replaced with cheese whey, PaE production was 10.8 U/ml after 3 days of cultivation. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Growth of cultured corneal endothelial cells onto a vitreous carbon matrix.

PubMed

Wickham, M G; Cleveland, P H; Binder, P S; Akers, P H

1983-01-01

Fourth passage cells of a rabbit corneal endothelial line were grown for 1 week in flasks containing pieces of a reticulated vitreous carbon matrix. The rate of cell growth in flasks containing the matrix was consistent with that in control flasks. Small fragments of the vitreous carbon material lying on the flask floor were covered by the monolayers as the cells grew to confluency. Vertical growth of cells onto larger pieces of the matrix proceeded in a staged fashion with maximum cell density on pieces of the matrix closest to the floor of the flask. As defined by scanning electron microscopy, cell growth occurred to a level at least 600 microns above the floor of the flask and the confluent monolayer. This novel culture procedure should be a model situation for study of many different aspects of the in vitro capabilities of corneal endothelial cells.

The Microbial Degradation of TCE (Trichloroethylene).

DTIC Science & Technology

1987-04-01

enrichment studies . All the sample flasks , including the controls, contained 14C in the 14C02 trap. The 14C measured in the control flask 14C02...layer compared to the controls. These data again suggested that TCE was being biologically modified. Those flasks flushed with air gave the first hard...only slightly soluble in water. All culture flasks were incubated at 250C for a minimum of a week. Results in the carbon and nitrogen source studies are

Atmospheric CO2 Concentrations from the Commonwealth Scientific and Industrial Research Organization (CSIRO) GASLAB Flask Sampling Network (March 1991 - December 2006)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO),; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2008-01-01

Individual measurements have been obtained from flask air samples returned to the CSIRO GASLAB. Typical sample storage times range from days to weeks for some sites (e.g. Cape Grim, Aircraft over Tasmania and Bass Strait) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for changes in sample CO2 mixing ratio during storage have shown significant drifts in some flask types over test periods of several months to years (Cooper et al., 1999). Corrections derived from the test results are applied to network data according to flask type. These measurements indicate a rise in annual average atmospheric CO2 concentration from 357.72 parts per million by volume (ppmv) in 1992 to 383.05 ppmv in 2006, or an increase in annual average of about 1.81 ppmv/year. These flask data may be compared with other flask measurements from the Scripps Institution of Oceanography, available through 2004 in TRENDS; both indicate an annual average increase of 1.72 ppmv/year throuth 2004. Differences may be attributed to different sampling times or days, different numbers of samples, and different curve-fitting techniques used to obtain monthly and annual average numbers from flask data. Measurement error in flask data is believed to be small (Masarie et al., 2001).

Oil on the water: Government regulation of a carcinogen in the twentieth-century Lancashire cotton spinning industry.

PubMed

Higgins, David; Tweedale, Geoffrey

2010-01-01

In the Lancashire cotton textile industry, mule spinners were prone to a chronic and sometimes fatal skin cancer (often affecting the groin). The disease had reached epidemic proportions by the 1920s, which necessitated action by the government, employers, and trade unions. In contrast to previous accounts, this article focuses on the government's reaction to mule spinners' cancer. Using official records in the National Archives, the slow introduction of health and safety measures by the government is explored in detail. Although obstructionism by the employers played a key role, one of the reasons for government inaction was the ambiguity of scientific research on engineering oils. On the other hand, prolonged scientific research suited a government policy that was framed around self regulation - a policy that had proved largely ineffective by the 1950s.

Preliminary report on the biological effects of space flight on the producing strain of a new immunosuppressant, Kanglemycin C.

PubMed

Zhou, Jianqin; Sun, Chenghang; Wang, Nanjin; Gao, Rongmei; Bai, Shuoke; Zheng, Huanrong; You, Xuefu; Li, Rongfeng

2006-08-01

Kanglemycin C (K-C) is a new immunosuppressant isolated from the culture broth of Nocardia mediterranei var. kanglensis 1747-64. To improve the productivity of K-C and to study the biological effects of space flight on its producing strain, spores from five K-C producing strains (U-10, U-15, U-7, M-13, gamma-33) mutated from the wild strain N. mediterranei var. kanglensis 1747-64 were carried into space by an unmanned spaceship, "Shenzhou III" (Divine Vessel III) on March 25, 2002. Comparatively, the strain U-7 was the highest K-C producing strain among the above five starting strains when cultivated in 500-ml Erlenmeyer flasks. After a 6 day and 18 h flight, the treated spores went through serial screening processes to screen for high-yield K-C mutant strains, using thin layer chromatography and high performance liquid chromatography (HPLC). The K-C yield produced by one mutant strain, designated as F-16, derived from the starting strain U-7 was increased by up to 200% when compared to that produced by the starting strain U-7 in 500-ml Erlenmeyer flasks after careful postflight HPLC analysis. Another mutant strain, designated as F-210, derived from the starting strain M-13 showed reduced productivity of K-C as well as exhibited changes in some morphological and physiological characteristics. For example, the broth color of the strain F-210 changed from yellow to purple after 96 h of culture, but that of the ground control strain M-13 remained yellow. Similarly, the mycelium morphological change from filamentous to coccoid of F-210 occurred later than that of ground control M-13. Examination of the survivability of postflight spores indicated that exposure to radiation, during the 162 h of space flight, plays a critical role in the survival rates of spores such that spores exposed to strong radiation exhibited lower survival rates than spores exposed to weak radiation.

Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2.

PubMed

Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

2016-06-01

Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield ( e.g . through genetic manipulation), or using low-cost substrates. In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L -1 ), yeast extracts (5 g.L -1 ), K 2 HPO 4 (6 g.L -1 ), NaH 2 PO 4 (7 g.L -1 ), NH 4 CL (0.7 g.L -1 ), and MgSO 4 (0.5 g.L -1 ). For evaluating the carbon and nitrogen sources' effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L -1 , respectively, and the maximum EPS production of 8.87 g.L -1 was achieved when glutamic acid (5 g.L -1 ) was employed as the nitrogen source. In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively.

Measurements of evaporated perfluorocarbon during partial liquid ventilation by a zeolite absorber.

PubMed

Proquitté, Hans; Rüdiger, Mario; Wauer, Roland R; Schmalisch, Gerd

2004-01-01

During partial liquid ventilation (PLV) the knowledge of the quantity of exhaled perfluorocarbon (PFC) allows a continuous substitution of the PFC loss to achieve a constant PFC level in the lungs. The aim of our in vitro study was to determine the PFC loss in the mixed expired gas by an absorber and to investigate the effect of the evaporated PFC on ventilatory measurements. To simulate the PFC loss during PLV, a heated flask was rinsed with a constant airflow of 4 L min(-1) and PFC was infused by different speeds (5, 10, 20 mL h(-1)). An absorber filled with PFC selective zeolites was connected with the flask to measure the PFC in the gas. The evaporated PFC volume and the PFC concentration were determined from the weight gain of the absorber measured by an electronic scale. The PFC-dependent volume error of the CO2SMO plus neonatal pneumotachograph was measured by manual movements of a syringe with volumes of 10 and 28 mL with a rate of 30 min(-1). Under steady state conditions there was a strong correlation (r2 = 0.999) between the infusion speed of PFC and the calculated PFC flow rate. The PFC flow rate was slightly underestimated by 4.3% (p < 0.01). However, this bias was independent from PFC infusion rate. The evaporated PFC volume was precisely measured with errors < 1%. The volume error of the CO2SMO-Plus pneumotachograph increased with increasing PFC content for both tidal volumes (p < 0.01). However for PFC flow rates up to 20 mL/h the error of the measured tidal volumes was < 5%. PFC selective zeolites can be used to quantify accurately the evaporated PFC volume during PLV. With increasing PFC concentrations in the exhaled air the measurement errors of ventilatory parameters have to be taken into account.

Ecofriendly Synthesis of nano Zero Valent Iron from Banana Peel Extract

NASA Astrophysics Data System (ADS)

Sunardi; Ashadi; Budi Rahardjo, Sentot; Inayati

2017-01-01

In this study, nano Zero Valent Iron (nZVI) were synthesized from banana peel extract (BPE) and ferrous sulfate. During the synthesis of nZVI both the precursor and the reducing agent were mixed in a clean sterilized flask in 1:1 proportion. For the reduction of Fe ions, 5 ml of filtered BPE was mixed to 5 ml of freshly prepared 0.001 M - 0.005 M aqueous of FeSO4 solution with constant stirring at room temperature. Within a particular time change in colour from brown to black color obtained by nanoparticles synthesis. A systematic characterization of nZVI was performed using UV-Vis. UV-visible absorption is used to investigate SPR. Characteristic surface plasmon absorption band was observed at 210 nm for the black colored nZVI synthesized from 0.001-0.005 M ferrous sulfate with BPE concentration 5 ml. It has been found that the optimum concentration for the synthesis of nZVI is 0.001M Fe2+ ions. There is small decrease in the intensity of SPR band from 0.001 to 0.005 M. The characterization size of nZVI was performed using TEM. The result shows that formation of particles size of nZVI was more 100 nm.

EVALUATION OF MIXING ENERGY IN FLASKS USED FOR DISPERSANT EFFECTIVENESS TESTING

EPA Science Inventory

A U.S. Environmental Protection Agency (EPA) laboratory screening protocol for dispersant effectiveness consists of placing water, oil, and a dispersant in a flask and mixing the contents on an orbital shaker. Two flasks are being investigated, a simple Erlenmeyer (used in EPA's...

Revisiting the round bottom flask rainbow experiment

NASA Astrophysics Data System (ADS)

Selmke, Markus; Selmke, Sarah

2018-01-01

A popular demonstration experiment in optics uses a round-bottom flask filled with water to project a circular rainbow on a screen with a hole through which the flask is illuminated. We show how the vessel's wall shifts the first- and second-order bows towards each other and consequently reduces the width of Alexander's dark band. We address the challenge this introduces in observing Alexander's dark band, and explain the importance of a sufficient distance between the flask and the screen. The wall-effect also introduces a splitting of the bows that can easily be misinterpreted.

Fidget physics

NASA Astrophysics Data System (ADS)

Marks, Julian; Bradfield, Philip

2017-08-01

Even without living with younger children, you can’t but be aware of the toy called a fidget spinner that is now ubiquitous and can be a nuisance when travelling on the bus with school children who continually fiddle with them.

Culture expansion of adipose derived stromal cells. A closed automated Quantum Cell Expansion System compared with manual flask-based culture.

PubMed

Haack-Sørensen, Mandana; Follin, Bjarke; Juhl, Morten; Brorsen, Sonja K; Søndergaard, Rebekka H; Kastrup, Jens; Ekblond, Annette

2016-11-16

Adipose derived stromal cells (ASCs) are a rich and convenient source of cells for clinical regenerative therapeutic approaches. However, applications of ASCs often require cell expansion to reach the needed dose. In this study, cultivation of ASCs from stromal vascular fraction (SVF) over two passages in the automated and functionally closed Quantum Cell Expansion System (Quantum system) is compared with traditional manual cultivation. Stromal vascular fraction was isolated from abdominal fat, suspended in α-MEM supplemented with 10% Fetal Bovine Serum and seeded into either T75 flasks or a Quantum system that had been coated with cryoprecipitate. The cultivation of ASCs from SVF was performed in 3 ways: flask to flask; flask to Quantum system; and Quantum system to Quantum system. In all cases, quality controls were conducted for sterility, mycoplasmas, and endotoxins, in addition to the assessment of cell counts, viability, immunophenotype, and differentiation potential. The viability of ASCs passage 0 (P0) and P1 was above 96%, regardless of cultivation in flasks or Quantum system. Expression of surface markers and differentiation potential was consistent with ISCT/IFATS standards for the ASC phenotype. Sterility, mycoplasma, and endotoxin tests were consistently negative. An average of 8.0 × 10 7 SVF cells loaded into a Quantum system yielded 8.96 × 10 7 ASCs P0, while 4.5 × 10 6 SVF cells seeded per T75 flask yielded an average of 2.37 × 10 6 ASCs-less than the number of SVF cells seeded. ASCs P1 expanded in the Quantum system demonstrated a population doubling (PD) around 2.2 regardless of whether P0 was previously cultured in flasks or Quantum, while ASCs P1 in flasks only reached a PD of 1.0. Manufacturing of ASCs in a Quantum system enhances ASC expansion rate and yield significantly relative to manual processing in T-flasks, while maintaining the purity and quality essential to safe and robust cell production. Notably, the use of the Quantum system entails significantly reduced working hours and thereby costs.

Some Interesting Thermodynamics of the Thermos Flask

ERIC Educational Resources Information Center

Berger, Roland

2007-01-01

When opening a thermos flask filled with coffee, one often "hears" the equalization of the pressure difference that appears to be present between the air cavity inside the flask and the surrounding room atmosphere. Recently we discussed this phenomenon while drinking coffee and guessed about the direction of the gas stream accompanying the…

Studies on Viral Disinfection: An Evaluation of Moist Heat Disinfection for HBV by Using A0 Concept Defined in ISO 15883-Washer-Disinfectors.

PubMed

Uetera, Yushi; Kawamura, Kunio; Kobayashi, Hiroyoshi; Saito, Yuhei; Yasuhara, Hiroshi; Saito, Ryoichi

2010-01-01

International Organization for Standardization (ISO) 15883 for washer-disinfectors has introduced the A(0) concept to allow comparison of the lethality of moist heat processes. The A(0) value is the equivalent disinfection time in seconds at 80 °C calculated on the basis of microbial killing kinetics when the disinfection temperature is over 65 °C. Hepatitis B virus (HBV), transmissible only to humans and chimpanzees, is an important heat-resistant, blood-borne pathogen. Therefore, it is mandatory to disinfect HBV thoroughly in the washer-disinfectors employed for surgical instruments. Additionally, it has become extremely difficult to use chimpanzees as experimental models or to perform human volunteer studies. Therefore, it is considered worthwhile to re-evaluate the reported data on the moist heat disinfection of HBV using the A(0) value. In the voluntary active immunization to humans in 1973, HBV serum (infectivity titer: 10(6.5) CID(50)/mL) underwent moist heat disinfection at 98 °C for 1 min in a flask over an electric burner (conservatively estimated A(0) value: 3786). Then, 0.1 mL was inoculated to each of 29 volunteers. No one revealed evidence of infection clinically or in the laboratory tests available at the time. In 1979, a more sensitive test appeared and revealed three sub-clinically infected volunteers. In the 1980s, there were two chimpanzee experimental models using HBV serum (infectivity titer: 10(5) CID(50)/mL). In one model, the serum underwent moist heat disinfection at 98 °C for 2 min in a thermostat bath (conservatively estimated A(0) value: 7571). One milliliter was inoculated to each of two chimpanzees, and both of them revealed no evidence of infection. In another model, the serum underwent moist heat disinfection using two conditions in a thermostat bath, respectively: at 103 °C for 90 s (A(0) value: 24865) and at 65 °C for 10 h (A(0) value: 1138). Ten milliliters of each sample were mixed. Then, the mixture was inoculated to each of two chimpanzees. Both of them revealed no evidence of infection. In the human volunteer study, the serum infectivity titer was more than 30 times (10(1.5) times) higher than that used in the two chimpanzee experimental models. Moreover, the serum was heated in the flask over an electric burner, which is considered less reliable than the thermostat baths to realize uniform heat distribution. It is assumed that these factors were predisposed to the result that a conservatively estimated A(0) value of 3786 failed to inactivate the HBV serum of 10(6.5) CID(50)/mL. In the two chimpanzee models, it was suggested that A(0) value not less than 1138 was able to inactivate the HBV serum of 10(5) CID(50)/mL.

Production and structural elucidation of exopolysaccharide from endophytic Pestalotiopsis sp. BC55.

PubMed

Mahapatra, Subhadip; Banerjee, Debdulal

2016-01-01

There is a little information on exopolysaccharide production by endophytic fungi. In this investigation endophytic Pestalotiopsis sp. BC55 was used for optimization of exopolysaccharide production. One variable at a time method and response surface methodology were adopted to find out the best culture conditions and medium compositions for maximum exopolysaccharide production. The organism produced maximum exopolysaccharide (4.320 ± 0.022 g/l EPS) in 250 ml Erlenmeyer flask containing 75 ml potato dextrose broth supplemented with (g%/l) glucose, 7.66; urea, 0.29; CaCl2, 0.05 with medium pH 6.93; after 3.76 days of incubation at 24°C. Exopolysaccharide [EPS (EP-I)] produced by this organism have Mw ∼2×10(5)Da with a melting point range of 122-124°C. Structural elucidation of the EPS (PS-I) was carried out after a series of experiments. Result indicated the presence of only (1→3)-linked β-d-glucopyranosyl moiety. The structure of the repeating unit was established as - →3)-β-d-Glcp-(1→. Copyright © 2015 Elsevier B.V. All rights reserved.

An effective and simplified scale-up strategy for acarbose fermentation based on the carbon source control.

PubMed

Li, Kun-tai; Wie, Sai-jin; Huang, Lin; Cheng, Xin

2012-02-01

The scale-up strategy for acarbose fermentation by Actinoplanes sp. A56 was explored in this paper. The results obtained in shake-flask cultivation demonstrated that the ratio of maltose and glucose had significant effects on the biosynthesis of acarbose, and the feeding medium containing 3:1 (mass ratio) of maltose and glucose was favorable for acarbose production. Then the correlation of the carbon source concentration with acarbose production was further investigated in 100-l fermenter, and the results showed that 7.5-8.0 g of total sugar/100 ml and 4.0-4.5 g of reducing sugar/100 ml were optimal for acarbose production. Based on the results in 100-l fermenter, an effective and simplified scale-up strategy was successfully established for acarbose fermentation in a 30-m(3) fermenter, by using total sugar and reducing sugar as the scale-up parameter. As a result, 4,327 mg of acarbose/l was obtained at 168 h of fermentation.

Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation

PubMed Central

Darah, I.; Sumathi, G.; Jain, K.; Hong, Lim Sheh

2011-01-01

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO3, 0.1% KH2PO4, 0.05% MgSO4 ·7H2O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 106 spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment. PMID:21826273

Involvement of Physical Parameters in Medium Improvement for Tannase Production by Aspergillus niger FETL FT3 in Submerged Fermentation.

PubMed

Darah, I; Sumathi, G; Jain, K; Hong, Lim Sheh

2011-01-01

Aspergillus niger FETL FT3, a local extracellular tannase producer strain that was isolated from one of dumping sites of tannin-rich barks of Rhizophora apiculata in Perak, Malaysia. This fungus was cultivated in 250 mL Erlenmeyer flask under submerged fermentation system. Various physical parameters were studied in order to maximize the tannase production. Maximal yield of tannase production, that is, 2.81 U per mL was obtained on the fourth day of cultivation when the submerged fermentation was carried out using liquid Czapek-Dox medium containing (percent; weight per volume) 0.25% NaNO(3), 0.1% KH(2)PO(4), 0.05% MgSO(4) ·7H(2)O, 0.05% KCl, and 1.0% tannic acid. The physical parameters used initial medium pH of 6.0, incubation temperature of 30°C, agitation speed of 200 rpm and inoculums size of 6 × 10(6) spores/ ml. This research has showed that physical parameters were influenced the tannase production by the fungus with 156.4 percent increment.

Cloning and strong expression of a Bacillus subtilis WL-3 mannanase gene in B. subtilis.

PubMed

Yoon, Ki-Hong; Lim, Byung-Lak

2007-10-01

A gene encoding the mannanase of Bacillus subtilis WL-3, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and the nucleotide sequence of a 2.7-kb DNA fragment containing the mannanase gene was subsequently determined. The mannanase gene, designated manA, consisted of 1,080 nucleotides encoding polypeptide of 360 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to glycosyl hydrolase family 26. The manA gene was strongly expressed in B. subtilis 168 by cloning the gene downstream of a strong B. subtilis promoter of plasmid pJ27Delta 88U. In flask cultures, the production of mannanase by recombinant B. subtilis 168 reached maximum levels of 300 units/ml and 450 units/ml in LB medium and LB medium containing 0.3% locust bean gum, respectively. Based on the zymogram of the mannanase, it was found that the mannanase produced by recombinant B. subtilis could be maintained stably without proteolytic degradation during the culture time.

Enhancement of docosahexaenoic acid production by Schizochytrium SW1 using response surface methodology

NASA Astrophysics Data System (ADS)

Nazir, Mohd Yusuf Mohd; Al-Shorgani, Najeeb Kaid Nasser; Kalil, Mohd Sahaid; Hamid, Aidil Abdul

2015-09-01

In this study, three factors (fructose concentration, agitation speed and monosodium glutamate (MSG) concentration) were optimized to enhance DHA production by Schizochytrium SW1 using response surface methodology (RSM). Central composite design was applied as the experimental design and analysis of variance (ANOVA) was used to analyze the data. The experiments were conducted using 500 mL flask with 100 mL working volume at 30°C for 96 hours. ANOVA analysis revealed that the process was adequately represented significantly by the quadratic model (p<0.0001) and two of the factors namely agitation speed and MSG concentration significantly affect DHA production (p<0.005). Level of influence for each variable and quadratic polynomial equation were obtained for DHA production by multiple regression analyses. The estimated optimum conditions for maximizing DHA production by SW1 were 70 g/L fructose, 250 rpm agitation speed and 12 g/L MSG. Consequently, the quadratic model was validated by applying of the estimated optimum conditions, which confirmed the model validity and 52.86% of DHA was produced.

The significance of fructose and MSG in affecting lipid and docosahexaenoic acid (DHA) production of Aurantiochytrium sp. SW1

NASA Astrophysics Data System (ADS)

Rahman, Shariffah Nurhidayah Syed Abdul; Kalil, Mohd Sahaid; Hamid, Aidil Abdul

2018-04-01

Optimization of fermentation medium for the production of docosahexaenoic acid (DHA) by Aurantiochytrium sp. SW1 was carried out. In this study, levels of fructose, monosodium glutamate (MSG) and sea salt were optimized for enhanced lipid and DHA production using response surface methodology (RSM). The design contains a total of 20 runs with 6 central points replication. Cultivation was carried out in 500 mL flasks containing 100 mL nitrogen limited medium at 30°C for 96h. Sequential model sum of squares (SS) revealed that the system was adequately represented by a quadratic model (p<0.0001). ANOVA results showed that fructose and MSG as a single factor has significant positive effect on the DHA content of SW1. The estimated optimal levels of the factors were 100 g/L fructose, 8 g/L MSG and 47% sea salt. Subsequent cultivation employing the suggested values confirmed that the predicted response values were experimentally achievable and reproducible, where 8.82 g/L DHA (51.34% g/g lipid) was achieved.

ARM-LBNL-NOAA Flask Sampler for Carbon Cycle Gases

DOE Data Explorer

Torn, Margaret

2008-01-15

Data from ccg-flasks are sampled at the ARM SGP site and analyzed by the NOAA Earth System Research Laboratory (ESRL) as part of the NOAA Cooperative Global Air Sampling Network. Surface samples are collected from a 60m tower at the SGP Central Facility, usually once per week on one afternoon. The aircraft samples are collected approximately weekly from a chartered aircraft, and the collection flight path is centered over the tower where the surface samples are collected. Samples are collected by the ARM/LBNL Carbon Project. CO2 flask data contains measurements of CO2 concentration and CO2 stable isotope ratios (13CO2 and C18OO) from flasks collected at the SGP site. The flask samples are collected at 2m, 4m, 25m, and 60m along the 60m tower.

A Self-Powered and Autonomous Fringing Field Capacitive Sensor Integrated into a Micro Sprinkler Spinner to Measure Soil Water Content.

PubMed

da Costa, Eduardo Ferreira; de Oliveira, Nestor E; Morais, Flávio J O; Carvalhaes-Dias, Pedro; Duarte, Luis Fernando C; Cabot, Andreu; Siqueira Dias, J A

2017-03-12

We present here the design and fabrication of a self-powered and autonomous fringing field capacitive sensor to measure soil water content. The sensor is manufactured using a conventional printed circuit board and includes a porous ceramic. To read the sensor, we use a circuit that includes a 10 kHz triangle wave generator, an AC amplifier, a precision rectifier and a microcontroller. In terms of performance, the sensor's capacitance (measured in a laboratory prototype) increases up to 5% when the volumetric water content of the porous ceramic changed from 3% to 36%, resulting in a sensitivity of S = 15.5 pF per unity change. Repeatability tests for capacitance measurement showed that the θ v sensor's root mean square error is 0.13%. The average current consumption of the system (sensor and signal conditioning circuit) is less than 1.5 μ A, which demonstrates its suitability for being powered by energy harvesting systems. We developed a complete irrigation control system that integrates the sensor, an energy harvesting module composed of a microgenerator installed on the top of a micro sprinkler spinner, and a DC/DC converter circuit that charges a 1 F supercapacitor. The energy harvesting module operates only when the micro sprinkler spinner is irrigating the soil, and the supercapacitor is fully charged to 5 V in about 3 h during the first irrigation. After the first irrigation, with the supercap fully charged, the system can operate powered only by the supercapacitor for approximately 23 days, without any energy being harvested.

A Self-Powered and Autonomous Fringing Field Capacitive Sensor Integrated into a Micro Sprinkler Spinner to Measure Soil Water Content

PubMed Central

da Costa, Eduardo Ferreira; de Oliveira, Nestor E.; Morais, Flávio J. O.; Carvalhaes-Dias, Pedro; Duarte, Luis Fernando C.; Cabot, Andreu; Siqueira Dias, J. A.

2017-01-01

We present here the design and fabrication of a self-powered and autonomous fringing field capacitive sensor to measure soil water content. The sensor is manufactured using a conventional printed circuit board and includes a porous ceramic. To read the sensor, we use a circuit that includes a 10 kHz triangle wave generator, an AC amplifier, a precision rectifier and a microcontroller. In terms of performance, the sensor’s capacitance (measured in a laboratory prototype) increases up to 5% when the volumetric water content of the porous ceramic changed from 3% to 36%, resulting in a sensitivity of S=15.5 pF per unity change. Repeatability tests for capacitance measurement showed that the θv sensor’s root mean square error is 0.13%. The average current consumption of the system (sensor and signal conditioning circuit) is less than 1.5 μA, which demonstrates its suitability for being powered by energy harvesting systems. We developed a complete irrigation control system that integrates the sensor, an energy harvesting module composed of a microgenerator installed on the top of a micro sprinkler spinner, and a DC/DC converter circuit that charges a 1 F supercapacitor. The energy harvesting module operates only when the micro sprinkler spinner is irrigating the soil, and the supercapacitor is fully charged to 5 V in about 3 h during the first irrigation. After the first irrigation, with the supercap fully charged, the system can operate powered only by the supercapacitor for approximately 23 days, without any energy being harvested. PMID:28287495

Preliminary Tests of Blowers of Three Designs Operating in Conjunction with a Wing-Duct Cooling System for Radial Engines, Special Report

NASA Technical Reports Server (NTRS)

Biermann, David; Valentine, E. Floyd

1939-01-01

This paper is one of several dealing with methods intended to reduce the drag of present-day radial engine installations and improve the cooling at zero and low air speeds, The present paper describes model wind-tunnel tests of blowers of three designs tested in conjunction with a wing-nacelle combination. The principle of operation involved consists of drawing cooling air into ducts located in the wing root at the point of maximum slipstream velocity, passing the air through the engine baffles from rear to front, and exhausting the air through an annular slot located between the propeller and the engine with the aid of a blower mounted on the spinner. The test apparatus consisted essentially of a stub wing having a 5-foot chord and a 15-foot span, an engine nacelle of 20 inches diameter enclosing a 25-horsepower electric motor, and three blowers mounted on propeller spinners. Two of the blowers utilize centrifugal force while the other uses the lift from airfoils to force the air out radially through the exit slot. Maximum efficiencies of over 70 percent were obtained for the system as a whole. Pressures were measured over the entire flight range which were in excess of those necessary to cool present-day engines, The results indicated that blowers mounted on propeller spinners could be built sufficiently powerful and efficient to warrant their use as the only, or chief, means of forcing air through the cooling system, so that cooling would be independent of the speed of the airplane.

Three-dimensional (3D) evaluation of liquid distribution in shake flask using an optical fluorescence technique.

PubMed

Azizan, Amizon; Büchs, Jochen

2017-01-01

Biotechnological development in shake flask necessitates vital engineering parameters e.g. volumetric power input, mixing time, gas liquid mass transfer coefficient, hydromechanical stress and effective shear rate. Determination and optimization of these parameters through experiments are labor-intensive and time-consuming. Computational Fluid Dynamics (CFD) provides the ability to predict and validate these parameters in bioprocess engineering. This work provides ample experimental data which are easily accessible for future validations to represent the hydrodynamics of the fluid flow in the shake flask. A non-invasive measuring technique using an optical fluorescence method was developed for shake flasks containing a fluorescent solution with a waterlike viscosity at varying filling volume (V L  = 15 to 40 mL) and shaking frequency ( n  = 150 to 450 rpm) at a constant shaking diameter (d o  = 25 mm). The method detected the leading edge (LB) and tail of the rotating bulk liquid (TB) relative to the direction of the centrifugal acceleration at varying circumferential heights from the base of the shake flask. The determined LB and TB points were translated into three-dimensional (3D) circumferential liquid distribution plots. The maximum liquid height (H max ) of the bulk liquid increased with increasing filling volume and shaking frequency of the shaking flask, as expected. The toroidal shapes of LB and TB are clearly asymmetrical and the measured TB differed by the elongation of the liquid particularly towards the torus part of the shake flask. The 3D liquid distribution data collected at varying filling volume and shaking frequency, comprising of LB and TB values relative to the direction of the centrifugal acceleration are essential for validating future numerical solutions using CFD to predict vital engineering parameters in shake flask.

CRYOGENIC DEWAR

DOEpatents

Chamberlain, W.H.; Maseck, H.E.

1964-01-28

This patent relates to a dewar for storing cryogenic gase and is of the type having aii inner flask surrounded by a vacuum jacket and having a vent spout through which evaporating gas escapes. Heretofore substantial gas loss has resulted from the radiation of heat towards the flask from the warmer outer elements of the dewar. In this invention, the mask is surrounded by a thermally conducting shield which is disposed in the vacuum space between the flask and the outer elements of the dewar. The shield contacts only the vent spout, which is cooled by the evaporating gas, and thus is maintained at a temperature very close to that of the flask itself. Accordingly, heat radiated toward the flask is intercepted and conducted to the evaporating gas rather than being re-radiated towards the hask. In a liquid helium dewar of typical configniration the mention reduces the boil-off rate by approximately one-half.(AEC)

Particle Trajectory and Icing Analysis of the E(sup 3) Turbofan Engine Using LEWICE3D Version 3

NASA Technical Reports Server (NTRS)

Bidwell, Colin S.

2011-01-01

Particle trajectory and ice shape calculations were made for the Energy Efficient Engine (E(sup 3)) using the LEWICE3D Version 3 software. The particle trajectory and icing computations were performed using the new "block-to-block" collection efficiency method which has been incorporated into the LEWICE3D Version 3 software. The E(sup 3) was developed by NASA and GE in the early 1980 s as a technology demonstrator and is representative of a modern high bypass turbofan engine. The E(sup 3) flow field was calculated using the NASA Glenn ADPAC turbomachinery flow solver. Computations were performed for the low pressure compressor of the E(sup 3) for a Mach 0.8 cruise condition at 11,887 m assuming a standard warm day for three drop sizes and two drop distributions typically used in aircraft design and certification. Particle trajectory computations were made for water drop sizes of 5, 20, and 100 microns. Particle trajectory and ice shape predictions were made for a 20 micron Langmuir-D distribution and for a 92 mm Super-cooled Large Droplet (SLD) distribution with and without splashing effects for a Liquid Water Content (LWC) of 0.3 g/cu m and an icing time of 30 min. The E3 fan and spinner combination proved to be an effective ice removal mechanism as they removed greater than 36 percent of the mass entering the inlet for the icing cases. The maximum free stream catch fraction for the fan and spinner combination was 0.60 while that on the elements downstream of the fan was 0.03. The non-splashing trajectory and collection efficiency results showed that as drop size increased impingement rates increased on the spinner and fan leaving less mass to impinge on downstream components. The SLD splashing case yielded more mass downstream of the fan than the SLD non-splashing case due to mass being splashed from the upstream inlet lip, spinner and fan components. The ice shapes generated downstream of the fan were either small or nonexistent due to the small available mass and evaporation except for the 92 m SLD splashing case. Relatively large ice shapes were predicted for internal guide vane #1 and rotor #1 for the 92 m SLD splashing case due to re-impingement of splashed mass.

Carbon and oxygen isotope ratios of ecosystem respiration along an Oregon conifer transect: preliminary observations based on small-flask sampling.

PubMed

Ehleringer, J. R.; Cook, C. S.

1998-01-01

Isotope ratio analyses of atmospheric CO(2) at natural abundance have significant potential for contributing to our understanding of photosynthetic and respiration processes in forest ecosystems. Recent advances in isotope ratio mass spectrometry allow for rapid, on-line analysis of small volumes of CO(2) in air, and open new research opportunities at the ecophysiological, whole-organism, and atmospheric levels. Among the immediate applications are the carbon and oxygen isotope ratio analyses of carbon dioxide in atmospheric air. Routine analysis of carbon dioxide in air volumes of approximately 50-300 &mgr;l is accomplished by linking a commercially available, trace gas condenser and gas chromatograph to an isotope ratio mass spectrometer operated in continuous-flow mode. Samples collected in the field are stored in either gas-tight syringes or 100-ml flasks. The small sample volume required makes it possible to subsample the air in flasks for CO(2) and then to sample the remaining air volume for the analysis of the isotopic composition of either methane or nitrous oxide. Reliable delta(13)C and delta(18)O values can be obtained from samples collected and stored for 1-3 days. Longer-term storage, on the order of weeks, is possible for delta(13)C measurements without drift in the isotope ratio signal, and should also be possible for delta(18)O measurements. When linked with an infrared gas analyzer, pump and flask sampling system, it is feasible to sample CO(2) extensively in remote forest locations. The air-sampling system was used to measure the isotope ratios of atmospheric CO(2) and to conduct a regression analysis of the relationship between these two parameters. From the regression, we calculated the delta(13)C of ecosystem respiration of four coniferous ecosystems along a precipitation gradient in central Oregon. The ecosystems along the coast-to-interior Oregon (OTTER) gradient are dominated by spruce-hemlock forests at the wet, coastal sites (> 200 cm precipitation annually) to juniper woodlands (20 cm precipitation) at the interior, dry end of the transect. The delta(13)C values of ecosystem respiration along this transect differed by only 1.3 per thousand (range of -25.2 to -23.9 per thousand ) during August at the peak of the summer drought. Following autumn rains in September, the delta(13)C of ecosystem respiration in the four stands decreased; overall the difference in the carbon isotope ratio of ecosystem respiration among sites increased to 3.9 per thousand (-26.8 to -22.9 per thousand ).

50 CFR 216.161 - Specified activity and incidental take levels by species.

Code of Federal Regulations, 2012 CFR

2012-10-01

...); rough-toothed dolphin (Steno bredanensis); common dolphin (Delphinus delphis), false killer whale... species: Minke whale (Balaenoptera acutorostrata), dwarf sperm whale (Kogia simus); pygmy sperm whale (K. breviceps); pilot whale (Globicephala macrorhynchus); Atlantic spotted dolphin (Stenella frontalis); spinner...

Determination of Se in biological samples by axial view inductively coupled plasma optical emission spectrometry after digestion with aqua regia and on-line chemical vapor generation

NASA Astrophysics Data System (ADS)

dos Santos, Éder José; Herrmann, Amanda Beatriz; de Caires, Suzete Kulik; Frescura, Vera Lúcia Azzolin; Curtius, Adilson José

2009-06-01

A simple and fast method for the determination of Se in biological samples, including food, by axial view inductively coupled plasma optical emission spectrometry using on-line chemical vapor generation (CVG-ICP OES) is proposed. The concentrations of HCl and NaBH 4, used in the chemical vapor generation were optimized by factorial analysis. Six certified materials (non-fat milk powder, lobster hepatopancreas, human hair, whole egg powder, oyster tissue, and lyophilised pig kidney) were treated with 10 mL of aqua regia in a microwave system under reflux for 15 min followed by additional 15 min in an ultrasonic bath. The solutions were transferred to a 100 mL volumetric flask and the final volume was made up with water. The Se was determined directly in these solutions by CVG-ICP OES, using the analytical line at 196.026 nm. Calibration against aqueous standards in 10% v/v aqua regia in the concentration range of 0.5-10.0 µg L - 1 Se(IV) was used for the analysis. The quantification limit, considering a 0.5 g sample weight in a final volume of 100 mL - 1 was 0.10 µg g - 1. The obtained concentration values were in agreement with the total certified concentrations, according to the t-test for a 95% confidence level.

Dextran Utilization During Its Synthesis by Weissella cibaria RBA12 Can Be Overcome by Fed-Batch Fermentation in a Bioreactor.

PubMed

Baruah, Rwivoo; Deka, Barsha; Kashyap, Niharika; Goyal, Arun

2018-01-01

Weissella cibaria RBA12 produced a maximum of 9 mg/ml dextran (with 90% efficiency) using shake flask culture under the optimized concentration of medium components viz. 2% (w/v) of each sucrose, yeast extract, and K 2 HPO 4 after incubation at optimized conditions of 20 °C and 180 rpm for 24 h. The optimized medium and conditions were used for scale-up of dextran production from Weissella cibaria RBA12 in 2.5-l working volume under batch fermentation in a bioreactor that yielded a maximum of 9.3 mg/ml dextran (with 93% efficiency) at 14 h. After 14 h, dextran produced was utilized by the bacterium till 18 h in its stationary phase under sucrose depleted conditions. Dextran utilization was further studied by fed-batch fermentation using sucrose feed. Dextran on production under fed-batch fermentation in bioreactor gave 35.8 mg/ml after 32 h. In fed-batch mode, there was no decrease in dextran concentration as observed in the batch mode. This showed that the utilization of dextran by Weissella cibaria RBA12 is initiated when there is sucrose depletion and therefore the presence of sucrose can possibly overcome the dextran hydrolysis. This is the first report of utilization of dextran, post-sucrose depletion by Weissella sp. studied in bioreactor.

Development of an ES-like cell culture system (RESC) from rohu, Labeo rohita (Ham.).

PubMed

Goswami, M; Lakra, W S; Yadav, Kamalendra; Jena, J K

2012-12-01

An embryonic stem (ES)-like cell culture system RESC from a commercially important freshwater carp, Labeo rohita, was developed using blastula stage embryos. The cells were cultured in Leibovitz-15 (L-15) medium in gelatin-coated cell culture flask supplemented with 15 % fetal bovine serum along with 10 ng ml(-1) basic fibroblast growth factor at 28 °C under feeder-free conditions. The ES-like cells were characterized by their unique morphology, alkaline phosphatase activity, embryoid body formation tendency, expression of transcription factor Oct4, and consistent chromosome count. The RESC cells when treated with retinoic acid differentiated into cells of different lineages. The RESC developed from mid-blastula embryos of L. rohita would be a useful tool for cellular differentiation and gene expression studies.

Effect of Oxygen-Supply Rates on Growth of Escherichia coli

PubMed Central

McDaniel, L. E.; Bailey, E. G.; Zimmerli, A.

1965-01-01

The effect of oxygen-supply rates on bacterial growth was studied in commercially available unbaffled and baffled flasks with the use of Escherichia coli in a synthetic medium as a test system. The amount of growth obtained depended on the oxygen-supply rate. Based on oxygen-absorption rates (OAR) measured by the rate of sulfite oxidation, equal OAR values in different types of flasks did not give equal amounts of growth. However, growth was essentially equal at the equal sulfite-oxidation rates when these were determined in the presence of killed whole cultures. Specific growth rates were reduced only at oxygen-supply rates much lower than those at which the total amount of growth was reduced. For the physical set-up used in this work and with the biological system employed, Bellco 598 flasks and flasks fitted with Biotech stainless-steel baffles gave satisfactory results at workable broth volumes; unbaffled and Bellco 600 flasks did not. PMID:14264837

Mercury - Its occurrence and economic trends

USGS Publications Warehouse

Bailey, Edgar H.; Smith, Roscoe M.

1964-01-01

price will be a predictable increase in production. If the price averages $300 a flask for 1 year or more, annual production may increase to a rate of 30,000-35,000 flasks in 1966 or 1967. If the price averages $400 a flask, production may increase to 45,000-50,000 flasks a year after a timelag of two or three years. New ore bodies would have to be found however, to attain these rates of production, and the mining of lower grade ores would be needed to sustain them for more than a few years. By 1966 the U.S. economy will require at least 67 000 flasks, and the domestic demand for mercury thereafter will continue to increase. The probability that domestic mines will continue to supply a fourth or more of the domestic consumption is dependent upon the price of mercury for the next few years. The outlook for the long-term supply is reassuring, but it is dependent, not only upon price, but upon continued progress in new techniques of discovery.

Sensor Access to the Cellular Microenvironment Using the Sensing Cell Culture Flask.

PubMed

Kieninger, Jochen; Tamari, Yaara; Enderle, Barbara; Jobst, Gerhard; Sandvik, Joe A; Pettersen, Erik O; Urban, Gerald A

2018-04-26

The Sensing Cell Culture Flask (SCCF) is a cell culture monitoring system accessing the cellular microenvironment in 2D cell culture using electrochemical microsensors. The system is based on microfabricated sensor chips embedded in standard cell culture flasks. Ideally, the sensor chips could be equipped with any electrochemical sensor. Its transparency allows optical inspection of the cells during measurement. The surface of the sensor chip is in-plane with the flask surface allowing undisturbed cell growth on the sensor chip. A custom developed rack system allows easy usage of multiple flasks in parallel within an incubator. The presented data demonstrates the application of the SCCF with brain tumor (T98G) and breast cancer (T-47D) cells. Amperometric oxygen sensors were used to monitor cellular respiration with different incubation conditions. Cellular acidification was accessed with potentiometric pH sensors using electrodeposited iridium oxide films. The system itself provides the foundation for electrochemical monitoring systems in 3D cell culture.

78 FR 33811 - Marine Mammals; File No. 17941

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-05

... educational photography on bottlenose (Tursiops truncatus) and spinner (Stenella longirostris) dolphins... Register (78 FR 17639) that a request for a permit to conduct commercial/educational photography had been... photography/filming projects. The first consists of helicopter flights over Florida Bay to film bottlenose...

Scrotal cancer in the north-west of England, 1962-68

PubMed Central

Lee, W. R.; Alderson, M. R.; Downes, Jean E.

1972-01-01

Lee, W. R., Alderson, M. R., and Downes, J. E. (1972).Brit. J. industr. Med.,29, 188-195. Scrotal cancer in the north-west of England, 1962-68. During the seven years 1962 to 1968 there were 103 cases of scrotal cancer reported to the Manchester Regional Cancer Registry. It was possible to obtain reasonably full occupational histories concerning 89 of these cases either from the men themselves or from close relatives. Fifty-one had been mule spinners, 40 of whom had worked in no other industry with a hazard of scrotal cancer, but the other 11 had. Another 19 men had never been mule spinners but had worked in occupations with a recognized hazard of this condition. Of the remainder, 14 had worked in occupations with a possible risk, although perhaps not generally recognized as such, and in five no clear cause was apparent. Because of the small numbers, it was possible to compare the observed and expected incidences only in broad occupational groups and these showed a statistically significant excess (P<0·001) only in textile workers and unskilled workers. However, our findings, taken with those of earlier workers, suggest that further attention might be given to four other groups, namely, automatic lathe operators, road workers, dye workers, and chain makers. Cancer of the scrotum tends to be a disease of the elderly. In 42% of men (other than mule spinners) scrotal cancer developed after the men had passed the age of retirement and therefore too late for routine medical examination at the workplace to be of any value. Fifty of the men had died, but scrotal cancer was registered as the underlying cause in only 19 and appeared on the death certificate in a further four. PMID:5063206

Technical note: Method for isolation of the bovine sweat gland and conditions for in vitro culture.

PubMed

Hamzaoui, S; Burger, C A; Collier, J L; Collier, R J

2018-05-01

Apocrine sweat glands in bovine skin are involved in thermoregulation. Human, horse, and sheep sweat gland epithelial cells have been isolated and grown in vitro. The present study was conducted to identify a method to isolate bovine sweat glands and culture apocrine bovine sweat gland epithelial cells in vitro. Mechanical shearing, collagenase digestion, centrifugation, and neutral red staining were used to identify and isolate the apocrine glands from skin. Bovine sweat glands in situ and after isolation comprised 2 major cell types consisting of a single layer of cuboidal epithelial cells resting on a layer of myoepithelial cells. In situ, the glands were embedded in a collagen matrix primarily comprising fibroblasts, and some of these cells were also present in the isolated material. The isolated material was transferred to complete medium (keratinocyte serum-free medium, bovine pituitary extract, and human recombinant epidermal growth factor + 2.5% fetal bovine serum) in a T 25 flask (Falcon, Franklin Lakes, NJ) with media film and then incubated at 37°C for 24 h. After sweat glands adhered to the bottom of the flask, an additional 2 mL of complete medium was added and the medium was changed every 3 d. Isolated apocrine sweat glands and bovine sweat gland epithelial cells were immunostained for cytokeratin and fibroblast specific protein, indicating fibroblast-free cultures. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Influence of nutritional and physicochemical variables on PHB production from raw glycerol obtained from a Colombian biodiesel plant by a wild-type Bacillus megaterium strain.

