Differential inhibitory and activating NK cell receptor levels and NK/NKT-like cell functionality in chronic and recovered stages of chikungunya.

Science.gov (United States)

Thanapati, Subrat; Ganu, Mohini A; Tripathy, Anuradha S

2017-01-01

The role of natural killer (NK; CD3-CD56+)/NKT (CD3+CD56+)-like cells in chikungunya virus (CHIKV) disease progression/recovery remains unclear. Here, we investigated the expression profiles and function of NK and NKT-like cells from 35 chronic chikungunya patients, 30 recovered individuals, and 69 controls. Percentage of NKT-like cells was low in chronic chikungunya patients. NKp30+, CD244+, DNAM-1+, and NKG2D+ NK cell percentages were also lower (MFI and/or percentage), while those of CD94+ and NKG2A+ NKT-like cells were higher (MFI and/or percentage) in chronic patients than in recovered subjects. IFN-γ and TNF-α expression on NKT-like cells was high in the chronic patients, while only IFN-γ expression on NK cells was high in the recovered individuals. Furthermore, percentage of perforin+NK cells was low in the chronic patients. Lower cytotoxic activity was observed in the chronic patients than in the controls. CD107a expression on NK and NKT-like cells post anti-CD94/anti-NKG2A blocking was comparable among the patients and controls. Upregulated inhibitory and downregulated activating NK receptor expressions on NK/NKT-like cells, lower perforin+ and CD107a+NK cells are likely responsible for inhibiting the NK and NKT-like cell function in the chronic stage of chikungunya. Therefore, deregulation of NKR expression might underlie CHIKV-induced chronicity.

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

KAUST Repository

Wittum, Rebecca

2017-12-07

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

Ethical dilemma of recovering organs before donor death.

Science.gov (United States)

Khalid, Umair; Khalil, Sarwat

2011-08-01

Organ transplant has had a momentous effect in improving global health over the years. However, there exists a sizeable discrepancy between the supply and demand of organs, especially in developing countries, where lack of expertise, financial constraints, and inadequate transplant facilities have been obstacles. According to current practice, donors must be dead before unpaired vital organs can be recovered. Equal health warrants needs-based health care for everyone. Recovering viable organs from donors while they are alive, but with death inevitable, may be significant to persons on waiting lists. Future policies in organ transplantation must be made after thorough consideration of all aspects of donation and dealing with the inequalities of health care. These pose a challenge for medicolegal and ethical organizations.

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

Science.gov (United States)

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

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

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

Directory of Open Access Journals (Sweden)

Maria-Judith Gonsalves

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

Dye Sensitized Solar Cells for Economically Viable Photovoltaic Systems.

Science.gov (United States)

Jung, Hyun Suk; Lee, Jung-Kun

2013-05-16

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

Promoting resuscitation of viable but nonculturable cells of Vibrio harveyi by a resuscitation-promoting factor-like protein YeaZ.

Science.gov (United States)

Li, Y; Chen, J; Zhao, M; Yang, Z; Yue, L; Zhang, X

2017-02-01

To demonstrate the resuscitation-promoting activities of recombinant YeaZ from Vibrio harveyi SF-1. The gene of resuscitation-promoting factor YeaZ was cloned from genomic DNA of V. harveyi SF-1. The gene was expressed in Escherichia coli, and the expressed protein was purified by Ni 2+ -affinity chromatography. A yeaZ mutant was constructed by using the suicide plasmid pNQ705 with homologous recombination. Disruption of yeaZ did not affect cell growth significantly in 2216 E broth at 28°C. The wild-type and mutant viable but nonculturable (VBNC) cells could be resuscitated by temperature upshift method. In addition, the recombinant YeaZ increased the culturable counts from 1·27 × 10 4  CFU per ml and 1·99 × 10 4 CFU per ml to 2·88 × 10 5  CFU per ml and 4·59 × 10 5 CFU per ml, respectively. After the VBNC cells of wild-type and mutant cells were maintained at 4°C for 120 days, no resuscitation was obtained by temperature upshift method, but addition of the recombinant YeaZ promoted the resuscitation of the wild-type and mutant cells, with the culturable cell counts of 1·13 × 10 3 and 1·44 × 10 3 CFU per ml, respectively. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. The lethal dose 50% of the yeaZ null mutant was more than 10-fold higher than that of the wild-type cells. The recombinant YeaZ could efficiently promote resuscitation of the wild-type and mutant cells of V. harveyi from VBNC to culturable state. The protein also promoted resuscitation of the VBNC wild-type and mutant cells, which were maintained at 4°C for 120 days and not recovered by temperature upshift method. Disruption of yeaZ decreased the virulence of V. harveyi in zebrafish. Here, we show clear evidence of a resuscitation-promoting factor YeaZ of V. harveyi and the roles in resuscitation of the VBNC cells and its pathogenicity. © 2016 The Society for Applied Microbiology.

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

Science.gov (United States)

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

2012-12-01

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

PMA-PhyloChip DNA Microarray to Elucidate Viable Microbial Community Structure

Science.gov (United States)

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

2011-01-01

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

Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Constructs

Energy Technology Data Exchange (ETDEWEB)

Michael Cornforth

2012-03-26

Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. Specific aims apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. The project includes research complementary to NASA/HRP space radiation project.

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

Science.gov (United States)

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

2012-10-01

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

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

Science.gov (United States)

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

2016-09-01

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

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

Science.gov (United States)

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

2013-01-01

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

Selective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide.

Science.gov (United States)

Tian, Qian; Feng, Jian-Jun; Hu, Jie; Zhao, Wen-Jun

2016-10-14

In recent years, use of the DNA-intercalating dye propidium monoazide (PMA) in real-time PCR has been reported as a novel method to detect viable bacteria in different types of samples, such as food, environmental, and microbiological samples. In this study, viable cells of Acidovorax citrulli, the causal agent of bacterial seedling blight and fruit blotch, were selectively detected and differentiated from dead cells by real-time fluorescent polymerase chain reaction amplification after the bacterial solution was treated with the DNA-binding dye PMA. The primers and TaqMan probe were based on the A. citrulli genome (Aave_1909, Gene ID: 4669443) and were highly specific for A. citrulli. The detection threshold of this assay was 10 3 colony-forming units per mL (CFU/mL) in pure cell suspensions containing viable and dead cells and infected watermelon seeds. Application of this assay enables the selective detection of viable cells of A. citrulli and facilitates monitoring of the pathogen in watermelon and melon seeds.

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

Energy Technology Data Exchange (ETDEWEB)

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

2012-04-12

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

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

Directory of Open Access Journals (Sweden)

Mohammad Salma

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

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

Science.gov (United States)

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

2002-01-01

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

Introduce of Viable But Nonculturable Bacteria

Directory of Open Access Journals (Sweden)

Mehdi Hassanshahian

2008-03-01

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

Survival of Lymphatic Cells after X-Irradiation in Mice

Energy Technology Data Exchange (ETDEWEB)

Vos, O. [Medical Biological Laboratory, National Defense Research Organization TNO, Ruswuk, Z.H. (Netherlands)

1967-07-15

Lymphatic tissues are generally classified among the most radiosensitive tissues of the body. The main reason for this is that histologically extensive destruction is found within a few hours after irradiation. We tried to estimate the degree of cellular degeneration by making cell suspensions from lymph nodes and thymus of mice at different times after X-irradiation with 800 R or at 24 h after radiation with different doses. The numbers of normal viable cells we obtained were expressed as percentages of the cells recovered from unirradiated control mice.

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

Science.gov (United States)

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

2017-11-27

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

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

Directory of Open Access Journals (Sweden)

Carlos Alfaro

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

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

International Nuclear Information System (INIS)

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

2013-01-01

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

Cultured bovine granulosa cells rapidly lose important features of their identity and functionality but partially recover under long-term culture conditions.

Science.gov (United States)

Yenuganti, Vengala Rao; Vanselow, Jens

2017-05-01

Cell culture models are essential for the detailed study of molecular processes. We analyze the dynamics of changes in a culture model of bovine granulosa cells. The cells were cultured for up to 8 days and analyzed for steroid production and gene expression. According to the expression of the marker genes CDH1, CDH2 and VIM, the cells maintained their mesenchymal character throughout the time of culture. In contrast, the levels of functionally important transcripts and of estradiol and progesterone production were rapidly down-regulated but showed a substantial up-regulation from day 4. FOXL2, a marker for granulosa cell identity, was also rapidly down-regulated after plating but completely recovered towards the end of culture. In contrast, expression of the Sertoli cell marker SOX9 and the lesion/inflammation marker PTGS2 increased during the first 2 days after plating but gradually decreased later on. We conclude that only long-term culture conditions (>4 days) allow the cells to recover from plating stress and to re-acquire characteristic granulosa cell features.

Separation of viable and non-viable tomato (Solanum lycopersicum L.) seeds using single seed near-infrared spectroscopy

DEFF Research Database (Denmark)

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

2017-01-01

Single seed near-infrared (NIR) spectroscopy is a non-destructive technology commonly used for predicting lipids, proteins, carbohydrates and water content of agricultural products. The aim of the current study is to investigate the prospects of NIR spectroscopy in classifying viable and non...... identified as important for classification of viable and non-viable tomato seeds by iPLS-DA. The sensitivity i.e. ability to correctly identify the positive samples and specificity i.e. ability to reject the negative samples of the (iPLS-DA) model on identified spectral regions for prediction of viable......-viable tomato seeds of two cultivars using chemometrics. The data exploration were performed by principal component analysis (PCA). Subsequently, viable and non-viable seeds were classified by partial least squares-discriminant analysis (PLS-DA) and interval PLS-DA (iPLS-DA). The indication of clustering...

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

Directory of Open Access Journals (Sweden)

Chieh-Hsien Lai

2017-07-01

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

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

Science.gov (United States)

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

2017-07-01

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

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

Digital Repository Service at National Institute of Oceanography (India)

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

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

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

Science.gov (United States)

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

2017-12-04

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

Failure of RNA synthesis to recover after UV irradiation: an early defect in cells from individuals with Cockayne's syndrome and xeroderma pigmentosum

International Nuclear Information System (INIS)

Mayne, L.V.; Lehmann, A.R.

1982-01-01

Previous work has shown that in cells from the ultraviolet-sensitive genetic disorder, Cockayne's syndrome, DNA synthesis fails to recover after ultraviolet irradiation, despite the fact that these cells have no detectable defect in either excision or daughter-strand repair pathways. We now show that Cockayne cells, as well as cells from a number of patients with xeroderma pigmentosum, are sensitive to the lethal effects of UV irradiation in stationary phase under conditions in which no DNA is synthesized after irradiation. Furthermore, in normal and defective human fibroblasts, RNA synthesis is depressed after UV irradiation. In normal (dividing) cells, RNA synthesis recovers very rapidly, but this recovery does not occur in Cockayne cells, and it is reduced or absent in xeroderma pigmentosum cells from different complementation groups. Qualitatively, similar results are obtained with cells in stationary phase. The recovery of RNA synthesis in the various defective cell strains is not correlated with the overall extent of excision repair, but there is some correlation between recovery of RNA synthesis and cell survival after ultraviolet irradiation. These results implicate recovery of RNA synthesis as an important early response to ultraviolet irradiation

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

Science.gov (United States)

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

2008-01-01

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

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

Directory of Open Access Journals (Sweden)

Jeremy eDesneux

2016-03-01

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

Long-term persistence of robust antibody and cytotoxic T cell responses in recovered patients infected with SARS coronavirus.

Directory of Open Access Journals (Sweden)

Taisheng Li

2006-12-01

Full Text Available Most of the individuals infected with SARS coronavirus (SARS-CoV spontaneously recovered without clinical intervention. However, the immunological correlates associated with patients' recovery are currently unknown. In this report, we have sequentially monitored 30 recovered patients over a two-year period to characterize temporal changes in SARS-CoV-specific antibody responses as well as cytotoxic T cell (CTL responses. We have found persistence of robust antibody and CTL responses in all of the study subjects throughout the study period, with a moderate decline one year after the onset of symptoms. We have also identified two potential major CTL epitopes in N proteins based on ELISPOT analysis of pooled peptides. However, despite the potent immune responses and clinical recovery, peripheral lymphocyte counts in the recovered patients have not yet been restored to normal levels. In summary, our study has, for the first time, characterized the temporal and dynamic changes of humoral and CTL responses in the natural history of SARS-recovered individuals, and strongly supports the notion that high and sustainable levels of immune responses correlate strongly with the disease outcome. Our findings have direct implications for future design and development of effective therapeutic agents and vaccines against SARS-CoV infection.

Viable Species of Flamella (Amoebozoa: Variosea) Isolated from Ancient Arctic Permafrost Sediments.

Science.gov (United States)

Shmakova, Lyubov; Bondarenko, Natalya; Smirnov, Alexey

2016-02-01

Six viable strains of amoebae belonging to the genus Flamella (Amoebozoa, Variosea) were isolated from permafrost sediments sampled in the Russian Arctic region. Two of them are from late Pleistocene permafrost in North-East Siberia, and four--from Holocene and late Pleistocene in North-West Siberia. Light- and electron-microscopic study and molecular phylogeny show that these isolates represent two new species belonging to the genus Flamella. Both species are cyst-forming. This is a remarkable case of high resistance of protozoan cysts, allowing them to survive and recover an amoebae population after a very long, geologically significant period of rest; a "snapshot" of evolution in time. This study directly shows for the first time that amoeba cysts can be conserved not only for years and decades but for many thousand years and then recover, contributing to the formation of an active microbial community. We propose to name the new species as Flamella pleistocenica n.sp. and Flamella beringiania n.sp. Phylogenetic analysis shows that the genus Flamella is a robust and potentially species-rich group of Variosea. Copyright © 2015 Elsevier GmbH. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Ching Giap Tan

2014-09-01

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

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

Science.gov (United States)

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

2018-03-09

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

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

International Nuclear Information System (INIS)

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

1997-01-01

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

Biocatalytically active silCoat-composites entrapping viable Escherichia coli.

Science.gov (United States)

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

2014-02-01

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

Stereotaxic implantation of dispersed cell suspensions into brain. A systematic appraisal of cell placement and survival

International Nuclear Information System (INIS)

Plunkett, R.J.; Weber, R.J.; Oldfield, E.H.

1988-01-01

The application of several recent advances in cell biology, brain implantation, and cell-mediated tumor immunotherapy requires successful and reproducible placement of viable cell suspensions into brain. Stereotaxic implantation is being used to inject cytotoxic lymphocytes into gliomas and to replace dopaminergic cells in parkinsonian models. Systematic assessment of the factors that influence success in implantation of cell suspensions into solid tissues is needed. A model was developed for investigation of stereotaxic implantation using radiolabeled rat lymphokine-activated killer (LAK) cells. Anesthetized rats received microliter injections of cell suspension into the right caudate nucleus. The injection volume, cell concentration, infusion rate, and needle size were varied systematically. The animals were sacrificed 1 hour after injection; the brain was removed and sectioned, and the radioactivity was counted. Three aliquots of the suspension were injected into counting tubes for control analysis. Recovery of radioactivity was expressed as the percent of mean counts per minute (cpm) in the right frontal lobe/mean cpm in the three control tubes. To assess the viability of implanted cells, the right frontal region was mechanically dissociated in media and centrifuged, and the pellet and supernatant were counted. By using small needles and slow infusion of volumes of 10 microliters or less, 85% to 90% of the radioactivity was recovered in the caudate nucleus. At least half of the implanted cells were viable. Consistent, accurate implantation of dispersed cells into brain over a range of volumes, cell concentrations, infusion rates, and needle sizes was achieved

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

Science.gov (United States)

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

2017-09-27

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

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

Science.gov (United States)

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

2014-11-01

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

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

Science.gov (United States)

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

2014-02-01

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

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

Directory of Open Access Journals (Sweden)

Pontus Thulin

2006-03-01

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

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

Directory of Open Access Journals (Sweden)

2006-01-01

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

Revisiting the single cell protein application of Cupriavidus necator H16 and recovering bioplastic granules simultaneously.

Directory of Open Access Journals (Sweden)

Balakrishnan Kunasundari

Full Text Available Cupriavidus necator H16 (formerly known as Hydrogenomonas eutropha was famous as a potential single cell protein (SCP in the 1970s. The drawback however was the undesirably efficient accumulation of non-nutritive polyhydroxybutyrate (PHB storage compound in the cytoplasm of this bacterium. Eventually, competition from soy-based protein resulted in SCP not receiving much attention. Nevertheless, C. necator H16 remained in the limelight as a producer of PHB, which is a material that resembles commodity plastics such as polypropylene. PHB is a 100% biobased and biodegradable polyester. Although tremendous achievements have been attained in the past 3 decades in the efficient production of PHB, this bioplastic is still costly. One of the main problems has been the recovery of PHB from the cell cytoplasm. In this study, we showed for the first time that kilogram quantities of PHB can be easily recovered in the laboratory without the use of any solvents and chemicals, just by using the cells as SCP. In addition, the present study also demonstrated the safety and tolerability of animal model used, Sprague Dawley given lyophilized cells of C. necator H16. The test animals readily produced fecal pellets that were whitish in color, as would be expected of PHB granules. The pellets were determined to contain about 82-97 wt% PHB and possessed molecular mass of around 930 kg/mol. The PHB granules recovered biologically possessed similar molecular mass compared to chloroform extracted PHB [950 kg/mol]. This method now allows the production and purification of substantial quantities of PHB for various experimental trials. The method reported here is easy, does not require expensive instrumentation, scalable and does not involve extensive use of solvents and strong chemicals.

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

Czech Academy of Sciences Publication Activity Database

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

2017-01-01

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

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

Science.gov (United States)

Liu, Yarui; Mustapha, Azlin

2014-01-17

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

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

Science.gov (United States)

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

2015-01-01

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

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

Science.gov (United States)

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

2018-03-14

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

Methods for recovering metals from electronic waste, and related systems

Science.gov (United States)

Lister, Tedd E; Parkman, Jacob A; Diaz Aldana, Luis A; Clark, Gemma; Dufek, Eric J; Keller, Philip

2017-10-03

A method of recovering metals from electronic waste comprises providing a powder comprising electronic waste in at least a first reactor and a second reactor and providing an electrolyte comprising at least ferric ions in an electrochemical cell in fluid communication with the first reactor and the second reactor. The method further includes contacting the powders within the first reactor and the second reactor with the electrolyte to dissolve at least one base metal from each reactor into the electrolyte and reduce at least some of the ferric ions to ferrous ions. The ferrous ions are oxidized at an anode of the electrochemical cell to regenerate the ferric ions. The powder within the second reactor comprises a higher weight percent of the at least one base metal than the powder in the first reactor. Additional methods of recovering metals from electronic waste are also described, as well as an apparatus of recovering metals from electronic waste.