PubMed

Moreno, Paalo; Yañez, Camilo; Cardozo, Nilo Sérgio Medeiros; Escalante, Humberto; Combariza, Marianny Y; Guzman, Carolina

2015-12-25

Biodegradable polymers are currently viable alternatives to traditional synthetic polymers. For instance, polyhydroxybutyrate (PHB) is intracellularly produced and accumulated by Bacillus species, among others. This study reports several wild-type Bacillus strains with the ability to accumulate PHB using raw glycerol from biodiesel production as the sole carbon source. Out of 15 strains from different sources, B. megaterium B2 was selected as the most promising strain for further statistical optimization of the medium composition. Plackett-Burman and central composite designs were used to establish key variables and optimal culture conditions for PHB production using both 250-mL shake flasks and a 7.5-L bioreactor. Temperature and concentrations of glycerol and Na2HPO4 are the experimental variables with the most significant influence on PHB production by B2. After 14 hours of fermentation in shake flasks with optimized medium, B2 produced 0.43 g/L of PHB with a 34% accumulation in the cells. In contrast, under the same conditions, a maximum PHB concentration of 1.20 g/L in the bioreactor was reached at 11 hours. These values correspond to a 48% and 314% increase in PHB production compared to the initial culture conditions. These results suggest the potential of B2 as a PHB producer using raw glycerol, which is an inexpensive, abundant and readily available carbon source. Copyright © 2015 Elsevier B.V. All rights reserved.

Oxygen consumption rate and mitochondrial density in human melanoma monolayer cultures and multicellular spheroids.

PubMed

Hystad, M E; Rofstad, E K

1994-05-15

Rate of oxygen consumption per cell has been shown in previous studies to decrease with increasing depth in the viable rim of multicellular spheroids initiated from rodent cells, human colon-carcinoma cells, and human glioma cells, due to progressive accumulation of quiescent cells during spheroid growth. The purpose of our work was to determine oxygen-consumption profiles in human melanoma spheroids. Monolayer cultures of 4 lines (BEX-c, COX-c, SAX-c, and WIX-c) and spheroid cultures of 2 lines (BEX-c and WIX-c) were subjected to investigation. Spheroids were initiated from monolayer cell cultures and grown in spinner flasks. Rate of oxygen consumption was measured with a Clarke-type electrode. Mitochondrial density was determined by stereological analysis of transmission electron micrographs. Thickness of viable rim and cell packing density were assessed by light microscopy of central spheroid sections. Cell-cycle distribution was determined by analysis of DNA histograms measured by flow cytometry. Cell volume was measured by an electronic particle counter. Rate of oxygen consumption per cell differed by a factor of approximately 1.8 between the 4 cell lines and was positively correlated to total volume of mitochondria per cell. Rate of oxygen consumption per cell and total volume of mitochondria per cell were equal for monolayer cell cultures, 600-microns spheroids and 1,200-microns spheroids of the same line. Mitochondrial density and location in the cell did not differ between cells at the spheroid surface, in the middle of the viable rim and adjacent to the central necrosis. Cell-cycle distribution, cell volume, and cell-packing density in the outer and inner halves of the viable rim were not significantly different. Consequently, the rate of oxygen consumption per cell in inner regions of the viable rim was probably equal to that at the spheroid surface, suggesting that oxygen diffusion distances may be shorter in some melanomas than in many other tumor types.

Wind Turbine - SWiFT southeast - WTGa1 - Reviewed Data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Herges, Thomas

Scaled Wind Farm Technology (SWiFT) Facility meteorological tower (MET), turbine, and Technical University of Denmark (DTU) SpinnerLidar data acquired on 20161216 UTC during a neutral atmospheric boundary layer inflow at a single focus distance of 2.5 D (D=27 m).

Surface Meteorological Station - SWiFT southwest - METa1 - Reviewed Data

DOE Data Explorer

Herges, Thomas

2017-10-23

Scaled Wind Farm Technology (SWiFT) Facility meteorological tower (MET), turbine, and Technical University of Denmark (DTU) SpinnerLidar data acquired on 20161216 UTC during a neutral atmospheric boundary layer inflow at a single focus distance of 2.5 D (D=27 m).

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Science.gov Websites

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A spinner magnetometer for large Apollo lunar samples.

PubMed

Uehara, M; Gattacceca, J; Quesnel, Y; Lepaulard, C; Lima, E A; Manfredi, M; Rochette, P

2017-10-01

We developed a spinner magnetometer to measure the natural remanent magnetization of large Apollo lunar rocks in the storage vault of the Lunar Sample Laboratory Facility (LSLF) of NASA. The magnetometer mainly consists of a commercially available three-axial fluxgate sensor and a hand-rotating sample table with an optical encoder recording the rotation angles. The distance between the sample and the sensor is adjustable according to the sample size and magnetization intensity. The sensor and the sample are placed in a two-layer mu-metal shield to measure the sample natural remanent magnetization. The magnetic signals are acquired together with the rotation angle to obtain stacking of the measured signals over multiple revolutions. The developed magnetometer has a sensitivity of 5 × 10 -7 Am 2 at the standard sensor-to-sample distance of 15 cm. This sensitivity is sufficient to measure the natural remanent magnetization of almost all the lunar basalt and breccia samples with mass above 10 g in the LSLF vault.

A spinner magnetometer for large Apollo lunar samples

NASA Astrophysics Data System (ADS)

Uehara, M.; Gattacceca, J.; Quesnel, Y.; Lepaulard, C.; Lima, E. A.; Manfredi, M.; Rochette, P.

2017-10-01

We developed a spinner magnetometer to measure the natural remanent magnetization of large Apollo lunar rocks in the storage vault of the Lunar Sample Laboratory Facility (LSLF) of NASA. The magnetometer mainly consists of a commercially available three-axial fluxgate sensor and a hand-rotating sample table with an optical encoder recording the rotation angles. The distance between the sample and the sensor is adjustable according to the sample size and magnetization intensity. The sensor and the sample are placed in a two-layer mu-metal shield to measure the sample natural remanent magnetization. The magnetic signals are acquired together with the rotation angle to obtain stacking of the measured signals over multiple revolutions. The developed magnetometer has a sensitivity of 5 × 10-7 Am2 at the standard sensor-to-sample distance of 15 cm. This sensitivity is sufficient to measure the natural remanent magnetization of almost all the lunar basalt and breccia samples with mass above 10 g in the LSLF vault.

Experimental aerodynamic performance of advanced 40 deg-swept 10-blade propeller model at Mach 0.6 to 0.85

NASA Technical Reports Server (NTRS)

Mitchell, Glenn A.

1988-01-01

A propeller designated as SR-6, designed with 40 deg of sweep and 10 blades to cruise at Mach 0.8 at an altitude of 10.7 km (35,000 ft), was tested in the NASA Lewis Research Center's 8- by 6-Foot Wind Tunnel. This propeller was one of a series of advanced single rotation propeller models designed and tested as part of the NASA Advanced Turboprop Project. Design-point net efficiency was almost constant to Mach 0.75 but fell above this speed more rapidly than that of any previously tested advanced propeller. Alternative spinners that further reduced the near-hub interblade Mach numbers and relieved the observed hub choking improved performance above Mach 0.75. One spinner attained estimated SR-6 Design-point net deficiencies of 80.6 percent at Mach 0.75 and 79.2 percent at Mach 0.8, higher than the measured performance of any previously tested advanced single-rotation propeller at these speeds.

Optimization of Carbon and Nitrogen Sources for Extracellular Polymeric Substances Production by Chryseobacterium indologenes MUT.2

PubMed Central

Khani, Mojtaba; Bahrami, Ali; Chegeni, Asma; Ghafari, Mohammad Davoud; Mansouran Zadeh, ALi

2016-01-01

Background Bacterial Extracellular Polymeric Substances (EPS) are environmental friendly and versatile polymeric materials that are used in a wide range of industries such as: food, textile, cosmetics, and pharmaceuticals. To make the production process of the EPS cost-effective, improvements in the production yield is required which could be implemented through application of processes such as optimized culture conditions, and development of the strains with higher yield (e.g. through genetic manipulation), or using low-cost substrates. Objectives In this work, the effects of carbon and nitrogen sources were studied in order to improve the EPS production by the submerged cultivation of Chryseobacterium indologenes MUT.2. Materials and Methods The mesophilic microorganism Chryseobacterium indologenes MUT.2, was grown and maintained in the Luria Bertani agar. The initial basal medium contained: glucose (20 g.L-1), yeast extracts (5 g.L-1), K2HPO4 (6 g.L-1), NaH2PO4 (7 g.L-1), NH4CL (0.7 g.L-1), and MgSO4 (0.5 g.L-1). For evaluating the carbon and nitrogen sources’ effect on the fermentation performance, cultures were prepared in 500 mL flasks filled with 300 mL of the medium. The single-factor experiments based on statistics was employed to evaluate and optimize the carbon and nitrogen sources for EPS production in the liquid culture medium of Chryseobacterium indologenes MUT.2. Results The preferred carbon-sources, sucrose and glucose, commonly gave the highest EPS production of 8.32 and 6.37 g.L-1, respectively, and the maximum EPS production of 8.87 g.L-1 was achieved when glutamic acid (5 g.L-1) was employed as the nitrogen source. Conclusions In this work, the culture medium for production of EPS by Chryseobacterium indologenes MUT.2 was optimized. Compared to the basal culture medium in shake-flasks and stirred tank bioreactor, the use of optimized culture medium has resulted in a 53% and 73% increase in the EPS production, respectively. PMID:28959321

Design of an efficient medium for heterologous protein production in Yarrowia lipolytica: case of human interferon alpha 2b.

PubMed

Gasmi, Najla; Ayed, Atef; Nicaud, Jean-Marc; Kallel, Héla

2011-05-20

The non conventional yeast Yarrowia lipolytica has aroused a strong industrial interest for heterologous protein production. However most of the studies describing recombinant protein production by this yeast rely on the use of complex media, such media are not convenient for large scale production particularly for products intended for pharmaceutical applications. In addition medium composition can also affect the production yield. Hence it is necessary to design an efficient medium for therapeutic protein expression by this host. Five different media, including four minimal media and a complex medium, were assessed in shake flasks for the production of human interferon alpha 2b (hIFN α2b) by Y. lipolytica under the control of POX2 promoter inducible with oleic acid. The chemically defined medium SM4 formulated by Invitrogen for Pichia pastoris growth was the most suitable. Using statistical experimental design this medium was further optimized. The selected minimal medium consisting in SM4 supplemented with 10 mg/l FeCl₃, 1 g/l glutamate, 5 ml/l PTM1 (Pichia Trace Metals) solution and a vitamin solution composed of myo-inositol, thiamin and biotin was called GNY medium. Compared to shake flask, bioreactor culture in GNY medium resulted in 416-fold increase of hIFN α2b production and 2-fold increase of the biological activity. Furthermore, SM4 enrichment with 5 ml/l PTM1 solution contributed to protect hIFN α2b against the degradation by the 28 kDa protease identified by zymography gel in culture supernatant. The screening of the inhibitory effect of the trace elements present in PTM1 solution on the activity of this protease was achieved using a Box-Behnken design. Statistical data analysis showed that FeCl₃ and MnSO₄ had the most inhibitory effect. We have designed an efficient medium for large scale production of heterologous proteins by Y. lipolytica. The optimized medium GNY is suitable for the production of hIFN α2b with the advantage that no complex nitrogen sources with non-defined composition were required.

New use of broomcorn millets for production of granular cultures of aphid-pathogenic fungus Pandora neoaphidis for high sporulation potential and infectivity to Myzus persicae.

PubMed

Hua, Li; Feng, Ming-Guang

2003-10-24

Glutinous broomcorn millets from the crop Panicum miliaceum were first used as substrate to produce granular cultures of Pandora neoaphidis, an obligate fungal pathogen specific to aphids. Carrying a water content of 36.5% after being steamed in a regular autoclaving procedure, millet grains of each 15 g (dry weight) in a 100-ml flask were mixed with 3 ml modified Sabouraud dextrose broth containing half a mashed colony of P. neoaphidis grown on egg yolk milk agar and then incubated at 20 degrees C and a light/dark cycle of 12 h/12 h for 21 days. Based on individually monitoring conidial production potential of 20 millet grains sampled from an arbitrarily taken flask at 3-day intervals, the millet cultures incubated for 6-15 days were capable of producing 16.8-23.4 x 10(4) conidia per millet grain with conidial ejection lasting for up to 6 days. The cultured millet grains individually produced significantly more conidia than apterous adults of Myzus persicae killed by P. neoaphidis (8.4 x 10(4) conidia per cadaver) and sporulated twice longer. The modeling of time-dose-mortality data from bioassays on M. persicae apterae exposed to conidial showers from the cultured millet grains and the mycelial mats produced in liquid culture resulted in similar estimates of LC(50) (millets: 21.4, 7.3, and 4.9 conidia mm(-2) on days 5-7 after exposure; mycelial mats: 22.1, 10.6, and 7.7 conidia mm(-2)) although the LT(50) estimated at a given conidial concentration was slightly smaller for the millet cultures than for the mycelial mats. This indicates that the millet grains cultured with P. neoaphidis produced conidia as infective as or slightly more infective to M. persicae than those from the mycelial mats. Based on the sporulation potential, infectivity, and ease and cost of the millet cultures, the method developed in this study highly improved in vitro cultures of P. neoaphidis and may adapt to culturing other entomophthoralean fungi for microbial control of insect pests.

New Generation Flask Sampling Technology Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Smith, James R.

Scientists are turning their focus to the Arctic, site of one of the strongest climate change signals. A new generation of technologies is required to function within that harsh environment, chart evolution of its trace gases and provide new kinds of information for models of the atmosphere. Our response to the solicitation tracks how global atmospheric monitoring was launched more than a half century ago; namely, acquisition of discrete samples of air by flask and subsequent analysis in the laboratory. AOS is proposing to develop a new generation of flask sampling technology. It will enable the new Arctic programs tomore » begin with objective high density sampling of the atmosphere by UAS. The Phase I program will build the prototype flask technology and show that it can acquire and store mol fractions of CH4 and CO2 and value of δ13C with good fidelity. A CAD model will be produced for the entire platform including a package with 100 flasks and the airframe with auto-pilot, electronic propulsion and ground-to-air communications. A mobile flask analysis station will be prototyped in Phase I and designed to final form in Phase II. It expends very small sample per analysis and will interface directly to the flask package integrated permanently into the UAS fuselage. Commercial Applications and Other Benefits: • The New Generation Flask Sampling Technology able to provide a hundred or more samples of air per UAS mission. • A mobile analysis station expending far less sample than the existing ones and small enough to be stationed at the remote sites of Arctic operations. • A new form of validation for continuous trace gas observations from all platforms including the small UAS. • Further demonstration to potential customers of the AOS capabilities to invent, build, deploy and exploit entire platforms for observations of Earth’s atmosphere and ocean. Key Words: Flask Sampler, Mobile Analysis Station, Trace Gas, CO2, CH4, δC13, UAS, Baseline Airborne Observatory, Arctic, Climate Change. Summary for Members of Congress: The air, land and sea of the Arctic combine to produce a large climate change signal. AOS is proposing to develop unmanned airborne technologies able to begin prompt, objective observations of the signal’s atmospheric component.« less

Efficient Extracellular Expression of Phospholipase D in Escherichia Coli with an Optimized Signal Peptide

NASA Astrophysics Data System (ADS)

Yang, Leyun; Xu, Yu; Chen, Yong; Ying, Hanjie

2018-01-01

New secretion vectors containing the synthetic signal sequence (OmpA’) was constructed for the secretory production of recombinant proteins in Escherichia coli. The E. coli Phospholipase D structural gene (Accession number:NC_018658) fused to various signal sequence were expressed from the Lac promoter in E. coli Rosetta strains by induction with 0.4mM IPTG at 28°C for 48h. SDS-PaGe analysis of expression and subcellular fractions of recombinant constructs revealed the translocation of Phospholipase D (PLD) not only to the medium but also remained in periplasm of E. coli with OmpA’ signal sequence at the N-terminus of PLD. Thus the study on the effects of various surfactants on PLD extracellular production in Escherichia coli in shake flasks revealed that optimal PLD extracellular production could be achieved by adding 0.4% Triton X-100 into the medium. The maximal extracellular PLD production and extracellular enzyme activity were 0.23mg ml-1 and 16U ml-1, respectively. These results demonstrate the possibility of efficient secretory production of recombinant PLD in E. coli should be a potential industrial applications.

Lead removal by Spirulina platensis biomass.

PubMed

Al-Homaidan, Ali A; Al-Abbad, Aljawharah F; Al-Hazzani, Amal A; Al-Ghanayem, Abdullah A; Alabdullatif, Jamila A

2016-01-01

In this investigation, we report on the biosorption of Pb (II) from aqueous solutions by the nonliving biomass of the micro-alga (cyanobacterium) Spirulina platensis. Propagation of the micro-alga was carried out in outside oblong raceway ponds. The biomass was cleaned, dried and used for the investigation. The effects of pH, adsorbent dose, temperature, initial concentration of Pb (II), and contact time on the adsorption of lead by the dry biomass were studied. The experiments were carried out in 250 ml conical flasks containing 100 ml of test solutions using an orbital incubator at 150 rpm. Concentrations of the metal before and after the experiments were measured using Atomic Absorption Spectrophotometer. Very high levels of Pb (II) removal (>91%) were obtained. The optimum conditions for maximal adsorption by S. platensis were found to be pH 3; 2 g of adsorbent dose; incubation at 26°C; 100 mg/l of lead initial concentration and 60 minutes of contact time. The experimental data fitted well with Freundlich isotherm equation with R(2) values greater than 0.97. Based on our results, we recommend the utilization of S. platensis biomass for heavy metal removal from aqueous solutions.

Bioprocess exploration for thermostable α-amylase production of a deep-sea thermophile Geobacillus sp. in high-temperature bioreactor.

PubMed

Jiang, Tao; Huang, Mengmeng; He, Hao; Lu, Jian; Zhou, Xiangshan; Cai, Menghao; Zhang, Yuanxing

2016-08-17

Geobacillus sp. 4j, a deep-sea high-salt thermophile, was found to produce thermostable α-amylase. In this work, culture medium and conditions were first optimized to enhance the production of thermostable α-amylase by statistical methodologies. The resulting extracellular production was increased by five times and reached 6.40 U/ml. Then, a high-temperature batch culture of the thermophile in a 15 l in-house-designed bioreactor was studied. The results showed that a relatively high dissolved oxygen (600 rpm and 15 l/min) and culture temperature of 60°C facilitated both cell growth and α-amylase production. Thus, an efficient fermentation process was established with initial medium of pH 6.0, culture temperature of 60°C, and dissolved oxygen above 20%. It gave an α-amylase production of 79 U/ml and productivity of 19804 U/l·hr, which were 10.8 and 208 times higher than those in shake flask, respectively. This work is useful for deep-sea high-salt thermophile culture, where efforts are lacking presently.

Scale-up from shake flasks to bioreactor, based on power input and Streptomyces lividans morphology, for the production of recombinant APA (45/47 kDa protein) from Mycobacterium tuberculosis.

PubMed

Gamboa-Suasnavart, Ramsés A; Marín-Palacio, Luz D; Martínez-Sotelo, José A; Espitia, Clara; Servín-González, Luis; Valdez-Cruz, Norma A; Trujillo-Roldán, Mauricio A

2013-08-01

Culture conditions in shake flasks affect filamentous Streptomyces lividans morphology, as well the productivity and O-mannosylation of recombinant Ala-Pro-rich O-glycoprotein (known as the 45/47 kDa or APA antigen) from Mycobacterium tuberculosis. In order to scale up from previous reported shake flasks to bioreactor, data from the literature on the effect of agitation on morphology of Streptomyces strains were used to obtain gassed volumetric power input values that can be used to obtain a morphology of S. lividans in bioreactor similar to the morphology previously reported in coiled/baffled shake flasks by our group. Morphology of S. lividans was successfully scaled-up, obtaining similar mycelial sizes in both scales with diameters of 0.21 ± 0.09 mm in baffled and coiled shake flasks, and 0.15 ± 0.01 mm in the bioreactor. Moreover, the specific growth rate was successfully scaled up (0.09 ± 0.02 and 0.12 ± 0.01 h(-1), for bioreactors and flasks, respectively), and the recombinant protein productivity measured by densitometry, as well. More interestingly, the quality of the recombinant glycoprotein measured as the amount of mannoses attached to the C-terminal of APA was also scaled- up; with up to five mannose residues in cultures carried out in shake flasks; and six in the bioreactor. However, final biomass concentration was not similar, indicating that although the process can be scaled-up using the power input, others factors like oxygen transfer rate, tip speed or energy dissipation/circulation function can be an influence on bacterial metabolism.

Quantifying the Erlenmeyer flask deformity

PubMed Central

Carter, A; Rajan, P S; Deegan, P; Cox, T M; Bearcroft, P

2012-01-01

Objective Erlenmeyer flask deformity is a common radiological finding in patients with Gaucher′s disease; however, no definition of this deformity exists and the reported prevalence of the deformity varies widely. To devise an easily applied definition of this deformity, we investigated a cohort of knee radiographs in which there was consensus between three experienced radiologists as to the presence or absence of Erlenmeyer flask morphology. Methods Using the presence or absence of Erlenmeyer flask morphology as a benchmark, we measured the diameter of the femur at the level of the physeal scar and serially at defined intervals along the metadiaphysis. Results A measured ratio in excess of 0.57 between the diameter of the femoral shaft 4 cm from the physis to the diameter of the physeal baseline itself on a frontal radiograph of the knee predicted the Erlenmeyer flask deformity with 95.6% sensitivity and 100% specificity in our series of 43 independently diagnosed adults with Gaucher′s disease. Application of this method to the distal femur detected the Erlenmeyer flask deformity reproducibly and was simple to carry out. Conclusion Unlike diagnostic assignments based on subjective review, our simple procedure for identifying the modelling deformity is based on robust quantitative measurement: it should facilitate comparative studies between different groups of patients, and may allow more rigorous exploration of the pathogenesis of the complex osseous manifestations of Gaucher′s disease to be undertaken. PMID:22010032

Modeling of growth and laccase production by Pycnoporus sanguineus.

PubMed

Saat, Muhammad Naziz; Annuar, Mohamad Suffian Mohamad; Alias, Zazali; Chuan, Ling Tau; Chisti, Yusuf

2014-05-01

Production of extracellular laccase by the white-rot fungus Pycnoporus sanguineus was examined in batch submerged cultures in shake flasks, baffled shake flasks and a stirred tank bioreactor. The biomass growth in the various culture systems closely followed a logistic growth model. The production of laccase followed a Luedeking-Piret model. A modified Luedeking-Piret model incorporating logistic growth effectively described the consumption of glucose. Biomass productivity, enzyme productivity and substrate consumption were enhanced in baffled shake flasks relative to the cases for the conventional shake flasks. This was associated with improved oxygen transfer in the presence of the baffles. The best results were obtained in the stirred tank bioreactor. At 28 °C, pH 4.5, an agitation speed of 600 rpm and a dissolved oxygen concentration of ~25 % of air saturation, the laccase productivity in the bioreactor exceeded 19 U L(-1 )days(-1), or 1.5-fold better than the best case for the baffled shake flask. The final concentration of the enzyme was about 325 U L(-1).

Delta 13C in CO2 at Alert, NWT, Canada (June 1991 - December 2001)

DOE Data Explorer

Allison, C. E. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Francey, R. J. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2003-04-01

Measurements have been made on air collected in flasks at Alert, Canada, through the CSIRO GASLAB worldwide network. Flasks are filled with air at Alert and returned to the CSIRO GASLAB for analysis; typical sample storage times for flasks collected at Alert range from a few weeks to a few months. No significant effect on the stable carbon isotopic composition, δ13C, has been detected as a consequence of the sample storage time.

50 CFR 218.181 - Permissible methods of taking.

Code of Federal Regulations, 2010 CFR

2010-10-01

... and the indicated number of times: (1) Level B Harassment: (i) Sperm whale (Physeter macrocephalus)—10...); (vii) Spinner dolphin (S. longirostris)—115 (an average of 23 annually); (viii) Melon-headed whale (Peponocephala electra)—10 (an average of 2 annually); (ix) Short-finned pilot whale (Globicephala macrorhynchus...

50 CFR 218.181 - Permissible methods of taking.

Code of Federal Regulations, 2012 CFR

2012-10-01

... and the indicated number of times: (1) Level B Harassment: (i) Sperm whale (Physeter macrocephalus)—10...); (vii) Spinner dolphin (S. longirostris)—115 (an average of 23 annually); (viii) Melon-headed whale (Peponocephala electra)—10 (an average of 2 annually); (ix) Short-finned pilot whale (Globicephala macrorhynchus...

50 CFR 218.181 - Permissible methods of taking.

Code of Federal Regulations, 2011 CFR

2011-10-01

... and the indicated number of times: (1) Level B Harassment: (i) Sperm whale (Physeter macrocephalus)—10...); (vii) Spinner dolphin (S. longirostris)—115 (an average of 23 annually); (viii) Melon-headed whale (Peponocephala electra)—10 (an average of 2 annually); (ix) Short-finned pilot whale (Globicephala macrorhynchus...

A Passion for Teaching.

ERIC Educational Resources Information Center

Levine, Sarah L., Ed.; McVay, Scott, Ed.

This book, celebrating teachers and teaching, contains the artistry and wisdom of 42 teachers who have remained passionate about classroom teaching for many years. Chapter 1, "Beginnings," includes "The First Day of School" (Richard A. Lawson), "Sustaining the Wonder of Teaching" and "Harvest Home," (Bettye T. Spinner), "The Call to Teach,"…

A "Virtual Spin" on the Teaching of Probability

ERIC Educational Resources Information Center

Beck, Shari A.; Huse, Vanessa E.

2007-01-01

This article, which describes integrating virtual manipulatives with the teaching of probability at the elementary level, puts a "virtual spin" on the teaching of probability to provide more opportunities for students to experience successful learning. The traditional use of concrete manipulatives is enhanced with virtual coins and spinners from…

Development of a method for efficient cost-effective screening of Aspergillus niger mutants having increased production of glucoamylase.

PubMed

Zhu, Xudong; Arman, Bessembayev; Chu, Ju; Wang, Yonghong; Zhuang, Yingping

2017-05-01

To develop an efficient cost-effective screening process to improve production of glucoamylase in Aspergillus niger. The cultivation of A. niger was achieved with well-dispersed morphology in 48-deep-well microtiter plates, which increased the throughput of the samples compared to traditional flask cultivation. There was a close negative correlation between glucoamylase and its pH of the fermentation broth. A novel high-throughput analysis method using Methyl Orange was developed. When compared to the conventional analysis method using 4-nitrophenyl α-D-glucopyranoside as substrate, a correlation coefficient of 0.96 by statistical analysis was obtained. Using this novel screening method, we acquired a strain with an activity of 2.2 × 10 3  U ml -1 , a 70% higher yield of glucoamylase than its parent strain.

Development of a simple and low cost microbioreactor for high-throughput bioprocessing.

PubMed

Rahman, Pattanathu K S M; Pasirayi, Godfrey; Auger, Vincent; Ali, Zulfiqur

2009-02-01

A simple microbioreactor for high-throughput bioprocessing made from low cost polymer polytetrafluoroethylene (PTFE) tubes with a working volume of 1.5 ml is described. We have developed a microfluidic system that handles a small population of cells of a model microorganism, Pseudomonas aeruginosa DS10-129. Under the conditions of the microbioreactor, the organism produced extracellular secondary metabolites by using nutrient broth modified with glycerol. Pyocyanins were isolated from the fermented medium as a metabolite of interest. Antibiotic properties of pyocyanin were effective against a number of microorganisms such as Staphylococcus aureus, S. epidermis, Bacillus subtilis, Micrococcus luteus and Saccharomyces cerevisiae. Batch fermentation of the model organism in the microbioreactor was compared to shake-flask and conventional bench fermenter methods. Results obtained from the microbioreactor compared favourably with the conventional processes.

Pilot-scale production and liquid formulation of Rhodotorula minuta, a potential biocontrol agent of mango anthracnose.

PubMed

Patiño-Vera, M; Jiménez, B; Balderas, K; Ortiz, M; Allende, R; Carrillo, A; Galindo, E

2005-01-01

To develop a pilot-plant fermentation process for the production of the yeast Rhodotorula minuta, to be used as a biocontrol agent of mango anthracnose, using a low-cost culture medium. To develop a stable liquid formulation that preserve high viability of the yeast stored at 4 degrees C. Keeping constant the volumetric power input, a fermentation process was scaled-up from shake flasks to a 100 l bioreactor. Preharvest applications of the yeast resulted in postharvest anthracnose severity equal or lower than that observed with a chemical fungicide. Glycerol was added to the formulation as water activity reducer and xanthan gum as a viscosity-enhancing agent. Yeast initial concentration of 10(10) CFU ml(-1) resulted in 4-5 orders of magnitude decrease after 1 month of storage at 4 degrees C, whereas when it was formulated at 10(9) CFU ml(-1), the decrease was of two orders of magnitude in 6 months. The fermentation process was successfully scaled-up using a low-cost culture medium. Postharvest anthracnose severity could be considerably reduced using this yeast. Formulating the yeast at 10(9) CFU ml(-1) and adding glycerol (20%) and xanthan (5 g l(-1)) avoided both contamination and yeast sedimentation and it was able to preserve up to 10(7) CFU ml(-1) after 6 months at 4 degrees C. The yeast R. minuta is reported as a novel antagonistic micro-organism against the pathogen Colletotrichum gloeosporioides. Pilot plant production of this yeast allowed us to conduct field tests in commercial orchards during three harvest seasons. Yeast suspensions applied to mango trees reduced the fruit anthracnose severity in levels similar or better than chemical fungicides.

Evaluation of calcium alginate beads for Ce, La and Nd preconcentration from groundwater prior to ICP OES analysis.

PubMed

Arantes de Carvalho, Gabriel G; Kondaveeti, Stalin; Petri, Denise F S; Fioroto, Alexandre M; Albuquerque, Luiza G R; Oliveira, Pedro V

2016-12-01

Analytical methods for the determination of rare earth elements (REE) in natural waters by plasma spectrochemical techniques often require sample preparation procedures for analytes preconcentration as well as for removing matrix constituents, that may interfere on the analytical measurements. In the present work, calcium alginate (CA) beads were used for the first time aiming at Ce, La and Nd preconcentration from groundwater samples for further determination by inductively coupled plasma optical emission spectrometry (ICP OES). Test samples were analyzed in batch mode by transferring a 40mL test portion (pH=5±0.2) into a 50mL polyethylene flask containing 125mg CA beads. After 15min contact, the analytes were quantitatively extracted from the loaded CA beads with 2.0mL of 1.0molL -1 HCl solution for further determination by ICP OES, using Ce (II) 456.236, La (II) 379.478 and Nd (II) 430.358nm emission lines. The proposed approach is a reliable alternative for REE single-stage preconcentration from aqueous samples, as it provided accurate results based on the addition and recovery analysis of groundwater. The results obtained by the proposed method were also compared with those from reference method based on inductively coupled plasma mass spectrometry (ICP-MS) and no significant differences were observed after applying the Student's t-test at 95% confidence level. Copyright © 2016 Elsevier B.V. All rights reserved.

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2010 CFR

2010-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2011 CFR

2011-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2014 CFR

2014-04-01

.... At the end of this time, disconnect the flask and evaporate the methyl alcohol on the steam bath... transfer the isooctane extract to a 250-milliliter separatory funnel. Dissolve the crystals in the flask...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2012 CFR

2012-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

21 CFR 172.250 - Petroleum naphtha.

Code of Federal Regulations, 2013 CFR

2013-04-01

... at room temperature, with intermittent swirling. At the end of this time, disconnect the flask and.... Dissolve the crystals in the flask with about 25 milliliters of distilled water and pour this also into the...

Atmospheric Hydrogen (H2) Concentrations from the CSIRO GASLAB Flask Sampling Network (1992 - 2001)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Atmospheric Research, Aspendale, Victoria, Australia

2003-01-01

Air samples from nine sites were collected from the CSIRO GASLAB Flask Sampling Network for the purpose of monitoring the atmospheric hydrogen (H2) concentrations. The listed data were obtained from flask air samples returned to the CSIRO GASLAB for analysis. Typical sample storage times ranged from days to weeks for some sites (e.g., Cape Grim) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for any change in sample H22 mixing ratio during storage have shown no consistent and systematic drift in these flask types over test periods of several months to years (Cooper et al., 1999). An annual cycle of H2 is evident, reflecting the seasonal nature of some of the major sources and sinks (Novelli et al., 1999).

A comparison of the effects of two methods of acclimation of aerobic biodegradability

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watson, H.M.

1993-11-01

The acclimation or adaptation of microorganisms to organic chemicals is an important factor influencing both the rate and the extent of biodegradation. In this study two acclimation procedures were evaluated in terms of their effectiveness in enhancing biodegradation, their relative ease of use in the laboratory, and the implications for biodegradability testing. In the single-flask procedure, microorganisms were acclimated for 2 to 7 d in a single acclimation flask at constant or increasing concentrations of the test chemical without transfer of microorganisms. In the second procedure, the enrichment procedure, microorganisms were acclimated in a series of flasks over a 21-dmore » period by making adaptive transfers to increasing concentrations of the test chemical. Acclimated microorganisms from each procedure were used as the source of inoculum for subsequent biodegradation tests in which carbon dioxide evolution was measured. Six chemicals were tested: quinoline, p-nitrophenol, N-methylaniline, N,N-dimethylaniline, acrylonitrile, and 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate. Microorganisms acclimated in the single-flask procedure were much more effective than those acclimated in the enrichment procedure in degrading the test chemicals. The single-flask procedure is more convenient to use, and it permits monitoring of the time needed for acclimation. The results from these studies have implications for the methodology used in biodegradation test systems and suggest caution before adopting a multiple-flask, enrichment acclimation procedure before the performance of standardized tests for aerobic biodegradability.« less

Sampling, storage, and analysis of C2-C7 non-methane hydrocarbons from the US National Oceanic and Atmospheric Administration Cooperative Air Sampling Network glass flasks.

PubMed

Pollmann, Jan; Helmig, Detlev; Hueber, Jacques; Plass-Dülmer, Christian; Tans, Pieter

2008-04-25

An analytical technique was developed to analyze light non-methane hydrocarbons (NMHC), including ethane, propane, iso-butane, n-butane, iso-pentane, n-pentane, n-hexane, isoprene, benzene and toluene from whole air samples collected in 2.5l-glass flasks used by the National Oceanic and Atmospheric Administration, Earth System Research Laboratory, Global Monitoring Division (NOAA ESRL GMD, Boulder, CO, USA) Cooperative Air Sampling Network. This method relies on utilizing the remaining air in these flasks (which is at below-ambient pressure at this stage) after the completion of all routine greenhouse gas measurements from these samples. NMHC in sample aliquots extracted from the flasks were preconcentrated with a custom-made, cryogen-free inlet system and analyzed by gas chromatography (GC) with flame ionization detection (FID). C2-C7 NMHC, depending on their ambient air mixing ratios, could be measured with accuracy and repeatability errors of generally < or =10-20%. Larger deviations were found for ethene and propene. Hexane was systematically overestimated due to a chromatographic co-elution problem. Saturated NMHC showed less than 5% changes in their mixing ratios in glass flask samples that were stored for up to 1 year. In the same experiment ethene and propene increased at approximately 30% yr(-1). A series of blank experiments showed negligible contamination from the sampling process and from storage (<10 pptv yr(-1)) of samples in these glass flasks. Results from flask NMHC analyses were compared to in-situ NMHC measurements at the Global Atmospheric Watch station in Hohenpeissenberg, Germany. This 9-months side-by-side comparison showed good agreement between both methods. More than 94% of all data comparisons for C2-C5 alkanes, isoprene, benzene and toluene fell within the combined accuracy and precision objectives of the World Meteorological Organization Global Atmosphere Watch (WMO-GAW) for NMHC measurements.

A mini-scale mass production and separation system for secretory heterologous proteins by perfusion culture of recombinant Pichia pastoris using a shaken ceramic membrane flask.

PubMed

Ohashi, R; Mochizuki, E; Suzuki, T

1999-01-01

The perfusion culture technique using a shaken ceramic membrane flask (SCM flask) was applied to the production of a secretory heterologous protein. A recombinant methylotrophic yeast strain, Pichia pastoris, was cultured aerobically on a reciprocal shaker using an SCM flask. High-level production of human serum albumin (HSA) was attempted by increasing both the cell concentration and the expression level of the recombinant gene. In the two-stage culture method, the cell concentration was first raised to 17 g/l by feeding glycerol, after which the expression of HSA was induced by feeding methanol. However, the concentration of HSA in the effluent filtrate was as low as 0.15 g/l, while the cell concentration continued to increase. In contrast, HSA was effectively produced by feeding methanol from an early stage of the culture. In this case, the HSA concentration reached 0.24 and 0.46 g/l, respectively, using the growth-associated production method without and with aeration into the head space of the SCM flask. The results showed that supplying sufficient oxygen together with the growth-associated induction method are effective for obtaining high-level expression of the methanol-inducible recombinant gene of P. pastoris. An HSA concentration in the filtrate of 1.5 g/l was finally achieved when the cell concentration was increased to 53 g/l by supplying oxygen-enriched gas to the SCM flask. The yield and productivity of HSA reached 2.6-fold and 10-fold those obtained in an ordinary fed-batch culture using a shake flask, and these levels were readily achieved by continuous replenishment of the culture supernatant. The achievements made in this study should contribute to the development of a handy bioreactor system for mini-scale mass production of target proteins with separation at high purity.

The Safe and Efficient Evaporation of a Solvent from Solution

NASA Astrophysics Data System (ADS)

Mahon, Andrew R.

1997-02-01

The process of evaporating a solvent from a solution can cause problems for many students. By using a water-vacuum aspirator, backflashes of water can flood the sample tube and be detrimental to the experiment. This type of apparatus can also cause problems by drawing the solution it is evaporating back into the vacuum hose, causing the student to lose part or all of the products of their experiment. Macroscale and Microscale Organic Experiments, 2nd edition (1), suggested two techniques to dissolve solvents from a mixture. It suggested blowing a stream of air over the solution from a Pasteur pipet, or attaching a Pasteur pipet to an aspirator and drawing air over the surface of the liquid. Again, the danger of blowing air over the solution leaves the risk of splattering the solution, and drawing air over the surface of the liquid as described further endangers the products of the experiment through the risk of sucking the products up into the pipet aspirator. In an effort to eliminate these problems, a new technique has been developed. By inverting an ordinary 200-mL vacuum flask and pulling a steady current of air from the vacuum apparatus through it, any type of small container can be placed under it, allowing the solvent to be evaporated in a steady, mistake-free manner . By evaporating the solvent from the container that the products will be submitted in, no sample is lost through the process of transferring it from a vacuum flask or beaker to the final container.

Hundreds of Area Residents Visit the National Lab Booth at the Annual In The Street Festival | Poster

Cancer.gov

Light-up yo-yos, brightly colored portion plates, and a fast spinner game lured hundreds of area residents to the Frederick National Lab booth at this year’s In The Street festival, where they also heard a message from the lab: Stay healthy through healthy habits.

Scribble Scrabble: Ready-in-a-Minute Math Games.

ERIC Educational Resources Information Center

Sharp, Richard M.; And Others

Mathematics research has discovered two elements that are consistently linked to student achievement: parental involvement and manipulative materials. This book offers a rich assortment of spinner, matrix, and card learning games which can be used in the classroom or at home. The concepts and skills range from whole number operations to geometry.…

The mechanism of ethanol treatment on inhibiting lettuce enzymatic browning and microbial growth

USDA-ARS?s Scientific Manuscript database

Tissue browning of fresh-cut lettuce greatly affects its quality and consumers’ appreciation. This study investigated the effects of ethanol treatment on enzymatic browning and natural microflora growth of lettuce stem discs. After treated with 20% ethanol for 2 min and then drained by a spinner, le...

Ecology and Energy Action Pack.

ERIC Educational Resources Information Center

McDonald's Corp., Oak Brook, IL.

One of five McDonald's Action Packs, these elementary school-level instructional materials are for use as an introduction to existing units of study, supplements to a textbook, or a source of special projects for environmental education. Contents include these six units: Make Your Own Ecology Mini-spinner, Let's Look at a Food Chain, Drip the…

PASSIVE ACOUSTIC MONITORING OF ODONTOCETES IN THE VICINITY OF PUULOA UNDERWATER DETONATION TRAINING RANGE, HAWAII RANGE COMPLEX, OAHU

DTIC Science & Technology

2016-09-25

marine soundscape , is the quietest time of the day (Heenehan 2016; Navy 2016) and therefore they may not be vocalizing frequently. Based on...Heenehan, H. L. (2016). Soundscape Ecology of Hawaiian Spinner Dolphin Resting Bays (Doctoral dissertation, Duke University). Hotchkin, C. F., Shoemaker

Development of a prototype sensor to detect plastic contamination in seed cotton at the gin

USDA-ARS?s Scientific Manuscript database

US cotton is considered to have some of the lowest levels of contamination in the world. That reputation is expected by foreign and domestic mills. Despite this reputation, U.S. spinners have recently experienced some serious contamination issues with US cotton. Of particular concern are plastic con...

Body movements during the off-ice execution of back spins in figure skating.

PubMed

Mapelli, Andrea; Rodano, Renato; Fiorentini, Angelo; Giustolisi, Andrea; Sidequersky, Fernanda V; Sforza, Chiarella

2013-10-01

Using an optoelectronic motion capture system, we quantitatively assessed the arrangement of body segments and the displacement of the horizontal projection of the center of mass (CM) in seven skaters performing off-ice back spins on a rotating device (spinner). The position of the CM at the beginning of the spins was not a determining factor, but its rapid stabilization towards the center of the spinner, together with the achievement of a stable arrangement of trunk and limbs, was crucial to get the dynamic equilibrium, necessary for a lasting performance. At full spinning, however, there was an indicative variety of individual body postures. A final deceleration, associable with the loss of body equilibrium, was detected in the last spin of most of skaters. In conclusion, the current investigation demonstrated that the off-ice execution of back spin, a critical movement of ice skating, can be measured in laboratory, thus providing quantitative information to both the skaters and the coaches. The analysis is not invasive, and it may be proposed also for longitudinal evaluations of skating and postural training. Copyright © 2013 Elsevier Ltd. All rights reserved.

A baseline maritime satellite communication system

NASA Technical Reports Server (NTRS)

Durrani, S. H.; Mcgregor, D. N.

1974-01-01

This paper describes a baseline system for maritime communications via satellite during the 1980s. The system model employs three geostationary satellites with global coverage antennas. Access to the system is controlled by a master station; user access is based on time-ordered polling or random access. Each Thor-Delta launched satellite has an RF power of 100 W (spinner) or 250 W (three-axis stabilized), and provides 10 equivalent duplex voice channels for up to 1500 ships with average waiting times of approximately 2.5 minutes. The satellite capacity is bounded by the available bandwidth to 50 such channels, which can serve up to 10,000 ships with an average waiting time of 5 minutes. The ships must have peak antenna gains of approximately 15.5 dB or 22.5 dB for the two cases (10 or 50 voice channels) when a spinner satellite is used; the required gains are 4 dB lower if a three-axis stabilized satellite is used. The ship antenna requirements can be reduced by 8 to 10 dB by employing a high-gain multi-beam phased array antenna on the satellite.

Evaluation of panel code predictions with experimental results of inlet performance for a 17-inch ducted prop/fab simulator operating at Mach 0.2

NASA Technical Reports Server (NTRS)

Boldman, D. R.; Iek, C.; Hwang, D. P.; Jeracki, R. J.; Larkin, M.; Sorin, G.

1991-01-01

An axisymmetric panel code was used to evaluate a series of ducted propeller inlets. The inlets were tested in the Lewis 9 by 15 Foot Low Speed Wind Tunnel. Three basic inlets having ratios of shroud length to propeller diameter of 0.2, 0.4, and 0.5 were tested with the Pratt and Whitney ducted prop/fan simulator. A fourth hybrid inlet consisting of the shroud from the shortest basic inlet coupled with the spinner from the largest basic inlet was also tested. This later configuration represented the shortest overall inlet. The simulator duct diameter at the propeller face was 17.25 inches. The short and long spinners provided hub-to-tip ratios of 0.44 at the propeller face. The four inlets were tested at a nominal free stream Mach number of 0.2 and at angles of attack from 0 degrees to 35 degrees. The panel code method incorporated a simple two-part separation model which yielded conservative estimates of inlet separation.

Occurrence of Anisakis (Nematoda: Anisakidae) larvae in unusual hosts in Southern hemisphere.

PubMed

Shamsi, Shokoofeh; Briand, Marine J; Justine, Jean-Lou

2017-12-01

Nematodes belonging to the genus Anisakis are important parasites due to their abundance in seafood and health impacts on humans. In the present study Anisakis larvae were found in a number of uncommon hosts including the Grey petrel, Procellaria cinerea, the Little penguin, Eudyptula minor, Blue-lipped sea krait, Laticauda laticaudata and Spinner shark, Carcharhinus brevipinna. Morphological examination showed nematodes in these animals are Anisakis larval type I. Genetic characterisation suggested that the larva from one Grey petrel was Anisakis berlandi, whereas the other larvae from the second Grey petrel and from the little penguin were Anisakis pegreffii. A number of larvae found in Blue-lipped sea krait and Spinner shark were identified as Anisakis typica. This is the first report of infective stage of Anisakis larvae parasitising hosts other than teleost fish. Understanding of the extent of infection and the pathogenicity of anisakid nematodes in hosts found in the present study is important in the conservation studies and management plans of these hosts. Copyright © 2017 Elsevier B.V. All rights reserved.

Liquid films on shake flask walls explain increasing maximum oxygen transfer capacities with elevating viscosity.

PubMed

Giese, Heiner; Azizan, Amizon; Kümmel, Anne; Liao, Anping; Peter, Cyril P; Fonseca, João A; Hermann, Robert; Duarte, Tiago M; Büchs, Jochen

2014-02-01

In biotechnological screening and production, oxygen supply is a crucial parameter. Even though oxygen transfer is well documented for viscous cultivations in stirred tanks, little is known about the gas/liquid oxygen transfer in shake flask cultures that become increasingly viscous during cultivation. Especially the oxygen transfer into the liquid film, adhering on the shake flask wall, has not yet been described for such cultivations. In this study, the oxygen transfer of chemical and microbial model experiments was measured and the suitability of the widely applied film theory of Higbie was studied. With numerical simulations of Fick's law of diffusion, it was demonstrated that Higbie's film theory does not apply for cultivations which occur at viscosities up to 10 mPa s. For the first time, it was experimentally shown that the maximum oxygen transfer capacity OTRmax increases in shake flasks when viscosity is increased from 1 to 10 mPa s, leading to an improved oxygen supply for microorganisms. Additionally, the OTRmax does not significantly undermatch the OTRmax at waterlike viscosities, even at elevated viscosities of up to 80 mPa s. In this range, a shake flask is a somehow self-regulating system with respect to oxygen supply. This is in contrary to stirred tanks, where the oxygen supply is steadily reduced to only 5% at 80 mPa s. Since, the liquid film formation at shake flask walls inherently promotes the oxygen supply at moderate and at elevated viscosities, these results have significant implications for scale-up. © 2013 Wiley Periodicals, Inc.

THE FEDERAL INTEGRATED BIOTREATMENT RESEARCH CONSORTIUM (FLASK TO FIELD)

EPA Science Inventory

The Federal Integrated Biotreatment Research Consortium (Flask to Field) represented a 7-year concerted effort by several research laboratories to develop bioremediation technologies for contaminated DoD sites. The consortium structure consisted of a director and four thrust are...

The Oxygen Flask Method

ERIC Educational Resources Information Center

Boulton, L. H.

1973-01-01

Discusses application of Schoniger's method of quantitative organic elemental analysis in teaching of qualitative analysis of the halogens, nitrogen, sulphur, and phosphorus. Indicates that the oxygen flask method is safe and suitable for both high school and college courses because of simple apparatus requirements. (CC)

Methods of Sensing Land Pollution from Sanitary Landfills

NASA Technical Reports Server (NTRS)

Nosanov, Myron Ellis; Bowerman, Frank R.

1971-01-01

Major cities are congested and large sites suitable for landfill development are limited. Methane and other gases are produced at most sanitary landfills and dumps. These gases may migrate horizontally and vertically and have caused fatalities. Monitoring these gases provides data bases for design and construction of safe buildings on and adjacent to landfills. Methods of monitoring include: (1) a portable combustible gas indicator; and (2) glass flasks valved to allow simultaneous exhaust of the flask and aspiration of the sample into the flask. Samples are drawn through tubing from probes as deep as twenty-five feet below the surface.

Conducting Miller-Urey Experiments

NASA Technical Reports Server (NTRS)

Parker, Eric Thomas; Cleaves, Henderson James; Burton, Aaron S.; Glavin, Daniel P.; Dworkin, Jason; Zhou, Manshui; Bada, Jeffrey L.; Fernandez, Facundo M.

2014-01-01

In 1953, Stanley Miller reported the production of biomolecules from simple gaseous starting materials, using apparatus constructed to simulate the primordial Earth's atmosphere-ocean system. Miller introduced 200 ml of water, 100 mmHg of H2, 200mmHg of CH4, and 200mmHg of NH3 into the apparatus, then subjected this mixture, under reflux, to an electric discharge for a week, while the water was simultaneously heated. The purpose of this manuscript is to provide the reader with a general experimental protocol that can be used to conduct a Miller-Urey type spark discharge experiment, using a simplified 3 L reaction flask. Since the experiment involves exposing inflammable gases to a high voltage discharge, it is worth highlighting important steps that reduce the risk of explosion. The general procedures described in this work can be extrapolated to design and conduct a wide variety of electric discharge experiments simulating primitive planetary environments.

Synthesis of Prebiotic Caramels Catalyzed by Ion-Exchange Resin Particles: Kinetic Model for the Formation of Di-d-fructose Dianhydrides.

PubMed

Ortiz Cerda, Imelda-Elizabeth; Thammavong, Phahath; Caqueret, Vincent; Porte, Catherine; Mabille, Isabelle; Garcia Fernandez, José Manuel; Moscosa Santillan, Mario; Havet, Jean-Louis

2018-02-21

Caramel enriched in di-d-fructose dianhydrides (DFAs, a family of prebiotic cyclic fructodisaccharides) is a functional food with beneficial properties for health. The aim of this work was to study the conversion of fructose into DFAs catalyzed by acid ion-exchange resin, in order to establish a simplified mechanism of the caramelization reaction and a kinetic model for DFA formation. Batch reactor experiments were carried out in a 250 mL spherical glass flask and afforded up to 50% DFA yields. The mechanism proposed entails order 2 reactions that describe fructose conversion on DFAs or formation of byproducts such as HMF or melanoidines. A third order 1 reaction defines DFA transformation into fructosyl-DFAs or fructo-oligosaccharides. The influence of fructose concentration, resin loading and temperature was studied to calculate the kinetic parameters necessary to scale up the process.

Downscaling the in vitro test of fungal bioremediation of polycyclic aromatic hydrocarbons: methodological approach.

PubMed

Drevinskas, Tomas; Mickienė, Rūta; Maruška, Audrius; Stankevičius, Mantas; Tiso, Nicola; Mikašauskaitė, Jurgita; Ragažinskienė, Ona; Levišauskas, Donatas; Bartkuvienė, Violeta; Snieškienė, Vilija; Stankevičienė, Antanina; Polcaro, Chiara; Galli, Emanuela; Donati, Enrica; Tekorius, Tomas; Kornyšova, Olga; Kaškonienė, Vilma

2016-02-01

The miniaturization and optimization of a white rot fungal bioremediation experiment is described in this paper. The optimized procedure allows determination of the degradation kinetics of anthracene. The miniaturized procedure requires only 2.5 ml of culture medium. The experiment is more precise, robust, and better controlled comparing it to classical tests in flasks. Using this technique, different parts, i.e., the culture medium, the fungi, and the cotton seal, can be analyzed. A simple sample preparation speeds up the analytical process. Experiments performed show degradation of anthracene up to approximately 60% by Irpex lacteus and up to approximately 40% by Pleurotus ostreatus in 25 days. Bioremediation of anthracene by the consortium of I. lacteus and P. ostreatus shows the biodegradation of anthracene up to approximately 56% in 23 days. At the end of the experiment, the surface tension of culture medium decreased comparing it to the blank, indicating generation of surfactant compounds.

Modeling and optimization of fermentation variables for enhanced production of lactase by isolated Bacillus subtilis strain VUVD001 using artificial neural networking and response surface methodology.

PubMed

Venkateswarulu, T C; Prabhakar, K Vidya; Kumar, R Bharath; Krupanidhi, S

2017-07-01

Modeling and optimization were performed to enhance production of lactase through submerged fermentation by Bacillus subtilis VUVD001 using artificial neural networks (ANN) and response surface methodology (RSM). The effect of process parameters namely temperature (°C), pH, and incubation time (h) and their combinational interactions on production was studied in shake flask culture by Box-Behnken design. The model was validated by conducting an experiment at optimized process variables which gave the maximum lactase activity of 91.32 U/ml. Compared to traditional activity, 3.48-folds improved production was obtained after RSM optimization. This study clearly shows that both RSM and ANN models provided desired predictions. However, compared with RSM (R 2  = 0.9496), the ANN model (R 2  = 0.99456) gave a better prediction for the production of lactase.

Enhancement in lipid content of Chlorella sp. MJ 11/11 from the spent medium of thermophilic biohydrogen production process.

PubMed

Ghosh, Supratim; Roy, Shantonu; Das, Debabrata

2017-01-01

The present study investigates the effect of spent media of acetogenic dark fermentation for mixotrophic algal cultivation for biodiesel production. Mixotrophic growth conditions were optimized in culture flask (250mL) using Chlorella sp. MJ 11/11. Maximum lipid accumulation (58% w/w) was observed under light intensity, pH, nitrate and phosphate concentration of 100μmolm -2 s -1 , 7, 2.7mM and 1.8mM, respectively. Air lift (1.4L) and flat panel (1.4L) reactors were considered for algal cultivation. Air lift showed significant improvement in biomass and lipid production as compared to flat panel reactor. The results could help in development of sustainable technology involving acetogenic hydrogen production integrated with sequential mitigation of spent media by algal cultivation for improved energy recovery. Copyright © 2016 Elsevier Ltd. All rights reserved.

Isolation and identification of a novel Candida sp. H2 producing D-arabitol and optimization of D-arabitol production.

PubMed

Song, Weibin; Lin, Yanqing; Hu, Haiyan; Xie, Zhipeng; Zhang, Jianguo

2011-03-01

To isolate a new osmophilic yeast for producing D-arabitol and research its optimal fermentation conditions for highest yield of D-arabitol from glucose. The isolated strain was characterized by electron microscopy, Biolog (GN) test, G + C content measurement and 26S rDNA D1/D2 domain sequences analysis. The purified fermentation product was identified by IR, 1H-NMR, 13C-NMR, MS and optical rotation analysis. Then the fermentation conditions for D-arabitol production were optimized. A new osmophilic yeast was isolated and identified as Candida sp. H2. Through the single factor experiment,the optimum conditions of 250 g/L glucose,10 g/L yeast extract, initial pH 6.0, 35 degrees C of culture temperature, 200 r/min of agitation, 200 mL medium in a 1000 mL flask of broth content, 1% (v/v) of inoculum size, 96 h of fermentation time were achieved. Based on the conditions above,weight yield of 35% (86.55 g D-arabitol from 250 g glucose) was obtained and 10% higher than the conditions not optimized. Candida sp. H2 was a novel strain for producing D-arabitol and valuable for further study.

[Isolation, identification and fermentation optimization of Bacillus tequilensis PanD37 producing L-aspartate α- decarboxylase].

PubMed

Feng, Zhibin; Zhang, Juan; Chen, Guozhong; Cha, Yaping; Liu, Jinjie; Ge, Yihe; Cheng, Shiwei; Yu, Botao

2016-01-04

We screened bacteria producing L-aspartate α-decarboxylase from grapery soil and optimized the fermentation conditions. L-aspartate α-decarboxylase producing bacteria were screened by color-changing circle and liquid secondary screening culture media. Combination of morphological, physiological and biochemical characteristics and 16S rRNA sequence analysis were used to identify the bacteria. Fermentation conditions were optimized by single factor test and orthogonal experiment. Strain PanD37 showed high L-aspartate α-decarboxylase producing property and was identified as Bacillus tequilensis. The optimum fermentation conditions of PanD37 were liquid volume of 50 mL in 500 mL flask, 220 r/min at 35 °C, inoculation amount of 5% for 28 h with a medium of 22.5 g/L sucrose, 7.5 g/L fumaric acid, 20 g/L peptone, 6 g/L L-aspartic acid, 2 g/L Triton X-100, at initial pH of 7.0. Under the optimal fermentation conditions, the highest L-aspartate α-decarboxylase activity reached 44.57 U/mL, which was 2.57 folds higher than that obtained before optimization. Strain PanD37 was identified as Bacillus tequilensiswhich was capable of highly producing L-aspartate α-decarboxylase under the optimal fermentation conditions.

Ethanol from Sugar Cane: Flask Experiments Using the EX-FERM Technique

PubMed Central

Rolz, Carlos; de Cabrera, Sheryl

1980-01-01

Alcohol production at the laboratory scale from sugar cane pieces by the EX-FERM technique was studied with 37 strains of Saccharomyces spp. The EX-FERM process is novel in that it employs the simultaneous extraction and fermentation of the sucrose in a cane-water suspension. Two types of cane treatments were used: chips and shredded pith, either fresh or dried. A mother culture of the yeast was prepared in enriched cane juice and then added to the cane-water mixture. After static fermentation for 40 h at 30°C, the cane was removed, and fresh cane was added to the yeast-alcohol broth. After an additional 24 h, the cane was again removed and the liquor was analyzed. After the first 40-h cycle, sugar consumption was above 99% with 10 of the 37 yeast strains tested, and ethanol reached levels of 1.29 to 4.00 g per 100 ml, depending on the yeast strain. The final ethanol concentration reached 4.27 to 5.37 g per 100 ml, and sugar consumption was above 98% in three cases during a second EX-FERM cycle employing previously air-dried chips and pith. Product yields were within accepted values. Cane treatment did not appear to affect the results at this level. PMID:16345626

Optimization of the procedure for counting the eggs of Fasciola gigantica in bovine faeces.

PubMed

Suhardono; Roberts, J A; Copeman, D B

2006-07-01

This paper describes a method for counting eggs of F. gigantica in bovine faeces that optimizes the proportion of eggs recovered and the repeatability of estimates. The method uses 3 g of faeces suspended in 0.05% Tween 20. The suspension is passed through three 6 cm diameter sieves in tandem to remove fibrous debris, with respective apertures of 1 mm, 450 microm, and either 266 or 200 microm. The filtrate is allowed to sediment for 3 min in a conical flask; the sediment is recovered, then resuspended in 200 ml of 0.05% Tween 20 and allowed to sediment. After 3 min the sediment is washed in a sieve with an aperture of 53 microm, which retains the eggs. Eggs suspended in 15 ml of 1% methylene blue are counted using a dissecting microscope. Use of Tween 20 instead of water as the suspending agent for faeces gave a significant threefold increased the proportion of eggs recovered and reduced variability between repeated counts. This method is able to detect about one-third of the eggs present. It was concluded that the high proportion of F. gigantica eggs lost may be due to the presence of hydrophobic and covalent bonds on the eggs that bind them to debris, with which they are discarded.

[Determination of olive oil content in olive blend oil by headspace gas chromatography-mass spectrometry].

PubMed

Jiang, Wanfeng; Zhang, Ning; Zhang, Fengyan; Yang, Zhao

2017-07-08

A method for the determination of the content of olive oil in olive blend oil by headspace gas chromatography-mass spectrometry (SH-GC/MS) was established. The amount of the sample, the heating temperature, the heating time, the amount of injection, the injection mode and the chromatographic column were optimized. The characteristic compounds of olive oil were found by chemometric method. A sample of 1.0 g was placed in a 20 mL headspace flask, and heated at 180℃ for 2700 s. Then, 1.0 mL headspace gas was taken into the instrument. An HP-88 chromatographic column was used for the separation and the analysis was performed by GC/MS. The results showed that the linear range was 0-100%(olive oil content). The linear correlation coefficient ( r 2 ) was more than 0.995, and the limits of detection were 1.26%-2.13%. The deviations of olive oil contents in the olive blend oil were from -0.65% to 1.02%, with the relative deviations from -1.3% to 6.8% and the relative standard deviations from 1.18% to 4.26% ( n =6). The method is simple, rapid, environment friendly, sensitive and accurate. It is suitable for the determination of the content of olive oil in olive blend oil.

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture

PubMed Central

Freund, Nathaniel W.; Croughan, Matthew S.

2018-01-01

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production—termed Lactate Supplementation and Adaptation (LSA) technology—through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications. PMID:29382079

A Simple Method to Reduce both Lactic Acid and Ammonium Production in Industrial Animal Cell Culture.

PubMed

Freund, Nathaniel W; Croughan, Matthew S

2018-01-28

Fed-batch animal cell culture is the most common method for commercial production of recombinant proteins. However, higher cell densities in these platforms are still limited due to factors such as excessive ammonium production, lactic acid production, nutrient limitation, and/or hyperosmotic stress related to nutrient feeds and base additions to control pH. To partly overcome these factors, we investigated a simple method to reduce both ammonium and lactic acid production-termed Lactate Supplementation and Adaptation (LSA) technology-through the use of CHO cells adapted to a lactate-supplemented medium. Using this simple method, we achieved a reduction of nearly 100% in lactic acid production with a simultaneous 50% reduction in ammonium production in batch shaker flasks cultures. In subsequent fed-batch bioreactor cultures, lactic acid production and base addition were both reduced eight-fold. Viable cell densities of 35 million cells per mL and integral viable cell days of 273 million cell-days per mL were achieved, both among the highest currently reported for a fed-batch animal cell culture. Investigating the benefits of LSA technology in animal cell culture is worthy of further consideration and may lead to process conditions more favorable for advanced industrial applications.

Stimulating Mathematical Thinking through Domino Games

ERIC Educational Resources Information Center

Gough, John

2015-01-01

Most readers would be familiar with the standard domino set which is played with rectangular domino tiles. The domino set, sometimes called a deck or pack, consists of 28 dominoes, colloquially nicknamed bones, cards, tiles, stones, or spinners. A domino set is a generic gaming device, similar to playing cards or dice, in that a variety of games…

Developmental Stages in Receptive Grammar Acquisition: A Processability Theory Account

ERIC Educational Resources Information Center

Buyl, Aafke; Housen, Alex

2015-01-01

This study takes a new look at the topic of developmental stages in the second language (L2) acquisition of morphosyntax by analysing receptive learner data, a language mode that has hitherto received very little attention within this strand of research (for a recent and rare study, see Spinner, 2013). Looking at both the receptive and productive…

The Shape of Things to Come: The Computational Pictograph as a Bridge from Combinatorial Space to Outcome Distribution

ERIC Educational Resources Information Center

Abrahamson, Dor

2006-01-01

This snapshot introduces a computer-based representation and activity that enables students to simultaneously "see" the combinatorial space of a stochastic device (e.g., dice, spinner, coins) and its outcome distribution. The author argues that the "ambiguous" representation fosters student insight into probability. [Snapshots are subject to peer…

Dispersant Effectiveness Of Heavy Fuel Oils Using The Baffled Flask Test

EPA Science Inventory

Dispersants have been widely used as a primary response measure for marine oil spills around the world. Recently, the U.S. Environmental Protection Agency (EPA) developed an improved laboratory dispersant testing protocol, called the Baffled Flask Test (BFT). The BFT protocol w...

Renewable Molecular Flasks with NADH Models: Combination of Light-Driven Proton Reduction and Biomimetic Hydrogenation of Benzoxazinones.

PubMed

Zhao, Liang; Wei, Jianwei; Lu, Junhua; He, Cheng; Duan, Chunying

2017-07-17

Using small molecules with defined pockets to catalyze chemical transformations resulted in attractive catalytic syntheses that echo the remarkable properties of enzymes. By modulating the active site of a nicotinamide adenine dinucleotide (NADH) model in a redox-active molecular flask, we combined biomimetic hydrogenation with in situ regeneration of the active site in a one-pot transformation using light as a clean energy source. This molecular flask facilitates the encapsulation of benzoxazinones for biomimetic hydrogenation of the substrates within the inner space of the flask using the active sites of the NADH models. The redox-active metal centers provide an active hydrogen source by light-driven proton reduction outside the pocket, allowing the in situ regeneration of the NADH models under irradiation. This new synthetic platform, which offers control over the location of the redox events, provides a regenerating system that exhibits high selectivity and efficiency and is extendable to benzoxazinone and quinoxalinone systems. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Corning HYPERFlask® for viral amplification and production of diagnostic reagents.

PubMed

Kearney, Brian J; Voorhees, Matthew A; Williams, Priscilla L; Olschner, Scott P; Rossi, Cynthia A; Schoepp, Randal J

2017-04-01

Viral preparations are essential components in diagnostic research and development. The production of large quantities of virus traditionally is done by infecting numerous tissue culture flasks or roller bottles, which require large incubators and/or roller bottle racks. The Corning HYPERFlask ® is a multilayer flask that uses a gas permeable film to provide gas exchange between the cells and culture medium and the atmospheric environment. This study evaluated the suitability of the HYPERFlask for production of Lassa, Ebola, Bundibugyo, Reston, and Marburg viruses and compared it to more traditional methods using tissue culture flasks and roller bottles. The HYPERFlask produced cultures were equivalent in virus titer and indistinguishable in immunodiagnostic assays. The use of the Corning HYPERFlask for viral production is a viable alternative to traditional tissue culture flasks and roller bottles. HYPERFlasks allow for large volumes of virus to be produced in a small space without specialized equipment. Copyright © 2016. Published by Elsevier B.V.

Strategies for Enhancing the Accumulation and Retention of Extracellular Matrix in Tissue-Engineered Cartilage Cultured in Bioreactors

PubMed Central

Shahin, Kifah; Doran, Pauline M.

2011-01-01

Production of tissue-engineered cartilage involves the synthesis and accumulation of key constituents such as glycosaminoglycan (GAG) and collagen type II to form insoluble extracellular matrix (ECM). During cartilage culture, macromolecular components are released from nascent tissues into the medium, representing a significant waste of biosynthetic resources. This work was aimed at developing strategies for improving ECM retention in cartilage constructs and thus the quality of engineered tissues produced in bioreactors. Human chondrocytes seeded into polyglycolic acid (PGA) scaffolds were cultured in perfusion bioreactors for up to 5 weeks. Analysis of the size and integrity of proteoglycans in the constructs and medium showed that full-sized aggrecan was being stripped from the tissues without proteolytic degradation. Application of low (0.075 mL min−1) and gradually increasing (0.075–0.2 mL min−1) medium flow rates in the bioreactor resulted in the generation of larger constructs, a 4.0–4.4-fold increase in the percentage of GAG retained in the ECM, and a 4.8–5.2-fold increase in GAG concentration in the tissues compared with operation at 0.2 mL min−1. GAG retention was also improved by pre-culturing seeded scaffolds in flasks for 5 days prior to bioreactor culture. In contrast, GAG retention in PGA scaffolds infused with alginate hydrogel did not vary significantly with medium flow rate or pre-culture treatment. This work demonstrates that substantial improvements in cartilage quality can be achieved using scaffold and bioreactor culture strategies that specifically target and improve ECM retention. PMID:21858004

Stress fermentation strategies for the production of hyperthermostable superoxide dismutase from Thermus thermophilus HB27: effects of ions.

PubMed

Zhu, Hu; Liu, Jianguo; Qu, Jianbo; Gao, Xinliang; Pan, Tao; Cui, Zhanfeng; Zhao, Xiubo; Lu, Jian R

2013-11-01

In this study, we explored how ammonium and metal ion stresses affected the production of recombinant hyperthermostable manganese superoxide dismutase (Mn-SOD). To improve Mn-SOD production, fed-batch culture in shake flasks and bioreactor fermentation were undertaken to examine the effects of [Formula: see text] and Mn(2+) feeding. Under the optimized feeding time and concentrations of [Formula: see text] and Mn(2+), the maximal SOD activity obtained from bioreactor fermentation reached some 480 U/ml, over 4 times higher than that in batch cultivation (113 U/ml), indicating a major enhancement of the concentration of Mn-SOD in the scale-up of hyperthermostable Mn-SOD production. In contrast, when the fed-batch culture with appropriate [Formula: see text] and Mn(2+) feeding was carried out in the same 5-L stirred tank bioreactor, a maximal SOD concentration of some 450 U/ml was obtained, again indicating substantial increase in SOD activity as a result of [Formula: see text] and Mn(2+) feeding. The isoelectric point (pI) of the sample was found to be 6.2. It was highly stable at 90 °C and circular dichroism measurements indicated a high α-helical content of 70 % as well, consistent with known SOD properties. This study indicates that [Formula: see text] and Mn(2+) play important roles in Mn-SOD expression. Stress fermentation strategies established in this study are useful for large-scale efficient production of hyperthermostable Mn-SOD and may also be valuable for the scale-up of other extremozymes.

Preliminary report on part of the Oat Hill quicksilver mine, Mayacmas district, Napa County, California

USGS Publications Warehouse

Fix, Philip Forsyth

1955-01-01

Oat Hill quicksilver mine, located in the Mayacmas district of northern California, and credited with having produced more than 160,000 flasks of quicksilver, was sampled cooperatively by the Buray of Mines and Geological Survey during 1944. 28 diamond drill holes totaling 8,120 feet were drilled by the Bureau of Mines in four of the six principal veins to sample virgin low-grade reserves and stope fill, and reserves in the other two veins were estimated from existing underground workings and by inferences from drill holes in nearby veins. The writer estimates a total of 10,220 flasks of quicksilver in indicated and inferred reserves totaling 320,000 tons. Indicated reserves minable under 1943 conditions are estimated at 1,960 flasks of quicksilver in 75,000 tons averaging 3.0 lbs Hg per ton. Inferred reserves minable under 1943 conditions are estimated at 4,640 flasks of quicksilver in 109,920 tons averaging about 3.2 lbs Hg per ton. Inferred reserves believed minable only under economic conditions much more favorable than even those of 1943 are estimated at 2,620 flasks of quicksilver in 135,080 tons averaging a little less than 1.5 lbs Hg per ton. About two-thirds of the indicated reserves are accessible in underground workings. All other reserves are estimated approximately without access underground. Several areas not sampled may possibly contain reserves.

Geochemistry of selected mercury mine-tailings in the Parkfield Mercury District, California

USGS Publications Warehouse

Rytuba, James J.; Kotlyar, Boris B.; Wilkerson, Gregg; Olson, Jerry

2001-01-01

The Parkfield mercury district is located in the southern part of the California Coast Range mercury mineral belt and contains three silica-carbonate-type mercury deposits that have had significant mercury production. Mercury was first produced in the district in 1873, but the main period of production occurred from 1915-1922. Total production from the district is about 5,000 flasks of mercury (a flask equals 76 pounds of mercury) with most production coming from the Patriquin mine (1,875 flasks), and somewhat less from the King (1,600 flasks) and Dawson (1,470 flasks) mines. Several other small prospects and mines occur in the district but only minor production has come from them. In 1969, Phelan Sulphur Company carried out mineral exploration at the King mine and announced the discovery of 55,000 tons of mercury ore with an average grade of 5.2 pounds per ton. The King mine is located on federal land administered by the U.S. Bureau of Land Management. Several other parcels of federal land are present adjacent to other mines and prospects in the Parkfield district. An environmental assessment of mine sites on and adjacent to federal land was carried out to determine the amount of mercury and other trace metals present in mine wastes and in sediments from streams impacted by past mining.

On-line prediction of the glucose concentration of CHO cell cultivations by NIR and Raman spectroscopy: Comparative scalability test with a shake flask model system.

PubMed

Kozma, Bence; Hirsch, Edit; Gergely, Szilveszter; Párta, László; Pataki, Hajnalka; Salgó, András

2017-10-25

In this study, near-infrared (NIR) and Raman spectroscopy were compared in parallel to predict the glucose concentration of Chinese hamster ovary cell cultivations. A shake flask model system was used to quickly generate spectra similar to bioreactor cultivations therefore accelerating the development of a working model prior to actual cultivations. Automated variable selection and several pre-processing methods were tested iteratively during model development using spectra from six shake flask cultivations. The target was to achieve the lowest error of prediction for the glucose concentration in two independent shake flasks. The best model was then used to test the scalability of the two techniques by predicting spectra of a 10l and a 100l scale bioreactor cultivation. The NIR spectroscopy based model could follow the trend of the glucose concentration but it was not sufficiently accurate for bioreactor monitoring. On the other hand, the Raman spectroscopy based model predicted the concentration of glucose in both cultivation scales sufficiently accurately with an error around 4mM (0.72g/l), that is satisfactory for the on-line bioreactor monitoring purposes of the biopharma industry. Therefore, the shake flask model system was proven to be suitable for scalable spectroscopic model development. Copyright © 2017 Elsevier B.V. All rights reserved.

Resource estimation from historical data: Mercury, a test case

USGS Publications Warehouse

Cargill, S.M.; Root, D.H.; Bailey, E.H.

1980-01-01

A simple technique based on historical records of tonnage and grade of ore produced provides a means for calculating how much of a mineral product will be available in the future at various average grades. Estimates made on this basis are independent of geologic considerations or changing economic and political factors, although they are based on mining history, which was largely determined by these factors. The relatively minor element, mercury, was used for the test case reported here, but the method has been found applicable to forecasts of resources for other mineral products. Mercury resources available in ore in which the average grade is as low as 0.1% are estimated to be 53 ??106kg (1.5 ??106flasks) for the United States and 1551 ??106kg (45 ??106flasks) for the world; this amount is more than adequate to meet predicted demand to the year 2000. The expectable price of mercury in 1978 dollars at this 0.1% grade is projected to be $58.75 per kg ($2,025 per flask), but at a 10% annual inflation rate, it would be more than $12,000 per flask. To satisfy just the projected U.S. demand for mercury by 2000, the price is calculated to be $48.96 per kg ($1,688 per flask) in 1978 dollars at an average annual grade of 0.12%. ?? 1980 Plenum Publishing Corporation.

THE BAFFLED FLASK TEST FOR DISPERSANT EFFECTIVENESS: A ROUND ROBIN EVALUATION OF REPRODUCIBILITY AND REPEATABILITY

EPA Science Inventory

After two previous investigations demonstrated that the Baffled Flask Test (BFT) was an effective and reproducible method for screening the effectiveness of dispersant products in the laboratory, the USEPA decided that before the new protocol cold be considered for replacement of...

EVALUATION OF MIXING ENERGY IN LABORATORY FLASKS USED FOR DISPERSANT EFFECTIVENESS TESTING

EPA Science Inventory

The evaluation of dispersant effectiveness used for oil spills is commonly done using tests conducted in laboratory flasks. The success of a test relies on replication of the conditions at sea. We used a hot wire anemometer to characterize the turbulence characteristics in the s...

Fluid Thinking.

ERIC Educational Resources Information Center

Clift, Philip A.

1992-01-01

Provides a student worksheet and set-up instructions for a science experiment to demonstrate the concept of density. Students are asked to explain the phenomenon in which a liquid of lower density placed in the bottom of two vertical flasks flows up to replace the liquid of higher density in the upper flask. (MDH)

Enhanced Biotransformation of Fluoranthene by Intertidally Derived Cunninghamella elegans under Biofilm-Based and Niche-Mimicking Conditions

PubMed Central

Mitra, Sayani; Pramanik, Arnab; Banerjee, Srijoni; Haldar, Saubhik; Gachhui, Ratan

2013-01-01

The aims of the investigation were to ascertain if surface attachment of Cunninghamella elegans and niche intertidal conditions provided in a bioreactor influenced biotransformation of fluoranthene by C. elegans. A newly designed polymethylmethacrylate (PMMA) conico-cylindrical flask (CCF) holding eight equidistantly spaced rectangular strips mounted radially on a circular disc allowed comparison of fluoranthene biotransformation between CCFs with a hydrophobic surface (PMMA-CCF) and a hydrophilic glass surface (GS-CCF) and a 500-ml Erlenmeyer flask (EF). Fluoranthene biotransformation was higher by 22-fold, biofilm growth was higher by 3-fold, and cytochrome P450 gene expression was higher by 2.1-fold when C. elegans was cultivated with 2% inoculum as biofilm culture in PMMA-CCF compared to planktonic culture in EF. Biotransformation was enhanced by 7-fold with 10% inoculum. The temporal pattern of biofilm progression based on three-channel fluorescence detection by confocal laser scanning microscopy demonstrated well-developed, stable biofilm with greater colocalization of fluoranthene within extracellular polymeric substances and filaments of the biofilm grown on PMMA in contrast to a glass surface. A bioreactor with discs rotating at 2 revolutions per day affording 6-hourly emersion and immersion mimicked the niche intertidal habitat of C. elegans and supported biofilm formation and transformation of fluoranthene. The amount of transformed metabolite was 3.5-fold, biofilm growth was 3-fold, and cytochrome P450 gene expression was 1.9-fold higher in the process mimicking the intertidal conditions than in a submerged process without disc rotation. In the CCF and reactor, where biofilm formation was comparatively greater, higher concentration of exopolysaccharides allowed increased mobilization of fluoranthene within the biofilm with consequential higher gene expression leading to enhanced volumetric productivity. PMID:24038685

Epigenetic Regulation of Autism-Associated Genes by Environmental Insults: Novel Associations

DTIC Science & Technology

2009-08-01

TITLE: Epigenetic Regulation of Autism -Associated Genes by Environmental Insults: Novel Associations PRINCIPAL INVESTIGATOR: Daryl Spinner Ph.D...SUBTITL Epigenetic regulation of Autism -associated genes by environmental insults: Novel associations 5a. CONTRACT NUMBER 5b. GRANT NUMBER...environmental-induced cases of autism . Accordingly, we established mouse embryonic cortical cultures of neurons and astrocytes, and exposed them to commonly

76 FR 18188 - Procurement List; Proposed Additions and Deletions

Federal Register 2010, 2011, 2012, 2013, 2014

2011-04-01

... objectives of the Javits-Wagner-O'Day Act (41 U.S.C. 46- 48c) in connection with the products and service....S. Coast Guard. Spinner, Salad NSN: M.R. 850. NPA: Industries for the Blind, Inc., West Allis, WI... alternatives which would accomplish the objectives of the Javits-Wagner-O'Day Act (41 U.S.C. 46- 48c) in...

Spiders (Arachnida: Araneae) Of Milbridge, Washington County, Maine

Treesearch

Daniel T. Jennings; Frank Jr. Graham

2007-01-01

An inventory or spiders associated with diverse habitats of Milbridge, a 6,290-ha area of the East Coastal BioPhysical Region, yielded 6,979 individuals of 19 families, 145 genera, and 302 species (4 unknown). Species richness per genus ranged from 1 to 13, with 88 genera represented by a single species. Total species composition favored web spinners over hunters;...

A Practical Home-Made Microcentrifuge for Teaching Purposes

ERIC Educational Resources Information Center

Moran, Paloma; Galindo, Juan

2011-01-01

We report a practical home-made microcentrifuge to be used for teaching purposes. It was made using a salad spinner and two polymerase chain reaction (PCR) tubes racks. It can accommodate 2 standard size 96-well plates or 24 strips or up to 192 microfuge tube. The centrifuge is hand-operated and is ideal for short spin down purposes. (Contains 3…

The Sakai Spinner: A Paperclip Top

ERIC Educational Resources Information Center

Corrao, Christian T.

2014-01-01

A challenge: Can you create a stable top from a single paper clip? Several interesting solutions to this problem were provided by Takao Sakai from Japan, the requirement of each being that the center of gravity be located on the vertical y-axis at the center of the top. In the simplest configuration, we see that there exists a single angle ?…

Generation of Neural Progenitor Spheres from Human Pluripotent Stem Cells in a Suspension Bioreactor.

PubMed

Yan, Yuanwei; Song, Liqing; Tsai, Ang-Chen; Ma, Teng; Li, Yan

2016-01-01

Conventional two-dimensional (2-D) culture systems cannot provide large numbers of human pluripotent stem cells (hPSCs) and their derivatives that are demanded for commercial and clinical applications in in vitro drug screening, disease modeling, and potentially cell therapy. The technologies that support three-dimensional (3-D) suspension culture, such as a stirred bioreactor, are generally considered as promising approaches to produce the required cells. Recently, suspension bioreactors have also been used to generate mini-brain-like structure from hPSCs for disease modeling, showing the important role of bioreactor in stem cell culture. This chapter describes a detailed culture protocol for neural commitment of hPSCs into neural progenitor cell (NPC) spheres using a spinner bioreactor. The basic steps to prepare hPSCs for bioreactor inoculation are illustrated from cell thawing to cell propagation. The method for generating NPCs from hPSCs in the spinner bioreactor along with the static control is then described. The protocol in this study can be applied to the generation of NPCs from hPSCs for further neural subtype specification, 3-D neural tissue development, or potential preclinical studies or clinical applications in neurological diseases.

Behavioural responses of spinner dolphins to human interactions.

PubMed

Fumagalli, Maddalena; Cesario, Amina; Costa, Marina; Harraway, John; Notarbartolo di Sciara, Giuseppe; Slooten, Elisabeth

2018-04-01

There is increasing evidence that whale and dolphin watching activities have detrimental effects on targeted cetacean populations. In Egypt, spinner dolphins regularly occur in the resting areas of Samadai, Satayah and Qubbat'Isa reefs. In-water human interactions with dolphins are regulated with a time-area closure system at Samadai, unregulated at Satayah and non-existent at Qubbat'Isa. This provided an ideal experimental setting to advance our understanding of the effects of tourism on a species highly sensitive to disturbances. Our study confirmed that the intensity and duration of interactions, and therefore, dolphin exposure to tourism, differed among the study sites. Compared with the Qubbat'Isa control site, behavioural reactions to boats and swimmers at the two tourism sites suggested that dolphin rest was disrupted, especially around the middle of the day and especially at Satayah, where dolphin tourism is unregulated. Our results indicate also that the dolphin protection measures at Samadai reduce the level of disturbance. We recommend that similar measures be implemented at other dolphin tourism locations, and that no new operations be initiated until the long-term impacts on dolphin populations are better understood. Our experience emphasizes the need to adopt precautionary approaches in research and management of whale and dolphin watching.

[Effects of naloxone on the expression of stem cell factor and C-kit receptor in combined oxygen-glucose deprivation of primary cultured human embryonic neuron in vitro].

PubMed

Zhu, Bo; Li, Lan-ying; Lü, Guo-yi; Xue, Yu-liang; Ye, Tie-hu

2010-04-01

To explore the effects of naloxone on the expression of c-kit receptor (c-kit R) and its ligand stem cell factor (SCF) in human embryo neuronal hypoxic injury. Serum-free cerebral cortical cultures prepared from embryonic human brains were deprived of both oxygen and glucose which would set up an environment more likely with that of in vivo ischemic injury. Neurons in 24-well culture plates were randomly divided into four groups: control group, hypoxia group, naloxone 0.5 microg/ml group and naloxone 10 microg/ml group. MTT assay and biological analysis were performed to study the cell death and the changes of extracellular concentrations of lactate dehydrogenase (LDH) after combined oxygen-glucose deprivation. Neurons in 25 ml culture flasks were also randomly allocated into four groups as previously described. Intracellular total RNA were extracted at different time points: pre-hypoxia, immediately after hypoxia, and 3, 6, 12, and 24 hours after reoxygenation. The changes of SCF/c-kit R mRNA expression in hypoxic neurons treated with different concentrations of naloxone pre and post oxygen-glucose deprivation were determined with RT-PCR. The cell vitality detected by MTT assay decreased significantly in hypoxia group and naloxone 0.5 microg/ml group when compared with control group (P<0.01), while no significant difference was found between naloxone 0.5 microg/ml group and hypoxia group or between naloxone 10 microg/ml group and control group. Extracellular concentration of LDH significantly increased in hypoxia group (P<0.05), while no difference was found between naloxone 0.5 microg/ml group and control group, between naloxone 0.5 microg/ml and hypoxia group, or between naloxone 10 microg/ml and control group (all P>0.05). Immediately after oxygen-glucose deprivation, the expression of SCF/c-kit R mRNA increased significantly (P<0.01). Among those the expression of SCF presented a distribution of double-peak value within 24 hours. After treated with different concentrations of naloxone, the peak value of each group were delayed to appear and went down with the increasing of naloxone concentration. The peak values in all treated groups were significantly different from that in control group (P<0.01). The expression of SCF/c-kit R mRNA increases at the early stage after combined oxygen-glucose deprivation. Naloxone 0.5 microg/ml can attenuate cell injuries and regulate the expression of SCF/c-kit R. Naloxone may protect neurons by modulating the expressions of some cytokines.

Theoretical and Practical Issues That Are Relevant When Scaling Up hMSC Microcarrier Production Processes

PubMed Central

Jossen, Valentin; Schirmer, Cedric; Mostafa Sindi, Dolman; Eibl, Regine; Kraume, Matthias; Pörtner, Ralf; Eibl, Dieter

2016-01-01

The potential of human mesenchymal stem cells (hMSCs) for allogeneic cell therapies has created a large amount of interest. However, this presupposes the availability of efficient scale-up procedures. Promising results have been reported for stirred bioreactors that operate with microcarriers. Recent publications focusing on microcarrier-based stirred bioreactors have demonstrated the successful use of Computational Fluid Dynamics (CFD) and suspension criteria (N S1u, N S1) for rapidly scaling up hMSC expansions from mL- to pilot scale. Nevertheless, one obstacle may be the formation of large microcarrier-cell-aggregates, which may result in mass transfer limitations and inhomogeneous distributions of stem cells in the culture broth. The dependence of microcarrier-cell-aggregate formation on impeller speed and shear stress levels was investigated for human adipose derived stromal/stem cells (hASCs) at the spinner scale by recording the Sauter mean diameter (d 32) versus time. Cultivation at the suspension criteria provided d 32 values between 0.2 and 0.7 mm, the highest cell densities (1.25 × 106 cells mL−1 hASCs), and the highest expansion factors (117.0 ± 4.7 on day 7), while maintaining the expression of specific surface markers. Furthermore, suitability of the suspension criterion N S1u was investigated for scaling up microcarrier-based processes in wave-mixed bioreactors for the first time. PMID:26981131

Process development for a recombinant Chinese hamster ovary (CHO) cell line utilizing a metal induced and amplified metallothionein expression system.

PubMed

Huang, Edwin P; Marquis, Christopher P; Gray, Peter P

2004-11-20

The suspension Chinese Hamster Ovary cell line, 13-10-302, utilizing the metallothionein (MT) expression system producing recombinant human growth hormone (hGH) was studied in a serum-free and cadmium-free medium at different fermentation scales and modes of operation. Initial experiments were carried out to optimize the concentration of metal addition to induce the MT promoter. Subsequently, the cultivation of the 13-10-302 cell line was scaled up from spinner flasks into bioreactors, and the cultivation duration was extended with fed-batch and perfusion strategies utilizing 180 microM zinc to induce the promoter controlling expression of recombinant hGH. It was shown that a fed-batch process could increase the maximum cell numbers twofold, from 3.3 to 6.3 x 10(6) cell/mL, over those obtained in normal batch fermentations, and this coupled with extended fermentation times resulted in a fourfold increase in final hGH titer, from 135 +/- 15 to 670 +/- 70 mg/L at a specific productivity q(hGH) value of 12 pg cell(-1)d(-1). The addition of sodium butyrate increased the specific productivity of hGH in cells to a value of approximately 48 pg cell(-1)d(-1), resulting in a final hGH titer of over a gram per liter during fed-batch runs. A BioSep acoustic cell recycler was used to retain the cells in the bioreactor during perfusion operation. It was necessary to maintain the specific feeding rates (SFR) above a value of 0.2 vvd/(10(6) cell/mL) to maintain the viability and productivity of the 13-10-302 cells; under these conditions the viable cell number increased to over 10(7) cell/mL and resulted in a volumetric productivity of over 120 mg(hGH) L(-1)d(-1). Process development described in this work demonstrates cultivation at various scales and sustained high levels of productivity under cadmium free condition in a CHO cell line utilizing an inducible metallothionein expression system. (c) 2004 Wiley Periodicals, Inc

Condensation of acetol and acetic acid vapor with sprayed liquid

USDA-ARS?s Scientific Manuscript database

A cellulose-derived fraction of biomass pyrolysis vapor was simulated by evaporating acetol and acetic acid (AA) from flasks on a hot plate. The liquid in the flasks was infused with heated nitrogen. The vapor/nitrogen stream was superheated in a tube oven and condensed by contact with a cloud of ...

PHYSICAL AND BIOLOGICAL PARAMETERS THAT DETERMINE THE FATE OF 'P'-CHLOROPHENOL IN LABORATORY TEST SYSTEMS

EPA Science Inventory

Shake flask and microcosm studies were conducted to determine the fate of parachlorophenol (p-CP) in water and sediment systems and the role of sediment and nonsediment surfaces in the biodegradation process. Biodegradation of p-CP in estuarine water samples in shake flasks was s...

A Volumetric Flask as a Projector

ERIC Educational Resources Information Center

Limsuwan, P.; Asanithi, P.; Thongpool, V.; Piriyawong, V.; Limsuwan, S.

2012-01-01

A lens based on liquid in the confined volume of a volumetric flask was presented as a potential projector to observe microscopic floating organisms or materials. In this experiment, a mosquito larva from a natural pond was selected as a demonstration sample. By shining a light beam from a laser pointer of any visible wavelength through the…

Process for casting hard-faced, lightweight camshafts and other cylindrical products

DOEpatents

Hansen, Jeffrey S.; Turner, Paul C.; Argetsinger, Edward R.; Wilson, Rick D.