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

Science.gov (United States)

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

2000-12-01

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

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

Science.gov (United States)

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

2018-01-01

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

Dissemination of Walker 256 carcinoma cells to rat skeletal muscle

International Nuclear Information System (INIS)

Ueoka, H.; Hayashi, K.; Namba, T.; Grob, D.

1986-01-01

After injection of 10 6 Walker 256 carcinoma cells labelled with 125 I-5-iodo-2'-deoxyuridine into the tail vein, peak concentration in skeletal muscle was 46 cells/g at 60 minutes, which was lower than 169202, 1665, 555, 198 and 133 cells/g, respectively, at 30 or 60 minutes in lung, liver, spleen, kidney and heart. Because skeletal muscle constitutes 37.4% of body weight, the total number of tumor cells was 2323 cells, which was much greater than in spleen, kidney and heart with 238, 271, and 85 cells, respectively, and only less than in lung and liver, at 222857 and 11700 cells, respectively. The total number in skeletal muscle became greater than in liver at 4 hours and than in lung at 24 hours. Ten minutes after injection of 7.5 x 10 6 Walker 256 carcinoma cells into the abdominal aorta of rats, a mean of 31 colony-forming cells were recovered from the gastrocnemius, while 106 cells were recovered from the lung after injection into the tail vein. These results indicate that a large number of viable tumor cells can be arrested in skeletal muscle through circulation. The rare remote metastasis of malignancies into skeletal muscle despite constantly circulating tumor cells does not appear to be due to poor dissemination of tumor cells into muscle but due to unhospitable environment of skeletal muscle

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

KAUST Repository

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

2014-01-01

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

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

KAUST Repository

Salam, Khaled W.

2014-08-23

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

Bio-electrosprayed multicellular zebrafish embryos are viable and develop normally

International Nuclear Information System (INIS)

Clarke, Jonathan D W; Jayasinghe, Suwan N

2008-01-01

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

Cannabidiol Reduces Leukemic Cell Size - But Is It Important?

Science.gov (United States)

Kalenderoglou, Nikoletta; Macpherson, Tara; Wright, Karen L

2017-01-01

The anti-cancer effect of the plant-derived cannabinoid, cannabidiol, has been widely demonstrated both in vivo and in vitro . However, this body of preclinical work has not been translated into clinical use. Key issues around this failure can be related to narrow dose effects, the cell model used and incomplete efficacy. A model of acute lymphoblastic disease, the Jurkat T cell line, has been used extensively to study the cannabinoid system in the immune system and cannabinoid-induced apoptosis. Using these cells, this study sought to investigate the outcome of those remaining viable cells post-treatment with cannabidiol, both in terms of cell size and tracking any subsequent recovery. The phosphorylation status of the mammalian Target of Rapamycin (mTOR) signaling pathway and the downstream target ribosomal protein S6, were measured. The ability of cannabidiol to exert its effect on cell viability was also evaluated in physiological oxygen conditions. Cannabidiol reduced cell viability incompletely, and slowed the cell cycle with fewer cells in the G2/M phase of the cell cycle. Cannabidiol reduced phosphorylation of mTOR, PKB and S6 pathways related to survival and cell size. The remaining population of viable cells that were cultured in nutrient rich conditions post-treatment were able to proliferate, but did not recover to control cell numbers. However, the proportion of viable cells that were gated as small, increased in response to cannabidiol and normally sized cells decreased. This proportion of small cells persisted in the recovery period and did not return to basal levels. Finally, cells grown in 12% oxygen (physiological normoxia) were more resistant to cannabidiol. In conclusion, these results indicate that cannabidiol causes a reduction in cell size, which persists post-treatment. However, resistance to cannabidiol under physiological normoxia for these cells would imply that cannabidiol may not be useful in the clinic as an anti-leukemic agent.

Workers' Self-management, Recovered Companies and the Sociology of Work.

Science.gov (United States)

Ozarow, Daniel; Croucher, Richard

2014-10-01

We analyse how far Argentina's worker-recovered companies (WRCs) have sustained themselves and their principles of equity and workers' self-management since becoming widespread following the country's 2001-2 economic crisis. Specialist Spanish-language sources, survey data and documents are analysed through four key sociological themes. We find that the number of WRCs has increased in Argentina, and that they represent a viable production model. Further, they have generally maintained their central principles and even flourished. This occurred despite the global economic crisis, legal and financial pressures to adopt capitalist practices and management structures, the risk of market absorption and state attempts to coopt, demobilise and depoliticise the movement. We argue that today they function as a much-needed international beacon of an alternative vision for labour and that integration of their experience has potential to revitalise the field.

Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Contracts. Final report

Energy Technology Data Exchange (ETDEWEB)

Cornforth, Michael N. [The University of Texas Medical Branch at Galveston, TX (United States)

2013-05-03

Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. The aims of this work apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. These aims are: to analyze by multi-flour fluorescence in situ hybridization (mFISH) the chromosomes in clonal descendents of individual human fibroblasts that were previously irradiated; to examine irradiated clones from Aim 1 for submicroscopic deletions by subjecting their DNA to comparative genomic hybridization (CGH) microarray analysis; and to flow-sort aberrant chromosomes from clones containing stable radiation-induced translocations and map the breakpoints to within an average resolution of 100 kb using the technique of 'array painting'.

Heritable Genetic Changes in Cells Recovered From Irradiated 3D Tissue Contracts. Final report

International Nuclear Information System (INIS)

Cornforth, Michael N.

2013-01-01

Combining contemporary cytogenetic methods with DNA CGH microarray technology and chromosome flow-sorting increases substantially the ability to resolve exchange breakpoints associated with interstitial deletions and translocations, allowing the consequences of radiation damage to be directly measured at low doses, while also providing valuable insights into molecular mechanisms of misrepair processes that, in turn, identify appropriate biophysical models of risk at low doses. The aims of this work apply to cells recovered from 3D tissue constructs of human skin and, for the purpose of comparison, the same cells irradiated in traditional 2D cultures. These aims are: to analyze by multi-flour fluorescence in situ hybridization (mFISH) the chromosomes in clonal descendents of individual human fibroblasts that were previously irradiated; to examine irradiated clones from Aim 1 for submicroscopic deletions by subjecting their DNA to comparative genomic hybridization (CGH) microarray analysis; and to flow-sort aberrant chromosomes from clones containing stable radiation-induced translocations and map the breakpoints to within an average resolution of 100 kb using the technique of 'array painting'

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

Science.gov (United States)

Senoh, Mitsutoshi; Hamabata, Takashi; Takeda, Yoshifumi

2015-08-01

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

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

DEFF Research Database (Denmark)

Damgaard, Christian; Magnussen, Karin; Enevold, Christian

2015-01-01

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

Fuel cells : a viable fossil fuel alternative

Energy Technology Data Exchange (ETDEWEB)

Paduada, M.

2007-02-15

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

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

Science.gov (United States)

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

2015-05-01

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

Managing Viable Knowledge

NARCIS (Netherlands)

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

2002-01-01

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

Cannabidiol Reduces Leukemic Cell Size – But Is It Important?

Science.gov (United States)

Kalenderoglou, Nikoletta; Macpherson, Tara; Wright, Karen L.

2017-01-01

The anti-cancer effect of the plant-derived cannabinoid, cannabidiol, has been widely demonstrated both in vivo and in vitro. However, this body of preclinical work has not been translated into clinical use. Key issues around this failure can be related to narrow dose effects, the cell model used and incomplete efficacy. A model of acute lymphoblastic disease, the Jurkat T cell line, has been used extensively to study the cannabinoid system in the immune system and cannabinoid-induced apoptosis. Using these cells, this study sought to investigate the outcome of those remaining viable cells post-treatment with cannabidiol, both in terms of cell size and tracking any subsequent recovery. The phosphorylation status of the mammalian Target of Rapamycin (mTOR) signaling pathway and the downstream target ribosomal protein S6, were measured. The ability of cannabidiol to exert its effect on cell viability was also evaluated in physiological oxygen conditions. Cannabidiol reduced cell viability incompletely, and slowed the cell cycle with fewer cells in the G2/M phase of the cell cycle. Cannabidiol reduced phosphorylation of mTOR, PKB and S6 pathways related to survival and cell size. The remaining population of viable cells that were cultured in nutrient rich conditions post-treatment were able to proliferate, but did not recover to control cell numbers. However, the proportion of viable cells that were gated as small, increased in response to cannabidiol and normally sized cells decreased. This proportion of small cells persisted in the recovery period and did not return to basal levels. Finally, cells grown in 12% oxygen (physiological normoxia) were more resistant to cannabidiol. In conclusion, these results indicate that cannabidiol causes a reduction in cell size, which persists post-treatment. However, resistance to cannabidiol under physiological normoxia for these cells would imply that cannabidiol may not be useful in the clinic as an anti-leukemic agent. PMID

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

Science.gov (United States)

Raymond, Yves; Champagne, Claude P

2015-04-01

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

Enhancing forward osmosis water recovery from landfill leachate by desalinating brine and recovering ammonia in a microbial desalination cell.

Science.gov (United States)

Iskander, Syeed Md; Novak, John T; He, Zhen

2018-05-01

In this work, a microbial desalination cell (MDC) was employed to desalinate the FO treated leachate for reduction of both salinity and chemical oxygen demand (COD). The FO recovered 51.5% water from a raw leachate and the recovery increased to 83.5% from the concentrated leachate after desalination in the MDC fed with either acetate or another leachate as an electron source and at a different hydraulic retention time (HRT). Easily-degraded substrate like acetate and a long HRT resulted in a low conductivity desalinated effluent. Ammonia was also recovered in the MDC cathode with a recovery efficiency varying from 11 to 64%, affected by current generation and HRT. Significant COD reduction, as high as 65.4%, was observed in the desalination chamber and attributed to the decrease of both organic and inorganic compounds via diffusion and electricity-driven movement. Copyright © 2018 Elsevier Ltd. All rights reserved.

Rapid cell separation with minimal manipulation for autologous cell therapies

Science.gov (United States)

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

2017-02-01

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

Workers’ Self-management, Recovered Companies and the Sociology of Work

Science.gov (United States)

Croucher, Richard

2014-01-01

We analyse how far Argentina’s worker-recovered companies (WRCs) have sustained themselves and their principles of equity and workers’ self-management since becoming widespread following the country’s 2001–2 economic crisis. Specialist Spanish-language sources, survey data and documents are analysed through four key sociological themes. We find that the number of WRCs has increased in Argentina, and that they represent a viable production model. Further, they have generally maintained their central principles and even flourished. This occurred despite the global economic crisis, legal and financial pressures to adopt capitalist practices and management structures, the risk of market absorption and state attempts to coopt, demobilise and depoliticise the movement. We argue that today they function as a much-needed international beacon of an alternative vision for labour and that integration of their experience has potential to revitalise the field. PMID:25419009

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

Directory of Open Access Journals (Sweden)

Yuexia Wang

2015-09-01

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

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

Science.gov (United States)

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

2015-09-01

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

Quantification of Campylobacter spp. in chicken rinse samples by using flotation prior to real-time PCR

DEFF Research Database (Denmark)

Wolffs, Petra; Norling, Börje; Hoorfar, Jeffrey

2005-01-01

Real-time PCR is fast, sensitive, specific, and can deliver quantitative data; however, two disadvantages are that this technology is sensitive to inhibition by food and that it does not distinguish between DNA originating from viable, viable nonculturable (VNC), and dead cells. For this reason......, real-time PCR has been combined with a novel discontinuous buoyant density gradient method, called flotation, in order to allow detection of only viable and VNC cells of thermotolerant campylobacters in chicken rinse samples. Studying the buoyant densities of different Campylobacter spp. showed...... enrichment and amounts as low as 2.6 X 10(3) CFU/ml could be quantified. Furthermore, subjecting viable cells and dead cells to flotation showed that viable cells were recovered after flotation treatment but that dead cells and/or their DNA was not detected. Also, when samples containing VNC cells mixed...

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

Science.gov (United States)

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

2016-01-01

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

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

Science.gov (United States)

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

Uranium material removing and recovering device

International Nuclear Information System (INIS)

Takita, Shin-ichi.

1997-01-01

A uranium material removing and recovering device for use in removing surplus uranium heavy metal (UO 2 ) generated in a uranium handling facility comprises a uranium material removing device and a uranium material recovering device. The uranium material removing device comprises an adsorbing portion filled with a uranium adsorbent, a control portion for controlling the uranium adsorbent of the uranium adsorbing portion by a controlling agent, a uranium adsorbing device connected thereto and a jetting device for jetting the adsorbing liquid to equipments deposited with uranium. The recovering device comprises a recovering apparatus for recovering uranium materials deposited with the adsorbent liquid removed by the jetting device and a recovering tank for storing the recovered uranium materials. The device of the present invention can remove surplus uranium simply and safely, mitigate body's load upon removing and recovering operations, facilitate the processing for the exchange of the adsorbent and reduces the radioactive wastes. (T.M.)

Hematologic and biochemistry values for black-faced spoonbills (Platalea minor) with and recovering from botulism.

Science.gov (United States)

Chou, Shih-Jen; Shieh, Yao-Ching; Yu, Chang-You

2008-07-01

Type C1 botulism outbreaks in Black-faced Spoonbills (Platalea minor) occurred in Taiwan from 2002 to 2003, and hematologic and biochemistry parameters from botulism-paralyzed birds and recovered birds were compared. Values for creatinine and uric acid were higher (Pbirds with botulism than in recovered birds. Lower white blood cell counts (Pbirds. Based on these observations, we suggest that hematologic and biochemistry analyses should be performed to assess the health condition of birds recovering from botulism.

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

Directory of Open Access Journals (Sweden)

Restu Syamsul Hadi

2014-08-01

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

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

Directory of Open Access Journals (Sweden)

Sean M. Devitt

2015-01-01

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

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

Directory of Open Access Journals (Sweden)

Angelovski Ljupco

2008-11-01

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

Effects of cell condition, pH, and temperature on lead, zinc, and copper sorption to Acidithiobacillus caldus strain BC13

International Nuclear Information System (INIS)

Aston, John E.; Apel, William A.; Lee, Brady D.; Peyton, Brent M.

2010-01-01

Research highlights: →At. caldus sorbs lead, zinc, and copper across a range of pH and temperature. →At. caldus shows a relatively high sorption capacity for zinc and copper at low pH. → Lead, zinc, and copper sorption decreases in tertiary mixtures. → Copper appears to sorb via a different mechanism(s) than lead or zinc. - Abstract: This study describes the effects of cell condition, pH, and temperature on lead, zinc, and copper sorption to Acidithiobacillus caldus strain BC13 with a Langmuir model. Copper exhibited the highest loading capacity, 4.76 ± 0.28 mmol g -1 , to viable cells at pH 5.5. The highest k L (binding-site affinity) observed was 61.2 ± 3.0 L mmol -1 to dehydrated cells at pH 4.0. The pHs that maximized loading capacities and binding-site affinities were generally between 4.0 and 5.5, where the sum of free-proton and complexed-metal concentrations was near a minimum. Of additional importance, lead, zinc, and copper sorbed to viable cells at pH values as low as 1.5. Previous studies with other acidithiobacilli did not measure viable-cell sorption below pH 4.0. In separate experiments, desorption studies showed that far less copper was recovered from viable cells than any other metal or cell condition, suggesting that uptake may play an important role in copper sorption by At. caldus strain BC13. To reflect an applied system, the sorption of metal mixtures was also studied. In these experiments, lead, zinc, and copper sorption from a tertiary mixture were 40.2 ± 4.3%, 28.7 ± 3.8%, and 91.3 ± 3.0%, respectively, of that sorbed in single-metal systems.

Culture of somatic cells isolated from frozen-thawed equine semen using fluorescence-assisted cell sorting.

Science.gov (United States)

Brom-de-Luna, Joao Gatto; Canesin, Heloísa Siqueira; Wright, Gus; Hinrichs, Katrin

2018-03-01

Nuclear transfer using somatic cells from frozen semen (FzSC) would allow cloning of animals for which no other genetic material is available. Horses are one of the few species for which cloning is commercially feasible; despite this, there is no information available on the culture of equine FzSC. After preliminary trials on equine FzSC, recovered by density-gradient centrifugation, resulted in no growth, we hypothesized that sperm in the culture system negatively affected cell proliferation. Therefore, we evaluated culture of FzSC isolated using fluorescence-assisted cell sorting. In Exp. 1, sperm were labeled using antibodies to a sperm-specific antigen, SP17, and unlabeled cells were collected. This resulted in high sperm contamination. In Exp. 2, FzSC were labeled using an anti-MHC class I antibody. This resulted in an essentially pure population of FzSC, 13-25% of which were nucleated. Culture yielded no proliferation in any of nine replicates. In Exp. 3, 5 × 10 3 viable fresh, cultured horse fibroblasts were added to the frozen-thawed, washed semen, then this suspension was labeled and sorted as for Exp. 2. The enriched population had a mean of five sperm per recovered somatic cell; culture yielded formation of monolayers. In conclusion, an essentially pure population of equine FzSC could be obtained using sorting for presence of MHC class I antigens. No equine FzSC grew in culture; however, the proliferation of fibroblasts subjected to the same processing demonstrated that the labeling and sorting methods, and the presence of few sperm in culture, were compatible with cell viability. Copyright © 2017 Elsevier B.V. All rights reserved.