1996-01-01

A process for casting a hard-faced cylindrical product such as an automobile camshaft includes the steps of: (a) preparing a composition formed from a molten base metal and an additive in particle form and having a hardness value greater than the hardness value of the base metal; (b) introducing the composition into a flask containing a meltable pattern of a cylindrical product such as an automobile camshaft to be manufactured and encased in sand to allow the composition to melt the pattern and assume the shape of the pattern within the sand; and (c) rotating the flask containing the pattern about the longitudinal axes of both the flask and the pattern as the molten base metal containing the additive in particle form is introduced into the flask to cause particles of the additive entrained in the molten base metal to migrate by centrifugal action to the radial extremities of the pattern and thereby provide a cylindrical product having a hardness value greater at it's radial extremities than at its center when the molten base metal solidifies.

Sensory Flask Cells in Sponge Larvae Regulate Metamorphosis via Calcium Signaling.

PubMed

Nakanishi, Nagayasu; Stoupin, Daniel; Degnan, Sandie M; Degnan, Bernard M

2015-12-01

The Porifera (sponges) is one of the earliest phyletic lineages to branch off the metazoan tree. Although the body-plan of sponges is among the simplest in the animal kingdom and sponges lack nervous systems that communicate environmental signals to other cells, their larvae have sensory systems that generate coordinated responses to environmental cues. In eumetazoans (Cnidaria and Bilateria), the nervous systems of larvae often regulate metamorphosis through Ca(2+)-dependent signal transduction. In sponges, neither the identity of the receptor system that detects an inductive environmental cue (hereafter "metamorphic cues") nor the signaling system that mediates settlement and metamorphosis are known. Using a combination of behavioral assays and surgical manipulations, we show here that specialized epithelial cells-referred to as flask cells-enriched in the anterior third of the Amphimedon queenslandica larva are most likely to be the sensory cells that detect the metamorphic cues. Surgical removal of the region enriched in flask cells in a larva inhibits the initiation of metamorphosis. The flask cell has an apical sensory apparatus with a cilium surrounded by an apical F-actin-rich protrusion, and numerous vesicles, hallmarks of eumetazoan sensory-neurosecretory cells. We demonstrate that these flask cells respond to metamorphic cues by elevating intracellular Ca(2+) levels, and that this elevation is necessary for the initiation of metamorphosis. Taken together, these analyses suggest that sponge larvae have sensory-secretory epithelial cells capable of converting exogenous cues into internal signals via Ca(2+)-mediated signaling, which is necessary for the initiation of metamorphosis. Similarities in the morphology, physiology, and function of the sensory flask cells in sponge larvae with the sensory/neurosecretory cells in eumetazoan larvae suggest this sensory system predates the divergence of Porifera and Eumetazoa. © The Author 2015. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology. All rights reserved. For permissions please email: journals.permissions@oup.com.

The O-mannosylation and production of recombinant APA (45/47 KDa) protein from Mycobacterium tuberculosis in Streptomyces lividans is affected by culture conditions in shake flasks.

PubMed

Gamboa-Suasnavart, Ramsés A; Valdez-Cruz, Norma A; Cordova-Dávalos, Laura E; Martínez-Sotelo, José A; Servín-González, Luis; Espitia, Clara; Trujillo-Roldán, Mauricio A

2011-12-20

The Ala-Pro-rich O-glycoprotein known as the 45/47 kDa or APA antigen from Mycobacterium tuberculosis is an immunodominant adhesin restricted to mycobacterium genus and has been proposed as an alternative candidate to generate a new vaccine against tuberculosis or for diagnosis kits. In this work, the recombinant O-glycoprotein APA was produced by the non-pathogenic filamentous bacteria Streptomyces lividans, evaluating three different culture conditions. This strain is known for its ability to produce heterologous proteins in a shorter time compared to M. tuberculosis. Three different shake flask geometries were used to provide different shear and oxygenation conditions; and the impact of those conditions on the morphology of S. lividans and the production of rAPA was characterized and evaluated. Small unbranched free filaments and mycelial clumps were found in baffled and coiled shake flasks, but one order of magnitude larger pellets were found in conventional shake flasks. The production of rAPA is around 3 times higher in small mycelia than in larger pellets, most probably due to difficulties in mass transfer inside pellets. Moreover, there are four putative sites of O-mannosylation in native APA, one of which is located at the carboxy-terminal region. The carbohydrate composition of this site was determined for rAPA by mass spectrometry analysis, and was found to contain different glycoforms depending on culture conditions. Up to two mannoses residues were found in cultures carried out in conventional shake flasks, and up to five mannoses residues were determined in coiled and baffled shake flasks. The shear and/or oxygenation parameters determine the bacterial morphology, the productivity, and the O-mannosylation of rAPA in S. lividans. As demonstrated here, culture conditions have to be carefully controlled in order to obtain recombinant O-glycosylated proteins with similar "quality" in bacteria, particularly, if the protein activity depends on the glycosylation pattern. Furthermore, it will be an interesting exercise to determine the effect of shear and oxygen in shake flasks, to obtain evidences that may be useful in scaling-up these processes to bioreactors. Another approach will be using lab-scale bioreactors under well-controlled conditions, and study the impact of those on rAPA productivity and quality.

Long-term greenhouse gas measurements from aircraft

NASA Astrophysics Data System (ADS)

Karion, A.; Sweeney, C.; Wolter, S.; Newberger, T.; Chen, H.; Andrews, A.; Kofler, J.; Neff, D.; Tans, P.

2013-03-01

In March 2009 the NOAA/ESRL/GMD Carbon Cycle and Greenhouse Gases Group collaborated with the US Coast Guard (USCG) to establish the Alaska Coast Guard (ACG) sampling site, a unique addition to NOAA's atmospheric monitoring network. This collaboration takes advantage of USCG bi-weekly Arctic Domain Awareness (ADA) flights, conducted with Hercules C-130 aircraft from March to November each year. Flights typically last 8 h and cover a large area, traveling from Kodiak up to Barrow, Alaska, with altitude profiles near the coast and in the interior. NOAA instrumentation on each flight includes a flask sampling system, a continuous cavity ring-down spectroscopy (CRDS) carbon dioxide (CO2)/methane (CH4)/carbon monoxide (CO)/water vapor (H2O) analyzer, a continuous ozone analyzer, and an ambient temperature and humidity sensor. Air samples collected in flight are analyzed at NOAA/ESRL for the major greenhouse gases and a variety of halocarbons and hydrocarbons that influence climate, stratospheric ozone, and air quality. We describe the overall system for making accurate greenhouse gas measurements using a CRDS analyzer on an aircraft with minimal operator interaction and present an assessment of analyzer performance over a three-year period. Overall analytical uncertainty of CRDS measurements in 2011 is estimated to be 0.15 ppm, 1.4 ppb, and 5 ppb for CO2, CH4, and CO, respectively, considering short-term precision, calibration uncertainties, and water vapor correction uncertainty. The stability of the CRDS analyzer over a seven-month deployment period is better than 0.15 ppm, 2 ppb, and 4 ppb for CO2, CH4, and CO, respectively, based on differences of on-board reference tank measurements from a laboratory calibration performed prior to deployment. This stability is not affected by variation in pressure or temperature during flight. We conclude that the uncertainty reported for our measurements would not be significantly affected if the measurements were made without in-flight calibrations, provided ground calibrations and testing were performed regularly. Comparisons between in situ CRDS measurements and flask measurements are consistent with expected measurement uncertainties for CH4 and CO, but differences are larger than expected for CO2. Biases and standard deviations of comparisons with flask samples suggest that atmospheric variability, flask-to-flask variability, and possible flask sampling biases may be driving the observed flask versus in situ CO2 differences rather than the CRDS measurements.

21 CFR 177.1970 - Vinyl chloride-lauryl vinyl ether copolymers.

Code of Federal Regulations, 2014 CFR

2014-04-01

... of the flask at 150 °F for 2 hours using a hot plate while also maintaining gentle mechanical agitation. Filter the contents of the flask rapidly through No. 42 Whatman filter paper with the aid of suction. Transfer the filtrate to flat glass dishes that are warmed on a hot plate and evaporate the...

Misconceptions in the Exploding Flask Demonstration Resolved through Students' Critical Thinking

ERIC Educational Resources Information Center

Spierenburg, Rick; Jacobse, Leon; de Bruin, Iris; van den Bos, Daan J.; Vis, Dominique M.; Juurlink, Ludo B. F.

2017-01-01

As it connects to a large set of important fundamental ideas in chemistry and analytical techniques discussed in high school chemistry curricula, we review the exploding flask demonstration. In this demonstration, methanol vapor is catalytically oxidized by a Pt wire catalyst in an open container. The exothermicity of reactions occurring at the…

Using PETRIMES to estimate mercury deposits in California

USGS Publications Warehouse

Lee, P.J.; Singer, D.A.

1994-01-01

In this article, we examine the use of an unconventional procedure, PETRIMES, to estimate mineral resources of mercury deposits in California. The study, which is based on the nonparametric discovery process model and Q-Q plots, suggests that a lognormal distribution is appropriate for the mercury deposits in California. The results of the assessment are summarized as follows: (1) the total number of mercury deposits in the population is approximately 165; (2) the median value of the largest undiscovered deposit size is 487 flasks; (3) the mean of the remaining mercury potential is 2,500 flasks; and (4) the population resource ranges from 1,040,000 to 4,300,000 flasks (at a 0.9 probability level). ?? 1994 Oxford University Press.

High speed turboprops for executive aircraft, potential and recent test results

NASA Technical Reports Server (NTRS)

Mikkelson, D. C.; Mitchell, G. A.

1980-01-01

Four high speed propeller models were designed and tested in an 8x6 foot wind tunnel in order to evaluate the potential of advanced propeller technology. Results from these tests show that the combination of: increased blade number, aerodynamically integrated propeller/nacelles, reduced blade thickness, spinner area ruling, and blade sweep are important in achieving high propeller efficiency at the high cruise speeds.

Volume to dissolve applied dose (VDAD) and apparent dissolution rate (ADR): tools to predict in vivo bioavailability from orally applied drug suspensions.

PubMed

Muenster, Uwe; Pelzetter, Christian; Backensfeld, Thomas; Ohm, Andreas; Kuhlmann, Thomas; Mueller, Hartwig; Lustig, Klemens; Keldenich, Jörg; Greschat, Susanne; Göller, Andreas H; Gnoth, Mark Jean

2011-08-01

Low solubility of drug candidates generated in research contributes to their elimination during subsequent development due to insufficient oral bioavailability (BA) of crystalline compound. Therefore, the purpose of the study was to identify critical in vitro solubility and dissolution parameter that would predict critical in vivo dissolution by means of in vitro-in vivo correlation. Thermodynamic solubility and apparent dissolution rate (ADR) were determined using the shake-flask method and mini-flow-through-cell, respectively. Oral BA studies in rats and humans were conducted from drug solution and suspension/tablets. Relative BA was calculated using F(rel) [%]=AUC(suspension)/AUC(solution)*100, representing a measure of in vivo dissolution. Roughly, F(rel) rat >50% translates into F(rel) human of >90%. Both, ADR and log volume to dissolve applied dose (VDAD), when plotted against F(rel) rat, revealed certain threshold levels, (ADR, ∼150-200 μg of compound dissolved under respective assay conditions; VDAD, ∼100-500 ml/kg) which translate into F(rel) in rats of >50%. Thus, assuming that F(rel)>50% in rats is indicative of sufficient in vivo dissolution in humans after oral application, drugs should exhibit a VDAD of ∼100-500 ml/kg or less in aqueous media to avoid insufficient or varying drug absorption. Copyright © 2011 Elsevier B.V. All rights reserved.

Detecting volatile compounds from Kraft lignin degradation in the headspace of microbial cultures by selected ion flow tube mass spectrometry (SIFT-MS).

PubMed

Gibson, Andrew; Malek, Lada; Dekker, Robert F H; Ross, Brian

2015-05-01

Selected Ion Flow Tube Mass Spectrometry (SIFT-MS) was used to quantify methanol and other volatile compounds in the headspace of one bacterial and 12 fungal lignin-degrading microbial cultures. Cultures were grown in 250 mL Erlenmeyer flasks capped with aluminum foil containing 40 mL of nutrient media using Kraft lignin (0.3% w/v) as the sole carbon source. Analysis was done using SIFT-MS with H3O(+) and NO(+) precursors. Product ions were identified with multiple ion mode (MIM). Full scan (FS) mode was used to identify other compounds of interest. Absidia cylindrospora, Ischnoderma resinosum and Pholiota aurivella increased headspace methanol concentration by 136 ppb, 1196 ppb and 278 ppb, respectively, while Flammulina velutipes and Laetiporus sulphureus decreased concentration below ambient levels. F. velutipes and L. sulphureus were found to produce products of methanol oxidation (formaldehyde and formic acid) and were likely metabolizing methanol. Some additional unidentified compounds generated by the fungal cultures are intriguing and will require further study. SIFT-MS can be used to quantify methanol and other volatile compounds in the headspace of microbial cultures and has the potential to be a rapid, sensitive, non-invasive tool useful in elucidating the mechanisms of lignin degradative pathways. Copyright © 2015 Elsevier B.V. All rights reserved.

Tannase sequence from a xerophilic Aspergillus niger Strain and production of the enzyme in Pichia pastoris.

PubMed

Fuentes-Garibay, José Antonio; Aguilar, Cristóbal Noé; Rodríguez-Herrera, Raúl; Guerrero-Olazarán, Martha; Viader-Salvadó, José María

2015-05-01

Tannin acyl hydrolases, or tannases (EC 3.1.1.20), are enzymes with potential biotechnological applications. In this work, we describe the gene and amino acid sequences of the tannase from Aspergillus niger GH1. In addition, we engineered Pichia pastoris strains to produce and secrete the enzyme, and the produced tannase was characterized biochemically. The nucleotide sequence of mature tannase had a length of 1,686 bp, and encodes a protein of 562 amino acids. A molecular model of mature A. niger GH1 tannase showed the presence of two structural domains, one with an α/β-hydrolase fold and one lid domain that covers the catalytic site, likely being residues Ser-196, Asp-448, and His-494 the putative catalytic triad, which are connected by a disulfide bond between the neighboring cysteines, Cys-195 and Cys-495. A 120-ml shake flask culture with a constructed recombinant P. pastoris strain showed extracellular tannase activity at 48 h induction of 0.57 U/ml. The produced tannase was N-glycosylated, consisted of two subunits, likely linked by a disulfide bond, and had an optimum pH of 5.0 and optimum temperature of 20 °C. These biochemical properties differed from those of native A. niger GH1 tannase. The recombinant tannase could be suitable for food and beverage applications.

Expression, purification and characterization of a phyAm-phyCs fusion phytase*

PubMed Central

Zou, Li-kou; Wang, Hong-ning; Pan, Xin; Tian, Guo-bao; Xie, Zi-wen; Wu, Qi; Chen, Hui; Xie, Tao; Yang, Zhi-rong

2008-01-01

The phyAm gene encoding acid phytase and optimized neutral phytase phyCs gene were inserted into expression vector pPIC9K in correct orientation and transformed into Pichia pastoris in order to expand the pH profile of phytase and decrease the cost of production. The fusion phytase phyAm-phyCs gene was successfully overexpressed in P. pastoris as an active and extracellular phytase. The yield of total extracellular fusion phytase activity is (25.4±0.53) U/ml at the flask scale and (159.1±2.92) U/ml for high cell-density fermentation, respectively. Purified fusion phytase exhibits an optimal temperature at 55 °C and an optimal pH at 5.5~6.0 and its relative activity remains at a relatively high level of above 70% in the range of pH 2.0 to 7.0. About 51% to 63% of its original activity remains after incubation at 75 °C to 95 °C for 10 min. Due to heavy glycosylation, the expressed fusion phytase shows a broad and diffuse band in SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis). After deglycosylation by endoglycosidase H (EndoHf), the enzyme has an apparent molecular size of 95 kDa. The characterization of the fusion phytase was compared with those of phyCs and phyAm. PMID:18600783

Adaptation and Molecular Characterization of Two Malaysian Very Virulent Infectious Bursal Disease Virus Isolates Adapted in BGM-70 Cell Line

PubMed Central

Lawal, Nafi'u; Arshad, Siti Suri

2017-01-01

Two Malaysian very virulent infectious bursal disease virus (vvIBDV) strains UPM0081 and UPM190 (also known as UPMB00/81 and UPM04/190, respectively) isolated from local IBD outbreaks were serially passaged 12 times (EP12) in specific pathogen free (SPF) chicken embryonated eggs (CEE) by chorioallantoic membrane (CAM) route. The EP12 isolate was further adapted and serially propagated in BGM-70 cell line up to 20 passages (P20). Characteristic cytopathic effects (CPEs) were subtly observed at P1 in both isolates 72 hours postinoculation (pi). The CPE became prominent at P5 with cell rounding, cytoplasmic vacuoles, granulation, and detachment from flask starting from day 3 pi, up to 7 days pi with titers of 109.50 TCID50/mL and log109.80 TCID50/mL for UPM0081 and UPM190, respectively. The CPE became subtle at P17 and disappeared by P18 and P19 for UPM0081 and UPM190, respectively. However, the presence of IBDV was confirmed by immunoperoxidase, immunofluorescence, and RT-PCR techniques. Phylogenetic analysis showed that these two isolates were of the vvIBDV. It appears that a single mutation of UPM190 and UPM0081 IBDV isolates at D279N could facilitate vvIBDV strain adaptability in CEE and BGM-70 cultures. PMID:29230245

Simplified production and concentration of HIV-1-based lentiviral vectors using HYPERFlask vessels and anion exchange membrane chromatography

PubMed Central

Kutner, Robert H; Puthli, Sharon; Marino, Michael P; Reiser, Jakob

2009-01-01

Background During the past twelve years, lentiviral (LV) vectors have emerged as valuable tools for transgene delivery because of their ability to transduce nondividing cells and their capacity to sustain long-term transgene expression in target cells in vitro and in vivo. However, despite significant progress, the production and concentration of high-titer, high-quality LV vector stocks is still cumbersome and costly. Methods Here we present a simplified protocol for LV vector production on a laboratory scale using HYPERFlask vessels. HYPERFlask vessels are high-yield, high-performance flasks that utilize a multilayered gas permeable growth surface for efficient gas exchange, allowing convenient production of high-titer LV vectors. For subsequent concentration of LV vector stocks produced in this way, we describe a facile protocol involving Mustang Q anion exchange membrane chromatography. Results Our results show that unconcentrated LV vector stocks with titers in excess of 108 transduction units (TU) per ml were obtained using HYPERFlasks and that these titers were higher than those produced in parallel using regular 150-cm2 tissue culture dishes. We also show that up to 500 ml of an unconcentrated LV vector stock prepared using a HYPERFlask vessel could be concentrated using a single Mustang Q Acrodisc with a membrane volume of 0.18 ml. Up to 5.3 × 1010 TU were recovered from a single HYPERFlask vessel. Conclusion The protocol described here is easy to implement and should facilitate high-titer LV vector production for preclinical studies in animal models without the need for multiple tissue culture dishes and ultracentrifugation-based concentration protocols. PMID:19220915

A simple and efficient method for generating Nurr1-positive neuronal stem cells from human wisdom teeth (tNSC) and the potential of tNSC for stroke therapy.

PubMed

Yang, Kuo-Liang; Chen, Mei-Fang; Liao, Chia-Hsin; Pang, Cheng-Yoong; Lin, Py-Yu

2009-01-01

We have isolated human neuronal stem cells from exfoliated third molars (wisdom teeth) using a simple and efficient method. The cultured neuronal stem cells (designated tNSC) expressed embryonic and adult stem cell markers, markers for chemotatic factor and its corresponding ligand, as well as neuron proteins. The tNSC expressed genes of Nurr1, NF-M and nestin. They were used to treat middle cerebral artery occlusion (MCAO) surgery-inflicted Sprague-Dawley (SD) rats to assess their therapeutic potential for stroke therapy. For each tNSC cell line, a normal human impacted wisdom tooth was collected from a donor with consent. The tooth was cleaned thoroughly with normal saline. The molar was vigorously shaken or vortexed for 30 min in a 50-mL conical tube with 15-20mL normal saline. The mixture of dental pulp was collected by centrifugation and cultured in a 25-cm(2) tissue culture flask with 4-5mL Medium 199 supplemented with 5-10% fetal calf serum. The tNSC harvested from tissue culture, at a concentration of 1-2x10(5), were suspended in 3 microL saline solution and injected into the right dorsolateral striatum of experimental animals inflicted with MCAO. Behavioral measurements of the tNSC-treated SD rats showed a significant recovery from neurologic dysfunction after MCAO treatment. In contrast, a sham group of SD rats failed to recover from the surgery. Immunohistochemistry analysis of brain sections of the tNSC-treated SD rats showed survival of the transplanted cells. These results suggest that adult neuronal stem cells may be procured from third molars, and tNSC thus cultivated have potential for treatment of stroke-inflicted rats.

Isolation of marine fungi Aspergillus sp. and its in vitro antifouling activity against marine bacteria.

PubMed

Thiyagarajan, Santhananmari; Bavya, Manoharan; Jamal, Alruwaili

2016-09-01

Biofouling is considered as a main issue of concern in aquatic environment causing severe economic loss and pollution. The aim of the present study was to isolate marine fungus antagonistic to biofouling bacteria and to define antifouling compounds present in it. Using standard plate method five predominant biofouling bacteria viz., Methylococcus sp., Flavobacterium sp., Marinococcus sp., Serratia sp. and Pseudomonas sp. were isolated from marine solid substances on Zobell's agar. Tolerance range of these bacteria to NaCl was 2-10%. Isolation of fungi from mangrove and estuarine sediments and their screening identified Aspergillus sp. EF4 as a potential isolate. This isolate caused inhibition of all the five test bacterial cultures measuring zone diameters respectively of 11, 16, 12, 13 and 11mm.? Subsequent to submerged fermentation using shaking flask method this fungus produced bioactive compounds within 5 days. The culture parameters optimized were raffinose as carbon source, yeast extract as lone nitrogen source, pH up to 9.0 and temperature up to 40?C. Antifouling compounds of culture filtrate were separated and detected by a three-step procedure involving thin layer chromatography, bioautography and preparative TLC. The in vitro assay involving glass slide-wooden stick-biofilm method revealed that these compounds could cause inhibition and destruction of bacteria to an extent of 2.16 x 104 CFU ml-1 and 2.46 x 104 CFU ml-1 respectively while growth of bacteria in control beaker was enumerated to be 4.41 x 104 CFU ml-1. High performance liquid chromatography of culture filtrate indicated probable principal antifouling compound as Fumonisin B2. Isolation of antagonistic marine fungus from Indian coast and detection of its antifouling compound would help in planning effective strategies for controlling biofouling in marine environment.

Production and characterization of active recombinant interleukin-12/eGFP fusion protein in stably-transfected DF1 chicken cells.

PubMed

Wu, Hsing Chieh; Chen, Yu San; Shen, Pin Chun; Shien, Jui Hung; Lee, Long Huw; Chiu, Hua Hsien

2015-01-01

The adjuvant activity of chicken interleukin-12 (chIL-12) protein has been described as similar to that of mammalian IL-12. Recombinant chIL-12 can be produced using several methods, but chIL-12 production in eukaryotic cells is lower than that in prokaryotic cells. Stimulating compounds, such as dimethyl sulfoxide (DMSO), can be added to animal cell cultures to overcome this drawback. In this study, we constructed a cell line, DF1/chIL-12 which stably expressed a fusion protein, chIL-12 and enhanced green fluorescent protein (eGFP) connected by a (G4 S)3 linker sequence. Fusion protein production was increased when cells were cultured in the presence of DMSO. When 1 × 10(6) DF1/chIL-12 cells were inoculated in a T-175 flask containing 30 mL of media, incubated for 15 h, and further cultivated in the presence of 4% DMSO for 48 h, the production of total fusion protein was mostly enhanced compared with the production of total fusion protein by using cell lysates induced with DMSO at other concentrations. The concentrations of the unpurified and purified total fusion proteins in cell lysates were 2,781 ± 2.72 ng mL(-1) and 2,207 ± 3.28 ng mL(-1) , respectively. The recovery rate was 79%. The fusion protein stimulated chicken splenocytes to produce IFN-γ, which was measured using an enzyme-linked immunosorbent assay, in the culture supernatant, indicating that treating DF1/chIL-12 cells with DMSO or producing chIL-12 in a fusion protein form does not have adverse effects on the bioactivity of chIL-12. © 2015 American Institute of Chemical Engineers.

Development of a circulation direct sampling and monitoring system for O2 and CO2 concentrations in the gas-liquid phases of shake-flask systems during microbial cell culture.

PubMed

Takahashi, Masato; Sawada, Yoshisuke; Aoyagi, Hideki

2017-08-23

Monitoring the environmental factors during shake-flask culture of microorganisms can help to optimise the initial steps of bioprocess development. Herein, we developed a circulation direct monitoring and sampling system (CDMSS) that can monitor the behaviour of CO 2 and O 2 in the gas-liquid phases and obtain a sample without interrupting the shaking of the culture in Erlenmeyer flasks capped with breathable culture plugs. Shake-flask culturing of Escherichia coli using this set-up indicated that a high concentration of CO 2 accumulated not only in the headspace (maximum ~100 mg/L) but also in the culture broth (maximum ~85 mg/L) during the logarithmic phase (4.5-9.0 h). By packing a CO 2 absorbent in the gas circulation unit of CDMSS, a specialised shake-flask culture was developed to remove CO 2 from the headspace. It was posited that removing CO 2 from the headspace would suppress increases in the dissolved CO 2 concentration in the culture broth (maximum ~15 mg/L). Furthermore, the logarithmic growth phase (4.5-12.0 h) was extended, the U.O.D. 580 and pH value increased, and acetic acid concentration was reduced, compared with the control. To our knowledge, this is the first report of a method aimed at improving the growth of E. coli cells without changing the composition of the medium, temperature, and shaking conditions.

Revisit: A Surprising Demonstration of Total Internal Reflection

ERIC Educational Resources Information Center

Lee, Jiwon; Cha, Yu Wha; Jung, Yeon Su; Oh, Eun Ju; Moon, Ye Lin; Kim, Jung Bog

2016-01-01

Melton demonstrated a surprising disappearance using total internal reflection. When he put a Florence flask filled with marbles into a water tank and looked straight down from directly above the flask, he was only able to see marbles above a certain water level. When he added more water into the tank above the top line of the marbles, all of the…

Wider-Opening Dewar Flasks for Cryogenic Storage

NASA Technical Reports Server (NTRS)

Ruemmele, Warren P.; Manry, John; Stafford, Kristin; Bue, Grant; Krejci, John; Evernden, Bent

2010-01-01

Dewar flasks have been proposed as containers for relatively long-term (25 days) storage of perishable scientific samples or other perishable objects at a temperature of 175 C. The refrigeration would be maintained through slow boiling of liquid nitrogen (LN2). For the purposes of the application for which these containers were proposed, (1) the neck openings of commercial off-the-shelf (COTS) Dewar flasks are too small for most NASA samples; (2) the round shapes of the COTS containers give rise to unacceptably low efficiency of packing in rectangular cargo compartments; and (3) the COTS containers include metal structures that are too thermally conductive, such that they cannot, without exceeding size and weight limits, hold enough LN2 for the required long-term-storage. In comparison with COTS Dewar flasks, the proposed containers would be rectangular, yet would satisfy the long-term storage requirement without exceeding size and weight limits; would have larger neck openings; and would have greater sample volumes, leading to a packing efficiency of about double the sample volume as a fraction of total volume. The proposed containers would be made partly of aerospace- type composite materials and would include vacuum walls, multilayer insulation, and aerogel insulation.

The radial transmission line as a broad-band shielded exposure system for microwave irradiation of large numbers of culture flasks.

PubMed

Moros, E G; Straube, W L; Pickard, W F

1999-01-01

The problem of simultaneously exposing large numbers of culture flasks at nominally equivalent incident power densities and with good thermal control is considered, and the radial transmission line (RTL) is proposed as a solution. The electromagnetic design of this structure is discussed, and an extensively bench-tested realization is described. Referred to 1 W of net forward power, the following specific absorption rate (SAR) data were obtained: at 835.62 MHz, 16.0+/-2.5 mW/kg (mean+/-SD) with range (11-22); at 2450 MHz, 245+/-50 mW/kg with range (130-323). Radio-frequency interference from an RTL driven at roughly 100 W is so low as to be compatible with a cellular base station only 500 m distant. To avoid potential confounding by temperature differences among as many as 144 T-75 flasks distributed over 9 RTLs (six irradiates and three shams), temperature within all flasks was controlled to 37.0+/-0.3 degrees C. Experience with over two years of trouble-free operation suggests that the RTL offers a robust, logistically friendly, and environmentally satisfactory solution to the problem of large-scale in vitro experiments in bioelectromagnetics.

A vacuum-operated pore-water extractor for estuarine and freshwater sediments

USGS Publications Warehouse

Winger, Parley V.; Lasier, Peter J.

1991-01-01

A vacuum-operated pore-water extractor for estuarine and freshwater sediments was developed and constructed from a fused-glass air stone attached with aquarium airline tubing to a 30 or 60 cc polypropylene syringe. Pore water is extracted by inserting the air stone into the sediment and creating a vacuum by retracting and bracing the syringe plunger. A hand-operated vacuum pump attached to a filtration flask was also evaluated as an alternative vacuum source. The volume and time to extract pore water varies with the number of devices and the sediment particle size. Extraction time is longer for fine sediments than for sandy sediments. Four liters of sediment generally yield between 500 and 1,500 mL of pore water. The sediment that surrounds and accumulates on the air stone acts as a filter, and, except for the first few milliliters, the collected pore water is clear. Because there is no exposure to air or avenue for escape, volatile compounds andin situ characteristics are retained in the extracted pore water.

Application of statistical experimental methodology to optimize bioremediation of n-alkanes in aquatic environment.

PubMed

Zahed, Mohammad Ali; Aziz, Hamidi Abdul; Mohajeri, Leila; Mohajeri, Soraya; Kutty, Shamsul Rahman Mohamed; Isa, Mohamed Hasnain

2010-12-15

Response surface methodology (RSM) was employed to optimize nitrogen and phosphorus concentrations for removal of n-alkanes from crude oil contaminated seawater samples in batch reactors. Erlenmeyer flasks were used as bioreactors; each containing 250 mL dispersed crude oil contaminated seawater, indigenous acclimatized microorganism and different amounts of nitrogen and phosphorus based on central composite design (CCD). Samples were extracted and analyzed according to US-EPA protocols using a gas chromatograph. During 28 days of bioremediation, a maximum of 95% total aliphatic hydrocarbons removal was observed. The obtained Model F-value of 267.73 and probability F<0.0001 implied the model was significant. Numerical condition optimization via a quadratic model, predicted 98% n-alkanes removal for a 20-day laboratory bioremediation trial using nitrogen and phosphorus concentrations of 13.62 and 1.39 mg/L, respectively. In actual experiments, 95% removal was observed under these conditions. Copyright © 2010 Elsevier B.V. All rights reserved.

An Innovative Very Low Thermal Power Waste Heat Recovery System for Thermal Control of Deep Space Missions: A Thermal Flask in Space

NASA Technical Reports Server (NTRS)

Bhandari, Pradeep

2015-01-01

Future missions to deep space, such as those to the outer planets (Jupiter, Saturn, etc.), which would rely on solar photovoltaic power, would need extremely large solar arrays to produce sufficient power for their operations because solar intensity is so low at those locations. Hence any additional power that would be needed for thermal control is extremely limited. Previous deep space missions like Juno (to Jupiter) required almost 200 W of electrical power for thermal control. This is prohibitively large for many future mission concepts, and leads to them needing very large solar arrays. For Saturn, where the solar flux is 1/4th the flux at Jupiter, this would entail an extremely large increase in the solar array size to accommodate the need for thermal survival power, which would be prohibitively large in size and mass, and very expensive. Hence there is a need to come up with a thermal architecture and design options that would not need such prohibitively large thermal power levels. One solution relies on harvesting the pre-existing waste heat from all the heat dissipation that would be present from operation of electronics, instruments, etc. for their own functionality. For example, for a generic Saturn mission, the various electronics would already dissipate about 200 Watts of heat that is simply "thrown away" to space from the spacecraft surfaces. The amount of thermal power that would be required for the safe thermal control of components within the spacecraft in deep space would be roughly of this magnitude for this class of spacecraft. So it makes good sense to try to harvest the waste heat and employ it to maintain the temperatures of all the components within their allowable limits. In particular, propulsion systems typically need to be kept above their freezing limits, around room temperature (15 C). Electronics needs to be kept typically above -40 C and batteries above -20 C. The next question becomes how to harvest this waste heat and direct it to the components that would need it for their survival. The proposed system utilizes a mechanically pumped, single phase fluid loop to pick up the waste heat from components attached to this loop's tubing and then directed to a thermal flask that has tubing attached to it. The thermal flask is cylindrically shaped and contains essentially all systems and components in the spacecraft within it, with the exception of the solar array, antennae, thrusters and various apertures of instruments, etc. to allow them an unobstructed view of space. Waste heat from the heat-dissipating components warms up the fluid and is carried to the flask surface and deposited on it via the fluid loop's flow. The entire flask is covered with Multi-Layered Insulation (MLI) to minimize the heat loss from the flask and allow it to remain warm. Hence the flask essentially creates a thermal environment within which the spacecraft components reside. The temperature of the components within the flask is then essentially the same as the temperature of the flask. This approach could be a very enabling feature for deep space missions. This paper describes the approach utilized for this thermal architecture, along with its mechanical and implementation aspects. Additionally it will compare and contrast this approach with the more conventional solutions utilized earlier.

HINCOF-1: a Code for Hail Ingestion in Engine Inlets

NASA Technical Reports Server (NTRS)

Gopalaswamy, N.; Murthy, S. N. B.

1995-01-01

One of the major concerns during hail ingestion into an engine is the resulting amount and space- and time-wise distribution of hail at the engine face for a given geometry of inlet and set of atmospheric and flight conditions. The appearance of hail in the capture streamtube is invariably random in space and time, with respect to size and momentum. During the motion of a hailstone through an inlet, a hailstone undergoes several processes, namely impact with other hailstones and material surfaces of the inlet and spinner, rolling and rebound following impact; heat and mass transfer; phase change; and shattering, the latter three due to friction and impact. Taking all of these factors into account, a numerical code, designated HINCOF-I, has been developed for determining the motion hailstones from the atmosphere, through an inlet, and up to the engine face. The numerical procedure is based on the Monte-Carlo method. The report presents a description of the code, along with several illustrative cases. The code can be utilized to relate the spinner geometry - conical or, more effective, elliptical - to the possible diversion of hail at the engine face into the bypass stream. The code is also useful for assessing the influence of various hail characteristics on the ingestion and distribution of hailstones over the engine face.

Resumption of traditional drive hunting of dolphins in the Solomon Islands in 2013.

PubMed

Oremus, Marc; Leqata, John; Baker, C Scott

2015-05-01

The 'drive hunting' of dolphins has a long history in the Solomon Islands, specifically at the island of Malaita. In 2010, the most active village, Fanalei, suspended hunting in exchange for financial compensation from an international non-governmental organization but resumed hunting again in early 2013. Here, we report on a visit to Fanalei in March 2013 to document the species and number of dolphins killed in the renewed hunting. Detailed records for the 2013 hunting, up to the time of our visit, included at least 1500 pantropical spotted dolphins (Stenella attenuata), 159 spinner dolphins (Stenella longirostris) and 15 'bottlenose' dolphins, probably Tursiops truncatus. Molecular identification confirmed two of the species, pantropical spotted and spinner dolphins. A summary of all available records from 1976 to 2013 documented a minimum total of 15 454 dolphins killed by the Fanalei villagers alone. We also found the local price of a dolphin tooth had increased from about US$0.14 (SBD$1) in 2004 to about US$0.70 (SBD$5) in 2013. The large number of dolphins killed and the apparent incentive for future hunting offered by the increasing commercial value of teeth, highlight an urgent need to monitor hunts and assess the abundance and trends in local populations.

Behavioural responses of spinner dolphins to human interactions

PubMed Central

Cesario, Amina; Costa, Marina; Harraway, John; Notarbartolo di Sciara, Giuseppe; Slooten, Elisabeth

2018-01-01

There is increasing evidence that whale and dolphin watching activities have detrimental effects on targeted cetacean populations. In Egypt, spinner dolphins regularly occur in the resting areas of Samadai, Satayah and Qubbat'Isa reefs. In-water human interactions with dolphins are regulated with a time-area closure system at Samadai, unregulated at Satayah and non-existent at Qubbat'Isa. This provided an ideal experimental setting to advance our understanding of the effects of tourism on a species highly sensitive to disturbances. Our study confirmed that the intensity and duration of interactions, and therefore, dolphin exposure to tourism, differed among the study sites. Compared with the Qubbat'Isa control site, behavioural reactions to boats and swimmers at the two tourism sites suggested that dolphin rest was disrupted, especially around the middle of the day and especially at Satayah, where dolphin tourism is unregulated. Our results indicate also that the dolphin protection measures at Samadai reduce the level of disturbance. We recommend that similar measures be implemented at other dolphin tourism locations, and that no new operations be initiated until the long-term impacts on dolphin populations are better understood. Our experience emphasizes the need to adopt precautionary approaches in research and management of whale and dolphin watching. PMID:29765660

40 CFR 63.2164 - If I monitor brew ethanol, what are my monitoring installation, operation, and maintenance...

Code of Federal Regulations, 2010 CFR

2010-07-01

... batch averages computed from two or more data points over each 1-hour period, except during periods when... performed. During these periods, a valid hour of data shall consist of at least one data point representing... volumetric flask and place the flask on a balance. Tare the balance. Weigh 2.3670 grams of the dry (anhydrous...

Development of Pattern Recognition Techniques for the Evaluation of Toxicant Impacts to Multispecies Systems

DTIC Science & Technology

1993-06-18

the exception. In the Standardized Aquatic Microcosm and the Mixed Flask Culture (MFC) microcosms, multivariate analysis and clustering methods...rule rather than the exception. In the Standardized Aquatic Microcosm and the Mixed Flask Culture (MFC) microcosms, multivariate analysis and...experiments using two microcosm protocols. We use nonmetric clustering, a multivariate pattern recognition technique developed by Matthews and Heame (1991

Optimization of CO2 Surface Flux using GOSAT Total Column CO2: First Results for 2009-2010

NASA Astrophysics Data System (ADS)

Basu, S.; Houweling, S.

2011-12-01

Constraining surface flux estimates of CO2 using satellite measurements has been one of the long-standing goals of the atmospheric inverse modeling community. We present the first results of inverting GOSAT total column CO2 measurements for obtaining global monthly CO2 flux maps over one year (June 2009 to May 2010). We use the SRON RemoTeC retrieval of CO2 for our inversions. The SRON retrieval has been shown to have no bias when compared to TCCON total column measurements, and latitudinal gradients of the retrieved CO2 are consistent with gradients deduced from the surface flask network [Butz et al, 2011]. This makes this retrieval an ideal candidate for atmospheric inversions, which are highly sensitive to spurious gradients. Our inversion system is analogous to the CarbonTracker (CT) data assimilation system; it is initialized with the prior CO2 fluxes of CT, and uses the same atmospheric transport model, i.e., TM5. The two major differences are (a) we add GOSAT CO2 data to the inversion in addition to flask data, and (b) we use a 4DVAR optimization system instead of a Kalman filter. We compare inversions using (a) only GOSAT total column CO2 measurements, (b) only surface flask CO2 measurements, and (c) the joint data set of GOSAT and surface flask measurements. We validate GOSAT-only inversions against the NOAA surface flask network and joint inversions against CONTRAIL and other aircraft campaigns. We see that inverted fluxes from a GOSAT-only inversion are consistent with fluxes from a stations-only inversion, reaffirming the low biases in SRON retrievals. From the joint inversion, we estimate the amount of added constraints upon adding GOSAT total column measurements to existing surface layer measurements.

Microgravity-Enhanced Stem Cell Selection

NASA Technical Reports Server (NTRS)

Claudio, Pier Paolo; Valluri, Jagan

2011-01-01

Stem cells, both embryonic and adult, promise to revolutionize the practice of medicine in the future. In order to realize this potential, a number of hurdles must be overcome. Most importantly, the signaling mechanisms necessary to control the differentiation of stem cells into tissues of interest remain to be elucidated, and much of the present research on stem cells is focused on this goal. Nevertheless, it will also be essential to achieve large-scale expansion and, in many cases, assemble cells in 3D as transplantable tissues. To this end, microgravity analog bioreactors can play a significant role. Microgravity bioreactors were originally conceived as a tool to study the cellular responses to microgravity. However, the technology can address some of the shortcomings of conventional cell culture systems; namely, the deficiency of mass transport in static culture and high mechanical shear forces in stirred systems. Unexpectedly, the conditions created in the vessel were ideal for 3D cell culture. Recently, investigators have demonstrated the capability of the microgravity bioreactors to expand hematopoietic stem cells compared to static culture, and facilitate the differentiation of umbilical cord stem cells into 3D liver aggregates. Stem cells are capable of differentiating into functional cells. However, there are no reliable methods to induce the stem cells to form specific cells or to gain enough cells for transplantation, which limits their application in clinical therapy. The aim of this study is to select the best experimental setup to reach high proliferation levels by culturing these cells in a microgravity-based bioreactor. In typical cell culture, the cells sediment to the bottom surface of their container and propagate as a one-cell-layer sheet. Prevention of such sedimentation affords the freedom for self-assembly and the propagation of 3D tissue arrays. Suspension of cells is easily achievable using stirred technologies. Unfortunately, in conventional bioreactors, stirring invokes deleterious forces that disrupt cell aggregation and results in cell death. First-generation rotating bioreactors provided rotation on the horizontal axis, which resulted in the suspension of cells without stirring, thus providing a suitable environment to propagate cells without sedimentation to a surface. The rotating wall bioreactors did not provide a way to remove air bubbles that were causing shear and disrupting 3D cultures. Johnson Space Center successfully engineered the hydrofocusing bioreactor (HFB) that resolved the problem of removing the air bubbles from the fluid medium of NASA's rotating-wall space bioreactors. The HFB uses the principle of hydrodynamic focusing that simultaneously produces a low-shear fluid culture environment and a variable hydrofocusing force that can control the movement, location, and removal of suspended cells, tissues, and air bubbles from the bioreactor. The HFB is a rotating, domeshaped cell culture vessel with a centrally located sampling port and an internal viscous spinner. The vessel and spinner can rotate at different speeds either in the same or opposite directions. Rotation of the vessel and viscous interaction at the spinner generate a hydrofocusing force. Adjusting the differential rotation rate between vessel and spinner controls the magnitude of the force.

Trophic interactions in the plankton of Malyi Sevan in July, 1984

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gulelmacher, B.L.; Simonyan, A.A.

1986-09-01

This paper describes some studies of the trophic interactions in the plankton in Lchashenskaya Bay in Malyi Sevan in July, 1984. The goal was to study the feeding characteristics of the major crustacean species and their influence on the phytoplankton. To study the effect of the planktonic crustaceans on the phytoplankton, a flask with an added 50 Acanthodiaptomus and a flask without crustaceans were exposed for 24 h after which a radioactive carbon isotope was added to them in the form of sodium carbonate. The radioactivity of the filters and working solutions was measured and autoradiographic preparations were made. Itmore » is shown that the more than threefold drop in the quantity of algae in a flask resulting from crustacean feeding did not lead to a significant decrease in primary production.« less

Atmospheric CO2 Records from Sites in the Scripps Institution of Oceanography (SIO) Air Sampling Network (1985 - 2007)

DOE Data Explorer

Keeling, R. F. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Piper, S. C. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Bollenbacher, A. F. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA); Walker , J. S. [Scripps Institution of Oceanography (SIO) University of California, La Jolla, California (USA)

2008-05-01

At Alert weekly air samples are collected in 5-L evacuated glass flasks exposed in triplicate. Flasks are returned to the SIO for CO2 determinations, which are made using an Applied Physics Corporation nondispersive infrared gas analyzer. In May 1983, the CO2-in-N2 calibration gases were replaced with CO2-in-air calibration gases, which are currently used (Keeling et al. 2002). Data are in terms of the Scripps "03A" calibration scale. On the basis of flask samples collected at Alert and analyzed by SIO, the annual average of the fitted monthly concentrations CO2 rose from 348.48 ppmv in 1986 to 384.84 ppmv in 2007. This represents an average annual growth rate of 1.73 ppmv per year at Alert.

Lithium-Ion Battery Failure: Effects of State of Charge and Packing Configuration

DTIC Science & Technology

2016-08-22

Naval Research Laboratory Washington, DC 20375-5320 NRL/MR/6180--16-9689 Lithium - Ion Battery Failure: Effects of State of Charge and Packing...PAGES 17. LIMITATION OF ABSTRACT Lithium - Ion Battery Failure: Effects of State of Charge and Packing Configuration Neil S. Spinner,* Katherine M. Hinnant...Steven G. Tuttle (202) 404-3419 Lithium - ion battery safety remains a significant concern, as battery failure leads to ejection of hazardous materials

Identification of a New Marine Bacterial Strain SD8 and Optimization of Its Culture Conditions for Producing Alkaline Protease

PubMed Central

Cui, Hongxia; Yang, Muyang; Wang, Liping; Xian, Cory J.