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

Science.gov (United States)

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

2018-01-01

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

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

Directory of Open Access Journals (Sweden)

Muhammed Majeed

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

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

Science.gov (United States)

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

2018-01-01

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

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

Science.gov (United States)

Zhong, Junliang; Zhao, Xihong

2018-01-01

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

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

Directory of Open Access Journals (Sweden)

Bee Luan Khoo

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

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

Science.gov (United States)

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

2017-12-01

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

Leishmania donovani-reactive Th1- and Th2-like T-cell clones from individuals who have recovered from visceral leishmaniasis

DEFF Research Database (Denmark)

Kemp, M; Kurtzhals, J A; Bendtzen, K

1993-01-01

analyzed in a panel of L. donovani-reactive CD4+ human T-cell clones generated from individuals who had recovered from VL after antimonial treatment. Two of the T-cell clones produced large amounts of IL-4 without production of IFN-gamma, seven clones produced both IFN-gamma and IL-4, and eight produced...... by interleukin-4 (IL-4)-producing Th2 cells, or cure may result by Th1 cells secreting gamma interferon (IFN-gamma). The present study examined the potential of human T cells to generate Th1 or Th2 responses to L. donovani. The profiles of IFN-gamma, IL-4, and lymphotoxin secretion after antigen stimulation were...... only IFN-gamma. This is the first report of a Th1- and Th2-type response in human leishmaniasis. These results suggest that in analogy with murine models, there is a dichotomy in the human T-cell response to L. donovani infections. Preferential activation of IL-4-producing Th2-like cells may...

A monetary comparison of energy recovered from microbial fuel cells and microbial electrolysis cells fed winery or domestic wastewaters

Energy Technology Data Exchange (ETDEWEB)

Cusick, Roland D.; Kiely, Patrick D.; Logan, Bruce E. [Department of Civil and Environmental Engineering, H2E Center, Penn State University, University Park, PA 16802 (United States)

2010-09-15

Microbial fuel (MFCs) and electrolysis cells (MECs) can be used to recover energy directly as electricity or hydrogen from organic matter. Organic removal efficiencies and values of the different energy products were compared for MFCs and MECs fed winery or domestic wastewater. TCOD removal (%) and energy recoveries (kWh/kg-COD) were higher for MFCs than MECs with both wastewaters. At a cost of 4.51/kg-H{sub 2} for winery wastewater and 3.01/kg-H{sub 2} for domestic wastewater, the hydrogen produced using MECs cost less than the estimated merchant value of hydrogen (6/kg-H{sub 2}). 16S rRNA clone libraries indicated the predominance of Geobacter species in anodic microbial communities in MECs for both wastewaters, suggesting low current densities were the result of substrate limitations. The results of this study show that energy recovery and organic removal from wastewater are more effective with MFCs than MECs, but that hydrogen production from wastewater fed MECs can be cost effective. (author)

Co-infusion of autologous adipose tissue derived neuronal differentiated mesenchymal stem cells and bone marrow derived hematopoietic stem cells, a viable therapy for post-traumatic brachial plexus injury: A case report

Directory of Open Access Journals (Sweden)

Umang G Thakkar

2014-08-01

Full Text Available Stem cell therapy is emerging as a viable approach in regenerative medicine. A 31-year-old male with brachial plexus injury had complete sensory-motor loss since 16 years with right pseudo-meningocele at C5-D1 levels and extra-spinal extension up to C7-D1, with avulsion on magnetic resonance imaging and irreversible damage. We generated adipose tissue derived neuronal differentiated mesenchymal stem cells (N-AD-MSC and bone marrow derived hematopoietic stem cells (HSC-BM. Neuronal stem cells expressed β-3 tubulin and glial fibrillary acid protein which was confirmed on immunofluorescence. On day 14, 2.8 ml stem cell inoculum was infused under local anesthesia in right brachial plexus sheath by brachial block technique under ultrasonography guidance with a 1.5-inch-long 23 gauge needle. Nucleated cell count was 2 × 10 4 /μl, CD34+ was 0.06%, and CD45-/90+ and CD45-/73+ were 41.63% and 20.36%, respectively. No untoward effects were noted. He has sustained recovery with re-innervation over a follow-up of 4 years documented on electromyography-nerve conduction velocity study.

Cellular and soluble components decrease the viable pathogen counts in milk from dairy cows with subclinical mastitis.

Science.gov (United States)

Koshiishi, Tomoko; Watanabe, Masako; Miyake, Hajime; Hisaeda, Keiichi; Isobe, Naoki

2017-08-10

The present study was undertaken to clarify the factors that reduce the viable pathogen count in milk collected from the udders of subclinical mastitic cows during preservation. Milk was centrifuged to divide somatic cells (cellular components, precipitates) and antimicrobial peptides (soluble components, supernatants without fat layer); each fraction was cultured with bacteria, and the number of viable bacteria was assessed prior to and after culture. In 28.8% of milk samples, we noted no viable bacteria immediately after collection; this value increased significantly after a 5-hr incubation of milk with cellular components but not with soluble components (48.1 and 28.8%, respectively). After culture with cellular components, the numbers of bacteria (excluding Staphylococcus aureus and Streptococcus uberis) and yeast decreased dramatically, although the differences were not statistically significant. After cultivation with soluble components, only yeasts showed a tendency toward decreased mean viability, whereas the mean bacterial counts of S. uberis and T. pyogenes tended to increase after 5-hr preservation with soluble components. These results suggest that most pathogens in high somatic cell count (SCC) milk decreased during preservation at 15 to 25°C, due to both the cellular components and antimicrobial components in the milk. Particularly, the cellular components more potently reduced bacterial counts during preservation.

How to recover more value from small pine trees: Essential oils and resins

International Nuclear Information System (INIS)

Kelkar, Vasant M.; Geils, Brian W.; Becker, Dennis R.; Overby, Steven T.; Neary, Daniel G.

2006-01-01

In recent years, the young dense forests of northern Arizona have suffered extreme droughts, wildfires, and insect outbreaks. Improving forest health requires reducing forest density by cutting many small-diameter trees with the consequent production of large volumes of residual biomass. To offset the cost of handling this low-value timber, additional marketing options for current operations are urgently needed to recover more value as wood products, energy, and chemicals. Northern Arizona forests are predominantly composed of ponderosa pine (Pinus ponderosa) which, besides producing abundant timber, can also yield many useful chemicals such as essential oils and resins. We describe a case study to assess the opportunities, constraints, and information required to integrate recovery of essential oils into forest and mill operations as might be used in northern Arizona. Preliminary results support the proposition there is an available, large supply of biomass with high concentrations of essential oils. The chemistry and process engineering for recovering these essential oils by distillation are well known. The potential output and uses also appear attractive given the substantial United States market for such products. However, less is known of the capability of essential oils extracted from ponderosa pine to compete with products imported from other countries. A more detailed assessment of product uses and further analysis of viable markets and environmental benefits are justified. (author)

Characterization of cells recovered from the xenotransplanted NG97 human-derived glioma cell line subcultured in a long-term in vitro

Directory of Open Access Journals (Sweden)

Heinrich Juliana K

2008-10-01

Full Text Available Abstract Background In order to elucidate tumoral progression and drug resistance, cultured cell lines are valuable tools applied on tumor related assays provided they are well established and characterized. Our laboratory settled the NG97 cell line derived from a human astrocytoma grade III, which started to develop and express important phenotypical characteristics of an astrocytoma grade IV after injection in the flank of nude mice. Astrocytomas are extremely aggressive malignancies of the Central Nervous System (CNS and account for 46% of all primary malignant brain tumors. Progression to worse prognosis occurs in 85% of the cases possibly due to changes in cell tumor microenvironment and through biological pathways that are still unclear. Methods This work focused on characterizing the NG97 cell line specifically after being recovered from the xenotransplant, who maintained their undifferentiated characteristics along the following 60th passages in vitro. These cells were subcultivated to evaluate the possible contribution of these undifferentiated characteristics to the malignant progression phenotype. These characteristics were the expression of molecules involved in the processes of migration, dedifferentiation and chromosomal instability. Results Results showed that NG97(ht had an decrease in doubling time through sub cultivation, which was characterized by a converse modulation between the expression of glial fibrillary acidic protein (GFAP and vimentin. In addition, β1 integrins were present in intermediate levels while α5 integrins had a high expression profile as well as fibronectin and laminin. Cytogenetic analysis of NG97(ht revealed several chromosomal abnormalities, 89% of the cells showed to be hyperdiploid and the modal number was assigned to be 63. Several acrocentric chromosomes were visualized and at least 30 figures were attributed to be murine. These findings suggest a possible fusion between the original NG97 cells

Characterization of cells recovered from the xenotransplanted NG97 human-derived glioma cell line subcultured in a long-term in vitro

International Nuclear Information System (INIS)

Machado, Camila ML; Boetcher-Luiz, Fátima; Verinaud, Liana; Ikemori, Rafael Y; Zorzeto, Tatiana Q; Nogueira, Ana CMA; Barbosa, Suse DS; Savino, Wilson; Schenka, André A; Vassallo, José; Heinrich, Juliana K

2008-01-01

In order to elucidate tumoral progression and drug resistance, cultured cell lines are valuable tools applied on tumor related assays provided they are well established and characterized. Our laboratory settled the NG97 cell line derived from a human astrocytoma grade III, which started to develop and express important phenotypical characteristics of an astrocytoma grade IV after injection in the flank of nude mice. Astrocytomas are extremely aggressive malignancies of the Central Nervous System (CNS) and account for 46% of all primary malignant brain tumors. Progression to worse prognosis occurs in 85% of the cases possibly due to changes in cell tumor microenvironment and through biological pathways that are still unclear. This work focused on characterizing the NG97 cell line specifically after being recovered from the xenotransplant, who maintained their undifferentiated characteristics along the following 60 th passages in vitro. These cells were subcultivated to evaluate the possible contribution of these undifferentiated characteristics to the malignant progression phenotype. These characteristics were the expression of molecules involved in the processes of migration, dedifferentiation and chromosomal instability. Results showed that NG97(ht) had an decrease in doubling time through sub cultivation, which was characterized by a converse modulation between the expression of glial fibrillary acidic protein (GFAP) and vimentin. In addition, β1 integrins were present in intermediate levels while α5 integrins had a high expression profile as well as fibronectin and laminin. Cytogenetic analysis of NG97(ht) revealed several chromosomal abnormalities, 89% of the cells showed to be hyperdiploid and the modal number was assigned to be 63. Several acrocentric chromosomes were visualized and at least 30 figures were attributed to be murine. These findings suggest a possible fusion between the original NG97 cells with stromal murine cells in the xenotransplant. In

Expression of adhesion molecules, chemokines and matrix metallo- proteinases (MMPs) in viable and degenerating stage of Taenia solium metacestode in swine neurocysticercosis.

Science.gov (United States)

Singh, Satyendra K; Singh, Aloukick K; Prasad, Kashi N; Singh, Amrita; Singh, Avinash; Rai, Ravi P; Tripathi, Mukesh; Gupta, Rakesh K; Husain, Nuzhat

2015-11-30

Neurocysticercosis (NCC) is a parasitic infection of central nervous system (CNS). Expression of adhesion molecules, chemokines and matrix metalloproteinases (MMPs) were investigated on brain tissues surrounding viable (n=15) and degenerating cysticerci (n=15) of Taenia solium in swine by real-time RT-PCR and ELISA. Gelatin gel zymography was performed for MMPs activity. ICAM-1 (intercellular adhesion molecule-1), E-selectin, MIP-1α (macrophage inflammatory protein-1α), Eotaxin-1 and RANTES (regulated on activation, normal T cell expressed and secreted) were associated with degenerating cysticerci (cysts). However, VCAM-1 (vascular cell adhesion molecule-1), MCP-1 (monocyte chemotactic protein-1), MMP-2 and MMP-9 were associated with both viable and degenerating cysts. In conclusion, viable and degenerating cysticerci have different immune molecule profiles and role of these molecules in disease pathogenesis needs to be investigated. Copyright © 2015 Elsevier B.V. All rights reserved.

Regulation of Viable and Optimal Cohorts

Energy Technology Data Exchange (ETDEWEB)

Aubin, Jean-Pierre, E-mail: aubin.jp@gmail.com [VIMADES (Viabilité, Marchés, Automatique, Décisions) (France)

2015-10-15

This study deals with the evolution of (scalar) attributes (resources or income in evolutionary demography or economics, position in traffic management, etc.) of a population of “mobiles” (economic agents, vehicles, etc.). The set of mobiles sharing the same attributes is regarded as an instantaneous cohort described by the number of its elements. The union of instantaneous cohorts during a mobile window between two attributes is a cohort. Given a measure defining the number of instantaneous cohorts, the accumulation of the mobile attributes on a evolving mobile window is the measure of the cohort on this temporal mobile window. Imposing accumulation constraints and departure conditions, this study is devoted to the regulation of the evolutions of the attributes which are1.viable in the sense that the accumulations constraints are satisfied at each instant;2.and, among them, optimal, in the sense that both the duration of the temporal mobile window is maximum and that the accumulation on this temporal mobile window is the largest viable one. This value is the “accumulation valuation” function. Viable and optimal evolutions under accumulation constraints are regulated by an “implicit Volterra integro-differential inclusion” built from the accumulation valuation function, solution to an Hamilton–Jacobi–Bellman partial differential equation under constraints which is constructed for this purpose.

Sorption and precipitation of Mn2+ by viable and autoclaved Shewanella putrefaciens: Effect of contact time

KAUST Repository

Chubar, Natalia

2013-01-01

The sorption of Mn(II) by viable and inactivated cells of Shewanella putrefaciens, a non-pathogenic, facultative anaerobic, gram-negative bacterium characterised as a Mn(IV) and Fe(III) reducer, was studied under aerobic conditions, as a function of pH, bacterial density and metal loading. During a short contact time (3-24h), the adsorptive behaviour of live and dead bacteria toward Mn(II) was sufficiently similar, an observation that was reflected in the studies on adsorption kinetics at various metal loadings, effects of pH, bacteria density, isotherms and drifting of pH during adsorption. Continuing the experiment for an additional 2-30days demonstrated that the Mn(II) sorption by suspensions of viable and autoclaved cells differed significantly from one another. The sorption to dead cells was characterised by a rapid equilibration and was described by an isotherm. In contrast, the sorption (uptake) to live bacteria exhibited a complex time-dependent uptake. This uptake began as adsorption and ion exchange processes followed by bioprecipitation, and it was accompanied by the formation of polymeric sugars (EPS) and the release of dissolved organic substances. FTIR, EXAFS/XANES and XPS demonstrated that manganese(II) phosphate was the main precipitate formed in 125ml batches, which is the first evidence of the ability of microbes to synthesise manganese phosphates. XPS and XANES spectra did not detect Mn(II) oxidation. Although the release of protein-like compounds by the viable bacteria increased in the presence of Mn2+ (and, by contrast, the release of carbohydrates did not change), electrochemical analyses did not indicate any aqueous complexation of Mn(II) by the organic ligands. © 2012 Elsevier Ltd.

Identification of Viable Helicobacter pylori in Drinking Water Supplies by Cultural and Molecular Techniques.

Science.gov (United States)

Santiago, Paula; Moreno, Yolanda; Ferrús, M Antonía

2015-08-01

Helicobacter pylori is one of the most common causes of chronic bacterial infection in humans, directly related to peptic ulcer and gastric cancer. It has been suggested that H. pylori can be acquired through different transmission routes, including water. In this study, culture and qPCR were used to detect and identify the presence of H. pylori in drinking water. Furthermore, the combined techniques PMA-qPCR and DVC-FISH were applied for detection of viable cells of H. pylori. Among 24 drinking water samples, 16 samples were positive for the presence of H. pylori, but viable cells were only detected in six samples. Characteristic colonies, covered by a mass of bacterial unspecific growth, were observed on selective agar plates from an only sample, after enrichment. The mixed culture was submitted to DVC-FISH and qPCR analysis, followed by sequencing of the amplicons. Molecular techniques confirmed the growth of H. pylori on the agar plate. Our results demonstrate for the first time that H. pylori can survive and be potentially infective in drinking water, showing that water distribution systems could be a potential route for H. pylori transmission. © 2015 John Wiley & Sons Ltd.

Recovering valuable metals from recycled photovoltaic modules.

Science.gov (United States)

Yi, Youn Kyu; Kim, Hyun Soo; Tran, Tam; Hong, Sung Kil; Kim, Myong Jun

2014-07-01

Recovering valuable metals such as Si, Ag, Cu, and Al has become a pressing issue as end-of-life photovoltaic modules need to be recycled in the near future to meet legislative requirements in most countries. Of major interest is the recovery and recycling of high-purity silicon (> 99.9%) for the production of wafers and semiconductors. The value of Si in crystalline-type photovoltaic modules is estimated to be -$95/kW at the 2012 metal price. At the current installed capacity of 30 GW/yr, the metal value in the PV modules represents valuable resources that should be recovered in the future. The recycling of end-of-life photovoltaic modules would supply > 88,000 and 207,000 tpa Si by 2040 and 2050, respectively. This represents more than 50% of the required Si for module fabrication. Experimental testwork on crystalline Si modules could recover a > 99.98%-grade Si product by HNO3/NaOH leaching to remove Al, Ag, and Ti and other metal ions from the doped Si. A further pyrometallurgical smelting at 1520 degrees C using CaO-CaF2-SiO2 slag mixture to scavenge the residual metals after acid leaching could finally produce > 99.998%-grade Si. A process based on HNO3/NaOH leaching and subsequent smelting is proposed for recycling Si from rejected or recycled photovoltaic modules. Implications: The photovoltaic industry is considering options of recycling PV modules to recover metals such as Si, Ag, Cu, Al, and others used in the manufacturing of the PV cells. This is to retain its "green" image and to comply with current legislations in several countries. An evaluation of potential resources made available from PV wastes and the technologies used for processing these materials is therefore of significant importance to the industry. Of interest are the costs of processing and the potential revenues gained from recycling, which should determine the viability of economic recycling of PV modules in the future.

Five years post whiplash injury: Symptoms and psychological factors in recovered versus non-recovered

Directory of Open Access Journals (Sweden)

Stålnacke Britt-Marie

2010-07-01

Full Text Available Abstract Background Few studies have focused on the differences between persons who are recovered after whiplash injury and those who suffer from persistent disability. The primary aim of this study was therefore to examine differences in symptoms, psychological factors and life satisfaction between subjects classified as recovered and those with persistent disability five years after whiplash injury based on the Neck Disability Index (NDI. Methods A set of questionnaires was answered by 158 persons (75 men, 83 women to assess disability (NDI, pain intensity (VAS, whiplash-related symptoms (Rivermead Post-Concussion Symptoms Questionnaire, RPQ, post-traumatic stress (Impact of Event Scale, IES, depression (Beck's depression inventory, BDI and life satisfaction (LiSat-11. The participants were divided into three groups based on the results of the NDI: recovered (34.8%, mild disability (37.3% and moderate/severe disability (27.3%. Results The moderate/severe group reported significantly higher VAS, BDI and IES scores and lower level of physical health and psychological health compared to the mild and the recovered groups. Less significant differences were reported between the mild and the recovered groups. Conclusions The group with the highest disability score reported most health problems with pain, symptoms, depression, post-traumatic stress and decreased life satisfaction. These findings indicate that classifying these subjects into subgroups based on disability levels makes it possible to optimize the management and treatment after whiplash injury.