2015-01-01

While much attention has been given to marine microorganisms for production of enzymes, which in general are relatively more stable and active compared to those from plants and animals, studies on alkaline protease production from marine microorganisms have been very limited. In the present study, the alkaline protease producing marine bacterial strain SD8 isolated from sea muds in the Geziwo Qinhuangdao sea area of China was characterized and its optimal culture conditions were investigated. Strain SD8 was initially classified to belong to genus Pseudomonas by morphological, physiological and biochemical characterizations, and then through 16S rDNA sequence it was identified to be likely Pseudomonas hibiscicola. In addition, the culture mediums, carbon sources and culture conditions of strain SD8 were optimized for maximum production of alkaline protease. Optimum enzyme production (236U/mL when cultured bacteria being at 0.75 mg dry weight/mL fermentation broth) was obtained when the isolate at a 3% inoculum size was grown in LB medium at 20 mL medium/100mL Erlenmeyer flask for 48h culture at 30°C with an initial of pH 7.5. This was the first report of strain Pseudomonas hibiscicola secreting alkaline protease, and the data for its optimal cultural conditions for alkaline protease production has laid a foundation for future exploration for the potential use of SD8 strain for alkaline protease production. PMID:26716833

The Flask Security Architecture: System Support for Diverse Security Policies

DTIC Science & Technology

2006-01-01

Flask microkernel -based operating sys­ tem, that successfully overcomes these obstacles to pol- icy flexibility. The cleaner separation of mechanism and...other object managers in the system to en- force those access control decisions. Although the pro­ totype system is microkernel -based, the security...mecha­ nisms do not depend on a microkernel architecture and will easily generalize beyond it. The resulting system provides policy flexibility. It sup

Biofilm lifestyle enhances diesel bioremediation and biosurfactant production in the Antarctic polyhydroxyalkanoate producer Pseudomonas extremaustralis.

PubMed

Tribelli, Paula M; Di Martino, Carla; López, Nancy I; Raiger Iustman, Laura J

2012-09-01

Diesel is a widely distributed pollutant. Bioremediation of this kind of compounds requires the use of microorganisms able to survive and adapt to contaminated environments. Pseudomonas extremaustralis is an Antarctic bacterium with a remarkable survival capability associated to polyhydroxyalkanoates (PHAs) production. This strain was used to investigate the effect of cell growth conditions--in biofilm versus shaken flask cultures--as well as the inocula characteristics associated with PHAs accumulation, on diesel degradation. Biofilms showed increased cell growth, biosurfactant production and diesel degradation compared with that obtained in shaken flask cultures. PHA accumulation decreased biofilm cell attachment and enhanced biosurfactant production. Degradation of long-chain and branched alkanes was observed in biofilms, while in shaken flasks only medium-chain length alkanes were degraded. This work shows that the PHA accumulating bacterium P. extremaustralis can be a good candidate to be used as hydrocarbon bioremediation agent, especially in extreme environments.

Atmospheric CO2 Concentrations from Aircraft for 1972-1981, CSIRO Monitoring Program

DOE Data Explorer

Beardsmore, David J. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Victoria, Australia; Pearman, Graeme I. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Victoria, Australia

2012-01-01

From 1972 through 1981, air samples were collected in glass flasks from aircraft at a variety of latitudes and altitudes over Australia, New Zealand, and Antarctica. The samples were analyzed for CO2 concentrations with nondispersive infrared gas analysis. The resulting data contain the sampling dates, type of aircraft, flight number, flask identification number, sampling time, geographic sector, distance in kilometers from the listed distance measuring equipment (DME) station, station number of the radio navigation distance measuring equipment, altitude of the aircraft above mean sea level, sample analysis date, flask pressure, tertiary standards used for the analysis, analyzer used, and CO2 concentration. These data represent the first published record of CO2 concentrations in the Southern Hemisphere expressed in the WMO 1981 CO2 Calibration Scale and provide a precise record of atmospheric CO2 concentrations in the troposphere and lower stratosphere over Australia and New Zealand.

Atmospheric Carbon Dioxide Record from Flask Measurements at Lampedusa Island (May 1992- December 2000)

DOE Data Explorer

Chamard, Paolo [ENEA, GEM-CLIM, Rome, Italy; Ciattaglia, Luigi [ENEA, GEM-CLIM, Rome, Italy; di Sarra, Alcide [ENEA, GEM-CLIM, Rome, Italy; Monteleone, Francesco [ENEA, GEM-CLIM, Rome, Italy

2001-01-01

Lampedusa Station rests on a rocky seashore on the eastern tip of Lampedusa Island, located south of Sicily in the central Mediterranean Sea. Air samples at Lampedusa Station are collected each Friday in two pairs of 2-L glass flasks. Flasks are evacuated and then pressurized to 280-300 kPa with ambient air at the sampling site. Determinations of CO2 are made by using a Siemens Ultramat 5E nondispersive infrared gas analyzer. Water vapor is removed during the sampling process by means of a chemical trap (Magnesium Perchlorate). During the measurement, residual water vapor is removed by means of a cold trap at -70°C. From the inception of the monitoring program at Lampedusa Station, CO2-in-synthetic air calibration gases have been used. Values are relative to the dry air WMO mole fraction scale.

Use of saline waste water from demineralization of cheese whey for cultivation of Schizochytrium limacinum PA-968 and Japonochytrium marinum AN-4.

PubMed

Humhal, Tomas; Kastanek, Petr; Jezkova, Zuzana; Cadkova, Anna; Kohoutkova, Jana; Branyik, Tomas

2017-03-01

Saline waste water from demineralization of cheese whey was used as the main component of waste saline medium (WSM) for cultivation of thraustochytrids. The suitability of WSM for cultivation of Schizochytrium limacinum PA-968 and Japonochytrium marinum AN-4 was evaluated by comparison with cultivation on nutrient medium (NM) in shake flask and fermenter cultures. Biomass productivities achieved in WSM for the thraustochytrids were comparable with those in NM for both shake flask and fermenter cultures. The maximum total lipid content (56.71% dry cell weight) and docosahexaenoic acid productivity (0.86 g/L/day) were achieved by J. marinum AN-4 grown on WSM in shake flask and fermenter cultures, respectively. A cost estimate of WSM suggests that this medium could result in lower production costs for thraustochytrid biomass and lipids and contribute to the effective reduction in saline diary process waste water.

Contrasted accumulation patterns of persistent organic pollutants and mercury in sympatric tropical dolphins from the south-western Indian Ocean

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dirtu, Alin C.; Department of Chemistry, “Al. I. Cuza” University of Iasi, 700506 Iasi; Malarvannan, Govindan

Due to their high trophic position and long life span, small cetaceans are considered as suitable bioindicators to monitor the presence of contaminants in marine ecosystems. Here, we document the contamination with persistent organic pollutants (POPs) and total mercury (T-Hg) of spinner (Stenella longirostris, n =21) and Indo-Pacific bottlenose dolphins (Tursiops aduncus, n=32) sampled from the coastal waters of La Réunion (south-western Indian Ocean). In addition, seven co-occurring teleost fish species were sampled and analyzed as well. Blubber samples from living dolphins and muscle from teleosts were analyzed for polychlorinated biphenyls (PCBs), DDT and metabolites (DDTs), chlordanes (CHLs), hexachlorocyclohexanes (HCHs),more » hexachlorobenzene (HCB), and polybrominated diphenyl ethers (PBDEs). Methoxylated PBDEs (MeO-PBDEs), reported as having a natural origin, were also analyzed. T-Hg levels were measured in blubber and skin biopsies of the two dolphin species. Stable isotopes δ{sup 13}C and δ{sup 15}N values were determined in skin of the dolphins and in the muscle of teleosts. For PCBs, HCHs and T-Hg, concentrations were significantly higher in T. aduncus than in S. longirostris. For other POP levels, intra-species variability was high. MeO-PBDEs were the dominant compounds (55% of the total POPs) in S. longirostris, while PCBs dominated (50% contribution) in T. aduncus. Other contaminants showed similar profiles between the two species. Given the different patterns of POPs and T-Hg contamination and the δ{sup 15}N values observed among analyzed teleosts, dietary and foraging habitat preferences most likely explain the contrasted contaminant profiles observed in the two dolphin species. Levels of each class of contaminants were significantly higher in males than females. Despite their spatial and temporal overlap in the waters of La Réunion, S. longirostris and T. aduncus are differently exposed to contaminant accumulation. - Highlights: • POPs and total Hg were measured in spinner and bottlenose dolphins from La Réunion. • Levels of PCBs, PBDEs and T-Hg were lower in spinner than in bottlenose dolphins. • Contrasted contaminant profiles were observed between the two dolphin species. • Profiles are most likely explained by dietary and foraging habitat preferences.« less

Studies of Cell-Mediated Immunity Against Immune Disorders Using Synthetic Peptides and Rotating Bioreactor System

NASA Technical Reports Server (NTRS)

Sastry, Jagannadha K.

1997-01-01

Our proposed experiments included: (1) immunzing mice with synthetic peptides; (2) preparing spleen and lymph node cells; (3) growing them under conventional conditions as well as in the rotatory vessel in appropriate medium reconstituting with synthetic peptides and/or cytokines as needed; and (4) comparing at regular time intervals the specific CTL activity as well as helper T-cell activity (in terms of both proliferative responses and cytokine production) using established procedures in my laboratory. We further proposed that once we demonstrated the merit of rotatory vessel technology to achieve desired results, these studies would be expanded to include immune cells from non-human primates (rhesus monkeys and chimpanzees) and also humans. We conducted a number of experiments to determine CTL induction by the synthetic peptides corresponding to antigenic proteins in HIV and HPV in different mouse strains that express MHC haplotypes H-2b or H-2d. We immunized mice with 100 ug of the synthetic peptide, suspended in sterile water, and emulsified in CFA (1:1). The immune lymph node cells obtained after 7 days were restimulated by culturing in T25 flask, HARV-10, or STLV-50, in the presence of the peptide at 20 ug/ml. The results from the 5'Cr-release assay consistently revealed complete abrogation of CTL activity of cells grown in the bioreactors (both HARV and STLV), while significant antigen-specific CTL activity was observed with cells cultured in tissue culture flasks. Thus, overall the data we generated in this study proved the usefulness of the NASA-developed developed technology for understanding the known immune deficiency during space travel. Additionally, this ex vivo microgravity technology since it mimics effectively the in vivo situation, it is also useful in understanding immune disorders in general. Thus, our proposed studies in TMC-NASA contract round II application benefit from data generated in this TMC-NASA contract round I study.

Long-term greenhouse gas measurements from aircraft

NASA Astrophysics Data System (ADS)

Karion, A.; Sweeney, C.; Wolter, S.; Newberger, T.; Chen, H.; Andrews, A.; Kofler, J.; Neff, D.; Tans, P.

2012-10-01

In March 2009 the NOAA/ESRL/GMD Carbon Cycle and Greenhouse Gases Group collaborated with the US Coast Guard (USCG) to establish the Alaska Coast Guard (ACG) sampling site, a unique addition to NOAA's atmospheric monitoring network. This collaboration takes advantage of USCG bi-weekly Arctic Domain Awareness (ADA) flights, conducted with Hercules C-130 aircraft from March to November each year. NOAA has installed window-replacement inlet plates on two USCG C-130 aircraft and deploys a pallet with NOAA instrumentation on each ADA flight. Flights typically last 8 h and cover a very large area, traveling from Kodiak, AK in the south up to Barrow, AK in the north, and making altitude profiles near the coast as well as in the interior. NOAA instrumentation on each flight includes: a flask sampling system, a continuous CO2/CH4/CO/H2O analyzer, a continuous ozone analyzer, and an ambient temperature and humidity sensor. GPS time and location from the aircraft's navigation system are also collected. Air samples collected in flight are analyzed at NOAA/ESRL for the major greenhouse gases and a variety of halocarbons and hydrocarbons that influence climate, stratospheric ozone, and air quality. Instruments on this aircraft are designed and deployed to be able to collect air samples and data autonomously, so that NOAA personnel visit the site only for installation at the beginning of each season. We present an assessment of the cavity ring-down spectroscopy (CRDS) CO2/CH4/CO/H2O analyzer performance operating on an aircraft over a three-year period. We describe the overall system for making accurate greenhouse gas measurements using a CRDS analyzer on an aircraft with minimal operator interaction. Short and long-term stability of the CRDS analyzer over a seven-month deployment period is better than 0.15 ppm, 2 ppb, and 5 ppb for CO2, CH4, CO respectively, considering differences of on-board reference tank measurements from a laboratory calibration performed prior to deployment. This stability is not affected by variation in pressure or temperature during flight. Biases and standard deviations of comparisons with flask samples suggest that atmospheric variability, flask-to-flask variability, and possible flask sampling biases may be driving biases in the comparison between flasks and in-situ CRDS measurements.

Metabolic Profiling of Geobacter sulfurreducens during Industrial Bioprocess Scale-Up.

PubMed

Muhamadali, Howbeer; Xu, Yun; Ellis, David I; Allwood, J William; Rattray, Nicholas J W; Correa, Elon; Alrabiah, Haitham; Lloyd, Jonathan R; Goodacre, Royston

2015-05-15

During the industrial scale-up of bioprocesses it is important to establish that the biological system has not changed significantly when moving from small laboratory-scale shake flasks or culturing bottles to an industrially relevant production level. Therefore, during upscaling of biomass production for a range of metal transformations, including the production of biogenic magnetite nanoparticles by Geobacter sulfurreducens, from 100-ml bench-scale to 5-liter fermentors, we applied Fourier transform infrared (FTIR) spectroscopy as a metabolic fingerprinting approach followed by the analysis of bacterial cell extracts by gas chromatography-mass spectrometry (GC-MS) for metabolic profiling. FTIR results clearly differentiated between the phenotypic changes associated with different growth phases as well as the two culturing conditions. Furthermore, the clustering patterns displayed by multivariate analysis were in agreement with the turbidimetric measurements, which displayed an extended lag phase for cells grown in a 5-liter bioreactor (24 h) compared to those grown in 100-ml serum bottles (6 h). GC-MS analysis of the cell extracts demonstrated an overall accumulation of fumarate during the lag phase under both culturing conditions, coinciding with the detected concentrations of oxaloacetate, pyruvate, nicotinamide, and glycerol-3-phosphate being at their lowest levels compared to other growth phases. These metabolites were overlaid onto a metabolic network of G. sulfurreducens, and taking into account the levels of these metabolites throughout the fermentation process, the limited availability of oxaloacetate and nicotinamide would seem to be the main metabolic bottleneck resulting from this scale-up process. Additional metabolite-feeding experiments were carried out to validate the above hypothesis. Nicotinamide supplementation (1 mM) did not display any significant effects on the lag phase of G. sulfurreducens cells grown in the 100-ml serum bottles. However, it significantly improved the growth behavior of cells grown in the 5-liter bioreactor by reducing the lag phase from 24 h to 6 h, while providing higher yield than in the 100-ml serum bottles. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Cooling of Airplane Engines at Low Air Speeds

NASA Technical Reports Server (NTRS)

Theodorsen, Theodore; Brevoort, M J; Stickle, George W

1937-01-01

Report presents the results of a comprehensive experimental study carried out at full scale in the NACA 20-foot wind tunnel, the general purpose of which is to furnish information in regard to the functioning of the power plant and propeller unit under different conditions. This report deals particularly with the problem of the cooling of an airplane engines on the ground. The influence of different nose forms, skirts, flaps, propellers, spinners, and special blowers has been investigated.

Design of a Geothermal Downhole Magnetic Flowmeter

DOE Office of Scientific and Technical Information (OSTI.GOV)

Glowka, Dave A.; Normann, Randy A.

2015-06-15

This paper covers the development of a 300°C geothermal solid-state magnetic flowmeter (or magmeter) to support in situ monitoring of future EGS (enhanced geothermal system) production wells. Existing flowmeters are simple mechanical spinner sensors. These mechanical sensors fail within as little as 10 hrs, while a solid-state magmeter has the potential for months/years of operation. The design and testing of a magnetic flow sensor for use with existing high-temperature electronics is presented.

Observations of Gas Emissions from Cascade Range Volcanoes (USA) using a Portable Real-Time Sensor Package and Evacuated Flasks

NASA Astrophysics Data System (ADS)

Kelly, P. J.; Werner, C. A.; Evans, W.; Ingebritsen, S.; Tucker, D.

2012-12-01

Degassing from most Cascade Range Volcanoes, USA, is characterized by low-temperature hydrothermal emissions. It is important to monitor these emissions as part of a comprehensive monitoring strategy yet access is often difficult and most features are sampled by the USGS only once per year at best. In an effort to increase the sampling frequency of major gas species and in preparation for building permanent, autonomous units, we built a portable sensor package capable of measuring H2O, CO2, SO2, and H2S in volcanic gas plumes. Here we compare results from the portable sensor package with gas analyses from direct samples obtained using a titanium tube and evacuated glass flasks collected at the same time. The sensor package is housed in a small, rugged case, weighs 5 kg, and includes sensors for measuring H2O (0-16 parts per thousand), CO2 (0-5000 ppmv), SO2 (0-100 ppm), and H2S (0-20 ppm) gases. Additional temperature and pressure sensors, a micro air pump, datalogger, and an internal battery are also incorporated. H2O and CO2 are measured using an infrared spectrometer (Licor 840) and sulfur-containing gases are measured using electrochemical sensors equipped with filters to mitigate cross-sensitivities. Data are collected at a 1 Hz sampling rate and can be recorded and displayed in real-time using a netbook computer or can be saved to the onboard datalogger. The data display includes timeseries of H2O, CO2, SO2, and H2S mixing ratios, the four-component bulk composition of the plume, and automated calculation of gas ratios commonly used in volcanic gas monitoring, such as H2O/CO2, CO2/SO2, and CO2/H2S . In the Cascade Range, the sensor package has been tested at Mt. Baker, Mt. St. Helens, Mt. Hood, and in Lassen Volcanic National Park. In each case, the instrument was placed 5 to 30 meters from the fumarole or fumarole field and emissions were sampled for 5 to 30 minutes. No SO2 was detected at any location. At Mt. Hood the sensor package yielded average CO2/H2S ratios from 10 to 16 in fumarole plumes versus flask CO2/H2S ratios (n = 2) of 13 and 16 on 9 July 2011, and on 28 July 2012 the sensor package yielded an average CO2/H2S ratio of 12 versus flask ratios (n = 2) of 13 (both sets of flask samples obtained in the Crater Rock area). At Mt. Baker, the sensor package yielded average CO2/H2S ratios from 19 to 22 whereas flask ratios (n = 3) were higher, from 25 to 32 (both fumarole-plume and flask samples obtained in the Sherman Crater area) on 22 July 2011. The mismatch falls slightly outside expected analytical uncertainty for the sensor package (about 20% relative for CO2/H2S ratios). However, flask samples collected in Sherman Crater in 2006 and 2007 (n = 5) yielded CO2/H2S ratios from 18 to 29, which nearly spans the range of observations in 2011. Therefore, one explanation for the small mismatch between the results of the sensor package and direct samples is that the sensor package measures bulk plume compositions that may integrate emissions from several chemically distinct fumaroles and the direct samples better represent the composition of discrete vents. Overall, the sensor package and evacuated flask data show good agreement and demonstrate that the real-time technique is a viable means for monitoring major volcanic gas species.

Navigation and vessel inspection circular No. 3-94. International maritime organization code for the safe carriage of irradiated nuclear fuel, plutonium and high-level radioactive wastes in flasks on board ships (IMO resolution a.748(18)). Final report

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1994-05-26

The Circular calls the attention of Coast Guard field units, marine surveyors, shippers and carriers of nuclear materials to the International Maritime Organization (IMO) Code for the Safe Carriage of Irradiated Nuclear Fuel, Plutonium and High-Level Radioactive Wastes in Flasks on Board Ships (IMO Resolution A.748(18)).

Application of an Online-Biomass Sensor in an Optical Multisensory Platform Prototype for Growth Monitoring of Biotechnical Relevant Microorganism and Cell Lines in Single-Use Shake Flasks

PubMed Central

Ude, Christian; Schmidt-Hager, Jörg; Findeis, Michael; John, Gernot Thomas; Scheper, Thomas; Beutel, Sascha

2014-01-01

In the context of this work we evaluated a multisensory, noninvasive prototype platform for shake flask cultivations by monitoring three basic parameters (pH, pO2 and biomass). The focus lies on the evaluation of the biomass sensor based on backward light scattering. The application spectrum was expanded to four new organisms in addition to E. coli K12 and S. cerevisiae [1]. It could be shown that the sensor is appropriate for a wide range of standard microorganisms, e.g., L. zeae, K. pastoris, A. niger and CHO-K1. The biomass sensor signal could successfully be correlated and calibrated with well-known measurement methods like OD600, cell dry weight (CDW) and cell concentration. Logarithmic and Bleasdale-Nelder derived functions were adequate for data fitting. Measurements at low cell concentrations proved to be critical in terms of a high signal to noise ratio, but the integration of a custom made light shade in the shake flask improved these measurements significantly. This sensor based measurement method has a high potential to initiate a new generation of online bioprocess monitoring. Metabolic studies will particularly benefit from the multisensory data acquisition. The sensor is already used in labscale experiments for shake flask cultivations. PMID:25232914

Application of multivariate analysis and mass transfer principles for refinement of a 3-L bioreactor scale-down model--when shake flasks mimic 15,000-L bioreactors better.

PubMed

Ahuja, Sanjeev; Jain, Shilpa; Ram, Kripa

2015-01-01

Characterization of manufacturing processes is key to understanding the effects of process parameters on process performance and product quality. These studies are generally conducted using small-scale model systems. Because of the importance of the results derived from these studies, the small-scale model should be predictive of large scale. Typically, small-scale bioreactors, which are considered superior to shake flasks in simulating large-scale bioreactors, are used as the scale-down models for characterizing mammalian cell culture processes. In this article, we describe a case study where a cell culture unit operation in bioreactors using one-sided pH control and their satellites (small-scale runs conducted using the same post-inoculation cultures and nutrient feeds) in 3-L bioreactors and shake flasks indicated that shake flasks mimicked the large-scale performance better than 3-L bioreactors. We detail here how multivariate analysis was used to make the pertinent assessment and to generate the hypothesis for refining the existing 3-L scale-down model. Relevant statistical techniques such as principal component analysis, partial least square, orthogonal partial least square, and discriminant analysis were used to identify the outliers and to determine the discriminatory variables responsible for performance differences at different scales. The resulting analysis, in combination with mass transfer principles, led to the hypothesis that observed similarities between 15,000-L and shake flask runs, and differences between 15,000-L and 3-L runs, were due to pCO2 and pH values. This hypothesis was confirmed by changing the aeration strategy at 3-L scale. By reducing the initial sparge rate in 3-L bioreactor, process performance and product quality data moved closer to that of large scale. © 2015 American Institute of Chemical Engineers.

The Influence of Ecological Isolation on the Structural and Functional Stability of Complex Microbial Communities

NASA Technical Reports Server (NTRS)

Franklin, R. B.; Garland, J. L.; Mills, A. L.

2005-01-01

To help understand how the behavior of microorganisms and microbial communities in insular space habitats may differ from the behavior of these groups on Earth, long-term incubations (100+ days) were conducting using wastewater bioreactors (batch fed) designed to mimic "closed" and "open" ecological systems. The issue of immigration was considered, and the goal of the research was to determine whether the stability of microbial communities in space is reduced due to their prolonged isolation. Bioreactors were established by inoculating flasks of sterile synthetic wastewater with the microbial community obtained from a local treatment facility; each day, one-third of the medium in the flask was replaced with an equal volume of sterile artificial wastewater. Flasks were divided into two treatments: "closed" and "open" to recruitment of additional microorganisms. "Closed" flasks were maintained as described above, while the medium used to feed the "open" flasks was supplemented daily with a small amount of raw sewage (which provided a continuous source of new potential community members). Significant differences in microbial community structure and function developed in the two sets of communities, and the results suggest that the open community was more stable and better able to adjust to changing environmental conditions. Each community's resistance to environmental (temperature fluctuations) and biological stresses (starvation and invasion by an opportunistic pathogen Pseudomonas aeruginosa) was monitored. Experiments were also conducted to determine whether the effect of isolation changes depending on the microbial communities' initial diversity or composition; communities with a low(er) initial diversity were less stable. Overall, the results indicate that isolation will be an important factor influencing the activity of microbial communities on board spacecraft. A possible way of mitigating these effects would be to include communities with high initial diversity or to periodically re-inoculate the systems using diverse inocula transported from Earth.

Chemistries for Protection and Decontamination

DTIC Science & Technology

2008-03-01

t i o n Measurement of Oxygen Uptake, Supported Catalyst and CEES in Solvent-Free System TDA R e s e a r c h Tests with HD • At CUBRC (Buffalo...dual gas burettes for HD tests at CUBRC . The dual burette on the left was connected to a flask with the catalyst and HD (and to an empty flask as a...Hill, Zhen Luo, Daniel Hillesheim • ECBC: Larry Procell • CUBRC : Meg Stapleton, Rich Fitzpatrick • Battelle: John Ontiveros, Walter Miller • ARO

Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation.

PubMed

Wang, C L; Li, D F; Lu, W Q; Wang, Y H; Lai, C H

2004-01-01

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Penicillium sp. in solid-state fermentation (SSF). Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately. The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively. The range of pH 5.5-6.5 was favourable. About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production. Seventy-five hours of incubation was enough for maximum alpha-galactosidase production. Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays. Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp. MAFIC-6 indicated 185.2 U g(-1) in tray of SSF. The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up.

Comparison of extraction techniques of robenidine from poultry feed samples.

PubMed

Wilga, Joanna; Wasik, Agata Kot-; Namieśnik, Jacek

2007-10-31

In this paper, effectiveness of six different commonly applied extraction techniques for the determination of robenidine in poultry feed has been compared. The sample preparation techniques included shaking, Soxhlet, Soxtec, ultrasonically assisted extraction, microwave - assisted extraction and accelerated solvent extraction. Comparison of these techniques was done with respect to the recovery extraction, temperature and time, reproducibility and solvent consumption. Every single extract was subjected to clean - up using aluminium oxide column (Pasteur pipette filled with 1g of aluminium oxide), from which robenidine was eluted with 10ml of methanol. The eluate from the clean-up column was collected in a volumetric flask, and finally it was analysed by HPLC-DAD-MS. In general, all extraction techniques were capable of isolating of robenidine from poultry feed, but the recovery obtained using modern extraction techniques was higher than that obtained using conventional techniques. In particular, accelerated solvent extraction was more superior to other techniques, which highlights the advantages of this sample preparation technique. However, in routine analysis, shaking and ultrasonically assisted extraction is still the preferred method for the solution of robenidine and other coccidiostatics.

An Analytical Study of Icing Similitude for Aircraft Engine Testing. Revision

DTIC Science & Technology

1987-02-01

MODELING GEOMETRIES Component Cowl Spinner Fan Blade Fan Stator Exit Vane Probe Approximating Geometry NACA 0012 Airfoil Sphere NACA 0012...DOT/FAA/CT·86/35 AEDC·TR·86·26 An Analytical Study of Icing Similitude for Aircraft Engine Testing c. Scott Bartlett Sverdrup Technology, Inc...8217~,feCa.ORI A n AnalYtical Study )f Icin~ Similitude for Aircraft Engine Tes t tu~ 12. PERSONAL AUTHOR/S) B a r t l e t t , C. Scot t , Sverdrup

Low-Cost, High-Throughput 3-D Pulmonary Imager Using Hyperpolarized Contrast Agents and Low-Field MRI

DTIC Science & Technology

2017-10-01

Body MRI Experiments. Enc. Magn. Reson. 2007, DOI: 10.1002/9780470034590.emrstm0491. (59) Minard, K. R.; Wind , R. A. Solenoidal microcoil designPart II...Optimizing winding parameters for maximum signal-to-noise perform- ance. Concepts Magn. Reson. 2001, 13, 190−210. (60) Danieli, E.; Perlo, J...NMR spinner turbine was adjusted for the detection of the gas phase just above the liquid (Figure S2). Next, the displacement of HP propane from the

Effect of flask vibration time on casting integrity, Surface Penetration and Coating Inclusion in lost foam casting of Al-Si Alloy

DOE Office of Scientific and Technical Information (OSTI.GOV)

Karimian, Majid; Idris, M. H.; Ourdjini, A.

2011-01-17

The paper presents the result of an experimental investigation conducted on medium aluminum silicon alloy casting- LM6, using no-vacuum assisted lost foam casting process. The study is directed for establishing the relationship between the flask vibrations times developed for molded sample on the casting integrity, surface penetration and coating inclusion defects of the casting. Four different flask vibration times namely 180, 120, 90 and 60 sec. were investigated. The casting integrity was investigated in terms of fulfilling in all portions and edges. The surface penetration was measured using optical microscope whilst image analyzer was used to quantify the percentage ofmore » coating inclusion in the casting. The results show that vibration time has significant influence on the fulfilling as well as the internal integrity of the lost foam casting. It was found that the lower vibration time produced comparatively sound casing.« less

Shear stress enhances microcin B17 production in a rotating wall bioreactor, but ethanol stress does not

NASA Technical Reports Server (NTRS)

Gao, Q.; Fang, A.; Pierson, D. L.; Mishra, S. K.; Demain, A. L.

2001-01-01

Stress, including that caused by ethanol, has been shown to induce or promote secondary metabolism in a number of microbial systems. Rotating-wall bioreactors provide a low stress and simulated microgravity environment which, however, supports only poor production of microcin B17 by Escherichia coli ZK650, as compared to production in agitated flasks. We wondered whether the poor production is due to the low level of stress and whether increasing stress in the bioreactors would raise the amount of microcin B17 formed. We found that applying shear stress by addition of a single Teflon bead to a rotating wall bioreactor improved microcin B17 production. By contrast, addition of various concentrations of ethanol to such bioreactors (or to shaken flasks) failed to increase microcin B17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the bioreactor were much more resistant to the growth-inhibitory and production-inhibitory effects of ethanol than cells growing in shaken flasks.

Atmospheric CO2 From Flask Air Samples at 10 Sites in the Scripps Institution of Oceanography (SIO) Air Sampling Network (1957 - 2001) (issued 2004)

DOE Data Explorer

Keeling, Charles D. [Univ. of California, San Diego, CA (United States). Scripps Inst. of Oceanography; Whorf, Timothy P. [Univ. of California, San Diego, CA (United States). Scripps Inst. of Oceanography; Blasing, T. J. [Carbon Dioxide Information Analysis Center (CDIAC), Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (USA); Jones, Sonja [Carbon Dioxide Information Analysis Center (CDIAC), Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (USA)

2004-09-01

The Carbon Dioxide Research Group, Scripps Institution of Oceanography, University of California, San Diego, has provided this data set, which includes long-term measurements of near-surface atmospheric CO2 concentrations at 10 locations spanning latitudes 82°N to 90°S. Most of the data are based on replicated (collected at the same time and place) flask samples taken at intervals of approximately one week to one month and subsequently subjected to infrared analysis. Periods of record begin in various years, ranging from 1957 (for the South Pole station) to 1985 (for Alert, Canada), and all flask data records except for Christmas Island and Baring Head, New Zealand extend through year 2001. Christmas Island data end with August, 2001 and Baring Head data end with October 2001. Weekly averages of continuous data from Mauna Loa Observatory, Hawaii, are available back to March 1958. Similar weekly averages are also available for La Jolla, California, from November 1972 to October 1975, and for the South Pole from June 1960 to October 1963. These long-term records of atmospheric CO2 concentration complement the continuous records made by SIO, and also complement the long term flask records of the Climate Monitoring and Diagnostics Laboratory of the National Oceanic and Atmospheric Administration. All these data are useful for characterizing seasonal and geographical variations in atmospheric CO2 over several years, and for assessing results of global carbon models.

Large-scale production of lentiviral vector in a closed system hollow fiber bioreactor

PubMed Central

Sheu, Jonathan; Beltzer, Jim; Fury, Brian; Wilczek, Katarzyna; Tobin, Steve; Falconer, Danny; Nolta, Jan; Bauer, Gerhard

2015-01-01

Lentiviral vectors are widely used in the field of gene therapy as an effective method for permanent gene delivery. While current methods of producing small scale vector batches for research purposes depend largely on culture flasks, the emergence and popularity of lentiviral vectors in translational, preclinical and clinical research has demanded their production on a much larger scale, a task that can be difficult to manage with the numbers of producer cell culture flasks required for large volumes of vector. To generate a large scale, partially closed system method for the manufacturing of clinical grade lentiviral vector suitable for the generation of induced pluripotent stem cells (iPSCs), we developed a method employing a hollow fiber bioreactor traditionally used for cell expansion. We have demonstrated the growth, transfection, and vector-producing capability of 293T producer cells in this system. Vector particle RNA titers after subsequent vector concentration yielded values comparable to lentiviral iPSC induction vector batches produced using traditional culture methods in 225 cm2 flasks (T225s) and in 10-layer cell factories (CF10s), while yielding a volume nearly 145 times larger than the yield from a T225 flask and nearly three times larger than the yield from a CF10. Employing a closed system hollow fiber bioreactor for vector production offers the possibility of manufacturing large quantities of gene therapy vector while minimizing reagent usage, equipment footprint, and open system manipulation. PMID:26151065

Potential hazards of toxic metals found in toothpastes commonly used in Nigeria.

PubMed

Orisakwe, Orish Ebere; Okolo, Kenneth Obinna; Igweze, Zelinjo Nkeiruka; Ajaezi, Godwin Chukwuebuka; Udowelle, Nnaemeka Arinze

2016-01-01

Toothpastes have multi-functional configurations as oral care products. They can however constitute a pos- sible source, amongst others, of toxic metal exposure in public health. Indeed, the public health impact of personal hygiene and consumer products is largely unknown. To determine the level of toxic metals (lead, cadmium, cobalt, chromium, nickel) in toothpastes available in Nigeria, (home produced and imported), and assess the potential risk to the people. The samples of toothpastes commonly used in Nigeria were tested. Using a market basket protocol thirty five different brands of toothpaste were used. Samples were digest by addition of 10 mL mixture of conc. nitric and hydrochloric acids (HCl:HNO(3), 3:1), followed by heating to dryness. 20 mL deionized water was added, stirred and filtered. The filtrate was made up in standard volumetric flask and lead, cadmium, chromium, cobalt and nickel concentrations were determined using the atomic absorption spectrophotometry 205A. The daily intake of metals and target hazard quotient (THQ) were then calculated. Pepsodent and Flodent had the highest levels of lead at respectively 23.575 and 18.092 mg/kg while Colgate Herbal had the highest nickel of 18.535 mg/kg. The daily intake estimates of all imported toothpaste samples were below the stated upper limits (UL). All target hazard quotients were also found to be below one. Although the UL, THQ and daily intake rates were all normal, the high levels of lead in some of the tooth- pastes an important concern to public health suggesting that pre-marketing safety studies of toothpastes may be worthwhile for the regulatory authorities.

A Simplified Extemporaneously Prepared Potassium Chloride Oral Solution.

PubMed

Tannous, Elias; Tal, Yana; Amarny, Kamal

2016-01-01

Although commercial preparations of oral potassium supplements are usually available, there are times when our Medical Center is faced with situations in which the oral solution of potassium chloride is not available. This solution is necessary for our pediatric outpatients who cannot swallow tablets and need an oral solution. Moreover, there are no studies available which describe an extemporaneously prepared potassium chloride oral solution on which we can rely for assigning a beyond-use date. The aim of this study was to formulate an extemporaneous pediatric oral solution of potassium chloride and to determine the physical and chemical stability of this preparation. We prepared 1 mMoL/mL by withdrawing 25 mL of potassium chloride 14.9%. Ora-Sweet SF was added to 50 mL in a metered flask. The solution was kept refrigerated (2°C to 8°C). Samples were withdrawn to measure potassium concentration, pH, and microbial overgrowth. The test was performed by our biochemical laboratory. The oral solution of potassium chloride 1 mMoL/mL stored at 2°C to 8°C maintained at least 91% of the initial concentration for 28 days. There were no notable changes in pH, and the solution remained physically stable with no visual microbial growth. The oral solution of potassium chloride 1 mMoL/mL prepared in Ora-Sweet and stored at 2°C to 8°C in amber glass bottles is expected to remain stable for 28 days. Copyright© by International Journal of Pharmaceutical Compounding, Inc.

Genetic Circuit Performance under Conditions Relevant for Industrial Bioreactors

PubMed Central

Moser, Felix; Broers, Nicolette J.; Hartmans, Sybe; Tamsir, Alvin; Kerkman, Richard; Roubos, Johannes A.; Bovenberg, Roel; Voigt, Christopher A.

2014-01-01

Synthetic genetic programs promise to enable novel applications in industrial processes. For such applications, the genetic circuits that compose programs will require fidelity in varying and complex environments. In this work, we report the performance of two synthetic circuits in Escherichia coli under industrially relevant conditions, including the selection of media, strain, and growth rate. We test and compare two transcriptional circuits: an AND and a NOR gate. In E. coli DH10B, the AND gate is inactive in minimal media; activity can be rescued by supplementing the media and transferring the gate into the industrial strain E. coli DS68637 where normal function is observed in minimal media. In contrast, the NOR gate is robust to media composition and functions similarly in both strains. The AND gate is evaluated at three stages of early scale-up: 100 ml shake-flask experiments, a 1 ml MTP microreactor, and a 10 L bioreactor. A reference plasmid that constitutively produces a GFP reporter is used to make comparisons of circuit performance across conditions. The AND gate function is quantitatively different at each scale. The output deteriorates late in fermentation after the shift from exponential to constant feed rates, which induces rapid resource depletion and changes in growth rate. In addition, one of the output states of the AND gate failed in the bioreactor, effectively making it only responsive to a single input. Finally, cells carrying the AND gate show considerably less accumulation of biomass. Overall, these results highlight challenges and suggest modified strategies for developing and characterizing genetic circuits that function reliably during fermentation. PMID:23656232

Shape control and compartmentalization in active colloidal cells

PubMed Central

Spellings, Matthew; Engel, Michael; Klotsa, Daphne; Sabrina, Syeda; Drews, Aaron M.; Nguyen, Nguyen H. P.; Bishop, Kyle J. M.; Glotzer, Sharon C.

2015-01-01

Small autonomous machines like biological cells or soft robots can convert energy input into control of function and form. It is desired that this behavior emerges spontaneously and can be easily switched over time. For this purpose we introduce an active matter system that is loosely inspired by biology and which we term an active colloidal cell. The active colloidal cell consists of a boundary and a fluid interior, both of which are built from identical rotating spinners whose activity creates convective flows. Similarly to biological cell motility, which is driven by cytoskeletal components spread throughout the entire volume of the cell, active colloidal cells are characterized by highly distributed energy conversion. We demonstrate that we can control the shape of the active colloidal cell and drive compartmentalization by varying the details of the boundary (hard vs. flexible) and the character of the spinners (passive vs. active). We report buckling of the boundary controlled by the pattern of boundary activity, as well as formation of core–shell and inverted Janus phase-separated configurations within the active cell interior. As the cell size is increased, the inverted Janus configuration spontaneously breaks its mirror symmetry. The result is a bubble–crescent configuration, which alternates between two degenerate states over time and exhibits collective migration of the fluid along the boundary. Our results are obtained using microscopic, non–momentum-conserving Langevin dynamics simulations and verified via a phase-field continuum model coupled to a Navier–Stokes equation. PMID:26253763

Shape control and compartmentalization in active colloidal cells.

PubMed

Spellings, Matthew; Engel, Michael; Klotsa, Daphne; Sabrina, Syeda; Drews, Aaron M; Nguyen, Nguyen H P; Bishop, Kyle J M; Glotzer, Sharon C

2015-08-25

Small autonomous machines like biological cells or soft robots can convert energy input into control of function and form. It is desired that this behavior emerges spontaneously and can be easily switched over time. For this purpose we introduce an active matter system that is loosely inspired by biology and which we term an active colloidal cell. The active colloidal cell consists of a boundary and a fluid interior, both of which are built from identical rotating spinners whose activity creates convective flows. Similarly to biological cell motility, which is driven by cytoskeletal components spread throughout the entire volume of the cell, active colloidal cells are characterized by highly distributed energy conversion. We demonstrate that we can control the shape of the active colloidal cell and drive compartmentalization by varying the details of the boundary (hard vs. flexible) and the character of the spinners (passive vs. active). We report buckling of the boundary controlled by the pattern of boundary activity, as well as formation of core-shell and inverted Janus phase-separated configurations within the active cell interior. As the cell size is increased, the inverted Janus configuration spontaneously breaks its mirror symmetry. The result is a bubble-crescent configuration, which alternates between two degenerate states over time and exhibits collective migration of the fluid along the boundary. Our results are obtained using microscopic, non-momentum-conserving Langevin dynamics simulations and verified via a phase-field continuum model coupled to a Navier-Stokes equation.

Shape control and compartmentalization in active colloidal cells

DOE PAGES

Spellings, Matthew; Engel, Michael; Klotsa, Daphne; ...

2015-08-07

Small autonomous machines like biological cells or soft robots can convert energy input into control of function and form. It is desired that this behavior emerges spontaneously and can be easily switched over time. For this purpose, in this paper we introduce an active matter system that is loosely inspired by biology and which we term an active colloidal cell. The active colloidal cell consists of a boundary and a fluid interior, both of which are built from identical rotating spinners whose activity creates convective flows. Similarly to biological cell motility, which is driven by cytoskeletal components spread throughout themore » entire volume of the cell, active colloidal cells are characterized by highly distributed energy conversion. We demonstrate that we can control the shape of the active colloidal cell and drive compartmentalization by varying the details of the boundary (hard vs. flexible) and the character of the spinners (passive vs. active). We report buckling of the boundary controlled by the pattern of boundary activity, as well as formation of core–shell and inverted Janus phase-separated configurations within the active cell interior. As the cell size is increased, the inverted Janus configuration spontaneously breaks its mirror symmetry. The result is a bubble–crescent configuration, which alternates between two degenerate states over time and exhibits collective migration of the fluid along the boundary. Finally, our results are obtained using microscopic, non–momentum-conserving Langevin dynamics simulations and verified via a phase-field continuum model coupled to a Navier–Stokes equation.« less

English Translation of Seven Papers on Thermophysical Properties

DTIC Science & Technology

1980-09-01

capillary constant (eq. 18) *- a = surface tension (eq. 18) g = acceleration of gravity (eq. 18) v = atomic volume at the melting point - . .. M-16...Accuracyoft/"m Ex*Ceperiment C 1941 r1143 oil 181-195 0.4610.01 Specific gravity flask 1946 [20 - t, Ar (up to 332-160 0.517 1.01 Specific gravity ...i 65 arm) flask, 0.52 1ś No1 P, 1 Kb 16 319T 400-1125 0.503 1.021 Specific gravity 1970 163) was - 242.-917 0.304 0.95 Hydrostatic sun-pension. -. 3

Atmospheric CH4 Concentrations from the Commonwealth Scientific and Industrial Research Organization (CSIRO) GASLAB Flask Sampling Network (1984 - 2001)

DOE Data Explorer

Steele, L. P. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Krummel, P. B. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia; Langenfelds, R. L. [Commonwealth Scientific and Industrial Research Organization (CSIRO), Aspendale, Victoria, Australia

2003-01-01

The listed data were obtained from flask air samples returned to the CSIRO GASLAB for analysis. Typical sample storage times ranged from days to weeks for some sites (e.g., Cape Grim) to as much as one year for Macquarie Island and the Antarctic sites. Experiments carried out to test for any change in sample CH4 mixing ratio during storage have shown no drift to within detection limits over test periods of several months to years (Cooper et al., 1999).

Fluid Dynamics Appearing during Simulated Microgravity Using Random Positioning Machines

PubMed Central

Stern, Philip; Casartelli, Ernesto; Egli, Marcel

2017-01-01

Random Positioning Machines (RPMs) are widely used as tools to simulate microgravity on ground. They consist of two gimbal mounted frames, which constantly rotate biological samples around two perpendicular axes and thus distribute the Earth’s gravity vector in all directions over time. In recent years, the RPM is increasingly becoming appreciated as a laboratory instrument also in non-space-related research. For instance, it can be applied for the formation of scaffold-free spheroid cell clusters. The kinematic rotation of the RPM, however, does not only distribute the gravity vector in such a way that it averages to zero, but it also introduces local forces to the cell culture. These forces can be described by rigid body analysis. Although RPMs are commonly used in laboratories, the fluid motion in the cell culture flasks on the RPM and the possible effects of such on cells have not been examined until today; thus, such aspects have been widely neglected. In this study, we used a numerical approach to describe the fluid dynamic characteristic occurring inside a cell culture flask turning on an operating RPM. The simulations showed that the fluid motion within the cell culture flask never reached a steady state or neared a steady state condition. The fluid velocity depends on the rotational velocity of the RPM and is in the order of a few centimeters per second. The highest shear stresses are found along the flask walls; depending of the rotational velocity, they can reach up to a few 100 mPa. The shear stresses in the “bulk volume,” however, are always smaller, and their magnitude is in the order of 10 mPa. In conclusion, RPMs are highly appreciated as reliable tools in microgravity research. They have even started to become useful instruments in new research fields of mechanobiology. Depending on the experiment, the fluid dynamic on the RPM cannot be neglected and needs to be taken into consideration. The results presented in this study elucidate the fluid motion and provide insight into the convection and shear stresses that occur inside a cell culture flask during RPM experiments. PMID:28135286

Investigations on the Use of Multi-Species Flask Measurements for Sector-Specific Fossil Fuel CO2 Attribution to Aid Policymakers

NASA Astrophysics Data System (ADS)

Nathan, B.; Lauvaux, T.; Turnbull, J. C.; Sweeney, C.; Karion, A.; Richardson, S.; Miles, N.; Gurney, K. R.; Patarasuk, R.