Analysis of DNA evidence recovered from epithelial cells in penile swabs.

Science.gov (United States)

Drobnic, Katja

2003-06-01

In the rape case presented here, no semen, hair, or other biological evidence were left by the perpetuator at the crime scene or on the victim. The alleged assailant was arrested soon after the crime. A classical stain recovery technique using cotton swab moistened with sterile water was taken for recovering potential female epithelial cells and leukocytes deposited on the alleged assailant's penis during sexual assault. The organic method used for DNA extraction was quantified according to the slot-blot procedure and amplified at 9 and 15 polymorphic loci. Penile swab revealed a DNA profile of mixed origin. In addition to the suspect's DNA profile, DNA contribution from the victim was identified as a minor component in the mixture. Frequency of the profile resulted in a value of 5 x 10(-14) for the multiplex systems AmpFlSTR Plus and 2.5 x 10(-18) for the multiplex system PowerPlex 16, taking into account only non-overlapping alleles between the suspect and the victim from the minor component in the DNA mixture. Moreover, three additional alleles were observed at D21S11 locus by use of PowerPlex and STR SGM plus primers, which could not belong to the suspect. The victim's DNA profile showed the same three-banded genotype at this locus. The same pattern was detected when the victim's saliva or blood were used as reference samples. Our laboratory finding was consistent with the police report that the victim was a person with Down syndrome, a human genetic disease mainly resulting from trisomy (triplication) of the 21 chromosome.

Asouzu's Complementary Ontology as a Foundation for a Viable ...

African Journals Online (AJOL)

This paper on “Asouzu's Complementary Ontology as a foundation for a viable Ethic of the Environment”, posits that an ethic of the environment can be seen as viable if it considers the whole of reality as ontologically relevant. This point of view would free environmental ethics of anthropocentric bias and its attendant ...

Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments

International Nuclear Information System (INIS)

Patheja, Pooja; Sahu, Khageswar

2017-01-01

Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MφCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MφCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation.

Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments

Energy Technology Data Exchange (ETDEWEB)

Patheja, Pooja, E-mail: pooja.patheja8@gmail.com [Laser Biomedical Applications Section, Raja Ramanna Centre for Advanced Technology, Indore 452013, Madhya Pradesh (India); Homi Bhabha National Institute, Training School Complex, Anushaktinagar, Mumbai 400094, Maharashtra (India); Sahu, Khageswar [Laser Biomedical Applications Section, Raja Ramanna Centre for Advanced Technology, Indore 452013, Madhya Pradesh (India)

2017-06-15

Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MφCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MφCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation.

Induction of Viable but Nonculturable Salmonella in Exponentially Grown Cells by Exposure to a Low-Humidity Environment and Their Resuscitation by Catalase.

Science.gov (United States)

Morishige, Yuta; Koike, Atsushi; Tamura-Ueyama, Ai; Amano, Fumio

2017-02-01

Salmonella is a major cause of foodborne disease that sometimes occurs in massive outbreaks around the world. This pathogen is tolerant of low-humidity conditions. We previously described a method for induction of viable but nonculturable (VBNC) Salmonella enterica serovar Enteritidis by treatment with hydrogen peroxide (H 2 O 2 ) and subsequent resuscitation with 0.3 mM sodium pyruvate. Here, we report a new method for the induction of the VBNC state in Salmonella Enteritidis cells, one involving dehydration. Exposure of Salmonella Enteritidis cells to dehydration stress under poor nutritional conditions (0.9% [wt/vol] NaCl) and 10 to 20% relative humidity at room temperature decreased the presence of culturable population to 0.0067%, but respiratory and glucose uptake active populations were maintained at 0.46 and 1.12%, respectively, meaning that approximately 1% may have entered the VBNC state. Furthermore, these VBNC cells could be resuscitated to acquire culturability by incubation with catalase in M9 minimal medium without glucose in a manner dependent on the dose of catalase but not sodium pyruvate. These results suggest that a low-humidity environment could cause Salmonella Enteritidis cells to enter the VBNC state and the cells could then be resuscitated for growth by treatment with catalase, suggesting a potential risk of Salmonella Enteritidis to survive in low water activity foods in the VBNC state and to start regrowth for foodborne illness.

Different strategies for recovering metals from CARON process residue

International Nuclear Information System (INIS)

Cabrera, G.; Gomez, J.M.; Hernandez, I.; Coto, O.; Cantero, D.

2011-01-01

The capacity of Acidithiobacillus thiooxidans DMS 11478 to recover the heavy metals contained in the residue obtained from the CARON process has been evaluated. Different bioreactor configurations were studied: a two-stage batch system and two semi-continuous systems (stirred-tank reactor leaching and column leaching). In the two-stage system, 46.8% Co, 36.0% Mg, 26.3% Mn and 22.3% Ni were solubilised after 6 h of contact between the residue and the bacteria-free bioacid. The results obtained with the stirred-tank reactor and the column were similar: 50% of the Mg and Co and 40% of the Mn and Ni were solubilised after thirty one days. The operation in the column reactor allowed the solid-liquid ratio to be increased and the pH to be kept at low values (<1.0). Recirculation of the leachate in the column had a positive effect on metal removal; at sixty five days (optimum time) the solubilisation levels were as follows: 86% Co, 83% Mg, 72% Mn and Ni, 62% Fe and 23% Cr. The results corroborate the feasibility of the systems studied for the leaching of metals from CARON process residue and these methodologies can be considered viable for the recovery of valuable metals.

Recovering recyclable materials from shredder residue

Science.gov (United States)

Jody, Bassam J.; Daniels, Edward J.; Bonsignore, Patrick V.; Brockmeier, Norman F.

1994-02-01

Each year, about 11 million tons of metals are recovered in the United States from about 10 million discarded automobiles. The recovered metals account for about 75 percent of the total weight of the discarded vehicles. The balance of the material, known as shredder residue, amounts to about three million tons annually and is currently landfilled. The residue contains a diversity of potentially recyclable materials, including polyurethane foams, iron oxides, and certain thermoplastics. This article discusses a process under development at Argonne National Laboratory to separate and recover the recyclable materials from this waste stream. The process consists essentially of two stages. First, a physical separation is used to recover the foams and the metal oxides, followed by a chemical process to extract certain thermoplastics. The status of the technology and the process economics are reviewed here.

A Novel Application for Low Frequency Electrochemical Impedance Spectroscopy as an Online Process Monitoring Tool for Viable Cell Concentrations

Directory of Open Access Journals (Sweden)

Christoph Slouka

2016-11-01

Full Text Available New approaches in process monitoring during industrial fermentations are not only limited to classical pH, dO2 and offgas analysis, but use different in situ and online sensors based on different physical principles to determine biomass, product quality, lysis and far more. One of the very important approaches is the in situ accessibility of viable cell concentration (VCC. This knowledge provides increased efficiency in monitoring and controlling strategies during cultivations. Electrochemical impedance spectroscopy—EIS—is used to monitor biomass in a fermentation of E. coli BL21(DE3, producing a recombinant protein using a fed batch-based approach. Increases in the double layer capacitance (Cdl, determined at frequencies below 1 kHz, are proportional to the increase of biomass in the batch and fed batch phase, monitored in offline and online modes for different cultivations. A good correlation of Cdl with cell density is found and in order to get an appropriate verification of this method, different state-of-the-art biomass measurements are performed and compared. Since measurements in this frequency range are largely determined by the double layer region between the electrode and media, rather minor interferences with process parameters (aeration, stirring are to be expected. It is shown that impedance spectroscopy at low frequencies is a powerful tool for cultivation monitoring.

Macrophage conditioned medium induced cellular network formation in MCF-7 cells through enhanced tunneling nanotube formation and tunneling nanotube mediated release of viable cytoplasmic fragments.

Science.gov (United States)

Patheja, Pooja; Sahu, Khageswar

2017-06-15

Infiltrating macrophages in tumor microenvironment, through their secreted cytokines and growth factors, regulate several processes of cancer progression such as cancer cell survival, proliferation, invasion, metastasis and angiogenesis. Recently, intercellular cytoplasmic bridges between cancer cells referred as tunneling nanotubes (TNTs) have been recognized as novel mode of intercellular communication between cancer cells. In this study, we investigated the effect of inflammatory mediators present in conditioned medium derived from macrophages on the formation of TNTs in breast adenocarcinoma cells MCF-7. Results show that treatment with macrophage conditioned medium (MɸCM) not only enhanced TNT formation between cells but also stimulated the release of independently migrating viable cytoplasmic fragments, referred to as microplasts, from MCF-7 cells. Time lapse microscopy revealed that microplasts were released from parent cancer cells in extracellular space through formation of TNT-like structures. Mitochondria, vesicles and cytoplasm could be transferred from parent cell body to microplasts through connecting TNTs. The microplasts could also be resorbed into the parent cell body by retraction of the connecting TNTs. Microplast formation inhibited in presence cell migration inhibitor, cytochalasin-B. Notably by utilizing migratory machinery within microplasts, distantly located MCF-7 cells formed several TNT based intercellular connections, leading to formation of physically connected network of cells. Together, these results demonstrate novel role of TNTs in microplast formation, novel modes of TNT formation mediated by microplasts and stimulatory effect of MɸCM on cellular network formation in MCF-7 cells mediated through enhanced TNT and microplast formation. Copyright © 2017 Elsevier Inc. All rights reserved.

On the use of the serial dilution culture method to enumerate viable phytoplankton in natural communities of plankton subjected to ballast water treatment.

Science.gov (United States)

Cullen, John J; MacIntyre, Hugh L

2016-01-01

Discharge standards for ballast water treatment (BWT) systems are based on concentrations of living cells, for example, as determined with vital stains. Ultraviolet radiation (UV) stops the reproduction of microorganisms without killing them outright; they are living, but not viable, and ecologically as good as dead. Consequently, UV-treated discharge can be compliant with the intent of regulation while failing a live/dead test. An alternative evaluation of BWT can be proposed based on the assessment of viable, rather than living, cells in discharge water. In principle, the serial dilution culture-most probable number (SDC-MPN) method provides the appropriate measure for phytoplankton. But, the method has been criticized, particularly because it is thought that many phytoplankton species cannot be cultured. A review of the literature shows that although SDC-MPN has been used for more than 50 years-generally to identify and count phytoplankton species that cannot be preserved-its application to enumerate total viable phytoplankton seems to be new, putting past criticisms of the method in a different light. Importantly, viable cells need to grow only enough to be detected, not to be brought into sustained culture, and competition between species in a dilution tube is irrelevant as long as the winner is detectable. Thorough consideration of sources of error leads to recommendations for minimizing and quantifying uncertainties by optimizing growth conditions and conducting systematic comparisons. We conclude that with careful evaluation, SDC-MPN is potentially an effective method for assessing the viability of phytoplankton after BWT.

PROCESS OF RECOVERING URANIUM

Science.gov (United States)

Carter, J.M.; Larson, C.E.

1958-10-01

A process is presented for recovering uranium values from calutron deposits. The process consists in treating such deposits to produce an oxidlzed acidic solution containing uranium together with the following imparities: Cu, Fe, Cr, Ni, Mn, Zn. The uranium is recovered from such an impurity-bearing solution by adjusting the pH of the solution to the range 1.5 to 3.0 and then treating the solution with hydrogen peroxide. This results in the precipitation of uranium peroxide which is substantially free of the metal impurities in the solution. The peroxide precipitate is then separated from the solution, washed, and calcined to produce uranium trioxide.

Reduced infectivity of waterborne viable but nonculturable Helicobacter pylori strain SS1 in mice.

Science.gov (United States)

Boehnke, Kevin F; Eaton, Kathryn A; Fontaine, Clinton; Brewster, Rebecca; Wu, Jianfeng; Eisenberg, Joseph N S; Valdivieso, Manuel; Baker, Laurence H; Xi, Chuanwu

2017-08-01

Helicobacter pylori infection has been consistently associated with lack of access to clean water and proper sanitation, but no studies have demonstrated that the transmission of viable but nonculturable (VBNC) H. pylori can occur from drinking contaminated water. In this study, we used a laboratory mouse model to test whether waterborne VBNCH. pylori could cause gastric infection. We performed five mouse experiments to assess the infectivity of VBNCH. pylori in various exposure scenarios. VBNC viability was examined using Live/Dead staining and Biolog phenotype metabolism arrays. High doses of VBNCH. pylori in water were chosen to test the "worst-case" scenario for different periods of time. One experiment also investigated the infectious capabilities of VBNC SS1 using gavage. Further, immunocompromised mice were exposed to examine infectivity among potentially vulnerable groups. After exposure, mice were euthanized and their stomachs were examined for H. pylori infection using culture and PCR methodology. VBNC cells were membrane intact and retained metabolic activity. Mice exposed to VBNCH. pylori via drinking water and gavage were not infected, despite the various exposure scenarios (immunocompromised, high doses) that might have permitted infection with VBNCH. pylori. The positive controls exposed to viable, culturable H. pylori did become infected. While other studies that have used viable, culturable SS1 via gavage or drinking water exposures to successfully infect mice, in our study, waterborne VBNC SS1 failed to colonize mice under all test conditions. Future studies could examine different H. pylori strains in similar exposure scenarios to compare the relative infectivity of the VBNC vs the viable, culturable state, which would help inform future risk assessments of H. pylori in water. © 2017 The Authors. Helicobacter Published by John Wiley & Sons Ltd.

Co-infusion of autologous adipose tissue derived insulin-secreting mesenchymal stem cells and bone marrow derived hematopoietic stem cells: Viable therapy for type III.C. a diabetes mellitus

Directory of Open Access Journals (Sweden)

Umang G Thakkar

2014-12-01

Full Text Available Transition from acute pancreatitis to insulin-dependent diabetes mellitus (IDDM is a rare manifestation of primary hyperparathyroidism caused by parathyroid adenoma because of impaired glucose tolerance and suppresses insulin secretion. We report the case of a 26-year-old male with pancreatic diabetes caused by parathyroid adenoma induced chronic pancreatitis. He had serum C-peptide 0.12 ng/ml, glutamic acid decarboxylase antibody 5.0 IU/ml, and glycosylated hemoglobin (HbA1C 8.9%, and required 72 IU/day of biphasic-isophane insulin injection for uncontrolled hyperglycemia. We treated him with his own adipose tissue derived insulin-secreting mesenchymal stem-cells (IS-ADMSC along with his bone marrow derived hematopoietic stem cells (BM-HSC. Autologous IS-ADMSC + BM-HSC were infused into subcutaneous tissue, portal and thymic circulation without any conditioning. Over a follow-up of 27 months, the patient is maintaining fasting and postprandial blood sugar levels of 132 and 165 mg/dl, respectively, with HbA1C 6.8% and requiring 36 IU/day of biphasic-isophane insulin. Co-infusion of IS-ADMSC + BM-HSC offers a safe and viable therapy for type III.C.a Diabetes Mellitus.

The radiation response of cells recovering after chronic hypoxia

International Nuclear Information System (INIS)

Kwok, T.T.; Sutherland, R.M.

1989-01-01

Experiments were performed to study the influence of hypoxic pretreatment on the radiation response of A431 human squamous carcinoma cells. Reaeration for 10 min after chronic hypoxia (greater than 2 h) was found to enhance the radiosensitivity of A431 cells, and the maximal effect was seen for those cells reaerated after 12 h of hypoxia. The radiosensitivity enhancement for reaerated cells after 12 h of hypoxia was maximized by 5 min after the return to aerobic conditions and reached the control level by 12 h of reaeration. This enhanced radiosensitive state was characterized by a reduced shoulder region and increased slope of the radiation dose-response curve for cells in both the exponential and plateau phases of growth. There was a slight increase in the number of G1 and decrease in the number of S and G2 + M cells for both exponential- and plateau-phase cultures following 12 h hypoxic treatment. Although growth inhibition induced by 12 h of hypoxia was seen for cells in the exponential phase, there was no cell number change in the plateau-phase culture after hypoxia. Plating efficiency (PE) of cells in both growth phases was reduced by 30% after hypoxia. Furthermore, in the exponential-phase culture, the extent of reduction in PE after hypoxia was similar among cells in different phases of the cell cycle. Although S-phase cells in exponentially growing cultures were relatively more resistant to radiation than G1 and G2 + M cells, the cell age-response pattern was the same whether the cells had been aerobic or hypoxic before reaeration and irradiation. Furthermore, the enhancement ratio associated with reaeration after 12 h of hypoxia for these three subpopulations of cells was 1.3. Our results indicate that the increase in radiosensitivity due to reaeration after chronic hypoxia is unlikely to be related to the changes of cell cycle stage and growth phase during hypoxic treatment

Utilization of Cell-Transfer Technique for Molecular Testing on Hematoxylin-Eosin-Stained Sections: A Viable Option for Small Biopsies That Lack Tumor Tissues in Paraffin Block.

Science.gov (United States)

Wu, Howard H; Jovonovich, Stephen M; Randolph, Melissa; Post, Kristin M; Sen, Joyashree D; Curless, Kendra; Cheng, Liang

2016-12-01

- In some instances the standard method of doing molecular testing from formalin-fixed, paraffin-embedded block is not possible because of limited tissue. Tumor cell-enriched cell-transfer technique has been proven useful for performing immunocytochemistry and molecular testing on cytologic smears. - To establish the cell-transfer technique as a viable option for isolating tumor cells from hematoxylin-eosin (H&E)-stained slides. - Molecular testing was performed by using the cell-transfer technique on 97 archived H&E-stained slides from a variety of different tumors. Results were compared to the conventional method of molecular testing. - Polymerase chain reaction-based molecular testing via the cell-transfer technique was successfully performed on 82 of 97 samples (85%). This included 39 of 47 cases for EGFR, 10 of 11 cases for BRAF, and 33 of 39 cases for KRAS mutations. Eighty-one of 82 cell-transfer technique samples (99%) showed agreement with previous standard method results, including 4 mutations and 35 wild-type alleles for EGFR, 4 mutations and 6 wild-type alleles for BRAF, and 11 mutations and 21 wild-type alleles for KRAS. There was only 1 discrepancy: a cell-transfer technique with a false-negative >KRAS result (wild type versus G12C). - Molecular testing performed on H&E-stained sections via cell-transfer technique is useful when tissue from cell blocks and small surgical biopsy samples is exhausted and the only available material for testing is on H&E-stained slides.