2016-12-01

Part of the Indianapolis Flux (INFLUX) Experiment has, since 2010, involved recording atmospheric trace gas measurements using NOAA flask packages. The goal of these measurements is to better inform policymakers about the behaviors of greenhouse gas emissions in the Indianapolis urban environment. Radiocarbon dioxide (14CO2) measurements recorded from the flasks allow for delineation of the fossil-fuel carbon dioxide (CO2ff) signal from the total carbon dioxide (CO2) measurement. To give policymakers even more detailed information, we investigate whether the co-measured trace gases could be used as tracers for economic source sectors of CO2ff as predefined by the bottom-up data product Hestia. This is extensively tested using an Observation System Simulation Experiment (OSSE) combining both a top-down approach for all species—influence functions from the tower flask measurements—, and attempting to assign sources via spatial overlaps with the available bottom-up inventory CO2ff source sector definitions. A self-organizing map is implemented for the mathematical attribution of signals to sources, because it can compensate for nonlinear signals (i.e. tracer emissions that do not scale linearly with CO2ff emissions). It is determined that proper attribution is at least not feasible with such a complete lack of bottom-up spatial information about all non-CO2ff potential tracers. This unfeasibility is shown not to be resolved by a test of expanding the dataset with many more theoretical measurements than are realistically available. Here we alter the approach to include the missing prior information: bottom-up estimates of the emission fluxes for a suite of species. We develop these bottom-up emission fluxes from existing whole-city emission fluxes, species-specific source sector partitioning, and the spatial patterns from Hestia CO2ff source sectors. We validate the general approach using the whole-city species: CO2ff ratios derived from all tower flask measurements. Finally, using these tools, multi-species, sector-specific inversions are investigated in the Bayesian framework.

Atmospheric CO2 Records from Sites in the Atmospheric Environment Service Air Sampling Network (1975 and 1994)

DOE Data Explorer

Trivett, N. B.A. [Atmospheric Environment Service, Downsview, Ontario, Canada; Hudec, V. C. [Atmospheric Environment Service, Downsview, Ontario, Canada; Wong, C. S. [Marine Carbon Research Centre, Institute of Ocean Sciences, Sidney, British Columbia, Canada

1997-01-01

From the mid-1970s through the mid-1990s, air samples were collected for the purposes of monitoring atmospheric CO2 from four sites in the AES air sampling network. Air samples were collected approximately once per week, between 12:00 and 16:00 local time, in a pair of evacuated 2-L thick-wall borosilicate glass flasks. Samples were collected under preferred conditions of wind speed and direction (i.e., upwind of the main station and when winds are strong and steady). The flasks were evacuated to pressures of ~1 × 10-4 mbar or 0.01 Pa prior to being sent to the stations. The airwas not dried during sample collection. The flask data from Alert show an increase in the annual atmospheric CO2 concentration from 341.35 parts per million by volume (ppmv) in 1981 to 357.21 ppmv in 1991. For Cape St. James, Trivett and Higuchi (1989) reported that the mean annual rate of increase, obtained from the slope of a least-squares regression line through the annual averages, was 1.43 ppmv per year. In August 1992, the weather station at Cape St. James was automated; as a result, the flask sampling program was discontinued at this site. Estevan Point, on the West Coast of Vancouver Island, was chosen as a replacement station. Sampling at Estevan Point started in 1992; thus, the monthly and annual CO2record from Estevan Point is too short to show any long-term trends. The sampling site at Sable Island, off the coast of Nova Scotia, was established in 1975. The flask data from Sable Island show an increase in the annual atmospheric CO2 concentration from 334.49 parts per million by volume (ppmv) in 1977 (the first full year of data) to 356.02 ppmv in 1990. For Sable Island, Trivett and Higuchi (1989) reported that the mean annual rate of increase, obtained from the slope of a least-squares regression line through the annual averages, was 1.48 ppmv per year.

Pericellular oxygen monitoring with integrated sensor chips for reproducible cell culture experiments.

PubMed

Kieninger, J; Aravindalochanan, K; Sandvik, J A; Pettersen, E O; Urban, G A

2014-04-01

Here we present an application, in two tumour cell lines, based on the Sensing Cell Culture Flask system as a cell culture monitoring tool for pericellular oxygen sensing. T-47D (human breast cancer) and T98G (human brain cancer) cells were cultured either in atmospheric air or in a glove-box set at 4% oxygen, in both cases with 5% CO2 in the gas phase. Pericellular oxygen tension was measured with the help of an integrated sensor chip comprising oxygen sensor arrays. Obtained results illustrate variation of pericellular oxygen tension in attached cells covered by stagnant medium. Independent of incubation conditions, low pericellular oxygen concentration levels, usually associated with hypoxia, were found in dense cell cultures. Respiration alone brought pericellular oxygen concentration down to levels which could activate hypoxia-sensing regulatory processes in cultures believed to be aerobic. Cells in culture believed to experience conditions of mild hypoxia may, in reality, experience severe hypoxia. This would lead to incorrect assumptions and suggests that pericellular oxygen concentration readings are of great importance to obtain reproducible results when dealing with hypoxic and normoxic (aerobic) incubation conditions. The Sensing Cell Culture Flask system allows continuous monitoring of pericellular oxygen concentration with outstanding long-term stability and no need for recalibration during cell culture experiments. The sensor is integrated into the flask bottom, thus in direct contact with attached cells. No additional equipment needs to be inserted into the flask during culturing. Transparency of the electrochemical sensor chip allows optical inspection of cells attached on top of the sensor. © 2014 John Wiley & Sons Ltd.

Postural control under clinorotation in upside-down catfish, Synodontis nigriventris.

PubMed

Ohnishi, K; Takahashi, A; Koyama, M; Ohnishi, T

1996-12-01

The upside-down catfish Synodontis nigriventris has a unique habit of swimming and resting upside-down in free water. This behavior leads to the assumption that the catfish has a specific gravity information processing system. We examined the postural control behaviors in the catfish under clinorotation which is usually used for producing pseudo-microgravity. Synodontis nigriventris kept its body posture at a stable area of the rotated flask in which the catfish was kept, when it was clinorotated at the rate of 60 rpm. In contrast to Synodontis nigriventris, a related species, Corydoras paleatus, did not show such steady postural control. When the flask was rotated at a lower rate of 30 rpm or a higher rate of 100 rpm, Synodontis nigriventris as well as Corydoras paleatus showed a considerable disturbed control of body posture. In this condition, they were frequently rotated with the flask. These findings suggest that Synodontis nigriventris has a high ability to keep upside-down posture and the gravity sensation in this catfish is likely to contribute to its different postural control from that of many other fishes.

Lithium Battery Safety/Cell-to-Cell Failure Project FY14 Progress Report

DTIC Science & Technology

2015-03-06

Delacourt, “Thermal modeling of a cylindrical LiFePO4 /graphite lithium-ion battery.” J. Power Sources 195 (2010) 2961- 2968. 18. T. B. Bandhauer, S...Ren, J. Xie, H. He and F. Xu, “Failure study of commercial LiFePO4 cells in over-discharge conditions using electrochemical impedance spectroscopy...graphite/ LiFePO4 cell.” J. Power Sources 208 (2012) 296-305. 61. N. S. Spinner, C. T. Love, S. G. Tuttle, K. Swider-Lyons and S. L. Rose-Pehrsson

Active colloidal molecules

NASA Astrophysics Data System (ADS)

Löwen, Hartmut

2018-03-01

Like ordinary molecules are composed of atoms, colloidal molecules consist of several species of colloidal particles tightly bound together. If one of these components is self-propelled or swimming, novel “active colloidal molecules” emerge. Active colloidal molecules exist on various levels such as “homonuclear”, “heteronuclear” and “polymeric” and possess a dynamical function moving as propellers, spinners or rotors. Self-assembly of such active complexes has been studied a lot recently and this perspective article summarizes recent progress and gives an outlook to future developments in the rapidly expanding field of active colloidal molecules.

Molecular Engineering of Liquid Crystalline Polymers by Living Polymerization. 10. Influence of Molecular Weight on the Phase Transitions of Poly(Omega-((4-Cyano-4’-Biphenyl)oxy)alkyl Vinyl Ethers)s with Nonyl and Decanyl Alkyl Groups

DTIC Science & Technology

1990-10-16

washed with concentrated sulfuric acid , then with water, dried over anhydrous magnesium sulfate, refluxed over calcium hydride and freshly distilled...oxide, filtered, and fractionally distilled under reduced pressure. Trifluoromethane sulfonic acid (triflic acid , 98%, Aldrich) w s distilled under...flask. Then the flask was filled with argon, cooled to 0°C and the methylene chloride, dimethyl sulfide and triflic acid were added via a syringe. The

Effect of a new tension system, used in acrylic resin flasking, on the dimensional stability of denture bases.

PubMed

Consani, Rafael Leonardo Xediek; Domitti, Saide Sarckis; Consani, Simonides

2002-09-01

The pressure of final closure may be released when the flask is removed from the mechanical or pneumatic press and placed in the spring clamp. This release in pressure may result in dimensional changes that distort the denture base. The purpose of this study was to investigate differences between the dimensional stability of standardized simulated denture bases processed by traditional moist heat-polymerization and those processed by use of a new tension system. A metal master die was fabricated to simulate an edentulous maxillary arch without irregularities in the alveolar ridge walls. A silicone mold of this metallic die was prepared, and 40 stone casts were formed from the mold with type III dental stone. The casts were randomly assigned to 4 test groups (A-D) of 10 specimens each. A uniform denture base pattern was made on each stone cast with a 1.5-mm thickness of base-plate wax, measured with a caliper. The patterns were invested for traditional hot water processing. A polymethyl methacrylate dough was prepared and packed for processing. The flasks in groups A and B were closed with the traditional pressure technique and placed in spring clamps after final closure. The flasks in groups C and D were pressed between the metallic plates of the new tension system after the final closure. The group A and C flasks were immediately immersed in the water processing unit at room temperature (25 degrees +/- 2 degrees C). The unit was programmed to raise the temperature to 74 degrees C over 1 hour, and then maintained the temperature at 74 degrees C for 8 hours. The group B and D flasks were bench stored at room temperature (25 degrees +/- 2 degrees C) for 6 hours and were then subjected to the same moist heat polymerization conditions as groups A and C. All processed dentures were bench cooled for 3 hours. After recovery from the flasks, the base-cast sets were transversally sectioned into 3 parts (corresponding to 3 zones): (1) distal of the canines, (2) mesial of the first molars, and (3) mesial of the posterior palate). These areas had been previously established and standardized by use of a pattern denture in the sawing device to determine the sections in each base-cast set. Base-cast gaps were measured at 5 predetermined points on each section with an optical micrometer that had a tolerance of 0.001 mm. Collected data were analyzed with analysis of variance and Tukey's test. Denture bases processed with the new tension system exhibited significantly better base adaptation than those processed with traditional acrylic resin packing. Immediately after polymerization (Groups A and C), mean dimensional change values were 0.213 +/- 0.055 mm for the traditional packing technique and 0.173 +/- 0.050 mm for new tension system. After delayed polymerization (Groups B and D), the values were 0.216 +/- 0.074 mm for the traditional packing technique and 0.164 +/- 0.032 mm for new tension system. With both techniques, dimensional changes in the posterior palatal zone were greater (conventional = 0.286 +/- 0.038 mm; new system = 0.214 +/- 0.024 mm) than those elsewhere on the base-cast set. Within the limitations of this study, the new tension packing system was associated with decreased dimensional changes in the simulated maxillary denture bases processed with heat-polymerization.

Influence of Light Intensity on Growth and Pigment Production by Monascus ruber in Submerged Fermentation.

PubMed

Bühler, Rose Marie Meinicke; Müller, Bruna Luíse; Moritz, Denise Esteves; Vendruscolo, Francielo; de Oliveira, Debora; Ninow, Jorge Luiz

2015-07-01

To reduce environmental problems caused by glycerine accumulation and to make the production of biodiesel more profitable, crude glycerin without treatment was used as substrate for obtaining higher value-added bioproducts. Monascus ruber is a filamentous fungus that produces pigments, particularly red ones, which are used for coloring foods (rice wine and meat products). The interest in developing pigments from natural sources is increasing due to the restriction of using synthetic dyes. The effects of temperature, pH, microorganism morphology, aeration, nitrogen source, and substrates have been studied in the cultivation of M. ruber. In this work, it was observed that light intensity is also an important factor that should be considered for understanding the metabolism of the fungus. In M. ruber cultivation, inhibition of growth and pigment production was observed in Petri dishes and blaffed flasks exposed to direct illumination. Growth and pigment production were higher in Petri dishes and flasks exposed to red light and in the absence of light. Radial growth rate of M. ruber in plates in darkness was 1.50 mm day(-1) and in plates exposed to direct illumination was 0.59 mm day(-1). Maximum production of red pigments (8.32 UA) and biomass (8.82 g L(-1)) were obtained in baffled flasks covered with red film and 7.17 UA of red pigments, and 7.40 g L(-1) of biomass was obtained in flasks incubated in darkness. Under conditions of 1248 lux of luminance, the maximum pigment production was 4.48 UA, with production of 6.94 g L(-1) of biomass, indicating that the fungus has photoreceptors which influence the physiological responses.

Evaluating maturation and genetic modification of human dendritic cells in a new polyolefin cell culture bag system.

PubMed

Macke, Lars; Garritsen, Henk S P; Meyring, Wilhelm; Hannig, Horst; Pägelow, Ute; Wörmann, Bernhard; Piechaczek, Christoph; Geffers, Robert; Rohde, Manfred; Lindenmaier, Werner; Dittmar, Kurt E J

2010-04-01

Dendritic cells (DCs) are applied worldwide in several clinical studies of immune therapy of malignancies, autoimmune diseases, and transplantations. Most legislative bodies are demanding high standards for cultivation and transduction of cells. Closed-cell cultivating systems like cell culture bags would simplify and greatly improve the ability to reach these cultivation standards. We investigated if a new polyolefin cell culture bag enables maturation and adenoviral modification of human DCs in a closed system and compare the results with standard polystyrene flasks. Mononuclear cells were isolated from HLA-A*0201-positive blood donors by leukapheresis. A commercially available separation system (CliniMACS, Miltenyi Biotec) was used to isolate monocytes by positive selection using CD14-specific immunomagnetic beads. The essentially homogenous starting cell population was cultivated in the presence of granulocyte-macrophage-colony-stimulating factor and interleukin-4 in a closed-bag system in parallel to the standard flask cultivation system. Genetic modification was performed on Day 4. After induction of maturation on Day 5, mature DCs could be harvested and cryopreserved on Day 7. During the cultivation period comparative quality control was performed using flow cytometry, gene expression profiling, and functional assays. Both flasks and bags generated mature genetically modified DCs in similar yields. Surface membrane markers, expression profiles, and functional testing results were comparable. The use of a closed-bag system facilitated clinical applicability of genetically modified DCs. The polyolefin bag-based culture system yields DCs qualitatively and quantitatively comparable to the standard flask preparation. All steps including cryopreservation can be performed in a closed system facilitating standardized, safe, and reproducible preparation of therapeutic cells.

Process optimization involving critical evaluation of oxygen transfer, oxygen uptake and nitrogen limitation for enhanced biomass and lipid production by oleaginous yeast for biofuel application.

PubMed

Chopra, Jayita; Sen, Ramkrishna

2018-04-20

Lipid accumulation in oleaginous yeast is generally induced by nitrogen starvation, while oxygen saturation can influence biomass growth. Systematic shake flask studies that help in identifying the right nitrogen source and relate its uptake kinetics to lipid biosynthesis under varying oxygen saturation conditions are very essential for addressing the bioprocessing-related issues, which are envisaged to occur in the fermenter scale production. In the present study, lipid bioaccumulation by P. guilliermondii at varying C:N ratios and oxygen transfer conditions (assessed in terms of k L a) was investigated in shake flasks using a pre-optimized N-source and a two-stage inoculum formulated in a hybrid medium. A maximum lipid concentration of 10.8 ± 0.5 g L -1 was obtained in shake flask study at the optimal condition with an initial C:N and k L a of 60:1 and 0.6 min -1 , respectively, at a biomass specific growth rate of 0.11 h -1 . Translating these optimal shake flask conditions to a 3.7 L stirred tank reactor resulted in biomass and lipid concentrations of 16.74 ± 0.8 and 8 ± 0.4 g L -1 . The fatty acid methyl ester (FAME) profile of lipids obtained by gas chromatography was found to be suitable for biodiesel application. We strongly believe that the rationalistic approach-based design of experiments adopted in the study would help in achieving high cell density with improved lipid accumulation and also minimize the efforts towards process optimization during bioreactor level operations, consequently reducing the research and development-associated costs.

Hydrogen isotope correction for laser instrument measurement bias at low water vapor concentration using conventional isotope analyses: application to measurements from Mauna Loa Observatory, Hawaii.

PubMed

Johnson, L R; Sharp, Z D; Galewsky, J; Strong, M; Van Pelt, A D; Dong, F; Noone, D

2011-03-15

The hydrogen and oxygen isotope ratios of water vapor can be measured with commercially available laser spectroscopy analyzers in real time. Operation of the laser systems in relatively dry air is difficult because measurements are non-linear as a function of humidity at low water concentrations. Here we use field-based sampling coupled with traditional mass spectrometry techniques for assessing linearity and calibrating laser spectroscopy systems at low water vapor concentrations. Air samples are collected in an evacuated 2 L glass flask and the water is separated from the non-condensable gases cryogenically. Approximately 2 µL of water are reduced to H(2) gas and measured on an isotope ratio mass spectrometer. In a field experiment at the Mauna Loa Observatory (MLO), we ran Picarro and Los Gatos Research (LGR) laser analyzers for a period of 25 days in addition to periodic sample collection in evacuated flasks. When the two laser systems are corrected to the flask data, they are strongly coincident over the entire 25 days. The δ(2)H values were found to change by over 200‰ over 2.5 min as the boundary layer elevation changed relative to MLO. The δ(2)H values ranged from -106 to -332‰, and the δ(18)O values (uncorrected) ranged from -12 to -50‰. Raw data from laser analyzers in environments with low water vapor concentrations can be normalized to the international V-SMOW scale by calibration to the flask data measured conventionally. Bias correction is especially critical for the accurate determination of deuterium excess in dry air. Copyright © 2011 John Wiley & Sons, Ltd.

Establishing Regular Measurements of Halocarbons at Taunus Observatory

NASA Astrophysics Data System (ADS)

Schuck, Tanja; Lefrancois, Fides; Gallmann, Franziska; Engel, Andreas

2017-04-01

In late 2013 an ongoing whole air flask collection program has been started at the Taunus Observatory (TO) in central Germany. Being a rural site in close vicinity to the densely populated Rhein-Main area with the city of Frankfurt, Taunus Observatory allows to assess local and regional emissions but owed to its altitude of 825m also regularly experiences background conditions. With its large caption area halocarbon measurements at the site have the potential to improve the data base for estimation of regional and total European halogenated greenhouse gas emissions. At current, flask samples are collected weekly for analysis using a GC-MS system at Frankfurt University employing a quadrupole as well as a time-of-flight (TOF) mass spectrometer. The TOF instrument yields full scan mass information and allows for retrospective analysis of so far undetected non-target species. For quality assurance additional samples are collected approximately bi-weekly at the Mace Head Atmospheric Research Station (MHD) analyzed in Frankfurt following the same measurement procedure. Thus the TO time series can be linked to both, the in-situ AGAGE measurements and the NOAA flask sampling program at MHD. In 2017 it is planned to supplement the current flask sampling by employing an in-situ GC-MS system at the site, thus increasing the measurement frequency. We will present the timeseries of selected halocarbons recorded at Taunus Observatory. While there is good agreement of baseline mixing ratios between TO and MHD, measurements at TO are regularly influenced by elevated halocarbon mixing ratios. An analysis of HYSPLIT trajectories for the existing time series revealed significant differences in halocarbon mixing ranges depending on air mass origin.

Three-dimensional HepaRG model as an attractive tool for toxicity testing.

PubMed

Leite, Sofia B; Wilk-Zasadna, Iwona; Zaldivar, Jose M; Airola, Elodie; Reis-Fernandes, Marcos A; Mennecozzi, Milena; Guguen-Guillouzo, Christiane; Chesne, Christopher; Guillou, Claude; Alves, Paula M; Coecke, Sandra

2012-11-01

The culture of HepaRG cells as three dimensional (3D) structures in the spinner-bioreactor may represent added value as a hepatic system for toxicological purposes. The use of a cost-effective commercially available bioreactor, which is compatible with high-throughput cell analysis, constitutes an attractive approach for routine use in the drug testing industry. In order to assess specific aspects of the biotransformation capacity of the bioreactor-based HepaRG system, the induction of CYP450 enzymes (i.e., CYP1A2, 2B6, 2C9, and 3A4) and the activity of the phase II enzyme, uridine diphosphate glucuronoltransferase (UGT), were tested. The long-term functionality of the system was demonstrated by 7-week stable profiles of albumin secretion, CYP3A4 induction, and UGT activities. Immunofluorescence-based staining showed formation of tissue-like arrangements including bile canaliculi-like structures and polar distribution of transporters. The use of in silico models to analyze the in vitro data related to hepatotoxic activity of acetaminophen (APAP) demonstrated the advantage of the integration of kinetic and dynamic aspects for a better understanding of the in vitro cell behavior. The bioactivation of APAP and its related cytotoxicity was assessed in a system compatible to high-throughput screening. The approach also proved to be a good strategy to reduce the time necessary to obtain fully differentiated cell cultures. In conclusion, HepaRG cells cultured in 3D spinner-bioreactors are an attractive tool for toxicological studies, showing a liver-like performance and demonstrating a practical applicability for toxicodynamic approaches.

Effect of light with different wavelengths on Nostoc flagelliforme cells in liquid culture.

PubMed

Dai, Yu-Jie; Li, Jing; Wei, Shu-Mei; Chen, Nan; Xiao, Yu-Peng; Tan, Zhi-Lei; Jia, Shi-Ru; Yuan, Nan-Nan; Tan, Ning; Song, Yi-Jie

2013-04-01

The effects of lights with different wavelengths on the growth and the yield of extracellular polysaccharides of Nostoc flagelliforme cells were investigated in a liquid cultivation. N. flagelliforme cells were cultured for 16 days in 500 ml conical flasks containing BG11 culture medium under 27 micromol·m-2·s-1 of light intensity and 25 degrees C on a rotary shaker (140 rpm). The chlorophyll a, phycocyanin, allophycocyanin, and phycoerythrin contents in N. flagelliforme cells under the lights of different wavelengths were also measured. It was found that the cell biomass and the yield of polysaccharide changed with different wavelengths of light. The biomass and the yield of extracellular polysaccharides under the red or violet light were higher than those under other light colors. Chlorophyll a, phycocyanin, and allophycocyanin are the main pigments in N. flagelliforme cells. The results showed that N. flagelliforme, like other cyanobacteria, has the ability of adjusting the contents and relative ratio of its pigments with the light quality. As a conclusion, N. flagelliforme cells favor red and violet lights and perform the complementary chromatic adaptation ability to acclimate to the changes of the light quality in the environment.

Ethanol production from agricultural wastes using Saccharomyces cerevisiae.

PubMed

Irfan, Muhammad; Nadeem, Muhammad; Syed, Quratualain

2014-01-01

The main objective of this study was production of ethanol from three lignocellulosic biomasses like sugarcane bagasse, rice straw and wheat straw by Sacchromyces cervisae. All the three substrates were ground to powder form (2 mm) and pretreated with 3%H2O2 + 2% NaOH followed by steaming at 130 °C for 60 min. These substrates were hydrolyzed by commercial cellulase enzyme. The whole fermentation process was carried out in 500 mL Erlenmeyer flask under anaerobic conditions in submerged fermentation at 30 °C for three days of incubation period. FTIR analysis of the substrates indicated significant changes in the alteration of the structure occurred after pretreatment which leads to efficient saccharification. After pretreatment the substrates were hydrolyzed by commercial cellulase enzyme and maximum hydrolysis was observed in sugarcane bagasse (64%) followed by rice straw (40%) and wheat straw (34%). Among all these tested substrates, sugarcane bagasse (77 g/L) produced more ethanol as compared to rice straw (62 g/L) and wheat straw (44 g/L) using medium composition of (%) 0.25 (NH4)2SO4, 0.1 KH2PO4, 0.05 MgSO4, 0.25 Yeast extract by S. cervisae.

Ethanol production from agricultural wastes using Sacchromyces cervisae

PubMed Central

Irfan, Muhammad; Nadeem, Muhammad; Syed, Quratualain

2014-01-01

The main objective of this study was production of ethanol from three lignocellulosic biomasses like sugarcane bagasse, rice straw and wheat straw by Sacchromyces cervisae. All the three substrates were ground to powder form (2 mm) and pretreated with 3%H2O2 + 2% NaOH followed by steaming at 130 °C for 60 min. These substrates were hydrolyzed by commercial cellulase enzyme. The whole fermentation process was carried out in 500 mL Erlenmeyer flask under anaerobic conditions in submerged fermentation at 30 °C for three days of incubation period. FTIR analysis of the substrates indicated significant changes in the alteration of the structure occurred after pretreatment which leads to efficient saccharification. After pretreatment the substrates were hydrolyzed by commercial cellulase enzyme and maximum hydrolysis was observed in sugarcane bagasse (64%) followed by rice straw (40%) and wheat straw (34%). Among all these tested substrates, sugarcane bagasse (77 g/L) produced more ethanol as compared to rice straw (62 g/L) and wheat straw (44 g/L) using medium composition of (%) 0.25 (NH4)2SO4, 0.1 KH2PO4, 0.05 MgSO4, 0.25 Yeast extract by S. cervisae. PMID:25242928

Comparison of different rubber stoppers; the effect of sterilization on the number of particles released.

PubMed

Mannermaa, J P; Muttonen, E; Yliruusi, J; Juppo, A

1992-01-01

The effect of sterilization on the number of particles released from five different types of rubber stoppers, as well as on their surface roughness and elemental composition before and after sterilization is described. The stoppers were immersed in 200 ml of 0.9% sodium chloride solution in conical flasks. The number of particles released into the sodium chloride solution was measured by Coulter Counter. The surface roughness and the elemental composition of the stoppers were determined by SEM/EDX. All measurements were made both before and after sterilization at 121 degrees C to F0 15 mins. The number of particles released from a stopper during sterilization varies considerably between different stoppers and even between different batches of the same stopper. The only non-siliconized stopper in this study performed well. The absence of surface siliconization may have contributed to this performance. The scanning electron micrographs revealed well the differences in the surface roughness of the stoppers. The sterilization generally increases the surface roughness of the samples. The x-ray microanalysis revealed that the elemental composition of the stoppers may vary not only between different types of stoppers but also between different batches of the same stopper.

Response of sago pondweed, a submerged aquatic macrophyte, to herbicides in three laboratory culture systems

USGS Publications Warehouse

Fleming, W.J.; Ailstock, M.S.; Momot, J.J.; Norman, C.M.; Gorsuch, Joseph W.; Lower, William R.; Wang, Wun-cheng; Lewis, M.A.

1991-01-01

The phytotoxicity of atrazine, paraquat, glyphosate, and alachlor to sago pondweed (Potamogeton pectinatus), a submerged aquatic macrophyte, was tested under three types of laboratory culture conditions. In each case, tests were conducted in static systems, the test period was four weeks, and herbicide exposure was chronic, resulting from a single addition of herbicide to the test vessels at the beginning of the test period. The three sets of test conditions employed were(1) axenic cultures in 125-mL flasks containing a nutrient media and sucrose; (2) a microcosm system employing 18.9-L buckets containing a sand, shell, and peat substrate; and (3) an algae-free system employing O.95-L jars containing reconstituted freshwater and a nutrient agar substrate. The primary variable measured was biomass production. Plants grew well in all three test systems, with biomass of untreated plants increasing by a factor of about 5 to 6.5 during the four-week test period. Biomass production in response to herbicide exposure differed significantly among culture systems, which demonstrates the need for a standardized testing protocol for evaluating the effects of toxics on submerged aquatic plants.

Semi-continuous production of high-activity pectinases by immobilized Rhizopus oryzae using tobacco wastewater as substrate and their utilization in the hydrolysis of pectin-containing lignocellulosic biomass at high solid content.

PubMed

Zheng, Yu-Xi; Wang, Yuan-Liang; Pan, Jun; Zhang, Jian-Rong; Dai, Ya; Chen, Kun-Yan

2017-10-01

In this study, highly reactive endo- and exo-polygalacturonases (PGs) were produced from the tobacco industry wastewater using immobilized Rhizopus oryzae. Compared with free cells, immobilized cells increased enzyme activity 2.8-fold and reduced production time to 24h by shake-flask production. Moreover, the immobilized cells enabled the semi-continuous production of enzymes through repeated-batch mode for seven consecutive cycles in a scale-up bioreactor. During the first five cycles, the average endo-PG and exo-PG activities reached 307.5 and 242.6U/ml, respectively. The addition of crude enzyme for the hydrolysis of pectin-containing lignocellulosic biomass under high-gravity conditions increased glucose release 4.2-fold (115.4 vs. 29.0g/L), compared with hydrolysis using cellulase alone. This process achieves the efficient production of pectin-degrading enzymes, provides a cost-effective method for tobacco wastewater treatment, and offers the possibility to obtain fermentable sugars with high-titer from pectin-containing lignocellulosic biomass, which has important potential for the commercial production of bio-fuels. Copyright © 2017 Elsevier Ltd. All rights reserved.

[Production of infectious units of Isaria fumosorosea (Hypocreales: Cordycipitaceae) from different indigenous isolates of northeastern Mexico using 3 propagation strategies].

PubMed

Gandarilla-Pacheco, Fatima L; Morales-Ramos, Lilia H; Pereyra-Alférez, Benito; Elías-Santos, Myriam; Quintero-Zapata, Isela

The aim of this study was to evaluate the production of blastospores and conidia of different native isolates and a strain of Isaria fumosorosea using different propagation techniques. Two liquid culture media of casamino acids and peptone as nitrogen sources and glucose as carbon source for both media cultures were respectively used in the production of blastospores, while for the production of conidia, the fungi were grown in potato dextrose agar; from these cultures, solutions of conidia to a concentration of 1×10 6 per milliliter were prepared to inoculate flasks with Sabouraud dextrose broth for the liquid phase of the biphasic culture, also known as preculture. Subsequently, rice grain bags were inoculated with the preculture and the conidia solutions, which were incubated for 14 days for solid fermentation and biphasic culture, respectively. The HIB-23 isolate recorded a concentration of 4.90×10 8 blastospores/ml in the casamino acid medium, while a concentration of 2.15×10 8 blastospores/ml was obtained in the peptone collagen medium. For the Pfr-612 strain, the conidia production in solid-state fermentation was 1.58×10 9 conidia/g, and for HIB-30 in the biphasic culture of 9.00×10 6 conidia/g. Solid-state fermentation proved to be the most effective method with an average of 1.09×10 9 conidia/g, whereas the biphasic culture was the least effective method with 2.76×10 6 conidia/g; no significant difference was reported for the submerged production media. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Short communication: Eicosatrienoic acid and docosatrienoic acid do not promote vaccenic acid accumulation in mixed ruminal cultures.

PubMed

AbuGhazaleh, A A; Holmes, L D; Jacobson, B N; Kalscheur, K F

2006-11-01

Previous research found that docosahexaenoic acid (C22:6n-3) was a component of fish oil that promotes trans-C18:1 accumulation in ruminal cultures when incubated with linoleic acid. The objective of this study was to determine if eicosatrienoic acid (C20:3n-3) and docosatrienoic acid (C22:3n-3), n-3 fatty acids in fish oil, promote accumulation of trans-C18:1, vaccenic acid (VA) in particular, using cultures of mixed ruminal microorganisms. Treatments consisted of control, control plus 5 mg of C20:3n-3 (ETA), control plus 5 mg of C22:3n-3 (DTA), control plus 15 mg of linoleic acid (LA), control plus 5 mg of C20:3n-3 and 15 mg of linoleic acid (ETALA), and control plus 5 mg of C22:3n-3 and 15 mg of linoleic acid (DTALA). Treatments were incubated in triplicate in 125-mL flasks, and 5 mL of culture contents was taken at 0 and 24 h for fatty acid analysis by gas-liquid chromatography. After 24 h of incubation, the concentrations of trans-C18:1 (0.87, 0.88, and 0.99 mg/culture), and VA (0.52, 0.56, and 0.62 mg/culture) were similar for the control, ETA, and DTA cultures, respectively. The concentrations of trans-C18:1 (5.51, 5.41, and 5.36 mg/culture), and VA (4.78, 4.62, and 4.59 mg/culture) were also similar between LA, ETALA, and DTALA cultures, respectively. These data suggest that C20:3n-3 and C22:3n-3 are not the active components in fish oil that promote VA accumulation when incubated with linoleic acid.

[Optimization of fermentation conditions for cold-adapted amylase production by Micrococcus antarcticus and its enzymatic properties].

PubMed

Fan, Hong-xi; Liu, Ying; Liu, Zhi-pei

2009-08-15

By single factor experiments, the fermentation conditions for cold-adapted amylase production from Micrococcus antarcticus were determined as follows(medium g/L): Na2 HPO4 2.0, KH2PO4 1.0, MgSO4 x 7H2O 0.1, NaCl 5.0, (NH4)2SO4 2.5, maltose 5.0, trace element solution 5.0 mL, pH 8.0, 100 mL/Erlenmeyer flask (500 mL); cultivation was in a rotating shaker at 12 degrees C and 160 r/min for 64 h.Under those conditions,the highest total enzyme activity (2.6 U/mL) was obtained and increased by 10.8 fold compared with the original value of 0.24 U/mL before optimization. This amylase was purified by concentration with ultrafiltration membrane module, Hitrap Q anion exchange chromatography and Superdex 200 gel filtration chromatography. The optimal temperature and pH for the purified amylase were 30 degrees C and 6.0, respectively.It still showed high activity at low temperature 10-15 degrees C. It was sensitive to high temperature but was stable at pH 6.0-10.0 with at least 70% activity remained. These results indicated that it was a typical cold-adapted enzyme. The enzyme activity was stimulated by Ca2+, Mn2+, Co2+ and Mg2+; but inhibited by Zn2+, Ba2+, Ag+, Cu2+, Al3+, Fe2, Fe3+, Hg2+, EDTA and citrate. This cold-adapted amylase showed resistance to inactivation of 0.1% nonionic surfactants such as Tween 80, TrintonX-100, etc. Its Km was 0.90 mg/mL.

Efficacy of chlorine, acidic electrolyzed water and aqueous chlorine dioxide solutions to decontaminate Escherichia coli O157:H7 from lettuce leaves.

PubMed

Keskinen, Lindsey A; Burke, Angela; Annous, Bassam A

2009-06-30

This study compared the efficacy of chlorine (20-200 ppm), acidic electrolyzed water (50 ppm chlorine, pH 2.6), acidified sodium chlorite (20-200 ppm chlorite ion concentration, Sanova), and aqueous chlorine dioxide (20-200 ppm chlorite ion concentration, TriNova) washes in reducing populations of Escherichia coli O157:H7 on artificially inoculated lettuce. Fresh-cut leaves of Romaine or Iceberg lettuce were inoculated by immersion in water containing E. coli O157:H7 (8 log CFU/ml) for 5 min and dried in a salad spinner. Leaves (25 g) were then washed for 2 min, immediately or following 24 h of storage at 4 degrees C. The washing treatments containing chlorite ion concentrations of 100 and 200 ppm were the most effective against E. coli O157:H7 populations on Iceberg lettuce, with log reductions as high as 1.25 log CFU/g and 1.05 log CFU/g for TriNova and Sanova wash treatments, respectively. All other wash treatments resulted in population reductions of less than 1 log CFU/g. Chlorine (200 ppm), TriNova, Sanova, and acidic electrolyzed water were all equally effective against E. coli O157:H7 on Romaine, with log reductions of approximately 1 log CFU/g. The 20 ppm chlorine wash was as effective as the deionized water wash in reducing populations of E. coli O157:H7 on Romaine and Iceberg lettuce. Scanning electron microscopy indicated that E. coli O157:H7 that was incorporated into biofilms or located in damage lettuce tissue remained on the lettuce leaf, while individual cells on undamaged leaf surfaces were more likely to be washed away.

ARM Carbon Cycle Gases Flasks at SGP Site

DOE Data Explorer

Biraud, Sebastien

2013-03-26

Data from flasks are sampled at the Atmospheric Radiation Measurement Program ARM, Southern Great Plains Site and analyzed by the National Oceanic and Atmospheric Administration NOAA, Earth System Research Laboratory ESRL. The SGP site is included in the NOAA Cooperative Global Air Sampling Network. The surface samples are collected from a 60 m tower at the ARM SGP Central Facility, usually once per week in the afternoon. The aircraft samples are collected approximately weekly from a chartered aircraft, and the collection flight path is centered over the tower where the surface samples are collected. The samples are collected by the ARM and LBNL Carbon Project.

Measurements of Atmospheric Methane and 13C/12C of Atmospheric Methane from Flask Air Samples (1999)

DOE Data Explorer

Quay, Paul [School of Oceanography, University of Washington; Stutsman, Johnny [School of Oceanography, University of Washington

1999-01-01

This database offers precise measurements of atmospheric methane and 13C/12C in atmospheric methane from flask air samples collected at eight sites worldwide and aboard NOAA cruises in the Pacific Ocean. The eight sites include Olympic Peninsula, Washington; Cape Grim, Tasmania; Fraserdale, Ontario; Marshall Islands; Baring Head, New Zealand; Mauna Loa, Hawaii; Point Barrow, Alaska; and American Samoa. The measurements span the period 1988 to mid-1996. These data are useful for global methane budget analyses and for determining the atmospheric isotopic composition of methane. All isotopic measurements have been corrected for standard drift.

Spectral lights trigger biomass accumulation and production of antioxidant secondary metabolites in adventitious root cultures of Stevia rebaudiana (Bert.).

PubMed

Idrees, Muhammad; Sania, Bibi; Hafsa, Bibi; Kumari, Sana; Khan, Haji; Fazal, Hina; Ahmad, Ishfaq; Akbar, Fazal; Ahmad, Naveed; Ali, Sadeeq; Ahmad, Nisar

2018-05-30

Stevia rebaudiana (S. rebaudiana) is the most important therapeutic plant species and has been accepted as such worldwide. It has a tendency to accumulate steviol glycosides, which are 300 times sweeter than marketable sugar. Recently, diabetic patients commonly use this plant as a sugar substitute for sweet taste. In the present study, the effects of different spectral lights were investigated on biomass accumulation and production of secondary metabolites in adventitious root cultures of S. rebaudiana. For callus development, leaf explants were excised from seed-derived plantlets and inoculated on a Murashige and Skoog (MS) medium containing the combination of 2,4-dichlorophenoxy acetic acid (2, 4-D, 2.0mg/l) and 6-benzyladenine (BA, 2.0mg/l), while 0.5mg/l naphthalene acetic acid (NAA) was used for adventitious root culture. Adventitious root cultures were exposed to different spectral lights (blue, green, violet, red and yellow) for a 30-day period. White light was used as control. The growth kinetics was studied for 30days with 3-day intervals. In this study, the violet light showed the maximum accumulation of fresh biomass (2.495g/flask) as compared to control (1.63g/flask), while red light showed growth inhibition (1.025g/flask) as compared to control. The blue light enhanced the highest accumulation of phenolic content (TPC; 6.56mg GAE/g DW), total phenolic production (TPP; 101mg/flask) as compared to control (5.44mg GAE/g DW; 82.2mg GAE/g DW), and exhibited a strong correlation with dry biomass. Blue light also improved the accumulation of total flavonoid content (TFC; 4.33mg RE/g DW) and total flavonoid production (TFP; 65mg/flask) as compared to control. The violet light showed the highest DPPH inhibition (79.72%), while the lowest antioxidant activity was observed for control roots (73.81%). Hence, we concluded that the application of spectral lights is an auspicious strategy for the enhancement of the required antioxidant secondary metabolites in adventitious root cultures of S. rebaudiana and of other medicinal plants. Copyright © 2018 Académie des sciences. Published by Elsevier Masson SAS. All rights reserved.

Effect of temperature on growth and paralytic toxin profiles in isolates of Gymnodinium catenatum (Dinophyceae) from the Pacific coast of Mexico.

PubMed

Band-Schmidt, Christine J; Bustillos-Guzmán, José J; Hernández-Sandoval, Francisco E; Núñez-Vázquez, Erick J; López-Cortés, David J

2014-11-01

The effects of temperature on growth, cell toxicity, toxin content, and profile of paralytic shellfish toxins was determined in eight isolates of Gymnodinium catenatum from several localities along the Pacific Coast of Mexico. The isolates were cultivated in modified f/2 media with Se (10(-8) M), and a reduced concentration of Cu (10(-8) M), under a 12 h:12 h day-night cycle with an irradiance of 150 μE m(-2) s(-1). Isolates were progressively adapted for three generations to each of the temperatures (16, 19, 22, 24, 27, 30, and 33 °C). The cultures were grown in 125 mL Erlenmeyer flasks with 60 mL of media and harvested by filtration in late exponential growth. Toxins were analyzed by HPLC with a post-column oxidation and fluorescent detection (FLD). G. catenatum isolates tolerate temperatures between 16 and 33 °C, with maximum growth rates of 0.32 and 0.39 div day(-1) at 21 °C and 24 °C, respectively; maximum cell densities of 4700 and 5500 cells mL(-1) were obtained at 27 and 21 °C, respectively. No effect of toxicity per cell with temperature was observed, varying between 10.10 and 28.19 pgSXTeq cell(-1). Ten saxitoxin analogues were detected in all isolates, observing changes in the toxin profile with temperature. C1/2 toxins decreased from 80% mol at 16 °C to 20% mol at 33 °C, B1/2 toxins increased from 19% mol at 16 °C to 42% mol at 33 °C, and decarbamoyl toxins were more abundant at 21 °C. These results show that G. catenatum isolates from different regions of the Pacific coast of Mexico have a similar response to temperature and that this parameter can modify growth rate, cell density, and toxin profile of the species, particularly the decarbamoyl and sulfocarbamoyl toxins. Copyright © 2014 Elsevier Ltd. All rights reserved.

Orbital experiment ``Gravisensor'': phototropic reactions of the moss Physcomitrella patens to different types of LED lighting.

NASA Astrophysics Data System (ADS)

Nikitin, Vladimir; Berkovich, Yuliy A.; Skripnikov, Alexander; Zyablova, Natalya; Mukhoyan, Makar; Emelianov, Grigory

The experiment was conducted on Russian Biological Satelite Bion-M #1 19.04-19.05 2013. Five transparent plastic cultural flasks were placed in five light isolated sections of Biocont-B2 cylindrical container with inner diameter of 120 mm and height of 230 mm. In four sections the flasks could be illuminated by top or side LED with wavelength of 458 nm, 630 nm, 730 nm, and white (color temperature 5000° K, peaks 453, 559 nm). Photon flux in each variant was 15 umol/(m2c). In the fifth section the flask with the shoots was in conditions of constant dark. Each section was equipped with its own video camera module. Cameras, video recorder and lighting were managed by micro controller. 12 days before launch, 5 tips of the moss shoots were explanted at each of the five flasks on the agar medium with nutrient components and were cultivated under white fluorescent lamps at 12 hour photo period till the launch. After entering the orbit and during next 14 days of flight top LEDs were turned on above the flasks. Then for the following 14 days of flight the side LEDs of similar wavelength were turned on. The moss gametophores were cultivated at 12-h photoperiod. During the experiment on an hourly basis a video recording of the moss was performed. Similar equipment was used for ground control. After the experiment video files were used to produce separate time-lapse films for each flask using AviSynth program. In flight the shoots demonstrated the maximum growth speed with far red lighting and slower speed with white lighting. With blue and red lighting after switching to side light stimuli the growth of shoots almost stopped. In the dark the shoots continued to grow until the 13 day after launch of the satellite, then their growth stopped. In ground control the relation of growth rate with various LEDs remained basically the same, with the exception of side blue lighting, where the shoots demonstrated considerable vertical growth. In flight the angle of inclination towards the light source was maximal (about 90º) with white lighting, and somewhat smaller with 730 nm. Under red and blue light the angle of phototropic inclination was difficult to measure due to poor growth of the shoots.In ground control the growth rate under blue light was several times higher, than in flight and final degree of inclination of the shoot tip came to about 10º. In ground control under side red lighting the growth was weak, while demonstrating a pronounced phototropic bend of 90º. In ground control in the dark a vertical growth of one shoot was observed with the rate somewhat larger, than in flight variant. Data on the dynamics of inclination of experimental and control plants are presented. The acquired data will be used to analyse the mechanisms of phototropic growth changes of moss shoots.