Differential Effect of Viable Versus Necrotic Neutrophils on Mycobacterium tuberculosis Growth and Cytokine Induction in Whole Blood

Directory of Open Access Journals (Sweden)

David M. Lowe

2018-04-01

Full Text Available Neutrophils exert both positive and negative influences on the host response to tuberculosis, but the mechanisms by which these differential effects are mediated are unknown. We studied the impact of live and dead neutrophils on the control of Mycobacterium tuberculosis using a whole blood bioluminescence-based assay, and assayed supernatant cytokine concentrations using Luminex™ technology and ELISA. CD15+ granulocyte depletion from blood prior to infection with M. tuberculosis-lux impaired control of mycobacteria by 96 h, with a greater effect than depletion of CD4+, CD8+, or CD14+ cells (p < 0.001. Augmentation of blood with viable granulocytes significantly improved control of mycobacteria by 96 h (p = 0.001, but augmentation with necrotic granulocytes had the opposite effect (p = 0.01. Both augmentations decreased supernatant concentrations of tumor necrosis factor and interleukin (IL-12 p40/p70, but necrotic granulocyte augmentation also increased concentrations of IL-10, G-CSF, GM-CSF, and CCL2. Necrotic neutrophil augmentation reduced phagocytosis of FITC-labeled M. bovis BCG by all phagocytes, whereas viable neutrophil augmentation specifically reduced early uptake by CD14+ cells. The immunosuppressive effect of dead neutrophils required necrotic debris rather than supernatant. We conclude that viable neutrophils enhance control of M. tuberculosis in blood, but necrotic neutrophils have the opposite effect—the latter associated with induction of IL-10, growth factors, and chemoattractants. Our findings suggest a mechanism by which necrotic neutrophils may exert detrimental effects on the host response in active tuberculosis.

Recovering method for solid waste and facility therefor

International Nuclear Information System (INIS)

Omura, Yutaka

1998-01-01

When recovering solid wastes in a dry-type storage vessel, a crusher is hoisted down from a cask, and the crusher is operated to crush the solid wastes while holding them. The crushed wastes are temporarily stored at the upper portion of the crusher, and recovered as crushed wastes. In this case, the crusher is turned down, and a shielding vessel is laid the recover downwardly to temporary store the crushed wastes in the shielding vessel. Then, the crusher and the shielding vessel are turned 180deg to contain the crushed wastes into the shielding vessel. With such procedures, the stored solid wastes can be recovered reliably, the stored solid wastes can be reduced in the size, and efficiency of recovering operation can be improved. (T.M.)

Mitochondrial respiration in human viable platelets-Methodology and influence of gender, age and storage

DEFF Research Database (Denmark)

Sjövall, Fredrik; Ehinger, Johannes K H; Marelsson, Sigurður E

2013-01-01

Studying whole cell preparations with intact mitochondria and respiratory complexes has a clear benefit compared to isolated or disrupted mitochondria due to the dynamic interplay between mitochondria and other cellular compartments. Platelet mitochondria have a potential to serve as a source...... of human viable mitochondria when studying mitochondrial physiology and pathogenic mechanisms, as well as for the diagnostics of mitochondrial diseases. The objective of the present study was to perform a detailed evaluation of platelet mitochondrial respiration using high-resolution respirometry. Further...

Sodium aerosol recovering device

International Nuclear Information System (INIS)

Fujimori, Koji; Ueda, Mitsuo; Tanaka, Kazuhisa.

1997-01-01

A main body of a recovering device is disposed in a sodium cooled reactor or a sodium cooled test device. Air containing sodium aerosol is sucked into the main body of the recovering device by a recycling fan and introduced to a multi-staged metal mesh filter portion. The air about against each of the metal mesh filters, and the sodium aerosol in the air is collected. The air having a reduced sodium aerosol concentration circulates passing through a recycling fan and pipelines to form a circulation air streams. Sodium aerosol deposited on each of the metal mesh filters is scraped off periodically by a scraper driving device to prevent clogging of each of the metal filters. (I.N.)

Articular Cartilage Repair Using Marrow Stimulation Augmented with a Viable Chondral Allograft: 9-Month Postoperative Histological Evaluation

Directory of Open Access Journals (Sweden)

James K. Hoffman

2015-01-01

Full Text Available Marrow stimulation is frequently employed to treat focal chondral defects of the knee. However, marrow stimulation typically results in fibrocartilage repair tissue rather than healthy hyaline cartilage, which, over time, predisposes the repair to failure. Recently, a cryopreserved viable chondral allograft was developed to augment marrow stimulation. The chondral allograft is comprised of native viable chondrocytes, chondrogenic growth factors, and extracellular matrix proteins within the superficial, transitional, and radial zones of hyaline cartilage. Therefore, host mesenchymal stem cells that infiltrate the graft from the underlying bone marrow following marrow stimulation are provided with the optimal microenvironment to undergo chondrogenesis. The present report describes treatment of a trochlear defect with marrow stimulation augmented with this novel chondral allograft, along with nine month postoperative histological results. At nine months, the patient demonstrated complete resolution of pain and improvement in function, and the repair tissue consisted of 85% hyaline cartilage. For comparison, a biopsy obtained from a patient 8.2 months after treatment with marrow stimulation alone contained only 5% hyaline cartilage. These outcomes suggest that augmenting marrow stimulation with the viable chondral allograft can eliminate pain and improve outcomes, compared with marrow stimulation alone.

Coal liquefaction becomes viable

Energy Technology Data Exchange (ETDEWEB)

NONE

2005-11-15

In 2003 the May/June issue of CoalTrans International speculated that coal liquefaction would become viable due to falling coal prices. This has not proved the case but the sustained high oil price is sparking new interest. A survey by Energy Intelligence and Marketing Research during November 2005 revealed a growth in the number of projects under development or at the feasibility stage. The article reports projects in China, the USA, Australia, New Zealand, the Philippines and India. China is commissioning the first wave of large liquefaction plants. The key question is whether other countries, particularly the USA, will follow.

The lifetime of hypoxic human tumor cells

International Nuclear Information System (INIS)

Durand, Ralph E.; Sham, Edward

1998-01-01

Purpose: For hypoxic and anoxic cells in solid tumors to be a therapeutic problem, they must live long enough to be therapeutically relevant, or else be rapidly recruited into the proliferating compartment during therapy. We have, therefore, estimated lifetime and recruitment rate of hypoxic human tumor cells in multicell spheroids in vitro, or in xenografted tumors in SCID mice. Materials and Methods: Cell turnover was followed by flow cytometry techniques, using antibodies directed at incorporated halogenated pyrimidines. The disappearance of labeled cells was quantified, and verified to be cell loss rather than label dilution. Repopulation was studied in SiHa tumor xenografts during twice-daily 2.5-Gy radiation exposures. Results: The longevity of hypoxic human tumor cells in spheroids or xenografts exceeded that of rodent cell lines, and cell turnover was slower in xenografts than under static growth as spheroids. Human tumor cells remained viable in the hypoxic regions of xenografts for 4-10 days, compared to 3-5 days in spheroids, and 1-3 days for most rodent cells in spheroids. Repopulation was observed within the first few radiation treatments for the SiHa xenografts and, with accumulated doses of more than 10 Gy, virtually all recovered cells had progressed through at least one S-phase. Conclusion: Our results suggest an important difference in the ability of human vs. rodent tumor cells to withstand hypoxia, and raise questions concerning the increased longevity seen in vivo relative to the steady-state spheroid system

Primordial germ cell-mediated chimera technology produces viable pure-line Houbara bustard offspring: potential for repopulating an endangered species.

Directory of Open Access Journals (Sweden)

Ulrich Wernery

2010-12-01

Full Text Available The Houbara bustard (Chlamydotis undulata is a wild seasonal breeding bird populating arid sandy semi-desert habitats in North Africa and the Middle East. Its population has declined drastically during the last two decades and it is classified as vulnerable. Captive breeding programmes have, hitherto, been unsuccessful in reviving population numbers and thus radical technological solutions are essential for the long term survival of this species. The purpose of this study was to investigate the use of primordial germ cell-mediated chimera technology to produce viable Houbara bustard offspring.Embryonic gonadal tissue was dissected from Houbara bustard embryos at eight days post-incubation. Subsequently, Houbara tissue containing gonadal primordial germ cells (gPGCs was injected into White Leghorn chicken (Gallus gallus domesticus embryos, producing 83/138 surviving male chimeric embryos, of which 35 chimeric roosters reached sexual maturity after 5 months. The incorporation and differentiation of Houbara gPGCs in chimeric chicken testis were assessed by PCR with Houbara-specific primers and 31.3% (5/16 gonads collected from the injected chicken embryos showed the presence of donor Houbara cells. A total of 302 semen samples from 34 chimeric roosters were analyzed and eight were confirmed as germline chimeras. Semen samples from these eight roosters were used to artificially inseminate three female Houbara bustards. Subsequently, 45 Houbara eggs were obtained and incubated, two of which were fertile. One egg hatched as a male live born Houbara; the other was female but died before hatching. Genotyping confirmed that the male chick was a pure-line Houbara derived from a chimeric rooster.This study demonstrates for the first time that Houbara gPGCs can migrate, differentiate and eventually give rise to functional sperm in the chimeric chicken testis. This approach may provide a promising tool for propagation and conservation of endangered avian

Strengthening Agricultural Research Capacity for Viable Extension ...

African Journals Online (AJOL)

Strengthening Agricultural Research Capacity for Viable Extension Policies in Nigeria: An Exploration of Ricoeur's Hermeneutic Theory for Analysing Extension Research. ... Progressively more, researchers use hermeneutic philosophy to inform the conduct of interpretive research. Analogy between the philosophical ...

Thermally assisted acoustofluidic separation of extracellular vesicles from cells

Science.gov (United States)

Mirtaheri, Elnaz; Dolatmoradi, Ata; Pimentel, Krystine; Bhansali, Shekhar; El-Zahab, Bilal

2018-02-01

Extracellular vesicles (EVs) have been gaining increasing attention given their role in communicating information between cells. Composition-based isolation of EVs is particularly of high significance as the proteomic and lipidomic characterization of their cargo could provide valuable clues to the role of EVs in mediating the biology of various conditions. This has, however, proved to be challenging as EVs, despite their abundance, are very small and difficult to be differentiated from the other constituents of host media. In addition, currently available methods like ultracentrifugation and filtration are cumbersome and capable of achieving mostly size-based separations. In this work, we demonstrate the possibility of separating submicron EV-like vesicles from cancer cells using a thermally-assisted acoustophoretic device. In a system composed of MCF-7 breast cancer cells spiked with two different types of same-size vesicles, composition-based isolation of vesicles was shown to be realizable through opposite focusing of the system's components at the node and antinodes of the overlaid ultrasonic standing wave. By proper choice of temperature in the microchannel, we were able to achieve separations with purities exceeding 93%. Furthermore, cells recovered from the channel were shown to be viable after the separation.

Increased cell surface metallopeptidase activity in cells undergoing UV-induced apoptosis

International Nuclear Information System (INIS)

Piva, T.J.; Davern, C.M.; Ellem, K.A.O.

1999-01-01

Full text: We have previously shown that UVC irradiation activated a range of cell surface peptidases (CSP) in HeLa cell monolayer cultures 20 h post-irradiation (1). In cells undergoing apoptosis there is an increase in CSP activity compared to control viable cells in cultures which have been treated by a wide range of agents including UV-irradiation (2). In order to further understand the mechanism involved in this process, we induced apoptosis in HeLa cells using 500 Jm -2 UVB. The separation of viable, apoptotic and necrotic cells of irradiated HeLa cell cultures was made by FACS analysis and sorting. The three populations were distinguished by their staining with PI and Hoechst 33342 dyes. CSP activity was measured using the P9 assay developed in this laboratory (1-3). The viable fraction of the irradiated cells had a higher level of CSP activity compared to unirradiated controls. The level of CSP activity in the apoptotic fraction was higher than that of the viable fraction, however that of the necrotic fraction was significantly lower. This finding agreed with that seen in UVC-irradiated (50 Jm -2 ) cultures (2). In order to elucidate the mechanism by which CSP activity was increased in UVB-irradiated cells undergoing apoptosis, the cultures were treated with the following agents: bestatin, aminopeptidase inhibitor, DEVD, caspase 3 inhibitor, and 3-aminobenzamide (3AB), PARP activation inhibitor. Bestatin and DEVD did not affect the level of CSP activity in the different cell subpopulations following UVB-irradiation. Treatment with 3AB abolished the increased CSP activity seen in the viable and apoptotic fraction following UVB-irradiation. All treated cells had the same morphology as observed under EM. The degree of phosphatidylserine eversion on the cell membrane was similar as were the cleavage profiles of PARP and actin. Only DEVD-treated cells had reduced caspase 3 activity which confirmed that the activation of CSP activity in apoptotic cells is

Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

Science.gov (United States)

Moyne, Anne-Laure; Harris, Linda J.; Marco, Maria L.

2013-01-01

Leafy green produce has been associated with numerous outbreaks of foodborne illness caused by strains of Escherichia coli O157:H7. While the amounts of culturable E. coli O157:H7 rapidly decline after introduction onto lettuce in the field, it remains to be determined whether the reduction in cell numbers is due to losses in cell viability, cell injury and a subsequent inability to be detected by standard laboratory culturing methods, or a lack of adherence and hence rapid removal of the organism from the plants during application. To assess which of these options is most relevant for E. coli O157:H7 on leafy green produce, we developed and applied a propidium monoazide (PMA) real-time PCR assay to quantify viable (with PMA) and total (without PMA) E. coli O157:H7 cells on growth chamber and field-grown lettuce. E. coli O157:H7, suspended in 0.1% peptone, was inoculated onto 4-week-old lettuce plants at a level of approximately 106 CFU/plant. In the growth chamber at low relative humidity (30%), culturable amounts of the nontoxigenic E. coli O157:H7 strain ATCC 700728 and the virulent strain EC4045 declined 100 to 1000-fold in 24 h. Fewer E. coli O157:H7 cells survived when applied onto plants in droplets with a pipette compared with a fine spray inoculation. Total cells for both strains were equivalent to inoculum levels for 7 days after application, and viable cell quantities determined by PMA real-time PCR were approximately 104 greater than found by colony enumeration. Within 2 h after application onto plants in the field, the number of culturable E. coli ATCC 700728 was reduced by up to 1000-fold, whereas PCR-based assessments showed that total cell amounts were equivalent to inoculum levels. These findings show that shortly after inoculation onto plants, the majority of E. coli O157:H7 cells either die or are no longer culturable. PMID:23936235

Whole-Genome Sequencing and iPLEX MassARRAY Genotyping Map an EMS-Induced Mutation Affecting Cell Competition in Drosophila melanogaster

Directory of Open Access Journals (Sweden)

Chang-Hyun Lee

2016-10-01

Full Text Available Cell competition, the conditional loss of viable genotypes only when surrounded by other cells, is a phenomenon observed in certain genetic mosaic conditions. We conducted a chemical mutagenesis and screen to recover new mutations that affect cell competition between wild-type and RpS3 heterozygous cells. Mutations were identified by whole-genome sequencing, making use of software tools that greatly facilitate the distinction between newly induced mutations and other sources of apparent sequence polymorphism, thereby reducing false-positive and false-negative identification rates. In addition, we utilized iPLEX MassARRAY for genotyping recombinant chromosomes. These approaches permitted the mapping of a new mutation affecting cell competition when only a single allele existed, with a phenotype assessed only in genetic mosaics, without the benefit of complementation with existing mutations, deletions, or duplications. These techniques expand the utility of chemical mutagenesis and whole-genome sequencing for mutant identification. We discuss mutations in the Atm and Xrp1 genes identified in this screen.

Investigation of Removal Capacities of Biofilters for Airborne Viable Micro-Organisms

Science.gov (United States)

Soret, Rémi; Fanlo, Jean-Louis; Malhautier, Luc; Geiger, Philippe; Bayle, Sandrine

2018-01-01

New emerging issues appears regarding the possible aerosolization of micro-organisms from biofilters to the ambient air. Traditional bioaerosol sampling and cultural methods used in literature offer relative efficiencies. In this study, a new method revolving around a particle counter capable of detecting total and viable particles in real time was used. This counter (BioTrak 9510-BD) uses laser-induced fluorescence (LIF) technology to determine the biological nature of the particle. The concentration of viable particles was measured on two semi-industrial pilot scale biofilters in order to estimate the Removal Efficiency in viable particles (REvp) in stable conditions and to examine the influence of pollutant feeding and relative humidification of the gaseous effluent on the REvp. The REvp of biofilters reached near 80% and highlighted both the stability of that removal and the statistical equivalence between two identical biofilters. Pollutant deprivation periods of 12 h, 48 h and 30 days were shown to have no influence on the biofilters’ removal capacity, demonstrating the robustness and adaptation capacities of the flora. In contrast, a 90-day famine period turned the biofilters into emitters of viable particles. Finally, the humidification of the effluent was shown to negatively influence the removal capacity for viable particles, as drying off the air was shown to increase the REvp from 60 to 85%. PMID:29562709

Autolyse the cell in order to save it? Inducing, then blocking, autolysis as a strategy for delaying cell death in the probiotic Lactobacillus reuteri.

Science.gov (United States)

Zimmerman, Tahl; Gyawali, Rabin; Ibrahim, Salam

2017-10-01

To examine whether choline and its derivatives can be used to preserve viable cells of Lactobacillus reuteri in autolytic models. A phosphate-induced autolytic model in de Man, Rogosa and Sharpe medium (MRS) was used. Viable cell counts were determined by plated on MRS-agar. Choline and hemicholinium-3 (HC-3) significantly blocked autolysis of L. reuteri at 360 mM and 4 mM, respectively. Viable cell counts corroborated these observations. Importantly, autolytically induced cells treated with choline and hemicholinium-3 were significantly more viable then even non-induced cells. Over-production of a known autolytic protein, spirosin, was not attenuated in the presence of choline and hemicholinium-3. Inducing autolysis and then blocking it with choline and its analogs is a promising approach for retaining the viability of L. reuteri cells.