Development of 0.5-5 W, 10K Reverse Brayton Cycle Cryocoolers - Phase II Final Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Doty, F. D.; Boman, A.; Arnold, S.

2001-10-15

Miniature cryocoolers for the 8-30 K range are needed to provide 0.5-5 w of cooling to high sensitivity detectors (for long-wave-length IR, magnetism, mm-wave, X-ray, dark matter, and possibly y-ray detection) while maintaining low mass, ultra-low vibration, and good efficiency. This project presents a new approach to eliminating the problems normally encountered in efforts to build low-vibration, fieldable, miniature cryocoolers. Using the reverse Brayton Cycle (RBC), the approach applies and expands on existing spinner technology previously used only in Nuclear Magnetic Resonance (NMR) probes.

Full-Scale Tests of NACA Cowlings

NASA Technical Reports Server (NTRS)

Theodorsen, Theodore; Brevoort, M J; Stickle, George W

1937-01-01

A comprehensive investigation has been carried on with full-scale models in the NACA 20-foot wind tunnel, the general purpose of which is to furnish information in regard to the physical functioning of the composite propeller-nacelle unit under all conditions of take-off, taxiing, and normal flight. This report deals exclusively with the cowling characteristics under condition of normal flight and includes the results of tests of numerous combinations of more than a dozen nose cowlings, about a dozen skirts, two propellers, two sizes of nacelle, as well as various types of spinners and other devices.

Engineering of a Stable Whole-Cell Biocatalyst Capable of (S)-Styrene Oxide Formation for Continuous Two-Liquid-Phase Applications

PubMed Central

Panke, Sven; de Lorenzo, Víctor; Kaiser, Arnë; Witholt, Bernard; Wubbolts, Marcel G.

1999-01-01

Recombinant strains of Pseudomonas putida KT2440 carrying genetic expression cassettes with xylene oxygenase- and styrene monooxygenase-encoding genes on their chromosomes could be induced in shaking-flask experiments to specific activities that rivaled those of multicopy-plasmid-based Escherichia coli recombinants. Such strains maintained the introduced styrene oxidation activity in continuous two-liquid-phase cultures for at least 100 generations, although at a lower level than in the shaking-flask experiments. The data suggest that placement of target genes on the chromosome might be a suitable route for the construction of segregationally stable and highly active whole-cell biocatalysts. PMID:10584030

Atmospheric Carbon Dioxide Mixing Ratios from the NOAA CMDL Carbon Cycle Cooperative Global Air Sampling Network (2009)

DOE Data Explorer

Conway, Thomas [NOAA Climate Monitoring and Diagnostics Laboratory, Boulder, CO (USA); Tans, Pieter [NOAA Climate Monitoring and Diagnostics Laboratory, Boulder, CO (USA)

2009-01-01

The National Oceanic and Atmospheric Administration's Climate Monitoring and Diagnostics Laboratory (NOAA/CMDL) has measured CO2 in air samples collected weekly at a global network of sites since the late 1960s. Atmospheric CO2 mixing ratios reported in these files were measured by a nondispersive infrared absorption technique in air samples collected in glass flasks. All CMDL flask samples are measured relative to standards traceable to the World Meteorological Organization (WMO) CO2 mole fraction scale. These measurements constitute the most geographically extensive, carefully calibrated, internally consistent atmospheric CO2 data set available and are essential for studies aimed at better understanding the global carbon cycle budget.

Feasibility of human hair follicle-derived mesenchymal stem cells/CultiSpher(®)-G constructs in regenerative medicine.

PubMed

Li, Pengdong; Liu, Feilin; Wu, Chunling; Jiang, Wenyue; Zhao, Guifang; Liu, Li; Bai, Tingting; Wang, Li; Jiang, Yixu; Guo, Lili; Qi, Xiaojuan; Kou, Junna; Fan, Ruirui; Hao, Deshun; Lan, Shaowei; Li, Yulin; Liu, Jin Yu

2015-10-01

The use of human mesenchymal stem cells (hMSCs) in cell therapies has increased the demand for strategies that allow efficient cell scale-up. Preliminary data on the three-dimensional (3D) spinner culture describing the potential use of microcarriers for hMSCs culture scale-up have been reported. We exploited a rich source of autologous stem cells (human hair follicle) and demonstrated the robust in vitro long-term expansion of human hair follicle-derived mesenchymal stem cells (hHF-MSCs) by using CultiSpher(®)-G microcarriers. We analyzed the feasibility of 3D culture by using hHF-MSCs/CultiSpher(®)-G microcarrier constructs for its potential applicability in regenerative medicine by comparatively analyzing the performance of hHF-MSCs adhered to the CultiSpher(®)-G microspheres in 3D spinner culture and those grown on the gelatin-coated plastic dishes (2D culture), using various assays. We showed that the hHF-MSCs seeded at various densities quickly adhered to and proliferated well on the microspheres, thus generating at least hundreds of millions of hHF-MSCs on 1 g of CultiSpher(®)-G within 12 days. This resulted in a cumulative cell expansion of greater than 26-fold. Notably, the maximum and average proliferation rates in 3D culture were significantly greater than that of the 2D culture. However, the hHF-MSCs from both the cultures retained surface marker and nestin expression, proliferation capacity and differentiation potentials toward adipocytes, osteoblasts and smooth muscle cells and showed no significant differences as evidenced by Edu incorporation, cell cycle, colony formation, apoptosis, biochemical quantification and qPCR assays.

Enhanced Biosynthesis of Withanolides by Elicitation and Precursor Feeding in Cell Suspension Culture of Withania somnifera (L.) Dunal in Shake-Flask Culture and Bioreactor

PubMed Central

Sivanandhan, Ganeshan; Selvaraj, Natesan; Ganapathi, Andy; Manickavasagam, Markandan

2014-01-01

The present study investigated the biosynthesis of major and minor withanolides of Withania somnifera in cell suspension culture using shake-flask culture and bioreactor by exploiting elicitation and precursor feeding strategies. Elicitors like cadmium chloride, aluminium chloride and chitosan, precursors such as cholesterol, mevalonic acid and squalene were examined. Maximum total withanolides detected [withanolide A (7606.75 mg), withanolide B (4826.05 mg), withaferin A (3732.81 mg), withanone (6538.65 mg), 12 deoxy withanstramonolide (3176.63 mg), withanoside IV (2623.21 mg) and withanoside V (2861.18 mg)] were achieved in the combined treatment of chitosan (100 mg/l) and squalene (6 mM) along with 1 mg/l picloram, 0.5 mg/l KN, 200 mg/l L-glutamine and 5% sucrose in culture at 4 h and 48 h exposure times respectively on 28th day of culture in bioreactor. We obtained higher concentrations of total withanolides in shake-flask culture (2.13-fold) as well as bioreactor (1.66-fold) when compared to control treatments. This optimized protocol can be utilized for commercial level production of withanolides from suspension culture using industrial bioreactors in a short culture period. PMID:25089711

Culture of prostate epithelial cells of the rhesus monkey on extracellular matrix substrate: influence of steroids and insulin-like growth factors.

PubMed

Udayakumar, T S; Jeyaraj, D A; Rajalakshmi, M; Sharma, R S

1999-09-01

Rhesus monkey prostate epithelial cells from the cranial lobe were isolated and cultured in flasks coated either with collagen IV or laminin. The effects of stromal cell medium, androgens and growth factors on cell number, thymidine incorporation and secretory activity were assessed. The results indicate that dihydrotestosterone (DHT) and androstenedione have stimulatory influences on cell proliferation and secretion in coated flasks. DHT was more effective in increasing cell number but the induction of secretory activity was similar with both steroids. The combination of IGF-I and -II resulted in inducing better cell proliferation and secretory activity than the individual IGFs but, of the two IGFs, IGF-I was more effective than IGF-II. DHT with IGFs was more potent in inducing proliferation, differentiation and secretion than androstenedione. Even in the absence of steroids or growth factors, colony formation and confluence occurred in coated flasks but cell differentiation and secretion only to a limited extent. In conclusion, we were able to establish an in vitro primary culture of prostate epithelial cells from rhesus monkey using extracellular matrix proteins, steroids and growth factors as additional supplements. This culture system may be useful to study prostate cell physiology and to identify drugs that can inhibit cell proliferation.

Production of Laccase by Recombinant Yarrowia lipolytica from Molasses: Bioprocess Development Using Statistical Modeling and Increase Productivity in Shake-Flask and Bioreactor Cultures.

PubMed

Darvishi, Farshad; Moradi, Marzieh; Madzak, Catherine; Jolivalt, Claude

2017-03-01

Laccases are used in numerous applications, from green degradation of various xenobiotic compounds, waste detoxification, textile dye bleaching, and delignification of lignocellulose materials to biofuel production. In this study, the recombinant Yarrowia lipolytica YL4 strain carrying the white-rot fungus Trametes versicolor laccase IIIb gene was used for laccase production from beet molasses as an agro-industrial residue. Response surface methodology was used to statistical optimization of the production of laccase by Y. lipolytica using an industrial medium containing molasses which allows a six times increase in laccase activity compared to primary medium contains glucose after 144 h. In bioreactor cultivation after 48 h, laccase production reached to 3.7- and 22.5-fold more than optimized and primary media in shake-flask cultures, respectively. Laccase productivity in bioreactor (0.0937 U/h) was higher than shake-flask culture (0.0084 U/h). The present study provides valuable information about statistical optimization of bioprocess development for cost-effective production of laccase and other heterologous proteins in Y. lipolytica from beet molasses as sole carbon source, thus allowing the valorization and decreasing environmental pollution of this agro-industrial waste.

Microarray platform affords improved product analysis in mammalian cell growth studies

PubMed Central

Li, Lingyun; Migliore, Nicole; Schaefer, Eugene; Sharfstein, Susan T.; Dordick, Jonathan S.; Linhardt, Robert J.

2014-01-01

High throughput (HT) platforms serve as cost-efficient and rapid screening method for evaluating the effect of cell culture conditions and screening of chemicals. The aim of the current study was to develop a high-throughput cell-based microarray platform to assess the effect of culture conditions on Chinese hamster ovary (CHO) cells. Specifically, growth, transgene expression and metabolism of a GS/MSX CHO cell line, which produces a therapeutic monoclonal antibody, was examined using microarray system in conjunction with conventional shake flask platform in a non-proprietary medium. The microarray system consists of 60 nl spots of cells encapsulated in alginate and separated in groups via an 8-well chamber system attached to the chip. Results show the non-proprietary medium developed allows cell growth, production and normal glycosylation of recombinant antibody and metabolism of the recombinant CHO cells in both the microarray and shake flask platforms. In addition, 10.3 mM glutamate addition to the defined base media results in lactate metabolism shift in the recombinant GS/MSX CHO cells in the shake flask platform. Ultimately, the results demonstrate that the high-throughput microarray platform has the potential to be utilized for evaluating the impact of media additives on cellular processes, such as, cell growth, metabolism and productivity. PMID:24227746

Inverse modeling of CO2 sources and sinks using satellite observations of CO2 from TES and surface flask measurements

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nassar, Ray; Jones, DBA; Kulawik, SS

2011-01-01

We infer CO2 surface fluxes using satellite observations of mid-tropospheric CO2 from the Tropospheric Emission Spectrometer (TES) and measurements of CO2 from surface flasks in a time-independent inversion analysis based on the GEOS-Chem model. Using TES CO2 observations over oceans, spanning 40 S 40 N, we find that the horizontal and vertical coverage of the TES and flask data are complementary. This complementarity is demonstrated by combining the datasets in a joint inversion, which provides better constraints than from either dataset alone, when a posteriori CO2 distributions are evaluated against independent ship and aircraft CO2 data. In particular, the jointmore » inversion offers improved constraints in the tropics where surface measurements are sparse, such as the tropical forests of South America. Aggregating the annual surface-to-atmosphere fluxes from the joint inversion for the year 2006 yields 1.13 0.21 PgC for the global ocean, 2.77 0.20 PgC for the global land biosphere and 3.90 0.29 PgC for the total global natural flux (defined as the sum of all biospheric, oceanic, and biomass burning contributions but excluding CO2 emissions from fossil fuel combustion). These global ocean and global land fluxes are shown to be near the median of the broad range of values from other inversion results for 2006. To achieve these results, a bias in TES CO2 in the Southern Hemisphere was assessed and corrected using aircraft flask data, and we demonstrate that our results have low sensitivity to variations in the bias correction approach. Overall, this analysis suggests that future carbon data assimilation systems can benefit by integrating in situ and satellite observations of CO2 and that the vertical information provided by satellite observations of mid-tropospheric CO2 combined with measurements of surface CO2, provides an important additional constraint for flux inversions.« less

Dynamic collection and analysis of volatile organic compounds from the headspace of cell cultures.

PubMed

Baranska, A; Smolinska, A; Boots, A W; Dallinga, J W; van Schooten, F J

2015-10-15

Exhaled breath has proven to be a valuable source of information about human bodies. Subtle differences between volatile organic compounds (VOCs) formed endogenously can be detected and become a base for a potential monitoring tool for health and disease. Until now, there has been a lack of biological and mechanistic knowledge of the processes involved in the production of relevant VOCs. Among the possible sources of health-related and disease-related VOCs are microorganisms found in the respiratory tract and in the gut. Other VOCs in the body are produced by cells that are influenced by the disease, for instance, due to metabolic disorders and/or inflammation. To gain insight into the in vivo production of VOCs by human cells and thus the exhaled breath composition, in vitro experiments involving relevant cells should be studied because they may provide valuable information on the production of VOCs by the affected cells. To this aim we developed and validated a system for dynamically (continuously) collecting headspace air in vitro using a Caco-2 cell line. The system allows the application of different cell lines as well as different experimental setups, including varying exposure times and treatment options while preserving cell viability. Significant correlation (p  ⩽  0.0001) between collection outputs within each studied group confirmed high reproducibility of the collection system. An example of such an application is presented here. We studied the influence of oxidative stress on the VOC composition of the headspace air of Caco-2 cells. By comparing the VOC composition of air flushed through empty culture flasks (n  =  35), flasks with culture medium (n  =  35), flasks with medium and cells (n  =  20), flasks with medium and an oxidative stressor (H2O2) (n  =  20), and flasks with medium, stressor, and cells (n  =  20), we were able to separate the effects from the stressor on the cells from all other interactions. Measurements were performed with gas chromatography time-of-flight mass spectrometry. Multivariate data analysis allowed detection of significant altered compounds in the compared groups. We found a significant change (p  ⩽  0.001) of the composition of VOCs due to the stressing of the Caco-2 cells by H2O2. A total of ten VOCs showed either increased or decreased abundance in the headspace of the cell cultures due to the presence of the H2O2 stressor.

Secondary metabolism in simulated microgravity

NASA Technical Reports Server (NTRS)

Demain, A. L.; Fang, A.

2001-01-01

We have studied microbial secondary metabolism in a simulated microgravity (SMG) environment provided by NASA rotating-wall bioreactors (RWBs). These reactors were designed to simulate some aspects of actual microgravity that occur in space. Growth and product formation were observed in SMG in all cases studied, i.e., Bacillus brevis produced gramicidin S (GS), Streptomyces clavuligerus made beta-lactam antibiotics, Streptomyces hygroscopicus produced rapamycin, and Escherichia coli produced microcin B17 (MccB17). Of these processes, only GS production was unaffected by SMG; production of the other three products was inhibited. This was determined by comparison with performance in an RWB positioned in a different mode to provide a normal gravity (NG) environment. Carbon source repression by glycerol of the GS process, as observed in shaken flasks, was not observed in the RWBs, whether operated in the SMG or NG mode. The same phenomenon occurred in the case of MccB17 production, with respect to glucose repression. Thus, the negative effects of carbon source on GS and beta-lactam formation are presumably dependent on shear, turbulence, and/or vessel geometry, but not on gravity. Stimulatory effects of phosphate and the precursor L-lysine on beta-lactam antibiotic production, as observed in flasks, also occurred in SMG. An almost complete shift in the localization of produced MccB17 from cells to extracellular medium was observed when E. coli was grown in the RWB under SMG or NG. If a plastic bead was placed in the RWB, accumulation became cellular, as it is in shaken flasks, indicating that sheer stress favors a cellular location. In the case of rapamycin, the same type of shift was observed, but it was less dramatic, i.e., growth in the RWB under SMG shifted the distribution of produced rapamycin from 2/3 cellular:1/3 extracellular to 1/3 cellular:2/3 extracellular. Stress has been shown to induce or promote secondary metabolism in a number of other microbial systems. RWBs provide a low stress SMG environment, which, however, supports only poor production of MccB17, as compared to production in shaken flasks. We wondered whether the poor production in RWBs under SMG is due to the low level of stress, and whether increasing stress in the RWBs would raise the amount of MccB17 formed. We found that increasing shear stress by adding a single Teflon bead to the RWB improved MccB17 production. Although shear stress seems to have a marked positive effect on MccB17 production in SMG, addition of various concentrations of ethanol to RWBs (or to shaken flasks) failed to increase MccB17 production. Ethanol stress merely decreased production and, at higher concentrations, inhibited growth. Interestingly, cells growing in the RWB were much more resistant to the growth- and production-inhibitory effects of ethanol than cells growing in shaken flasks. With respect to S. hygroscopicus, addition of Teflon beads to the RWB reversed the inhibition of growth, but rapamycin production was still markedly inhibited, and the distribution did not revert back to a preferential cellular site.

Assessment of JVX Proprotor Performance Data in Hover and Airplane-Mode Flight Conditions

NASA Technical Reports Server (NTRS)

Acree, C. W., Jr.

2016-01-01

A 0.656-scale V-22 proprotor, the Joint Vertical Experimental (JVX) rotor, was tested at the NASA Ames Research Center in both hover and airplane-mode (high-speed axial flow) flight conditions, up to an advance ratio of 0.562 (231 knots). This paper examines the two principal data sets generated by those tests, and includes investigations of hub spinner tares, torque/thrust measurement interactions, tunnel blockage effects, and other phenomena suspected of causing erroneous measurements or predictions. Uncertainties in hover and high-speed data are characterized. The results are reported here to provide guidance for future wind tunnel tests, data processing, and data analysis.

[Functional condition of the spine in female workers in the cotton-processing industry evaluated on the basis of subjective and radiological studies].

PubMed

Split, W; Kotwica, S

1987-01-01

Studies involved 179 weaving and 173 spinning women. All of them underwent functional tests of the spine supported by X-ray tests. In approximately 45% of weavers and 29% of spinners functional disturbances in the lower segment of the spine were found. The incidence of the motorial disturbances of the spine was found to increase with age. The analysis of the impact of employment length upon the rate of motorial disturbances demonstrated that the disturbances occurred within the first years of work and reached the highest level after 10 years.

Low and high speed propellers for general aviation: Performance potential and recent wind tunnel test results

NASA Technical Reports Server (NTRS)

Jeracki, R. J.; Mitchell, G. A.

1981-01-01

The performance of lower speed, 5 foot diameter model general aviation propellers, was tested in the Lewis wind tunnel. Performance was evaluated for various levels of airfoil technology and activity factor. The difference was associated with inadequate modeling of blade and spinner losses for propellers round shank blade designs. Suggested concepts for improvement are: (1) advanced blade shapes (airfoils and sweep); (2) tip devices (proplets); (3) integrated propeller/nacelles; and (4) composites. Several advanced aerodynamic concepts were evaluated in the Lewis wind tunnel. Results show that high propeller performance can be obtained to at least Mach 0.8.

Effect of stover fraction and storage method on glucose production during enzymatic hydrolysis.

PubMed

Montross, M D; Crofcheck, C L

2004-05-01

One avenue for overcoming the economic challenges associated with the production of ethanol from renewable resources is to reduce the cost of the biomass feedstock. The balance between storage costs and benefits depend on the storage method and composition changes of individual stover fractions. Corn stover from bales stored inside and outside of a barn was separated into an interior and exterior layer after approximately 10 months of storage. The cobs, stalks, and leaves and husks were separated, dried, and ground through a 2 mm screen. Stover, sodium acetate (buffer), cellulase, and deionized water were added to 125 ml flasks. The mixture was held at 50 degrees C in an incubator and samples taken for glucose determination. The average glucose concentration after 60 h of hydrolysis from cobs, leaves and husks, and stalks was 10.5, 9.6, and 3.1 g/l, respectively. Cobs, leaves, and husks produced over 300% more glucose than stalks. Storage outside of the barn decreased the glucose production from individual stover components between 4% and 8%. The effect of stover fraction type on glucose production was significant, while the storage treatment effect was not significant. Fractionation of corn stover may be a method to increase the value of corn stover as a feedstock for glucose production.

Deglycosylation of isoflavones in isoflavone-rich soy germ flour by Aspergillus oryzae KACC 40247.

PubMed

Lee, Seon-Hwa; Seo, Min-Ho; Oh, Deok-Kun

2013-12-11

Aspergillus oryzae KACC 40247 was selected as an efficient daidzein-producing fungus from strains of the genus Aspergillus by using 5% (w/v) soy germ flour (SGF) as an isoflavone-glycoside-rich medium. The culture conditions, including SGF concentration, agitation speed, initial pH, temperature, and time, were optimized as follows: 7% (w/v) SGF, initial pH 6.0, 33 °C, 300 rpm, and 24 h in a 100 mL baffled flask. The determined amount of isoflavone aglycons in SGF using 50% ethyl acetate was the highest among the solvent systems tested and it was 3.7-fold higher than that using 70% methanol. Under the optimized conditions, the content and concentration of daidzein were 134 mg/g of SGF and 9.4 g/L, respectively, with a productivity of 391 ± 2.8 mg/L/h, and those of isoflavone aglycons were 165 mg/g of SGF and 11.5 g/L, respectively, with a productivity of 479 mg/L/h. Optimization of culture conditions increased the content, concentration, and productivity of isoflavone aglycons by 3.1-, 3.0-, and 3.7-fold, respectively. To our knowledge, this is the highest production of isoflavone aglycons reported to date.

Structural and functional characterization of recombinant napin-like protein of Momordica charantia expressed in methylotrophic yeast Pichia pastoris.

PubMed

Yadav, Shailesh Kumar R; Sahu, Tejram; Dixit, Aparna

2016-08-01

Napin and napin-like proteins belong to the 2S albumin seed storage family of proteins and have been shown to display a variety of biological activities. However, due to a high degree of polymorphism, purification of a single napin or napin-like protein exhibiting biological activity is extremely difficult. In the present study, we have produced the napin-like protein of Momordica charantia using the methylotrophic Pichia pastoris expression system. The recombinant napin-like protein (rMcnapin) secreted in the extracellular culture supernatant was enriched by ammonium sulfate precipitation, and purified using size exclusion chromatography at a yield of ∼290 mg/L of culture. Secondary structure analysis of the purified rMcnapin revealed it to be predominantly α-helical with minimal β strand content. CD spectroscopic and fluorescence spectroscopic analyses revealed the rMcnapin to be stable at a wide range of temperatures and pH. The rMcnapin exhibited antifungal activity against Trichoderma viride with an IC50 of ∼3.7 μg/ml and trypsin inhibitor activity with an IC50 of 4.2 μM. Thus, large amounts of homogenous preparations of the biologically active rMcnapin could be obtained at shake flask level, which is otherwise difficult from its natural source.

Rapid and qualitative fluorescence-based method for the assessment of PHA production in marine bacteria during batch culture.

PubMed

Elain, Anne; Le Fellic, Magali; Corre, Yves-Marie; Le Grand, Adélaïde; Le Tilly, Véronique; Audic, Jean-Luc; Bruzaud, Stéphane

2015-10-01

The expansion of polyhydroxyalkanoates (PHAs) into the biodegradable polymers market is mainly prevented by their production process which is still complicated with a low efficiency, resulting in relatively expensive products. In this study, we developed a method that used the lipophilic fluorescent probe Nile Red (1 mg l(-1) solution in DMSO) directly into the culture broth to stain the PHA inclusions inside bacterial cells followed by detection of the emitted fluorescence by both microscopic and spectrometric techniques. Epifluorescence microscopy provides a rapid tool to distinguish producing from non-producing bacterial species and the relative fluorescence intensity (FI) determined at the maximum of emission spectra in the wavelength region of 560-710 nm (λ(ex): 543 nm), allows a fast assessment of the cultural conditions that may enhance PHA production yield. During two-step cultivation in 500-ml flasks with glucose as the sole carbon source, the method aimed to select bacterial strains efficient for PHA synthesis among a marine collection. Subsequently, the NR assay was used to determine the C0/N0 ratio of the producing media that may improve the polymer yield as well as to follow the time course of fermentation. Characterization by GC-MS and DSC confirmed the production of the P(3-HB) homopolymer.

Soluble Prokaryotic Expression and Purification of Bioactive Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand.

PubMed

Do, Bich Hang; Nguyen, Minh Tan; Song, Jung-A; Park, Sangsu; Yoo, Jiwon; Jang, Jaepyeong; Lee, Sunju; So, Seoungjun; Yoon, Yejin; Kim, Inki; Lee, Kyungjin; Jang, Yeon Jin; Choe, Han

2017-12-28

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is considered as an antitumor agent owing to its ability to induce apoptosis of cancer cells without imparting toxicity toward most normal cells. TRAIL is produced in poor yield because of its insoluble expression in the cytoplasm of E. coli . In this study, we achieved soluble expression of TRAIL by fusing maltose-binding protein (MBP), b'a' domain of protein disulfide isomerase (PDIb'a'), or protein disulfide isomerase at the N-terminus of TRAIL. The TRAIL was purified using subsequent immobilized metal affinity chromatography and amylose-binding chromatography, with the tag removal using tobacco etch virus protease. Approximately 4.5 mg of pure TRAIL was produced from 125 ml flask culture with a purification yield of 71.6%. The endotoxin level of the final product was 0.4 EU/μg, as measured by the Limulus amebocyte lysate endotoxin assay. The purified TRAIL was validated and shown to cause apoptosis of HeLa cells with an EC₅₀ and Hill coefficient of 0.6 ± 0.03 nM and 2.41 ± 0.15, respectively. The high level of apoptosis in HeLa cells following administration of purified TRAIL indicates the significance and novelty of this method for producing high-grade and high-yield TRAIL.

Determination of mercury in agroindustrial samples by flow-injection cold vapor atomic absorption spectrometry using ion exchange and reductive elution.

PubMed

Gomes Neto, J A; Zara, L F; Rocha, J C; Santos, A; Dakuzaku, C S; Nóbrega, J A

2000-03-06

A flow-injection system with a Chelite-S(R) cationic resin packed minicolumn is proposed for the determination of trace levels of mercury in agroindustrial samples by cold vapor atomic absorption spectrometry. Improved sensitivity and selectivity are attained since mercuric ions are on-line concentrated whereas other potential interferents are discarded. With on-line reductive elution procedure, concentrated hydrochloric acid could be replaced by 10% w/v SnCl(2), in 6 M HCl as eluent. The reversed-intermittent stream either carries the atomic mercury to the flow cell in the forward direction or removes the residue from reactor/gas-liquid separator to a discarding flask in the opposite direction. Concentration and volume of reagent, acidity, flow rates, commutation times and potential interfering species were investigated. For 120 s preconcentration time, the proposed system handles about 25 samples h(-1) (50.0-500 ng l(-1)), consuming about 10 ml sample and 5 mg SnCl(2) per determination. The detection limit is 0.8 ng l(-1) and the relative standard deviation (RSD) (n=12) of a 76.7 ng l(-1) sample is about 5%. Results are in agreement with certified value of standard materials at 95% confidence level and good recoveries (97-128%) of spiked samples were found.

Raman gas sensing of modified Ag nanoparticle SERS

NASA Astrophysics Data System (ADS)

Myoung, NoSoung; Yoo, Hyung Keun; Hwang, In-Wook

2014-03-01

Recent progress in modified Surface Enhanced Raman Scattering (SERS) using Ag nanoparticles makes them promising optical technique for direct gas sensing of interest. However, SERS has been shown to provide sub ppb level detection of the compounds in the vapor phase. The major problem with the sensitivity scaling-up was in the development of fabrication technology for stability and reproducibility of SERS substrates. We report an optimization of 1-propanethiol coated multiple Ag nanoparticle layers on SiO2 substrate as well as new records of real-time, simultaneous vapor phase detection of toluene and 1-2 dichlorobenzene by the radiation of fiber optic coupled 785 nm diode laser and spectrograph. Multiple depositions of Ag NPs were loaded on SiO2 and soaked in 1-propanethiol solution for 24 hours to modify the surface into hydrophobic due to the characteristics of vapor phase of our interests. Raman bands at 1003 cm-1 and 1130 cm-1 for toluene and 12DCB, respectively were compared to 1089 cm-1 and each gas concentration in 1000 mL flask were calculated as a function of each vapor phase ratio. The saturation of toluene and 12DCB were limited only by 800 ppm and the detectable range was 0.6-800 ppm.

Evaluation of cysteine ethyl ester as efficient inducer for glutathione overproduction in Saccharomyces spp.

PubMed

Lorenz, Eric; Schmacht, Maximilian; Senz, Martin

2016-11-01

Economical yeast based glutathione (GSH) production is a process that is influenced by several factors like raw material and production costs, biomass production and efficient biotransformation of adequate precursors into the final product GSH. Nowadays the usage of cysteine for the microbial conversion into GSH is industrial state of practice. In the following study, the potential of different inducers to increase the GSH content was evaluated by means of design of experiments methodology. Investigations were executed in three natural Saccharomyces strains, S. cerevisiae, S. bayanus and S. boulardii, in a well suited 50ml shake tube system. Results of shake tube experiments were confirmed in traditional baffled shake flasks and finally via batch cultivation in lab-scale bioreactors under controlled conditions. Comprehensive studies showed that the usage of cysteine ethyl ester (CEE) for the batch-wise biotransformation into GSH led up to a more than 2.2 times higher yield compared to cysteine as inducer. Additionally, the intracellular GSH content could be significantly increased for all strains in terms of 2.29±0.29% for cysteine to 3.65±0.23% for CEE, respectively, in bioreactors. Thus, the usage of CEE provides a highly attractive inducing strategy for the GSH overproduction. Copyright © 2016 Elsevier Inc. All rights reserved.

Integrated ozone and biotreatment of pulp mill effluent and changes in biodegradability and molecular weight distribution of organic compounds.

PubMed

Bijan, Leila; Mohseni, Madjid

2005-10-01

The overall effectiveness of integrating ozonation with biological treatment on the biodegradability enhancement and recalcitrant organic matter (ROM) removal from pulp mill alkaline bleach plant effluent was investigated. Ozonation was performed in a semi-batch bubble column reactor at pH of 11 and 4.5. Batch biological treatment was conducted in shake flasks. Samples obtained during the treatments were monitored for BOD5, COD, TOC, and molecular weight distribution. At an ozone dosage of 0.7-0.8 mg O3/mL wastewater, integrated treatment showed about 30% higher TOC mineralization compared to individual ozonation or biotreatment. Ozone treatment enhanced the biodegradability of the effluent (monitored as 21% COD reduction and 13% BOD5 enhancement), allowing for a higher removal of pollutants. The conversion of high molecular weight (HMW) to low molecular weight (LMW) compounds was an important factor in the overall biodegradability enhancement of the alkaline effluent. The overall biodegradability of the LMW compounds did not change over the course of ozonation, but it increased from 5% to 50% (measured as COD removal) for the HMW portion. Ozonation at pH of 11 was more effective than that at pH of 4.5 in terms of generating more biodegradable compounds.

A preliminary study on performance of Saccharomyces cerevisiae n0 DY 7221 immobilized using grafted bioflocculant in bioethanol production

NASA Astrophysics Data System (ADS)

Suci, Windhu Griyasti; Margono, Kaavessina, Mujtahid

2018-02-01

Bioethanol has been well acknowledged to be developed as a biofuel and can be derived from renewable resources. Currently, the utilization of bioethanol as a fuel is more expensive than that of gasoline due to the high production cost. Researchers from industrial and academia have been doing some efforts to reduce it, namely: energy efficiency, exploring many potential renewable resources, increasing fermentation productivity, etc. We propose a novel immobilized Saccharomyces cerevisiae trapped in grafted bioflocculant. The flocculant was developed from polyacrylamide chains grafted into modified starches. This research aims to preliminary performance study of S. cerevisiae immobilized using our new developed method. The bioflocculant solution with the various concentration of 1%, 2%, and 2.5% v/v was dropped into 90 ml of developed inoculum to get flocs which would be used as a starter in fermentation process. The fermentation process was carried out in a shaken flask at 30oC and 150 rpm for 72 hours. The best result was obtained in the sample of 2.5% bioflocculant fraction, i.e. bioethanol 9.25% and enhanced productivity 3.6 times of free cell. These results indicate that flocculation method is a way of immobilizing yeast that needs to be further investigated.

Effect of cell passage on the susceptibility of BALB/3T3 clone A31-1-1 cells to 3-methylcholanthrene-induced morphological transformation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sheu, C.W.; Moreland, F.M.; Dunkel, V.C.

1987-01-01

The response of BALB/3T3 clone A31-1-1 cells to chemically induced morphological transformation was evaluated using 3-methylcholanthrene (MCA). Stock cultures were initiated from cryopreserved cells, grown in T25 flasks containing 5 ml of medium, and replated at subconfluency. Serially transferred cells were then subjected to transformation assay. After 24-hr seeding, cells were incubated 48 hr with MCA in a 5% CO/sub 2/ incubator. They were then rinsed and incubated for an additional 4 weeks with twice weekly medium change. Type III foci were scored after fixation and staining with Giemsa. With serial passage from the frozen state, cells of passages 3-14more » had a low level of spontaneous transformation; zero to 6 type III foci per 20 dishes were counted. In the MCA-treated cultures the number of transformed foci, however, increased with passage. Such passage-related sensitivity to MCA was demonstrated for cells cultured in two batches of sera: one from MA Bioproducts (Lot no. 2E052) and the other from Armour Pharmaceuticals (Lot no. Y65801). The passage-related increase in number of transformed foci was not related to doubling time, cloning efficiency, or MCA-induced growth inhibition.« less

Inventory Control.

ERIC Educational Resources Information Center

Byrum, David L., Ed.

1984-01-01

Describes an electronic thermometer using a precision temperature sensor (includes detailed schematic of circuits) and inexpensive ring holders for round-bottomed flasks. Also describes a method for reducing funnel breakage. (JN)

Process development for the production of 15β-hydroxycyproterone acetate using Bacillus megaterium expressing CYP106A2 as whole-cell biocatalyst.

PubMed

Kiss, Flora M; Lundemo, Marie T; Zapp, Josef; Woodley, John M; Bernhardt, Rita

2015-03-05

CYP106A2 from Bacillus megaterium ATCC 13368 was first identified as a regio- and stereoselective 15β-hydroxylase of 3-oxo-∆4-steroids. Recently, it was shown that besides 3-oxo-∆4-steroids, 3-hydroxy-∆5-steroids as well as di- and triterpenes can also serve as substrates for this biocatalyst. It is highly selective towards the 15β position, but the 6β, 7α/β, 9α, 11α and 15α positions have also been described as targets for hydroxylation. Based on the broad substrate spectrum and hydroxylating capacity, it is an excellent candidate for the production of human drug metabolites and drug precursors. In this work, we demonstrate the conversion of a synthetic testosterone derivative, cyproterone acetate, by CYP106A2 under in vitro and in vivo conditions. Using a Bacillus megaterium whole-cell system overexpressing CYP106A2, sufficient amounts of product for structure elucidation by nuclear magnetic resonance spectroscopy were obtained. The product was characterized as 15β-hydroxycyproterone acetate, the main human metabolite. Since the product is of pharmaceutical interest, our aim was to intensify the process by increasing the substrate concentration and to scale-up the reaction from shake flasks to bioreactors to demonstrate an efficient, yet green and cost-effective production. Using a bench-top bioreactor and the recombinant Bacillus megaterium system, both a fermentation and a transformation process were successfully implemented. To improve the yield and product titers for future industrial application, the main bottlenecks of the reaction were addressed. Using 2-hydroxypropyl-β-cyclodextrin, an effective bioconversion of 98% was achieved using 1 mM substrate concentration, corresponding to a product formation of 0.43 g/L, at a 400 mL scale. Here we describe the successful scale-up of cyproterone acetate conversion from shake flasks to bioreactors, using the CYP106A2 enzyme in a whole-cell system. The substrate was converted to its main human metabolite, 15β-hydroxycyproterone acetate, a highly interesting drug candidate, due to its retained antiandrogen activity but significantly lower progestogen properties than the mother compound. Optimization of the process led to an improvement from 55% to 98% overall conversion, with a product formation of 0.43 g/L, approaching industrial process requirements and a future large-scale application.

Non-invasive and non-destructive measurements of confluence in cultured adherent cell lines.

PubMed

Busschots, Steven; O'Toole, Sharon; O'Leary, John J; Stordal, Britta

2015-01-01

Many protocols used for measuring the growth of adherent monolayer cells in vitro are invasive, destructive and do not allow for the continued, undisturbed growth of cells within flasks. Protocols often use indirect methods for measuring proliferation. Microscopy techniques can analyse cell proliferation in a non-invasive or non-destructive manner but often use expensive equipment and software algorithms. In this method images of cells within flasks are captured by photographing under a standard inverted phase contract light microscope using a digital camera with a camera lens adaptor. Images are analysed for confluence using ImageJ freeware resulting in a measure of confluence known as an Area Fraction (AF) output. An example of the AF method in use on OVCAR8 and UPN251 cell lines is included. •Measurements of confluence from growing adherent cell lines in cell culture flasks is obtained in a non-invasive, non-destructive, label-free manner.•The technique is quick, affordable and eliminates sample manipulation.•The technique provides an objective, consistent measure of when cells reach confluence and is highly correlated to manual counting with a haemocytometer. The average correlation co-efficient from a Spearman correlation (n = 3) was 0.99 ± 0.008 for OVCAR8 (p = 0.01) and 0.99 ± 0.01 for UPN251 (p = 0.01) cell lines.

Scale-up laccase production from Trametes versicolor stimulated by vanillic acid.

PubMed

Wang, Ke-Feng; Hu, Jian-Hua; Guo, Chen; Liu, Chun-Zhao

2016-07-01

An efficient strategy for laccase production in Trametes versicolor cultures was developed using vanillic acid as the inducer. The optimized vanillic acid treatment strategy consisted of exposing 2-day-old mycelia cultures to 80 mg/L vanillic acid. After 4 days, laccase activity of 588.84 U/L was achieved in flasks which represented a 1.79-fold increase compared to the control. In 200-L airlift bioreactor, the maximal laccase activity reached up to 785.12 U/L using the optimized vanillic acid treatment strategy. The zymograms of culture supernatants revealed three bands with laccase activity, among which Lac1 and Lac2 were abundant laccase isoforms constitutively expressed, and Lac3 was an inducible isozyme by vanillic acid. The results of real-time quantitative PCR showed that the transcription level of lcc in T. versicolor cultures grown with vanillic acid for 7 days was about 5.64-fold greater than that without vanillic acid in flasks. In 200-L airlift bioreactor cultures of T. versicolor with addition of vanillic acid, the transcript level of lcc at day 7 was 2.62-fold higher than that in flasks with vanillic acid due to the good mass transfer and oxygen supply in the bioreactor system. This study provides a basis for understanding the induction mechanism of vanillic acid for laccase production and has good potential for industrial applications.

Study of plant phototropic responses to different LEDs illumination in microgravity

NASA Astrophysics Data System (ADS)

Zyablova, Natalya; Berkovich, Yuliy A.; Skripnikov, Alexander; Nikitin, Vladimir

2012-07-01

The purpose of the experiment planned for Russian BION-M #1, 2012, biosatellite is research of Physcomitrella patens (Hedw.) B.S.G. phototropic responses to different light stimuli in microgravity. The moss was chosen as small-size higher plant. The experimental design involves five lightproof culture flasks with moss gametophores fixed inside the cylindrical container (diameter 120 mm; height 240 mm). The plants in each flask are illuminated laterally by one of the following LEDs: white, blue (475 nm), red (625 nm), far red (730 nm), infrared (950 nm). The gametophores growth and bending are captured periodically by means of five analogue video cameras and recorder. The programmable command module controls power supply of each camera and each light source, commutation of the cameras and functioning of video recorder. Every 20 minutes the recorder is sequentially connecting to one of the cameras. This results in a clip, containing 5 sets of frames in a row. After landing time-lapse films are automatically created. As a result we will have five time-lapse films covering transformations in each of the five culture flasks. Onground experiments demonstrated that white light induced stronger gametophores phototropic bending as compared to red and blue stimuli. The comparison of time-lapse recordings in the experiments will provide useful information to optimize lighting assemblies for space plant growth facilities.

An innovative iterative thresholding algorithm for tumour segmentation and volumetric quantification on SPECT images: Monte Carlo-based methodology and validation.

PubMed

Pacilio, M; Basile, C; Shcherbinin, S; Caselli, F; Ventroni, G; Aragno, D; Mango, L; Santini, E

2011-06-01

Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) imaging play an important role in the segmentation of functioning parts of organs or tumours, but an accurate and reproducible delineation is still a challenging task. In this work, an innovative iterative thresholding method for tumour segmentation has been proposed and implemented for a SPECT system. This method, which is based on experimental threshold-volume calibrations, implements also the recovery coefficients (RC) of the imaging system, so it has been called recovering iterative thresholding method (RIThM). The possibility to employ Monte Carlo (MC) simulations for system calibration was also investigated. The RIThM is an iterative algorithm coded using MATLAB: after an initial rough estimate of the volume of interest, the following calculations are repeated: (i) the corresponding source-to-background ratio (SBR) is measured and corrected by means of the RC curve; (ii) the threshold corresponding to the amended SBR value and the volume estimate is then found using threshold-volume data; (iii) new volume estimate is obtained by image thresholding. The process goes on until convergence. The RIThM was implemented for an Infinia Hawkeye 4 (GE Healthcare) SPECT/CT system, using a Jaszczak phantom and several test objects. Two MC codes were tested to simulate the calibration images: SIMIND and SimSet. For validation, test images consisting of hot spheres and some anatomical structures of the Zubal head phantom were simulated with SIMIND code. Additional test objects (flasks and vials) were also imaged experimentally. Finally, the RIThM was applied to evaluate three cases of brain metastases and two cases of high grade gliomas. Comparing experimental thresholds and those obtained by MC simulations, a maximum difference of about 4% was found, within the errors (+/- 2% and +/- 5%, for volumes > or = 5 ml or < 5 ml, respectively). Also for the RC data, the comparison showed differences (up to 8%) within the assigned error (+/- 6%). ANOVA test demonstrated that the calibration results (in terms of thresholds or RCs at various volumes) obtained by MC simulations were indistinguishable from those obtained experimentally. The accuracy in volume determination for the simulated hot spheres was between -9% and 15% in the range 4-270 ml, whereas for volumes less than 4 ml (in the range 1-3 ml) the difference increased abruptly reaching values greater than 100%. For the Zubal head phantom, errors ranged between 9% and 18%. For the experimental test images, the accuracy level was within +/- 10%, for volumes in the range 20-110 ml. The preliminary test of application on patients evidenced the suitability of the method in a clinical setting. The MC-guided delineation of tumor volume may reduce the acquisition time required for the experimental calibration. Analysis of images of several simulated and experimental test objects, Zubal head phantom and clinical cases demonstrated the robustness, suitability, accuracy, and speed of the proposed method. Nevertheless, studies concerning tumors of irregular shape and/or nonuniform distribution of the background activity are still in progress.

Mercury recycling in the United States in 2000

USGS Publications Warehouse

Brooks, William E.; Matos, Grecia R.

2005-01-01

Reclamation and recycling of mercury from used mercury- containing products and treatment of byproduct mercury from gold mining is vital to the continued, though declining, use of this metal. Mercury is reclaimed from mercury-containing waste by treatment in multistep high-temperature retorts-the mercury is volatized and then condensed for purification and sale. Some mercury-containing waste, however, may be landfilled, and landfilled material represents loss of a recyclable resource and a threat to the environment. Related issues include mercury disposal and waste management, toxicity and human health, and regulation of mercury releases in the environment. End-users of mercury-containing products may face fines and prosecution if these products are improperly recycled or not recycled. Local and State environmental regulations require adherence to the Resource Conservation and Recovery Act and the Comprehensive Environmental Response, Compensation, and Liability Act to regulate generation, treatment, and disposal of mercury-containing products. In the United States, several large companies and a number of smaller companies collect these products from a variety of sources and then reclaim and recycle the mercury. Because mercury has not been mined as a principal product in the United States since 1992, mercury reclamation from fabricated products has become the main source of mercury. Principal product mercury and byproduct mercury from mining operations are considered to be primary materials. Mercury may also be obtained as a byproduct from domestic or foreign gold-processing operations. In the early 1990s, U.S. manufacturers used an annual average that ranged from 500 to 600 metric tons of recycled and imported mercury for fabrication of automobile convenience switches, dental amalgam, fluorescent lamps, medical uses and thermometers, and thermostats. The amount now used for fabrication is estimated to be 200 metric tons per year or less. Much of the data on mercury is estimated because it is a low-volume commodity and its production, use, and disposal is difficult to track. The prices and volumes of each category of mercury-containing material may change dramatically from year to year. For example, the average price of mercury was approximately $150 per flask from 2000 until 2003 and then rose sharply to $650 per flask in fall 2004 and approximately $850 per flask in spring 2005. Since 1927, the common unit for measuring and pricing mercury has been the flask in order to conform to the system used at Almaden, Spain (Meyers, 1951). One flask weighs 34.5 kilograms, and 29 flasks of mercury are contained in a metric ton. In the United States, the chlorine-caustic soda industry, which is the leading end-user of elemental mercury, recycles most of its mercury in-plant as home scrap. Annual purchases of replacement mercury by the chlorine-caustic soda industry indicate that some mercury may be lost through evaporation to the environment, put into a landfill as industrial waste, or trapped within pipes in the plant. Impending closure of domestic and foreign mercury-cell chlorine-caustic soda plants and the shift to nonmercury technology for chlorine-caustic soda production could ultimately result in a significant volume of elemental mercury for recycling, sale, or storage. Globally, mercury is widely used in artisanal, or small-scale, gold mining. Most of that mercury is lost to the environment and is not recycled. The recycling rate for mercury was not available owing to insufficient data in 2000, and the efficiency of mercury recycling was estimated to be 62 percent.