Post-Flight Microbial Analysis of Samples from the International Space Station Water Recovery System and Oxygen Generation System

Science.gov (United States)

Birmele, Michele N.

2011-01-01

The Regenerative, Environmental Control and Life Support System (ECLSS) on the International Space Station (ISS) includes the the Water Recovery System (WRS) and the Oxygen Generation System (OGS). The WRS consists of a Urine Processor Assembly (UPA) and Water Processor Assembly (WPA). This report describes microbial characterization of wastewater and surface samples collected from the WRS and OGS subsystems, returned to KSC, JSC, and MSFC on consecutive shuttle flights (STS-129 and STS-130) in 2009-10. STS-129 returned two filters that contained fluid samples from the WPA Waste Tank Orbital Recovery Unit (ORU), one from the waste tank and the other from the ISS humidity condensate. Direct count by microscopic enumeration revealed 8.38 x 104 cells per mL in the humidity condensate sample, but none of those cells were recoverable on solid agar media. In contrast, 3.32 x lOs cells per mL were measured from a surface swab of the WRS waste tank, including viable bacteria and fungi recovered after S12 days of incubation on solid agar media. Based on rDNA sequencing and phenotypic characterization, a fungus recovered from the filter was determined to be Lecythophora mutabilis. The bacterial isolate was identified by rDNA sequence data to be Methylobacterium radiotolerans. Additional UPA subsystem samples were returned on STS-130 for analysis. Both liquid and solid samples were collected from the Russian urine container (EDV), Distillation Assembly (DA) and Recycle Filter Tank Assembly (RFTA) for post-flight analysis. The bacterium Pseudomonas aeruginosa and fungus Chaetomium brasiliense were isolated from the EDV samples. No viable bacteria or fungi were recovered from RFTA brine samples (N= 6), but multiple samples (N = 11) from the DA and RFTA were found to contain fungal and bacterial cells. Many recovered cells have been identified to genus by rDNA sequencing and carbon source utilization profiling (BiOLOG Gen III). The presence of viable bacteria and fungi from WRS

Ergonomic analysis jobs in recovered factories.

Science.gov (United States)

Cuenca, Gabriela; Zotta, Gastón

2012-01-01

With the advent of the deep economic crisis in Argentina on 2001, the recovery of companies through to the creation of the Cooperatives Working Self-Management or Factories Recovered by its workers was constituted as one of the ways in which the salaried disobeyed the increasing unemployment. When the companies turn into recovered factories they tend to leave of side practices that have been seen like imposed by the previous organization and not understanding them as a primary condition for the execution of his tasks. Safety and ergonomics are two disciplines that are no longer considered relevant to the daily work. Therefore this investigation aims to revalue, undergo semantic to give back to a place in every organization analyzed. This research developed a self-diagnostic tool for working conditions, and the environment, present in the recovered factories.

Viable-but-Nonculturable Listeria monocytogenes and Salmonella enterica Serovar Thompson Induced by Chlorine Stress Remain Infectious

Directory of Open Access Journals (Sweden)

Callum J. Highmore

2018-04-01

Full Text Available The microbiological safety of fresh produce is monitored almost exclusively by culture-based detection methods. However, bacterial food-borne pathogens are known to enter a viable-but-nonculturable (VBNC state in response to environmental stresses such as chlorine, which is commonly used for fresh produce decontamination. Here, complete VBNC induction of green fluorescent protein-tagged Listeria monocytogenes and Salmonella enterica serovar Thompson was achieved by exposure to 12 and 3 ppm chlorine, respectively. The pathogens were subjected to chlorine washing following incubation on spinach leaves. Culture data revealed that total viable L. monocytogenes and Salmonella Thompson populations became VBNC by 50 and 100 ppm chlorine, respectively, while enumeration by direct viable counting found that chlorine caused a <1-log reduction in viability. The pathogenicity of chlorine-induced VBNC L. monocytogenes and Salmonella Thompson was assessed by using Caenorhabditis elegans. Ingestion of VBNC pathogens by C. elegans resulted in a significant life span reduction (P = 0.0064 and P < 0.0001, and no significant difference between the life span reductions caused by the VBNC and culturable L. monocytogenes treatments was observed. L. monocytogenes was visualized beyond the nematode intestinal lumen, indicating resuscitation and cell invasion. These data emphasize the risk that VBNC food-borne pathogens could pose to public health should they continue to go undetected.

9 CFR 113.26 - Detection of viable bacteria and fungi except in live vaccine.

Science.gov (United States)

2010-01-01

... 9 Animals and Animal Products 1 2010-01-01 2010-01-01 false Detection of viable bacteria and fungi... VECTORS STANDARD REQUIREMENTS Standard Procedures § 113.26 Detection of viable bacteria and fungi except... required to be free of viable bacteria and fungi, they shall also be tested as prescribed in this section...

Can Malin's gravitational-field equations be modified to obtain a viable theory of gravity to obtain a viable theory of gravity to obtain a viable theory of gravity

International Nuclear Information System (INIS)

Smalley, L.L.; Prestage, J.

1976-01-01

Malin's gravitational theory, which was recently shown by Lindblom and Nester to be incorrect, is modified by means of a recently proposed method for obtaining viable gravitational theories. The resulting self-consistent theory, which is in effect a Rastall-type modification of the Einstein theory, exhibits nonconservation of momentum, yet agrees with all experimental limits known to date within the PPN framework

Monotone viable trajectories for functional differential inclusions

Science.gov (United States)

Haddad, Georges

This paper is a study on functional differential inclusions with memory which represent the multivalued version of retarded functional differential equations. The main result gives a necessary and sufficient equations. The main result gives a necessary and sufficient condition ensuring the existence of viable trajectories; that means trajectories remaining in a given nonempty closed convex set defined by given constraints the system must satisfy to be viable. Some motivations for this paper can be found in control theory where F( t, φ) = { f( t, φ, u)} uɛU is the set of possible velocities of the system at time t, depending on the past history represented by the function φ and on a control u ranging over a set U of controls. Other motivations can be found in planning procedures in microeconomics and in biological evolutions where problems with memory do effectively appear in a multivalued version. All these models require viability constraints represented by a closed convex set.

Phospholipid fatty acid analysis as part of the Yucca Mountain Project. Final report

International Nuclear Information System (INIS)

Ringelberg, D.B.; White, D.C.

1996-01-01

In support of the Yucca Mountain subsurface microbial characterization project phospholipid fatty acid (PLFA) analyses for viable microbial biomass, community composition and nutritional status were performed. Results showed a positive correlation between a decrease in viable biomass and increase in depth with the lowest biomass values being obtained from the Topopah Spring geologic horizon. A plot of the ratio of non-viable (diglyceride fatty acids) to viable (PLFA) cells also showed the lowest values to derive from the Topopah Spring horizon. Estimations of microbial community composition, made from the patterns of PLFA recovered from the sediment samples, revealed similarities between samples collected within the same geologic horizons: Tiva Canyon, Pre-Pah Canyon and Topopah Spring. Results indicated the presence of mixed communities composed of gram positive, gram negative, actinomycete and obligate anaerobic bacteria. Culturable organisms, recovered from similar sediments, were representative of the same bacterial classifications although gram positive bacterial isolates typically outnumbered gram negative isolates. Within the gram negative bacterial community, corroborative indicators of physiological stress were apparent in the Topopah Spring horizon

Distinctive cellular immunity in genetically susceptible BALB/c mice recovered from Leishmania major infection or after subcutaneous immunization with killed parasites

International Nuclear Information System (INIS)

Liew, F.Y.; Dhaliwal, J.S.

1987-01-01

Genetically susceptible BALB/c mice are refractory to further infection after recovery from Leishmania major infection after a sublethal dose of gamma-irradiation. In contrast, mice immunized with killed promastigotes s.c. develop exacerbated lesions after infection. Both groups of mice produce only a low level of specific antibody and no detectable cytotoxic T cells, but do have a strong antigen-specific DTH, which is adoptively transferable with Lyt-1+2-, L3T4+ T cells. Kinetic and histological studies revealed that mice immunized s.c. developed Jones-Mote hypersensitivity, peaking at 15 hr. with little mononuclear cell infiltration at the site of antigen administration; whereas mice that had recovered from infection developed tuberculin-type of reactivity, peaking at 24 to 48 hr, with intense mononuclear cell infiltration. Splenic T cells from recovered mice, when injected into the footpads of normal recipients together with live promastigotes, were able to retard lesion development; whereas T cells from s.c. immunized mice, when similarly transferred, accelerated disease progression. Antigen-specific culture supernatant of spleen cells from recovered mice also activated normal resident peritoneal macrophages to kill intracellular L. major amastigotes and tumor cells. Culture supernatants of spleen cells from s.c. immunized or normal mice were devoid of such activities. Part of the macrophage-activating potential can be inhibited by antibody specific for IFN-gamma. These results therefore demonstrate that whereas the Jones-Mote reaction is correlated with disease exacerbation, the tuberculin-type of DTH may be protective. Furthermore, in vivo immunity is directly related to the capacity of T cells to produce macrophage-activating factor

Viable Techniques, Leontief’s Closed Model, and Sraffa’s Subsistence Economies

Directory of Open Access Journals (Sweden)

Alberto Benítez

2014-11-01

Full Text Available This paper studies the production techniques employed in economies that reproduce themselves. Special attention is paid to the distinction usually made between those that do not produce a surplus and those that do, which are referred to as first and second class economies, respectively. Based on this, we present a new definition of viable economies and show that every viable economy of the second class can be represented as a viable economy of the first class under two different forms, Leontief‘s closed model and Sraffa’s subsistence economies. This allows us to present some remarks concerning the economic interpretation of the two models. On the one hand, we argue that the participation of each good in the production of every good can be considered as a normal characteristic of the first model and, on the other hand, we provide a justification for the same condition to be considered a characteristic of the second model. Furthermore, we discuss three definitions of viable techniques advanced by other authors and show that they differ from ours because they admit economies that do not reproduce themselves completely.

Arginase treatment prevents the recovery of canine lymphoma and osteosarcoma cells resistant to the toxic effects of prolonged arginine deprivation.

Science.gov (United States)

Wells, James W; Evans, Christopher H; Scott, Milcah C; Rütgen, Barbara C; O'Brien, Timothy D; Modiano, Jaime F; Cvetkovic, Goran; Tepic, Slobodan

2013-01-01

Rapidly growing tumor cells require a nutrient-rich environment in order to thrive, therefore, restricting access to certain key amino acids, such as arginine, often results in the death of malignant cells, which frequently display defective cell cycle check-point control. Healthy cells, by contrast, become quiescent and remain viable under arginine restriction, displaying full recovery upon return to arginine-rich conditions. The use of arginase therapy to restrict available arginine for selectively targeting malignant cells is currently under investigation in human clinical trials. However, the suitability of this approach for veterinary uses is unexplored. As a prelude to in vivo studies in canine malignancies, we examined the in vitro effects of arginine-deprivation on canine lymphoid and osteosarcoma cell lines. Two lymphoid and 2 osteosarcoma cell lines were unable to recover following 6 days of arginine deprivation, but all remaining cell lines displayed full recovery upon return to arginine-rich culture conditions. These remaining cell lines all proved susceptible to cell death following the addition of arginase to the cultures. The lymphoid lines were particularly sensitive to arginase, becoming unrecoverable after just 3 days of treatment. Two of the osteosarcoma lines were also susceptible over this time-frame; however the other 3 lines required 6-8 days of arginase treatment to prevent recovery. In contrast, adult progenitor cells from the bone marrow of a healthy dog were able to recover fully following 9 days of culture in arginase. Over 3 days in culture, arginase was more effective than asparaginase in inducing the death of lymphoid lines. These results strongly suggest that short-term arginase treatment warrants further investigation as a therapy for lymphoid malignancies and osteosarcomas in dogs.

Development of a propidium monoazide-polymerase chain reaction assay for detection of viable Lactobacillus brevis in beer.

Science.gov (United States)

Ma, Yanlin; Deng, Yang; Xu, Zhenbo; Liu, Junyan; Dong, Jianjun; Yin, Hua; Yu, Junhong; Chang, Zongming; Wang, Dongfeng

The spoilage of beer by bacteria is of great concern to the brewer as this can lead to turbidity and abnormal flavors. The polymerase chain reaction (PCR) method for detection of beer-spoilage bacteria is highly specific and provides results much faster than traditional microbiology techniques. However, one of the drawbacks is the inability to differentiate between live and dead cells. In this paper, the combination of propidium monoazide (PMA) pretreatment and conventional PCR had been described. The established PMA-PCR identified beer spoilage Lactobacillus brevis based not on their identity, but on the presence of horA gene which we show to be highly correlated with the ability of beer spoilage LAB to grow in beer. The results suggested that the use of 30μg/mL or less of PMA did not inhibit the PCR amplification of DNA derived from viable L. brevis cells. The minimum amount of PMA to completely inhibit the PCR amplification of DNA derived from dead L. brevis cells was 2.0μg/mL. The detection limit of PMA-PCR assay described here was found to be 10 colony forming units (CFU)/reaction for the horA gene. Moreover, the horA-specific PMA-PCR assays were subjected to 18 reference isolates, representing 100% specificity with no false positive amplification observed. Overall the use of horA-specific PMA-PCR allows for a substantial reduction in the time required for detection of potential beer spoilage L. brevis and efficiently differentiates between viable and nonviable cells. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Primordial Germ Cell-Mediated Chimera Technology Produces Viable Pure-Line Houbara Bustard Offspring: Potential for Repopulating an Endangered Species

Science.gov (United States)

Wernery, Ulrich; Liu, Chunhai; Baskar, Vijay; Guerineche, Zhor; Khazanehdari, Kamal A.; Saleem, Shazia; Kinne, Jörg; Wernery, Renate

2010-01-01

Background The Houbara bustard (Chlamydotis undulata) is a wild seasonal breeding bird populating arid sandy semi-desert habitats in North Africa and the Middle East. Its population has declined drastically during the last two decades and it is classified as vulnerable. Captive breeding programmes have, hitherto, been unsuccessful in reviving population numbers and thus radical technological solutions are essential for the long term survival of this species. The purpose of this study was to investigate the use of primordial germ cell-mediated chimera technology to produce viable Houbara bustard offspring. Methodology/Principal Findings Embryonic gonadal tissue was dissected from Houbara bustard embryos at eight days post-incubation. Subsequently, Houbara tissue containing gonadal primordial germ cells (gPGCs) was injected into White Leghorn chicken (Gallus gallus domesticus) embryos, producing 83/138 surviving male chimeric embryos, of which 35 chimeric roosters reached sexual maturity after 5 months. The incorporation and differentiation of Houbara gPGCs in chimeric chicken testis were assessed by PCR with Houbara-specific primers and 31.3% (5/16) gonads collected from the injected chicken embryos showed the presence of donor Houbara cells. A total of 302 semen samples from 34 chimeric roosters were analyzed and eight were confirmed as germline chimeras. Semen samples from these eight roosters were used to artificially inseminate three female Houbara bustards. Subsequently, 45 Houbara eggs were obtained and incubated, two of which were fertile. One egg hatched as a male live born Houbara; the other was female but died before hatching. Genotyping confirmed that the male chick was a pure-line Houbara derived from a chimeric rooster. Conclusion This study demonstrates for the first time that Houbara gPGCs can migrate, differentiate and eventually give rise to functional sperm in the chimeric chicken testis. This approach may provide a promising tool for propagation

Investigation of mechanical properties of hydrate-bearing pressure core sediments recovered from the Eastern Nankai Trough using transparent acrylic cell triaxial testing system (TACTT-system)

Science.gov (United States)

Yoneda, J.; Masui, A.; Konno, Y.; Jin, Y.; Kida, M.; Suzuki, K.; Nakatsuka, Y.; Tenma, N.; Nagao, J.

2014-12-01

Natural gas hydrate-bearing pressure core sediments have been sheared in compression using a newly developed Transparent Acrylic Cell Triaxial Testing (TACTT) system to investigate the geophysical and geomechanical behavior of sediments recovered from the deep seabed in the Eastern Nankai Trough, the first Japanese offshore production test region. The sediments were recovered by hybrid pressure core system (hybrid PCS) and pressure cores were cut by pressure core analysis tools (PCATs) on board. These pressure cores were transferred to the AIST Hokkaido centre and trimmed by pressure core non-destructive analysis tools (PNATs) for TACTT system which maintained the pressure and temperature conditions within the hydrate stability boundary, through the entire process of core handling from drilling to the end of laboratory testing. An image processing technique was used to capture the motion of sediment in a transparent acrylic cell, and digital photographs were obtained at every 0.1% of vertical strain during the test. Analysis of the optical images showed that sediments with 63% hydrate saturation exhibited brittle failure, although nonhydrate-bearing sediments exhibited ductile failure. In addition, the increase in shear strength with hydrate saturation increase of natural gas hydrate is in agreement with previous data from synthetic gas hydrate. This research was financially supported by the Research Consortium for Methane Hydrate Resources in Japan (MH21 Research Consortium) that carries out Japan's Methane Hydrate R&D Program by the Ministry of Economy, Trade and Industry (METI).

The viable but non-culturable state in pathogenic Escherichia coli: A general review

Directory of Open Access Journals (Sweden)

Jennifer A. Pienaar

2016-05-01

Objectives: This review discusses various general aspects of the VBNC state, the mechanisms and possible public health impact of indicator and pathogenic E. coli entering into the VBNC state. Method: A literature review was conducted to ascertain the possibleimpact of E. coli entering into the VBNC state. Results: Escherichia coli enter into the VBNC state by means of several induction mechanisms. Various authors have found that E. coli can be resuscitated post-VBNC. Certain strains of pathogenic E. coli are still able to produce toxins in the VBNC state, whilst others are avirulent during the VBNC state but are able to regain virulence after resuscitation. Conclusion: Pathogenic and indicator E. coli entering into the VBNC state could have an adverse effect on public health if conventional detection methods are used, where the number of viable cells could be underestimated and the VBNC cells still produce toxins or could, at anytime, be resuscitated and become virulent again.