Antifoam addition to shake flask cultures of recombinant Pichia pastoris increases yield

PubMed Central

2011-01-01

Background Pichia pastoris is a widely-used host for recombinant protein production. Initial screening for both suitable clones and optimum culture conditions is typically carried out in multi-well plates. This is followed by up-scaling either to shake-flasks or continuously stirred tank bioreactors. A particular problem in these formats is foaming, which is commonly prevented by the addition of chemical antifoaming agents. Intriguingly, antifoams are often added without prior consideration of their effect on the yeast cells, the protein product or the influence on downstream processes such as protein purification. In this study we characterised, for the first time, the effects of five commonly-used antifoaming agents on the total amount of recombinant green fluorescent protein (GFP) secreted from shake-flask cultures of this industrially-relevant yeast. Results Addition of defined concentrations of Antifoam A (Sigma), Antifoam C (Sigma), J673A (Struktol), P2000 (Fluka) or SB2121 (Struktol) to shake-flask cultures of P. pastoris increased the total amount of recombinant GFP in the culture medium (the total yield) and in the case of P2000, SB2121 and J673A almost doubled it. When normalized to the culture density, the GFP specific yield (μg OD595-1) was only increased for Antifoam A, Antifoam C and J673A. Whilst none of the antifoams affected the growth rate of the cells, addition of P2000 or SB2121 was found to increase culture density. There was no correlation between total yield, specific yield or specific growth rate and the volumetric oxygen mass transfer coefficient (kLa) in the presence of antifoam. Moreover, the antifoams did not affect the dissolved oxygen concentration of the cultures. A comparison of the amount of GFP retained in the cell by flow cytometry with that in the culture medium by fluorimetry suggested that addition of Antifoam A, Antifoam C or J673A increased the specific yield of GFP by increasing the proportion secreted into the medium. Conclusions We show that addition of a range of antifoaming agents to shake flask cultures of P. pastoris increases the total yield of the recombinant protein being produced. This is not only a simple method to increase the amount of protein in the culture, but our study also provides insight into how antifoams interact with microbial cell factories. Two mechanisms are apparent: one group of antifoams (Antifoam A, Antifoam C and J673A) increases the specific yield of GFP by increasing the total amount of protein produced and secreted per cell, whilst the second (P2000 or SB2121) increases the total yield by increasing the density of the culture. PMID:21426555

How-To-Do-It: Plant Regeneration.

ERIC Educational Resources Information Center

Pietraface, William J.

1988-01-01

Describes a procedure for the growth of tobacco plants in flasks. Demonstrates plant tissue culture manipulation, totipotency, and plant regeneration in approximately 12 weeks. Discusses methods, materials, and expected results. (CW)

Cytotoxic effects of some animal and vegetable extracts and some chemicals on liver and colon carcinoma and myosarcoma.

PubMed

Bayazit, Vahdettin

2004-02-01

To study, the cytotoxic effects of some biological and chemical agents on G1, S, G2, M and G0 phases of liver and colon carcinomas and myosarcoma cells obtained with chemical carcinogens dimethylbenzanthracene (DMBA) and cadmium chloride. Eight rabbit livers, colon carcinoma and myosarcoma cell lines were obtained by injection of DMBA in the Biology Laboratory, of the University of Dumlupinar, Kutahya, Turkey between January 2001 and June 2003. All lines were grown at 37 degrees celsius and 5% carbon dioxide in sterile RPMI-1640 medium with 10% fetal bovine serum after addition of glutamate, penicillin (50 units/ml) and streptomycin (50 ug/ml) (complete medium). Cells were grown on standard tissue culture plastic flasks to 80% confluence and passed by trypsinization. Tortoise (Testudo graeca) shell, sponge (Geodia cydonium), medusa (Aurelia aurita), meat flies (Calliphora erythrocephala) larva, frog (Rana ridibunda) larva and juniper (Juniperus communis) berry extracts killed a large amount of the liver and colon carcinomas and the myosarcoma cells in G2, M and G0 phases (p<0.01). The mistletoe (Viscum album) extract had more effect in only the G0 phase (p<0.05). Genistein, genistin, glycitein, glycitin, daitzein and daitzin have significantly decreased in the cancer cells tests, particularly, genistein and daitzein caused the apoptotic effect in G2, M and G0 phases (p<0.01). Cesium chloride, a mixture of cesium chloride with magnesium chloride had the most effect on tumor cells (p<0.01). AzhexSi, Azhex-AzhepSi, Et-Azhex-AzhepSi, AzhepSi, Hexamine and DL 54 have been inhibited in various levels of the cancer cells (p<0.05, p<0.01). This data suggest that some biological extracts and chemicals tested may be useful chemotherapeutic agents to inhibit the growth of cancer cells. This study sheds some light for new anti cancerogenic experiments preventing various cancers on humans.

Sugarcane straw as a feedstock for xylitol production by Candida guilliermondii FTI 20037.

PubMed

Hernández-Pérez, Andrés Felipe; de Arruda, Priscila Vaz; Felipe, Maria das Graças de Almeida

2016-01-01

Sugarcane straw has become an available lignocellulosic biomass since the progressive introduction of the non-burning harvest in Brazil. Besides keeping this biomass in the field, it can be used as a feedstock in thermochemical or biochemical conversion processes. This makes feasible its incorporation in a biorefinery, whose economic profitability could be supported by integrated production of low-value biofuels and high-value chemicals, e.g., xylitol, which has important industrial and clinical applications. Herein, biotechnological production of xylitol is presented as a possible route for the valorization of sugarcane straw and its incorporation in a biorefinery. Nutritional supplementation of the sugarcane straw hemicellulosic hydrolyzate as a function of initial oxygen availability was studied in batch fermentation of Candida guilliermondii FTI 20037. The nutritional supplementation conditions evaluated were: no supplementation; supplementation with (NH4)2SO4, and full supplementation with (NH4)2SO4, rice bran extract and CaCl2·2H2O. Experiments were performed at pH 5.5, 30°C, 200rpm, for 48h in 125mL Erlenmeyer flasks containing either 25 or 50mL of medium in order to vary initial oxygen availability. Without supplementation, complete consumption of glucose and partial consumption of xylose were observed. In this condition the maximum xylitol yield (0.67gg(-1)) was obtained under reduced initial oxygen availability. Nutritional supplementation increased xylose consumption and xylitol production by up to 200% and 240%, respectively. The maximum xylitol volumetric productivity (0.34gL(-1)h(-1)) was reached at full supplementation and increased initial oxygen availability. The results demonstrated a combined effect of nutritional supplementation and initial oxygen availability on xylitol production from sugarcane straw hemicellulosic hydrolyzate. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Nutritional demands and metabolic characteristics of the DSIR-HA-1179 insect cell line during growth and infection with the Oryctes nudivirus.

PubMed

Pushparajan, Charlotte; Claus, Juan Daniel; Marshall, Sean D G; Visnovsky, Gabriel

2017-12-01

The DSIR-HA-1179 coleopteran cell line has been identified as a susceptible and permissive host for the in vitro replication of the Oryctes nudivirus, which can be used as a biopesticide against the coconut rhinoceros beetle, pest of palms. The major challenge to in vitro large-scale Oryctes nudivirus production is ensuring process economy. This rests, among other requisites, on the use of low-cost culture media tailored to the nutritional and metabolic needs of the cell line, both in uninfected and infected cultures. The aim of the present study was to characterize the nutritional demands and the metabolic characteristics of the DSIR-HA-1179 cell line during growth and subsequent infection with Oryctes nudivirus in the TC-100 culture medium. Serum-supplementation of the culture medium was found to be critical for cell growth, and addition of 10% fetal bovine serum v/v led to a maximum viable cell density (16.8 × 10 5 cells ml -1 ) with a population doubling time of 4.2 d. Nutritional and metabolic characterization of the cell line revealed a trend of glucose and glutamine consumption but minimal uptake of other amino acids, negligible production of lactate and ammonia, and the accumulation of alanine, both before and after infection. The monitoring of virus production kinetics showed that the TC-100 culture medium was nutritionally sufficient to give a peak yield of 7.38 × 10 7 TCID 50 ml -1 of OrNV at the 6th day post-infection in attached cultures of DSIR-HA-1179 cells in 25 cm 2 T-flasks. Knowledge of the cell line's nutritional demands and virus production kinetics will aid in the formulation of a low-cost culture medium and better process design for large-scale OrNV production in future.

Spontaneously Generating Life in Your Classroom? Pasteur, Spallanzani and Science Process.

ERIC Educational Resources Information Center

Byington, Scott

2001-01-01

Presents an experiment that tests for spontaneous generation, or abiogenesis. Observes microbial growth in nutrient broth under seven different flask environments. Includes instructions for the methods. (YDS)

Cross-section perimeter is a suitable parameter to describe the effects of different baffle geometries in shaken microtiter plates

PubMed Central

2014-01-01

Background Biotechnological screening processes are performed since more than 8 decades in small scale shaken bioreactors like shake flasks or microtiter plates. One of the major issues of such reactors is the sufficient oxygen supply of suspended microorganisms. Oxygen transfer into the bulk liquid can in general be increased by introducing suitable baffles at the reactor wall. However, a comprehensive and systematic characterization of baffled shaken bioreactors has never been carried out so far. Baffles often differ in number, size and shape. The exact geometry of baffles in glass lab ware like shake flasks is very difficult to reproduce from piece to piece due to the hard to control flow behavior of molten glass during manufacturing. Thus, reproducibility of the maximum oxygen transfer capacity in such baffled shake flasks is hardly given. Results As a first step to systematically elucidate the general effect of different baffle geometries on shaken bioreactor performance, the maximum oxygen transfer capacity (OTRmax) in baffled 48-well microtiter plates as shaken model reactor was characterized. This type of bioreactor made of plastic material was chosen, as the exact geometry of the baffles can be fabricated by highly reproducible laser cutting. As a result, thirty different geometries were investigated regarding their maximum oxygen transfer capacity (OTRmax) and liquid distribution during shaking. The relative perimeter of the cross-section area as new fundamental geometric key parameter is introduced. An empirical correlation for the OTRmax as function of the relative perimeter, shaking frequency and filling volume is derived. For the first time, this correlation allows a systematic description of the maximum oxygen transfer capacity in baffled microtiter plates. Conclusions Calculated and experimentally determined OTRmax values agree within ± 30% accuracy. Furthermore, undesired out-of-phase operating conditions can be identified by using the relative perimeter as key parameter. Finally, an optimum well geometry characterized by an increased perimeter of 10% compared to the unbaffled round geometry is identified. This study may also assist to comprehensively describe and optimize the baffles of shake flasks in future. PMID:25093039

Effects of Water Vapor on the Data Quality of the Stable Oxygen Isotopic Ratio of Atmospheric Carbon Dioxide

NASA Astrophysics Data System (ADS)

Evans, C. U.; White, J. W.; Vaughn, B.; Tans, P. P.; Pardo, L.

2007-12-01

The stable oxygen isotopic ratio of carbon dioxide can potentially track fundamental indicators of environmental change such as the balance between photosynthesis and respiration on regional to global scales. The Stable Isotope Laboratory (SIL) at the Institute of Arctic and Alpine Research (INSTAAR), University of Colorado at Boulder, has measured the stable isotopes of atmospheric carbon dioxide from more than 60 NOAA/Earth System Research Laboratory (ESRL) air flask-sampling sites since the early 1990s. If air is sampled without drying, oxygen can exchange between carbon dioxide and water in the flasks, entirely masking the desired signal. An attempt to investigate how water vapor is affecting the δ18O signal is accomplished by comparing the SIL measurements with specific humidity, calculated from the National Climatic Data Center (NCDC) global integrated surface hourly temperature and dew point database, at the time of sampling. Analysis of sites where samples have been collected initially without drying, and subsequently with a drying kit, in conjunction with the humidity data, has led to several conclusions. Samples that initially appear isotopically unaltered, in that their δ18O values are within the expected range, are being subtly influenced by the water vapor in the air. At Bermuda and other tropical to semi-tropical sites, the 'wet' sampling values have a seasonal cycle that is strongly anti-correlated to the specific humidity, while the 'dry' values have a seasonal cycle that is shifted earlier than the specific humidity cycle by 1-2 months. The latter phasing is expected given the seasonal phasing between climate over the ocean and land, while the former is consistent with a small, but measurable isotope exchange in the flasks. In addition, we note that there is a strong (r > 0.96) correlation between the average specific humidity and the percent of rejected samples for 'wet' sampling. This presents an opportunity for determining a threshold of specific humidity, below which air flask samples can be trusted. This approach may allow segregation of suspect and trusted data, and thus provide an improved record of oxygen isotopic ratios of carbon dioxide over the past two decades.

Organochlorine residues in odontocete species from the southeast coast of India.

PubMed

Karuppiah, S; Subramanian, A; Obbard, J P

2005-08-01

Blubber from bottle-nose dolphins, spinner dolphins, humpback dolphin (Tursiops truncatus, Stenella longirostris and Sousa chinensis) were collected from the Bay of Bengal (southeast coast of India) and analyzed for the organochlorine pesticides hexachlorocyclohexane (HCHs), p,p'-dichlorodiphenyl trichloroethane (DDTs), and polychlorinated biphenyls (PCBs). All nine specimens analyzed contained considerable levels of all the three chemical classes where DDT was in the range of 3330-23330 ng/g; HCHs in the range of 95-765 ng/g; and PCBs in the range of 210-1220 ng/g (wet weight basis). The reasons for this and the variations in the isomer pattern of HCHs and DDT and its metabolites in marine mammal tissues are discussed.

Video-to-film color-image recorder.

NASA Technical Reports Server (NTRS)

Montuori, J. S.; Carnes, W. R.; Shim, I. H.

1973-01-01

A precision video-to-film recorder for use in image data processing systems, being developed for NASA, will convert three video input signals (red, blue, green) into a single full-color light beam for image recording on color film. Argon ion and krypton lasers are used to produce three spectral lines which are independently modulated by the appropriate video signals, combined into a single full-color light beam, and swept over the recording film in a raster format for image recording. A rotating multi-faceted spinner mounted on a translating carriage generates the raster, and an annotation head is used to record up to 512 alphanumeric characters in a designated area outside the image area.

Reconditioning of semiconductor substrates to remove photoresist during semiconductor device fabrication

DOEpatents

Farino, Anthony J.

2004-01-27

A method for reconditioning the surface of a semiconductor substrate to remove an unwanted (i.e. defective) layer of photoresist is disclosed. The method adapts a conventional automated spinner which is used to rotate the substrate at high speed while a stream of a first solvent (e.g. acetone) is used to dissolve the photoresist. A stream of a second solvent (e.g. methanol) is then used to clean the substrate at a lower speed, with the substrate being allowed to dry with continued rotation. The method of the present invention can be used within a photolithography track so that the substrates need never leave the track for reconditioning.

In situ stress and fracture permeability along the Stillwater fault zone, Dixie Valley Nevada

USGS Publications Warehouse

Hickman, S.H.; Barton, C.A.; Zoback, M.D.; Morin, R.; Sass, J.; Benoit, R.

1997-01-01

Borehole televiewer and hydrologic logging and hydraulic fracturing stress measurements were carried out in a 2.7-km-deep geothermal production well (73B-7) drilled into the Stillwater fault zone. Precision temperature and spinner flowmeter logs were also acquired in well 73B-7, with and without simultaneously injecting water into the well. Localized perturbations to well-bore temperature and flow were used to identify hydraulically conductive fractures. Comparison of these data with fracture orientations from the televiewer log indicates that permeable fractures within and adjacent to the Stillwater fault zone are critically stressed, potentially active shear planes in the current west-northwest extensional stress regime at Dixie Valley.

A hollow definitive obturator fabrication technique for management of partial maxillectomy.

PubMed

Patil, Pravinkumar Gajanan; Patil, Smita Pravinkumar

2012-11-01

Maxillary obturator prosthesis is the most frequent treatment option for management of partial or total maxillectomy. Heavy weight of the obturators is often a dislocating factor. Hollowing the prosthesis to reduce its weight is the well established fact. The alternate technique to hollow-out the prosthesis has been described in this article which is a variation of previously described processing techniques. A pre-shaped wax-bolus was incorporated inside the flasks during packing of the heat-polymerized acrylic resin to automatically create the hollow space. The processing technique described is a single step flasking procedure to construct a closed-hollow-obturator prosthesis as a single unit. To best understand the technique, this article describes management of a patient who had undergone partial maxillectomy secondary to squamous cell carcinoma rehabilitated with a hollow-obturator prosthesis.

A hollow definitive obturator fabrication technique for management of partial maxillectomy

PubMed Central

Patil, Smita Pravinkumar

2012-01-01

Maxillary obturator prosthesis is the most frequent treatment option for management of partial or total maxillectomy. Heavy weight of the obturators is often a dislocating factor. Hollowing the prosthesis to reduce its weight is the well established fact. The alternate technique to hollow-out the prosthesis has been described in this article which is a variation of previously described processing techniques. A pre-shaped wax-bolus was incorporated inside the flasks during packing of the heat-polymerized acrylic resin to automatically create the hollow space. The processing technique described is a single step flasking procedure to construct a closed-hollow-obturator prosthesis as a single unit. To best understand the technique, this article describes management of a patient who had undergone partial maxillectomy secondary to squamous cell carcinoma rehabilitated with a hollow-obturator prosthesis. PMID:23236579

Morphological changes in human melanoma cells following irradiation with thermal neutrons.

PubMed

Barkla, D H; Allen, B J; Brown, J K; Mountford, M; Mishima, Y; Ichihashi, M

1989-01-01

Morphological changes in two human melanoma cell lines, MM96 and MM418, following irradiation with thermal neutrons, were studied using light and electron microscopy. The results show that the response of human malignant melanoma cells to neutron irradiation is both cell line dependent and dose dependent, and that in any given cell line, some cells are more resistant to irradiation than others, thus demonstrating heterogeneity in respect to radiosensitivity. Cells repopulating MM96 flasks after irradiation were morphologically similar to the cells of origin whereas in MM418 flasks cells differentiated into five morphologically distinct subgroups and showed increased melanization. The results also show that radiation causes distinctive morphological patterns of damage although ultrastructural changes unique to the high LET particles released from boron 10 neutron capture are yet to be identified.

[Analysis of false-positive reaction for bacterial detection of blood samples with the automated microbial detection system BacT/ALERT 3D].

PubMed

Zhu, Li-Wei; Yang, Xue-Mei; Xu, Xiao-Qin; Xu, Jian; Lu, Huang-Jun; Yan, Li-Xing

2008-10-01

This study was aimed to analyze the results of false positive reaction in bacterial detection of blood samples with BacT/ALERT 3D system, to evaluate the specificity of this system, and to decrease the false positive reaction. Each reaction flasks in past five years were processed for bacteria isolation and identification. When the initial cultures were positive, the remaining samples and the corresponding units were recultured if still available. 11395 blood samples were detected. It is worthy of note that the incubator temperature should be stabilized, avoiding fluctuation; when the cultures were alarmed, the reaction flasks showed be kept some hours for further incubation so as to trace a sharply increasing signal to support the judgement of true bacterial growth. The results indicated that 122 samples (1.07%) wee positive at initial culture, out of them 107 samples (88.7%) were found bacterial, and 15 samples (12.3%) were found nothing. The detection curves of positive samples resulted from bacterial growth showed ascent. In conclusion, maintenance of temperature stability and avoidance of temperature fluctuation in incubator could decrease the occurrence of false-positive reaction in detection process. The reaction flasks with positive results at initial culture should be recultured, and whether existence of a sharply ascending logarilhimic growth phase in bacterial growth curve should be further detected, which are helpful to distinguish false-positive reactions from true positive, and thus increase the specificity of the BacT/ALERT system.

Bench-scale evaluation of in situ bioremediation strategies for soil at a former manufactured gas plant site.

PubMed

Li, Jun; Pignatello, Joseph J; Smets, Barth F; Grasso, Domenico; Monserrate, Esteban

2005-03-01

We examined the biodegradation and desorption of a set of 15 polycyclic aromatic hydrocarbon (PAH) compounds in coal tar-contaminated soil at a former manufactured gas plant site to evaluate the feasibility of in situ bioremediation. Experiments were conducted in well-mixed aerobic soil suspensions containing various additives over a 93- to 106-d period. In general, both biotransformation and desorption decreased with PAH ring size, becoming negligible for the six-ring PAH compounds. Biodegradation by indigenous microorganisms was strongly accelerated by addition of inorganic nutrients (N, P, K, and trace metals). The rates of biotransformation of PAH compounds by indigenous microorganisms in nutrient-amended flasks outpaced their maximum (i.e., chelate-enhanced) rates of desorption to an infinite sink (Tenax) in sterilized systems run in parallel, suggesting that indigenous organisms facilitated desorption. Biodegradation by indigenous organisms in nutrient-amended flasks appeared to be unaffected by the addition of a site-derived bacterial enrichment culture, resulting in approximately 100-fold higher aromatic dioxygenase levels, and by the addition of 0.01 M chelating agent (citrate or pyrophosphate), although such chelating agents greatly enhanced desorption in microbially inactivated flasks. The strong ability of nutrients to enhance degradation of the bioavailable PAHs indicates that their persistence for many decades at this site likely results from nutrient-limited natural biodegradation, and it also suggests that an effective strategy for their bioremediation could consist simply of adding inorganic nutrients.

Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method

PubMed Central

Salwe, Sukeshani; Kothari, Sweta; Chowdhary, Abhay; Deshmukh, Ranjana A.

2018-01-01

12–14 days of culturing of bone marrow (BM) cells containing various growth factors is widely used method for generating dendritic cells (DCs) from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs' purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference (P < 0.05) between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population. PMID:29682352

First 2 years of Atmospheric CO2 measurements in the Estany Llong plain (2100 masl, Parc Nacional d'Aigüestortes i Estany de Sant Maurici, Pyrenees, Catalonia, Spain).

NASA Astrophysics Data System (ADS)

Curcoll, Roger; Recolons, Montserrat; Font, Anna; Agraz, Laura; Parga, Elena; Bacardit, Montse; Camarero, Lluís.; Pueyo, Salva; Rodó, Xavier; Morguí, Josep Anton

2010-05-01

Since April 2009, air samples are being taken bi-weekly at 10 GMT in the plain of the Estany Llong at 2100 masl. Estany Llong air sampling site (ELL, 42°34'29''N 0°57'17''E) is a remote site situated in the SW principal valley of the Parc Nacional d'Aigüestortes i Estany de Sant Maurici. New Flask-sampling equipment for Remote Mountain Sites was developed by the Institut Català de Ciències del Clima (IC3) to allow flask sampling in extreme weather conditions and carrying the sampling equipment for more than 10 km without damaging flasks. Dry Air analysis for CO2 are done at the Laboratory of IC3 using two coupled modified IRGA Licor-7000, where both pressure and flow are externally controlled. Far away from populated areas, ELL site acts as a remote site, but it is also responding to discrete events as snow melting, summer cattle breeding on pastures and trekking frequentation. Series of CO2 obtained are included as part of Long Term Ecological Research (LTER) at the Parc Nacional d'Aigüestortes i Estany de Sant Maurici. In the long term, these measurements show the mountain ecosystems contribution and geomorphologic influence on the CO2 budget of the air masses crossing a mountain range.

Atmospheric Methane Mixing Ratios--The NOAA/CMDL Global Cooperative Air Sampling Network\\, 1983-1993

DOE Data Explorer

Dlugokencky, E. J. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Lang, P. M. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Masarie, K. A. [National Oceanic and Atmospheric Administration, Boulder, Colorado (USA); Steele, L. P. [Commonwealth Scientific and Industrial Research Organisation, Aspendale, Victoria, Australia

1994-01-01

This data base presents atmospheric methane (CH4) mixing ratios from flask air samples collected over the period 1983-1993 by the National Oceanic and Atmospheric Administration, Climate Monitoring and Diagnostics Laboratory's (NOAA/CMDL's) global cooperative air sampling network. Air samples were collected approximately once per week at 44 fixed sites (37 of which were still active at the end of 1993). Samples were also collected at 5 degree latitude intervals along shipboard cruise tracks in the Pacific Ocean between North America and New Zealand (or Australia) and at 3 degree latitude intervals along cruise tracks in the South China Sea between Singapore and Hong Kong. The shipboard measurements were made approximately every 3 weeks per latitude zone by each of two ships in the Pacific Ocean and approximately once every week per latitude zone in the South China Sea. All samples were analyzed for CH4 at the NOAA/CMDL laboratory in Boulder, Colorado, by gas chromatography with flame ionization detection, and each aliquot was referenced to the NOAA/CMDL methane standard scale. In addition to providing the complete set of atmospheric CH4 measurements from flask air samples collected at the NOAA/CMDL network sites, this data base also includes files which list monthly mean mixing ratios derived from the individual flask air measurements. These monthly summary data are available for 35 of the fixed sites and 21 of the shipboard sampling sites.

Simultaneous Extraction Optimization and Analysis of Flavonoids from the Flowers of Tabernaemontana heyneana by High Performance Liquid Chromatography Coupled to Diode Array Detector and Electron Spray Ionization/Mass Spectrometry

PubMed Central

Sathishkumar, Thiyagarajan; Baskar, Ramakrishnan; Aravind, Mohan; Tilak, Suryanarayanan; Deepthi, Sri; Bharathikumar, Vellalore Maruthachalam

2013-01-01

Flavonoids are exploited as antioxidants, antimicrobial, antithrombogenic, antiviral, and antihypercholesterolemic agents. Normally, conventional extraction techniques like soxhlet or shake flask methods provide low yield of flavonoids with structural loss, and thereby, these techniques may be considered as inefficient. In this regard, an attempt was made to optimize the flavonoid extraction using orthogonal design of experiment and subsequent structural elucidation by high-performance liquid chromatography-diode array detector-electron spray ionization/mass spectrometry (HPLC-DAD-ESI/MS) techniques. The shake flask method of flavonoid extraction was observed to provide a yield of 1.2 ± 0.13 (mg/g tissue). With the two different solvents, namely, ethanol and ethyl acetate, tried for the extraction optimization of flavonoid, ethanol (80.1 mg/g tissue) has been proved better than ethyl acetate (20.5 mg/g tissue). The optimal conditions of the extraction of flavonoid were found to be 85°C, 3 hours with a material ratio of 1 : 20, 75% ethanol, and 1 cycle of extraction. About seven different phenolics like robinin, quercetin, rutin, sinapoyl-hexoside, dicaffeic acid, and two unknown compounds were identified for the first time in the flowers of T. heyneana. The study has also concluded that L16 orthogonal design of experiment is an effective method for the extraction of flavonoid than the shake flask method. PMID:25969771

Optimization of Ex Vivo Murine Bone Marrow Derived Immature Dendritic Cells: A Comparative Analysis of Flask Culture Method and Mouse CD11c Positive Selection Kit Method.

PubMed

Gosavi, Rahul Ashok; Salwe, Sukeshani; Mukherjee, Sandeepan; Dahake, Ritwik; Kothari, Sweta; Patel, Vainav; Chowdhary, Abhay; Deshmukh, Ranjana A

2018-01-01

12-14 days of culturing of bone marrow (BM) cells containing various growth factors is widely used method for generating dendritic cells (DCs) from suspended cell population. Here we compared flask culture method and commercially available CD11c Positive Selection kit method. Immature BMDCs' purity of adherent as well as suspended cell population was generated in the decreasing concentration of recombinant-murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF) in nontreated tissue culture flasks. The expression of CD11c, MHCII, CD40, and CD86 was measured by flow cytometry. We found significant difference ( P < 0.05) between the two methods in the adherent cells population but no significant difference was observed between the suspended cell populations with respect to CD11c+ count. However, CD11c+ was significantly higher in both adhered and suspended cell population by culture method but kit method gave more CD11c+ from suspended cells population only. On the other hand, using both methods, immature DC expressed moderate level of MHC class II molecules as well as low levels of CD40 and CD86. Our findings suggest that widely used culture method gives the best results in terms of yield, viability, and purity of BMDCs from both adherent and suspended cell population whereas kit method works well for suspended cell population.

Metals in Metal Salts: A Copper Mirror Demonstration

ERIC Educational Resources Information Center

Pike, Robert D.

2010-01-01

A simple lecture demonstration is described to show the latent presence of metal atoms in a metal salt. Copper(II) formate tetrahydrate is heated in a round-bottom flask forming a high-quality copper mirror.

75 FR 67714 - Notice of Intent To Suspend Certain Pesticide Registrations

Federal Register 2010, 2011, 2012, 2013, 2014

2010-11-03

.... August 30, 2008.. No data received. coefficient (n- octanol/water) shake flask method. 830.7570 Partition December 14, 2007. December 24, 2007. August 30, 2008.. No data received. coefficient (n- octanol/water...

Modifications to the algal growth inhibition test for use as a regulatory assay

DOE Office of Scientific and Technical Information (OSTI.GOV)

Geis, S.W.; Fleming, K.L.; Korthals, E.T.

2000-01-01

Biological assays using aquatic invertebrates and fish do not necessarily predict protection levels for primary producers such as algae and aquatic macrophytes. State regulatory programs may not be protecting the environment from many phytotoxic compounds. Recent modifications of the US Environmental Protection Agency's algal test were evaluated for their potential use as a regulatory assay. Primary goals of this investigation were to downsize the algal assay and to evaluate various methods of automation. Disposable microplates with 2-ml sample wells were evaluated as an alternative testing chamber for the 96-h growth inhibition test with Raphidocelis subcapitata (formerly known as Selenastrum capricornutum).more » The authors compared the standardized Erlenmeyer {reg_sign} flask test to the microplate test using CuCl{sub 2}, NaCl, phenol, ZnCl{sub 2}, and a surfactant. They noted improved control performance with the microplate test, whereas median inhibitory concentration values were similar for both methods. Other procedures they addressed included the use of EDTA, filtration of samples, and the effect of colored samples on algal growth. They also evaluated growth estimates by comparing manual cell counting to more automated growth estimates using fluorescence and absorbance endpoints. The use of fluorescence and absorbance measurements demonstrated reductions in replicate variability over manual counting and may offer time-saving alternatives for laboratory analysts.« less

Strategies for the production of high-content fructo-oligosaccharides through the removal of small saccharides by co-culture or successive fermentation with yeast.

PubMed

Nobre, C; Castro, C C; Hantson, A-L; Teixeira, J A; De Weireld, G; Rodrigues, L R

2016-01-20

Fructo-oligosaccharides (FOS) obtained by fermentation of sucrose may be purified at large-scale by continuous chromatography (Simulated Moving Bed: SMB). In order to improve the efficiency of the subsequent SMB purification, the optimization of the fermentative broth composition in salts and sugars was investigated. Fermentations conducted at reduced amount of salts, using Aureobasidium pullulans whole cells, yielded 0.63 ± 0.03 g of FOS per gram of initial sucrose. Additionally, a microbial treatment was proposed to reduce the amount of small saccharides in the mixture. Two approaches were evaluated, namely a co-culture of A. pullulans with Saccharomyces cerevisiae; and a two-step fermentation in which FOS were first synthesized by A. pullulans and then the small saccharides were metabolized by S. cerevisiae. Assays were performed in 100mL shaken flasks and further scaled-up to a 3 L working volume bioreactor. Fermentations in two-step were found to be more efficient than the co-culture ones. FOS were obtained with a purity of 81.6 ± 0.8% (w/w), on a dry weight basis, after the second-step fermentation with S. cerevisiae. The sucrose amount was reduced from 13.5 to 5.4% in total sugars, which suggests that FOS from this culture broth will be more efficiently separated by SMB. Copyright © 2015 Elsevier Ltd. All rights reserved.

Suppression of pyritic sulphur during flotation tests using the bacterium Thiobacillus ferrooxidans.

PubMed

Townsley, C C; Atkins, A S; Davis, A J

1987-07-01

Environmental concern about sulphur dioxide emissions has led to the examination of the possibility of removing pyritic sulphur from coal prior to combustion during froth flotation, a routine method for coal cleaning at the pit-head. The bacterium Thiobacillus ferrooxidans was effective in leaching 80% and 63% -53 mum pyrite at 2% and 6% pulp density in shake flasks in 240 and 340 h, respectively.The natural floatability of pyrite was significantly reduced in the Hallimond tube following 2.5 min of conditioning in membrane-filtered bacterial liquor prior to flotation. The suppression effect was greatly enhanced in the presence of Thiobacillus ferrooxidans. A bacterial suspension in pH 2.0 distilled water showed 85% suppression, whereas in spent growth liquor this value was 95%. The optimum bacterial density was 3.25 x 10(10) cells/g pyrite in 230-ml distilled water (2% pulp density) in the Hallimond tube. The degree of suppression by the cells was related to particle size but not to pH or temperature. The sulphur content of a synthetic coal/pyrite mixture was reduced from 10.9 to 2.1% by flotation after bacterial preconditioning. It is postulated that pyrite removal in coals which are cleaned by froth flotation could be significantly reduced using a bacterial preconditioning stage with a short residence time of 2.5 min.

[Using Excess Activated Sludge Treated 4-Chlorophenol Contained Waste Water to Cultivate Chlorella vulgaris].

PubMed

Wang, Lu; Chen, Xiu-rong; Yan, Long; He, Yi-xuan; Shi, Zhen-dong

2015-04-01

Using different rations of sludge extracts and supernate from 4-Chlorophenol (4-CP) simulated wastewater's excess sludge after centrifugation to cultivate the Chlorella vulgaris to achieve the goal of excess sludge utilization together with chlorella cultivating. The experiments were performed in 500 mL flasks with different rations of sludge extracts & BG-11 and supernate & BG-11 in a light growth chamber respectively. Number of algal cells, Chlorophyll, enzyme activity, oil and water total nitrogen (TN), total phosphorus (TP), total organic carbon (TOC), toxicity index were investigated. Result showed that the nutrition supplies and toxicity in the excess sludge were removed efficiently via Chlorella vulgaris, the removal rates of TN and TP were at least 40% and 90% respectively; After 10 days cultivation, the density growth of 50% sludge extracts was 20 times higher of the beginning while its chlorophyll content was lower than that of the blank group. Sludge extracts could promote the proliferation of algae, but were not conducive to the synthesis of chlorophyll. The quantity of SOD in per cell showed Chlorella vulgaris gave a positive response via stimulation from toxicant in sludge extracts and supernate. The best time for collecting chlorella vulgaris was the fifth day of cultivation, taking neutral oil accumulation as the evaluating indicator for its utilization combined with the removal of supplies and toxicity.

Ultrasound assisted dialysis of semi-permeable membrane devices for the simultaneous analysis of a wide number of persistent organic pollutants.

PubMed

Bustamante, Julen; Navarro, Patricia; Arana, Gorka; de Diego, Alberto; Madariaga, Juan Manuel

2013-09-30

A new procedure based on ultrasound assisted dialysis (UAD) for the simultaneous and quantitative extraction of a wide number of persistent organic pollutants (POPs) such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) or some other organochlorinated pesticides (OCPs) contained in semi-permeable membrane devices (SPMDs) has been developed. This extraction technique combines the advantages of the organic solvent dialysis (OSD) and the speed of the ultrasound assisted extraction. The extraction was performed in an ultrasound bath for 32 min placing the SPMD in a glass flask covered with 80 mL of hexane. This set-up is able to extract simultaneously up to 8 samples. The proposed method entails good repeatabilities (RSD 2-13%) and recoveries (around 100% for almost every analyte). Limits of detection were at ng SPMD(-1) level and enough for the determination of the target analytes in a slightly polluted aquatic environment, as it was tested by successfully comparing the OSD to the proposed methodology. Therefore, the results obtained show that the UAD can be a good alternative for the extraction of POPs in SPMDs as it requires short extraction times and solvent volumes, and provides a cleaner extract for the subsequent clean-up step. Moreover, it fits better than the OSD to the general requirements of Green Chemistry. Copyright © 2013 Elsevier B.V. All rights reserved.

Construction of a highly thermostable 1,3-1,4-β-glucanase by combinational mutagenesis and its potential application in the brewing industry.

PubMed

Niu, Chengtuo; Zhu, Linjiang; Hill, Annie; Alex Speers, R; Li, Qi

2017-01-01

To improve the thermostability and catalytic property of a mesophilic 1,3-1,4-β-glucanase by combinational mutagenesis and to test its effect in congress mashing. A mutant β-glucanase (rE-BglTO) constructed by combinational mutagenesis showed a 25 °C increase in optimal temperature (to 70 °C) a 19.5 °C rise in T 50 value and a 15.6 °C increase in melting temperature compared to wild-type enzyme. Its half-life values at 60 and 70 °C were 152 and 99 min, which were 370 and 800 % higher than those of wild-type enzyme. Besides, its specific activity and k cat value were 42,734 U mg -1 and 189 s -1 while its stability under acidic conditions was also improved. In flask fermentation, the catalytic activity of rE-BglTO reached 2381 U ml -1 , which was 63 % higher than that of wild-type enzyme. The addition of rE-BglTO in congress mashing decreased the filtration time and viscosity by 21.3 and 9.6 %, respectively. The mutant β-glucanase showed high catalytic activity and thermostability which indicated that rE-BglTO is a good candidate for application in the brewing industry.

In silico and in vitro prediction of gastrointestinal absorption from potential drug eremantholide C.

PubMed

Caldeira, Tamires G; Saúde-Guimarães, Dênia A; Dezani, André B; Serra, Cristina Helena Dos Reis; de Souza, Jacqueline

2017-11-01

Analysis of the biopharmaceutical properties of eremantholide C, sesquiterpene lactone with proven pharmacological activity and low toxicity, is required to evaluate its potential to become a drug. Preliminary analysis of the physicochemical characteristics of eremantholide C was performed in silico. Equilibrium solubility was evaluated using the shake-flask method, at 37.0 °C, 100 rpm during 72 h in biorelevant media. The permeability was analysed using parallel artificial membrane permeability assay, at 37.0 °C, 50 rpm for 5 h. The donor compartment was composed of an eremantholide C solution in intestinal fluid simulated without enzymes, while the acceptor compartment consisted of phosphate buffer. Physicochemical characteristics predicted in silico indicated that eremantholide C has a low solubility and high permeability. In-vitro data of eremantholide C showed low solubility, with values for the dose/solubility ratio (ml): 9448.82, 10 389.61 e 15 000.00 for buffers acetate (pH 4.5), intestinal fluid simulated without enzymes (pH 6.8) and phosphate (pH 7.4), respectively. Also, it showed high permeability, with effective permeability of 30.4 × 10 -6 cm/s, a higher result compared with propranolol hydrochloride (9.23 × 10 -6 cm/s). The high permeability combined with its solubility, pharmacological activity and low toxicity demonstrate the importance of eremantholide C as a potential drug candidate. © 2017 Royal Pharmaceutical Society.

A method for the solvent extraction of low-boiling-point plant volatiles.

PubMed

Xu, Ning; Gruber, Margaret; Westcott, Neil; Soroka, Julie; Parkin, Isobel; Hegedus, Dwayne

2005-01-01

A new method has been developed for the extraction of volatiles from plant materials and tested on seedling tissue and mature leaves of Arabidopsis thaliana, pine needles and commercial mixtures of plant volatiles. Volatiles were extracted with n-pentane and then subjected to quick distillation at a moderate temperature. Under these conditions, compounds such as pigments, waxes and non-volatile compounds remained undistilled, while short-chain volatile compounds were distilled into a receiving flask using a high-efficiency condenser. Removal of the n-pentane and concentration of the volatiles in the receiving flask was carried out using a Vigreux column condenser prior to GC-MS. The method is ideal for the rapid extraction of low-boiling-point volatiles from small amounts of plant material, such as is required when conducting metabolic profiling or defining biological properties of volatile components from large numbers of mutant lines.

Globally Averaged Atmospheric CFC-11 Concentrations: Monthly and Annual Data for the Period 1975-1992 (DB1010)

DOE Data Explorer

Khalil, M. A.K. [Oregon Graduate Institute of Science and Technology Portland, Oregon (USA); Rasmussen, R. A. [Oregon Graduate Institute of Science and Technology Portland, Oregon

1996-01-01

This data set presents globally averaged atmospheric concentrations of chlorofluorocarbon 11, known also as CFC-11 or F-11 (chemical name: trichlorofluoromethane; formula: CCl3F). The monthly global average data are derived from flask air samples collected at eight sites in six locations over the period August 1980-July 1992. The sites are Barrow (Alaska), Cape Meares (Oregon), Cape Kumukahi and Mauna Loa (Hawaii), Cape Matatula (American Samoa), Cape Grim (Tasmania), Palmer Station, and the South Pole (Antarctica). At each collection site, monthly averages were obtained from three flask samples collected every week. In addition to the monthly global averages available for 1980-992, this data set also contains annual global average data for 1975-1985. These annual global averages were derived from January measurements at the South Pole and in the Pacific Northwest of the United States (specifically, Washington state and the Oregon coast).

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin.

PubMed

Sar, Taner; Stark, Benjamin C; Yesilcimen Akbas, Meltem

2017-03-04

Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48-96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli.

Effective ethanol production from whey powder through immobilized E. coli expressing Vitreoscilla hemoglobin

PubMed Central

Sar, Taner; Stark, Benjamin C.; Yesilcimen Akbas, Meltem

2017-01-01

ABSTRACT Ethanol production from whey powder was investigated by using free as well as alginate immobilized E. coli and E. coli expressing Vitreoscilla hemoglobin (VHb) in both shake flask and fermenter cultures. Media with varying levels of whey (lactose contents of 3%, 5%, 8% or 15%) and yeast extract (0.3% or 0.5%) were evaluated with fermentation times of 48–96 h. Immobilization and VHb expression resulted in higher ethanol production with all media; the increases ranged from 2% to 89% for immobilization and from 2% to 182% for VHb expression. It was determined that growth medium containing 8% lactose with 0.5% yeast extract yielded the highest ethanol production for free or immobilized strains, with or without VHb expression, in both shake flask and fermenter cultures. Immobilization with alginate was found to be a promising process for ethanol production by VHb-expressing ethanologenic E. coli. PMID:27579556

Optimization of growth for the hyperthermophilic archaeon Aeropyrum pernix on a small-batch scale.

PubMed

Milek, Igor; Cigic, Blaz; Skrt, Mihaela; Kaletunç, Gönül; Ulrih, Natasa Poklar

2005-09-01

Growth of Aeropyrum pernix, the first reported aerobic neutrophilic hyperthermophilic archaeon, was investigated under different cultivation parameters. Different sources of seawater, pH, and the cultivation methods were tested with the aim to improve the biomass production. A 1-L glass flask fitted with a condenser and air diffuser was used as a bioreactor. The optimum conditions for maximizing A. pernix biomass were obtained when Na2S2O3.5H2O (1 g/L) with added marine broth 2216 at pH 7.0 (20 mmol HEPES buffer/L) was used as a growing medium in a 1-L flask. The biomass production was 0.45 g dry cell mass/L in 40 h under the optimum conditions, which is more than the 0.42 g dry cell mass/L in 60 h previously obtained.

Atmospheric CO2 Concentrations--The Canadian Background Air Pollution Monitoring Network (1993) (NDP-034)

DOE Data Explorer

Trivett, N. B. A. [Environment Canada, Atmospheric Environment Service, Downsview, Ontario, Canada; Hudec, V. C. [Environment Canada, Atmospheric Environment Service, Downsview, Ontario, Canada; Wong, C. S. [Marine Carbon Research Centre, Institute of Ocean Sciences, Sidney, British Columbia, Canada

1993-01-01

Flask air samples collected at roughly weekly intervals at three Canadian sites [Alert, Northwest Territories (July 1975 through July 1992); Sable Island, Nova Scotia (March 1975 through July 1992); and Cape St. James, British Columbia (May 1979 through July 1992)] were analyzed for CO2 concentration with the measurements directly traceable to the WMO primary CO2 standards. Each record includes the date, atmospheric CO2 concentration, and flask classification code. They provide an accurate record of CO2 concentration levels in Canada during the past two decades. Because these data are directly traceable to WMO standards, this record may be compared with records from other Background Air Pollution Monitoring Network (BAPMoN) stations. The data are in three files (one for each of the monitoring stations) ranging in size from 9.4 to 20.1 kB.