Recovering hydrocarbons with surfactants from lignin

Energy Technology Data Exchange (ETDEWEB)

Naae, D.G.; Whittington, L.E.; Ledoux, W.A.; Debons, F.E.

1988-11-29

This patent describes a method of recovering hydrocarbons from an underground hydrocarbon formation penetrated by at least one injection well and at least one production well, which comprises: injecting into the formation through an injection well a surfactant slug comprising about 0.1% to about 10% by weight of surfactants produced from lignin, the surfactants produced by placing lignin in contact with water, converting the lignin into low molecular weight lignin phenols by reducing the lignin in the presence of a reducing agent of carbon monoxide or hydrogen creating a reduction reaction mixture comprising oil soluble lignin phenols, the reduction occurring at a temperature greater than about 200/sup 0/C and a pressure greater than about 100 psi, recovering the oil soluble lignin phenols from the reduction mixture, and converting the lignin phenols into lignin surfactants by a reaction selected from the group consisting of alkoxylation, sulfonation, sulfation, aklylation, sulfomethylation, and alkoxysulfation; injecting into the formation through the injection well a drive fluid to push the surfactant slug towards a production well; and recovering hydrocarbons at the production well.

Prophage induction and cell division in E. coli. Pt. 3

International Nuclear Information System (INIS)

George, J.; Castellazzi, M.; Buttin, G.

1975-01-01

In E. coli K12, cell filamentation promoted by tif is enhanced by the lon mutation; in contrast, prophage induction and repair of UV-irradiated phage lambda, also promoted by tif, are not affected by lon. From a tif lon double mutant, 'revertants' having recovered the ability to divide at 41 0 were isolated, among which most (95%) had also lost heir Lon filamentous phenotype after ultraviolet (UV) irradiation. From these 95% of revertants 94% are suppressed for the whole Tif phenotype, by additional mutations that render them deficient in DNA repair, as judged from their high UV sensitivity; some have been characterized as recA mutants. 1% have recovered a control on cell division at 41% or after UV irradiation by means of secondary mutations altering neither the other phenotypic properties of tif and lon, nor the repair and recombination ability of the cells: in particular, this class of 'revertants' remains thermoinducible upon lysogenisation; the mutations which specifically supress filamentation have been mapped at two loci, sfiA and sfiB, cotransducible respectively with pyrD and leu. In the remaining 5% of revertants that still exhibit an UV-induced filamentous growth, 3% can be tentatively classified as true tif + revertants; 2% behave as tif thermodependent revertants, showing suppression of Tif (and Lon) phenotype only at 41 0 : 2 recAts have been identified in this class. Non-lysogenic tif lon sfi and tif sfi strains remain viable during prolonged growth at 41 0 . Under these conditions, tif expresses mutator properties, which can be conveniently analyzed in this sfi background. The action of tif, lon and sfi mutations is tentatively interpreted on the basis of a negative control of cell division specifically associated with DNA repair. (orig.) [de

Characterization and functional properties of gastric tissue-resident memory T cells from children, adults and the elderly

Directory of Open Access Journals (Sweden)

Jayaum S. Booth

2014-06-01

Full Text Available T cells are the main orchestrators of protective immunity in the stomach; however, limited information on the presence and function of the gastric T subsets is available mainly due to the difficulty in recovering high numbers of viable cells from human gastric biopsies. To overcome this shortcoming we optimized a cell isolation method that yielded high numbers of viable lamina propria mononuclear cells (LPMC from gastric biopsies. Classic memory T (TM subsets were identified in gastric LPMC and compared to peripheral blood mononuclear cells (PBMC obtained from children, adults and the elderly using an optimized 14 color flow cytometry panel. A dominant effector memory (TEM phenotype was observed in gastric LPMC CD4+ and CD8+ T cells in all age groups. We then evaluated whether these cells represented a population of gastric tissue-resident memory T (TRM cells by assessing expression of CD103 and CD69. The vast majority of gastric LPMC CD8+ T cells either co-expressed CD103/CD69 (>70% or expressed CD103 alone (~20%. Gastric LPMC CD4+ T cells also either co-expressed CD103/CD69 (>35% or expressed at least one of these markers. Thus, gastric LPMC CD8+ and CD4+ T cells had the characteristics of TRM cells. Gastric CD8+ and CD4+ TRM cells produced multiple cytokines (IFN-γ, IL-2, TNF-α, IL-17A, MIP-1β and up-regulated CD107a upon stimulation. However, marked differences were observed in their cytokine and multi-cytokine profiles when compared to their PBMC TEM counterparts. Furthermore, gastric CD8+ TRM and CD4+ TRM cells demonstrated differences in the frequency, susceptibility to activation and cytokine/multi-cytokine production profiles among the age groups. Most notably, children’s gastric TRM cells responded differently to stimuli than gastric TRM cells from adults or the elderly. In conclusion, we demonstrate the presence of gastric TRM which exhibit diverse functional characteristics in children, adults and the elderly.

Transuranium element recovering method for spent nuclear fuel

International Nuclear Information System (INIS)

Todokoro, Akio; Kihara, Yoshiyuki; Okada, Hisashi

1998-01-01

Spent fuels are dissolved in nitric acid, the obtained dissolution liquid is oxidized by electrolysis, and nitric acid of transuranium elements are precipitated together with nitric acid of uranium elements from the dissolution solution and recovered. Namely, the transuranium elements are oxidized to an atomic value level at which nitric acid can be precipitated by an oxidizing catalyst, and cooled to precipitate nitric acid of transuranium elements together with nitric acid of transuranium elements, accordingly, it is not necessary to use a solvent which has been used so far upon recovering transuranium elements. Since no solvent waste is generated, a recovery method taking the circumstance into consideration can be provided. Further, nitric acid of uranium elements and nitric acid of transuranium elements precipitated and recovered together are dissolved in nitric acid again, cooled and only uranium elements are precipitated selectively, and recovered by filtration. The amount of wastes can be reduced to thereby enabling to mitigate control for processing. (N.H.)

Growth Phase, Oxygen, Temperature and Starvation Affect the Development of Viable but Non-Culturable State of Vibrio cholerae

Directory of Open Access Journals (Sweden)

Bin eWu

2016-03-01

Full Text Available AbstractVibrio cholerae can enter into a viable but non-culturable (VBNC state in order to survive in unfavourable environments. In this study, we studied the roles of five physicochemical and microbiological factors or states, namely, different strains, growth phases, oxygen, temperature, and starvation, on the development of VBNC of V. cholerae in artificial sea water (ASW. Different strains of the organism, the growth phase, and oxygen levels affected the progress of VBNC development. It was found that the VBNC state was induced faster in V. cholerae serogroup O1 classical biotype strain O395 than in O1 El Tor biotype strains C6706 and N16961. When cells in different growth phases were used for VBNC induction, stationary-phase cells lost their culturability more quickly than exponential-phase cells, while induction of a totally non-culturable state took longer to achieve for stationary-phase cells in all three strains, suggesting that heterogeneity of cells should be considered. Aeration strongly accelerated the loss of culturability. During the development of the VBNC state, the culturable cell count under aeration conditions was almost 106-fold lower than under oxygen-limited conditions for all three strains. The other two factors, temperature and nutrients-rich environment, may prevent the induction of VBNC cells. At 22°C or 37°C in ASW, most of the cells rapidly died and the culturable cell count reduced from about 108 CFU/mL to 106–105 CFU/mL. The total cell counts showed that cells that lost viability were decomposed, and the viable cell counts were the same as culturable cell counts, indicating that the cells did not reach the VBNC state. VBNC state development was blocked when ASW was supplied with Luria-Bertani broth (LB, but it was not affected in ASW with M9, suggesting that specific nutrients in LB may prevent the development of VBNC state. These results revealed that the five factors evaluated in this study had different

Association of viable Mycobacterium leprae with Type 1 reaction in leprosy.

Science.gov (United States)

Save, Mrudula Prakash; Dighe, Anju Rajaram; Natrajan, Mohan; Shetty, Vanaja Prabhakaran

2016-03-01

The working hypothesis is that, viable Mycobacterium leprae (M. leprae) play a crucial role in the precipitation of Type 1 reaction (T1R) in leprosy. A total of 165 new multibacillary patients were studied. To demonstrate presence of viable M. leprae in reactional lesion (T1R+), three tests were used concurrently viz. growth in the mouse foot pad (MFP), immunohistochemical detection of M. leprae secretory protein Ag85, and 16s rRNA--using in situ RT-PCR. Mirror biopsies and non reactional lesions served as controls (T1R-). A significantly higher proportion of lesion biopsy homogenates obtained at onset, from T1R(+) cases have shown unequivocal growth in MFP, proving the presence of viable bacteria, as compared to T1R(-) (P leprae is a component/prerequisite and the secretory protein Ag 85, might be the trigger for precipitation of T1R.

Agglomeration and Deposition Behaviour of Solid Recovered Fuel

DEFF Research Database (Denmark)

Pedersen, Morten Nedergaard; Jensen, Peter Arendt; Nielsen, Mads

2015-01-01

Waste derived fuels such as Solid Recovered Fuel (SRF) are increasingly being used in the cement industry as a means to reduce cost [1]. SRF is produced by separating the combustible fraction from industrial or municipal solid waste (MSW). The recovered fraction has a higher content of combustibl...

Treating effluents; recovering coal, etc

Energy Technology Data Exchange (ETDEWEB)

Jones, F B; Bury, E

1920-02-18

Liquor obtained by scrubbing coal gas with sea-water or fresh water, and containing or having added to it finely-divided carbonaceous material in suspension, is subjected to a froth-flotation process to recover the carbonaceous matter and organic materials in the froth, and render the remaining liquor innocuous. Liquor obtained by scrubbing distillation gases, such as coal gas, may be used as a frothing-agent in a froth flotation process for the recovery of carbonaceous substances such as coal from materials containing them, thereby producing a froth containing the coal, etc., and also the organic materials from the liquor. In some cases the effluent may be diluted with sea-water, and, in recovering carbonaceous shales, there may be added to the liquor a small proportion of paraffin oil.

Cognitive bias in symptomatic and recovered agoraphobics.

Science.gov (United States)

Stoler, L S; McNally, R J

1991-01-01

Symptomatic agoraphobics, recovered agoraphobics, and normal control subjects completed a series of sentence stems that had either ambiguous or unambiguous meanings, and had either a potentially threatening or a nonthreatening connotation. The written completions made by subjects to these stems were classified as indicating either a biased (i.e. threat-related) or unbiased interpretation of the meaning of the stem, and if a biased interpretation was made, whether the subject indicated efforts at adaptive coping with the perceived threat. Results indicated that symptomatic agoraphobics exhibited strong biases for interpreting information as threatening, relative to normal control subjects. Moreover, recovered agoraphobics resembled symptomatic agoraphobics more than normal control subjects, thus indicating that cognitive biases may persist following cessation of panic attacks and reductions in avoidance behavior. However, recovered agoraphobics also exhibited tendencies to cope adaptively with perceived threats whereas symptomatic agoraphobics did not.

Whole-Genome Sequencing and iPLEX MassARRAY Genotyping Map an EMS-Induced Mutation Affecting Cell Competition in Drosophila melanogaster.

Science.gov (United States)

Lee, Chang-Hyun; Rimesso, Gerard; Reynolds, David M; Cai, Jinlu; Baker, Nicholas E

2016-10-13

Cell competition, the conditional loss of viable genotypes only when surrounded by other cells, is a phenomenon observed in certain genetic mosaic conditions. We conducted a chemical mutagenesis and screen to recover new mutations that affect cell competition between wild-type and RpS3 heterozygous cells. Mutations were identified by whole-genome sequencing, making use of software tools that greatly facilitate the distinction between newly induced mutations and other sources of apparent sequence polymorphism, thereby reducing false-positive and false-negative identification rates. In addition, we utilized iPLEX MassARRAY for genotyping recombinant chromosomes. These approaches permitted the mapping of a new mutation affecting cell competition when only a single allele existed, with a phenotype assessed only in genetic mosaics, without the benefit of complementation with existing mutations, deletions, or duplications. These techniques expand the utility of chemical mutagenesis and whole-genome sequencing for mutant identification. We discuss mutations in the Atm and Xrp1 genes identified in this screen. Copyright © 2016 Lee et al.

Single cell adhesion force measurement for cell viability identification using an AFM cantilever-based micro putter

Science.gov (United States)

Shen, Yajing; Nakajima, Masahiro; Kojima, Seiji; Homma, Michio; Kojima, Masaru; Fukuda, Toshio

2011-11-01

Fast and sensitive cell viability identification is a key point for single cell analysis. To address this issue, this paper reports a novel single cell viability identification method based on the measurement of single cell shear adhesion force using an atomic force microscopy (AFM) cantilever-based micro putter. Viable and nonviable yeast cells are prepared and put onto three kinds of substrate surfaces, i.e. tungsten probe, gold and ITO substrate surfaces. A micro putter is fabricated from the AFM cantilever by focused ion beam etching technique. The spring constant of the micro putter is calibrated using the nanomanipulation approach. The shear adhesion force between the single viable or nonviable cell and each substrate is measured using the micro putter based on the nanorobotic manipulation system inside an environmental scanning electron microscope. The adhesion force is calculated based on the deflection of the micro putter beam. The results show that the adhesion force of the viable cell to the substrate is much larger than that of the nonviable cell. This identification method is label free, fast, sensitive and can give quantitative results at the single cell level.

Sycamore produces viable seed after six years

Science.gov (United States)

A. F. Ike

1966-01-01

In the early stages of any tree improvement program it is desirable to know how soon progenies of selected parents can themselves be included in a breeding program. How soon will they produce viable pollen and seed? In the case of sycamore (Platanus occidentalis L.), the information is meager: the Woody- Plant Seed Manual lists the minimum commercial seedbearing age...

Arginase treatment prevents the recovery of canine lymphoma and osteosarcoma cells resistant to the toxic effects of prolonged arginine deprivation.

Directory of Open Access Journals (Sweden)

James W Wells

Full Text Available Rapidly growing tumor cells require a nutrient-rich environment in order to thrive, therefore, restricting access to certain key amino acids, such as arginine, often results in the death of malignant cells, which frequently display defective cell cycle check-point control. Healthy cells, by contrast, become quiescent and remain viable under arginine restriction, displaying full recovery upon return to arginine-rich conditions. The use of arginase therapy to restrict available arginine for selectively targeting malignant cells is currently under investigation in human clinical trials. However, the suitability of this approach for veterinary uses is unexplored. As a prelude to in vivo studies in canine malignancies, we examined the in vitro effects of arginine-deprivation on canine lymphoid and osteosarcoma cell lines. Two lymphoid and 2 osteosarcoma cell lines were unable to recover following 6 days of arginine deprivation, but all remaining cell lines displayed full recovery upon return to arginine-rich culture conditions. These remaining cell lines all proved susceptible to cell death following the addition of arginase to the cultures. The lymphoid lines were particularly sensitive to arginase, becoming unrecoverable after just 3 days of treatment. Two of the osteosarcoma lines were also susceptible over this time-frame; however the other 3 lines required 6-8 days of arginase treatment to prevent recovery. In contrast, adult progenitor cells from the bone marrow of a healthy dog were able to recover fully following 9 days of culture in arginase. Over 3 days in culture, arginase was more effective than asparaginase in inducing the death of lymphoid lines. These results strongly suggest that short-term arginase treatment warrants further investigation as a therapy for lymphoid malignancies and osteosarcomas in dogs.

Issues of organizational cybernetics and viability beyond Beer's viable systems model

Science.gov (United States)

Nechansky, Helmut

2013-11-01

The paper starts summarizing the claims of Beer's viable systems model to identify five issues any viable organizations has to deal with in an unequivocal hierarchical structure of five interrelated systems. Then the evidence is introduced for additional issues and related viable structures of organizations, which deviate from Beer's model. These issues are: (1) the establishment and (2) evolution of an organization; (3) systems for independent top-down control (like "Six Sigma"); (4) systems for independent bottom-up correction of performance problems (like "Kaizen"), both working outside a hierarchical structure; (5) pull production systems ("Just in Time") and (6) systems for checks and balances of top-level power (like boards and shareholder meetings). Based on that an evolutionary approach to organizational cybernetics is outlined, addressing the establishment of organizations and possible courses of developments, including recent developments in quality and production engineering, as well as problems of setting and changing goal values determining organizational policies.

Formas cocoides de Helicobacter pylori: viables o degenerativas

Directory of Open Access Journals (Sweden)

Felipe Cava

2003-06-01

Full Text Available De los trabajos presentados acerca de las formas cocoides de Helicobacter pylori se deduce una controversia mucho mayor que la resultante del mero estudio clínico de este microorganismo. Parece claro que existe una conversión tanto in vivo como in vitro de las formas espirales a las formas cocoides inducida por varios motivos, como cultivos prolongados, estrés físico y químico, y agentes antimicrobianos. En esta revisión repasamos los puntos de vista que han dividido a investigadores de esta área en dos grupos bien definidos: Los que consideran a estas formas cocoides como un producto no viable de degeneración celular y los que piensan que estas formas son estructuras viables,durmientes o de resistencia frente a condiciones ambientales adversas. Esta discrepancia conlleva a que interrogantes sobre la relación entre la transmisión de la enfermedad y estas formas cocoides permanezcan sin respuesta todavía.

Casticin impairs cell growth and induces cell apoptosis via cell cycle arrest in human oral cancer SCC-4 cells.

Science.gov (United States)

Chou, Guan-Ling; Peng, Shu-Fen; Liao, Ching-Lung; Ho, Heng-Chien; Lu, Kung-Wen; Lien, Jin-Cherng; Fan, Ming-Jen; La, Kuang-Chi; Chung, Jing-Gung

2018-02-01

Casticin, a polymethoxyflavone, present in natural plants, has been shown to have biological activities including anti-cancer activities. Herein, we investigated the anti-oral cancer activity of casticin on SCC-4 cells in vitro. Viable cells, cell cycle distribution, apoptotic cell death, reactive oxygen species (ROS) production, and Ca 2+ production, levels of ΔΨ m and caspase activity were measured by flow cytometric assay. Cell apoptosis associated protein expressions were examined by Western blotting and confocal laser microscopy. Results indicated that casticin induced cell morphological changes, DNA condensation and damage, decreased the total viable cells, induced G 2 /M phase arrest in SCC-4 cells. Casticin promoted ROS and Ca 2+ productions, decreases the levels of ΔΨ m , promoted caspase-3, -8, and -9 activities in SCC-4 cells. Western blotting assay demonstrated that casticin affect protein level associated with G2/M phase arrest and apoptosis. Confocal laser microscopy also confirmed that casticin increased the translocation of AIF and cytochrome c in SCC-4 cells. In conclusion, casticin decreased cell number through G 2 /M phase arrest and the induction of cell apoptosis through caspase- and mitochondria-dependent pathways in SCC-4 cells. © 2017 Wiley Periodicals, Inc.

Waste heat recovering device for reactors

International Nuclear Information System (INIS)

Sonoda, Masanobu; Shiraishi, Tadashi; Mizuno, Hiroyuki; Sekine, Yasuhiro.

1982-01-01

Purpose: To enable utilization of auxiliary-equipment-cooling water from a non-regenerative heat exchanger as a heat source, as well as prevent radioactive contamination. Constitution: A water warming device for recovering the heat of auxiliary equipment cooling water from a non-regenerative heat exchanger is disposed at the succeeding stage of the heat exchanger. Heat exchange is performed in the water warming device between the auxiliary equipment cooling water and a heat source water set to a higher pressure and recycled through the water warming device. The heat recovered from the auxiliary equipment cooling water is utilized in the heat source water for operating relevant equipments. (Aizawa, K.)

Extracellular delivery induced by ultrasound and microbubbles in cells

Science.gov (United States)

Hussein, Farah; Antonescu, Costin; Karshafian, Raffi

2017-03-01

Ultrasound and microbubble treatment (USMB) can enhance the intracellular uptake of molecules, which otherwise would be excluded from the cell, through USMB-mediated transient membrane disruption and through enhanced endocytosis. However, the effect of USMB on the outward movement of molecules from cells is not well understood. This study investigates the effects of USMB on the release of molecules from various cellular compartments including cytoplasm, lysosomes, and recycling endosomes. In vitro ARPE-19 (RPE henceforth) cells were loaded with Alexa fluor-labeled transferrin as a marker for recycling endosomes, LAMP-1 antibody was used to detect the fusion of lysosomes with the plasma membrane, GFP-transfected RPE cells were used to examine the release of GFP from the cytoplasm, and 7-AAD was used to assess cell viability. Subsequently, cells were exposed to USMB (106 cells/mL, 300 kPa peak negative pressure, 1 min treatment duration, and 20 µL/mL Definity microbubbles). Following USMB, the release of the fluorescent markers was examined at 1.5, 11.5, and 21.5 minutes from the start of USMB. The mean fluorescent intensity (MFI) of untreated and USMB treated samples were measured using flow cytometry. USMB increased the extracellular delivery of GFP molecules from the cytoplasm; the MFI in USMB treated GFP-transfected RPE cells decreased by 17% in viable cells and this MFI decreased by 70% in non-viable cells. This could be due to diffusion of GFP through the membrane disruptions induced by USMB. Additionally, the MFI of viable cells stained with LAMP-1 antibody increased by 50% and this increase was 15 folds in the non-viable cells indicating lysosome exocytosis as a mechanism for membrane repair. Furthermore, the MFI of cells loaded with fluorescent transferrin decreased by 22% after USMB treatment in viable cells, indicating a significant increase in transferrin recycling to the cell membrane. However, the increased recycling was not statistically significant

First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri.

Science.gov (United States)

Liu, Junyan; Li, Lin; Li, Bing; Peters, Brian M; Deng, Yang; Xu, Zhenbo; Shirtliff, Mark E

2017-06-01

This study aimed to investigate the spoilage capability of Lactobacillus lindneri during the induction and resuscitation of viable but nonculturable (VBNC) state. L. lindneri strain was identified by sequencing the PCR product (amplifying 16S rRNA gene) using ABI Prism 377 DNA Sequencer. During the VBNC state induction by low temperature storage and beer adaption, total, culturable, and viable cells were assessed by acridine orange direct counting, plate counting, and Live/Dead BacLight bacterial viability kit, respectively. Organic acids and diacetyl concentration were measured by reversed-phase high performance liquid chromatography and head dpace gas chromatography, respectively. VBNC state of L. lindneri was successfully induced by both beer adaption and low temperature storage, and glycerol frozen stock was the optimal way to maintain the VBNC state. Addition of catalase was found to be an effective method for the resuscitation of VBNC L. lindneri cells. Furthermore, spoilage capability remained similar during the induction and resuscitation of VBNC L. lindneri. This is the first report of induction by low temperature storage and resuscitation of VBNC L. lindneri strain, as well as the first identification of spoilage capability of VBNC and resuscitated L. lindneri cells. This study indicated that the potential colonization of L. lindneri strain in brewery environment, formation and resuscitation of VBNC state, as well as maintenance in beer spoilage capability, may be an important risk factor for brewery environment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Membrane Lipids as Indicators for Viable Bacterial Communities Inhabiting Petroleum Systems.

Science.gov (United States)

Gruner, Andrea; Mangelsdorf, Kai; Vieth-Hillebrand, Andrea; Horsfield, Brian; van der Kraan, Geert M; Köhler, Thomas; Janka, Christoph; Morris, Brandon E L; Wilkes, Heinz

2017-08-01

Microbial activity in petroleum reservoirs has been implicated in a suite of detrimental effects including deterioration of petroleum quality, increases in oil sulfur content, biofouling of steel pipelines and other infrastructures, and well plugging. Here, we present a biogeochemical approach, using phospholipid fatty acids (PLFAs), for detecting viable bacteria in petroleum systems. Variations within the bacterial community along water flow paths (producing well, topside facilities, and injection well) can be elucidated in the field using the same technique, as shown here within oil production plants in the Molasse Basin of Upper Austria. The abundance of PLFAs is compared to total cellular numbers, as detected by qPCR of the 16S rDNA gene, to give an overall comparison between the resolutions of both methods in a true field setting. Additionally, the influence of biocide applications on lipid- and DNA-based quantification was investigated. The first oil field, Trattnach, showed significant PLFA abundances and cell numbers within the reservoir and topside facilities. In contrast, the second field (Engenfeld) showed very low PLFA levels overall, likely due to continuous treatment of the topside facilities with a glutaraldehyde-based antimicrobial. In comparison, Trattnach is dosed once per week in a batch fashion. Changes within PLFA compositions across the flow path, throughout the petroleum production plants, point to cellular adaptation within the system and may be linked to shifts in the dominance of certain bacterial types in oil reservoirs versus topside facilities. Overall, PLFA-based monitoring provides a useful tool to assess the abundance and high-level taxonomic diversity of viable microbial populations in oil production wells, topside infrastructure, pipelines, and other related facilities.

Projecting recovered paper supply in the United States.

Science.gov (United States)

James L. Howard; Ken Skog; Peter J. Ince

2002-01-01

The use of recovered paper has an influence on the amount of timber harvested as well as the feasibility of expanded recycling programs. During the 1990’s markets and prices for recovered materials have fluctuated widely. There are a number of factors driving these market changes including economic conditions domestically and internationally. The objective of this...

Exploring continuous and integrated strategies for the up- and downstream processing of human mesenchymal stem cells.

Science.gov (United States)

Cunha, Bárbara; Aguiar, Tiago; Silva, Marta M; Silva, Ricardo J S; Sousa, Marcos F Q; Pineda, Earl; Peixoto, Cristina; Carrondo, Manuel J T; Serra, Margarida; Alves, Paula M

2015-11-10

The integration of up- and downstream unit operations can result in the elimination of hold steps, thus decreasing the footprint, and ultimately can create robust closed system operations. This type of design is desirable for the bioprocess of human mesenchymal stem cells (hMSC), where high numbers of pure cells, at low volumes, need to be delivered for therapy applications. This study reports a proof of concept of the integration of a continuous perfusion culture in bioreactors with a tangential flow filtration (TFF) system for the concentration and washing of hMSC. Moreover, we have also explored a continuous alternative for concentrating hMSC. Results show that expanding cells in a continuous perfusion operation mode provided a higher expansion ratio, and led to a shift in cells' metabolism. TFF operated either in continuous or discontinuous allowed to concentrate cells, with high cell recovery (>80%) and viability (>95%); furthermore, continuous TFF permitted to operate longer with higher cell concentrations. Continuous diafiltration led to higher protein clearance (98%) with lower cell death, when comparing to discontinuous diafiltration. Overall, an integrated process allowed for a shorter process time, recovering 70% of viable hMSC (>95%), with no changes in terms of morphology, immunophenotype, proliferation capacity and multipotent differentiation potential. Copyright © 2015 Elsevier B.V. All rights reserved.

Process for recovering yttrium and lanthanides from wet-process phosphoric acid

Energy Technology Data Exchange (ETDEWEB)

Janssen, J.A.; Weterings, C.A.

1983-06-28

Process for recovering yttrium and lanthanides from wet-process phosphoric acid by adding a flocculant to the phosphoric acid, separating out the resultant precipitate and then recovering yttrium and lanthanides from the precipitate. Uranium is recovered from the remaining phosphoric acid.

Recovering heavy rare earth metals from magnet scrap

Science.gov (United States)

Ott, Ryan T.; McCallum, Ralph W.; Jones, Lawrence L.

2017-08-08

A method of treating rare earth metal-bearing permanent magnet scrap, waste or other material in a manner to recover the heavy rare earth metal content separately from the light rare earth metal content. The heavy rare earth metal content can be recovered either as a heavy rare earth metal-enriched iron based alloy or as a heavy rare earth metal based alloy.

Detection of viable Helicobacter pylori inside free-living amoebae in wastewater and drinking water samples from Eastern Spain.

Science.gov (United States)

Moreno-Mesonero, Laura; Moreno, Yolanda; Alonso, José Luis; Ferrús, M Antonia

2017-10-01

Helicobacter pylori is one of the most concerning emerging waterborne pathogens. It has been suggested that it could survive in water inside free-living amoebae (FLA), but nobody has studied this relationship in the environment yet. Thus, we aimed to detect viable H. pylori cells from inside FLA in water samples. Sixty-nine wastewater and 31 drinking water samples were collected. FLA were purified and identified by PCR and sequencing. For exclusively detecting H. pylori inside FLA, samples were exposed to sodium hypochlorite and assayed by specific PMA-qPCR, DVC-FISH and culture. FLA were detected in 38.7% of drinking water and 79.7% of wastewater samples, even after disinfection. In wastewater, Acanthamoeba spp. and members of the family Vahlkampfiidae were identified. In drinking water, Acanthamoeba spp. and Echinamoeba and/or Vermamoeba were present. In 39 (58.2%) FLA-positive samples, H. pylori was detected by PMA-qPCR. After DVC-FISH, 21 (31.3%) samples harboured viable H. pylori internalized cells. H. pylori was cultured from 10 wastewater samples. To our knowledge, this is the first report that demonstrates that H. pylori can survive inside FLA in drinking water and wastewater, strongly supporting the hypothesis that FLA could play an important role in the transmission of H. pylori to humans. © 2017 Society for Applied Microbiology and John Wiley & Sons Ltd.

Electrochemical performance studies of MnO2 nanoflowers recovered from spent battery

International Nuclear Information System (INIS)

Ali, Gomaa A.M.; Tan, Ling Ling; Jose, Rajan; Yusoff, Mashitah M.; Chong, Kwok Feng

2014-01-01

Highlights: • MnO 2 is recovered from spent zinc–carbon batteries as nanoflowers structure. • Recovered MnO 2 nanoflowers show high specific capacitance. • Recovered MnO 2 nanoflowers show stable electrochemical cycling up to 900 cycles. • Recovered MnO 2 nanoflowers show low resistance in EIS data. - Abstract: The electrochemical performance of MnO 2 nanoflowers recovered from spent household zinc–carbon battery is studied by cyclic voltammetry, galvanostatic charge/discharge cycling and electrochemical impedance spectroscopy. MnO 2 nanoflowers are recovered from spent zinc–carbon battery by combination of solution leaching and electrowinning techniques. In an effort to utilize recovered MnO 2 nanoflowers as energy storage supercapacitor, it is crucial to understand their structure and electrochemical performance. X-ray diffraction analysis confirms the recovery of MnO 2 in birnessite phase, while electron microscopy analysis shows the MnO 2 is recovered as 3D nanostructure with nanoflower morphology. The recovered MnO 2 nanoflowers exhibit high specific capacitance (294 F g −1 at 10 mV s −1 ; 208.5 F g −1 at 0.1 A g −1 ) in 1 M Na 2 SO 4 electrolyte, with stable electrochemical cycling. Electrochemical data analysis reveal the great potential of MnO 2 nanoflowers recovered from spent zinc–carbon battery in the development of high performance energy storage supercapacitor system

Production of viable male unreduced gametes in Brassica interspecific hybrids is genotype specific and stimulated by cold temperatures

Directory of Open Access Journals (Sweden)

Cowling Wallace A

2011-06-01

Full Text Available Abstract Background Unreduced gametes (gametes with the somatic chromosome number may provide a pathway for evolutionary speciation via allopolyploid formation. We evaluated the effect of genotype and temperature on male unreduced gamete formation in Brassica allotetraploids and their interspecific hybrids. The frequency of unreduced gametes post-meiosis was estimated in sporads from the frequency of dyads or giant tetrads, and in pollen from the frequency of viable giant pollen compared with viable normal pollen. Giant tetrads were twice the volume of normal tetrads, and presumably resulted from pre-meiotic doubling of chromosome number. Giant pollen was defined as pollen with more than 1.5 × normal diameter, under the assumption that the doubling of DNA content in unreduced gametes would approximately double the pollen cell volume. The effect of genotype was assessed in five B. napus, two B. carinata and one B. juncea parents and in 13 interspecific hybrid combinations. The effect of temperature was assessed in a subset of genotypes in hot (day/night 30°C/20°C, warm (25°C/15°C, cool (18°C/13°C and cold (10°C/5°C treatments. Results Based on estimates at the sporad stage, some interspecific hybrid genotypes produced unreduced gametes (range 0.06 to 3.29% at more than an order of magnitude higher frequency than in the parents (range 0.00% to 0.11%. In nine hybrids that produced viable mature pollen, the frequency of viable giant pollen (range 0.2% to 33.5% was much greater than in the parents (range 0.0% to 0.4%. Giant pollen, most likely formed from unreduced gametes, was more viable than normal pollen in hybrids. Two B. napus × B. carinata hybrids produced 9% and 23% unreduced gametes based on post-meiotic sporad observations in the cold temperature treatment, which was more than two orders of magnitude higher than in the parents. Conclusions These results demonstrate that sources of unreduced gametes, required for the triploid

Expression of proposed implantation marker genes CDX2 and HOXB7 in the blastocyst does not distinguish viable from non-viable human embryos

DEFF Research Database (Denmark)

Kirkegaard, Kirstine; Hindkjær, Johnny Juhl; Ingerslev, Hans Jakob

2012-01-01

expression differs between viable and non-viable embryos in both human and non-humans, suggesting transcriptome analysis of trophectoderm (TE) as a novel method of improving embryo selection. Potential candidate marker genes have been identified with array studies on animal blastocysts. The aim of this study...... was to investigate the expression of selected genes in human blastocysts in relation to the outcome of implantation. Materials and methods: Embryos from 10 oatients undergoing in vitro fertilization treatment were included in the project. A single blastocyst was chosen for biopsy on the morning of day 5 after oocyte...... of 15 key genes associated with developmental competence in animals were evaluated in high quality human embryos with monogenic or chromosomal disorders from a pre-implantation genetic disorder program. Triplicate cDNA amplifications for quantitative (q) RT-PCR were performed using pre-designed gene...

Preserving and using germplasm and dissociated embryonic cells for conserving Caribbean and Pacific coral.

Science.gov (United States)

Hagedorn, Mary; Carter, Virginia; Martorana, Kelly; Paresa, Malia K; Acker, Jason; Baums, Iliana B; Borneman, Eric; Brittsan, Michael; Byers, Michael; Henley, Michael; Laterveer, Michael; Leong, Jo-Ann; McCarthy, Megan; Meyers, Stuart; Nelson, Brian D; Petersen, Dirk; Tiersch, Terrence; Uribe, Rafael Cuevas; Woods, Erik; Wildt, David

2012-01-01

Coral reefs are experiencing unprecedented degradation due to human activities, and protecting specific reef habitats may not stop this decline, because the most serious threats are global (i.e., climate change), not local. However, ex situ preservation practices can provide safeguards for coral reef conservation. Specifically, modern advances in cryobiology and genome banking could secure existing species and genetic diversity until genotypes can be introduced into rehabilitated habitats. We assessed the feasibility of recovering viable sperm and embryonic cells post-thaw from two coral species, Acropora palmata and Fungia scutaria that have diffferent evolutionary histories, ecological niches and reproductive strategies. In vitro fertilization (IVF) of conspecific eggs using fresh (control) spermatozoa revealed high levels of fertilization (>90% in A. palmata; >84% in F. scutaria; P>0.05) that were unaffected by tested sperm concentrations. A solution of 10% dimethyl sulfoxide (DMSO) at cooling rates of 20 to 30°C/min most successfully cryopreserved both A. palmata and F. scutaria spermatozoa and allowed producing developing larvae in vitro. IVF success under these conditions was 65% in A. palmata and 53% in F. scutaria on particular nights; however, on subsequent nights, the same process resulted in little or no IVF success. Thus, the window for optimal freezing of high quality spermatozoa was short (∼5 h for one night each spawning cycle). Additionally, cryopreserved F. scutaria embryonic cells had∼50% post-thaw viability as measured by intact membranes. Thus, despite some differences between species, coral spermatozoa and embryonic cells are viable after low temperature (-196°C) storage, preservation and thawing. Based on these results, we have begun systematically banking coral spermatozoa and embryonic cells on a large-scale as a support approach for preserving existing bio- and genetic diversity found in reef systems.

Preserving and using germplasm and dissociated embryonic cells for conserving Caribbean and Pacific coral.

Directory of Open Access Journals (Sweden)

Mary Hagedorn

Full Text Available Coral reefs are experiencing unprecedented degradation due to human activities, and protecting specific reef habitats may not stop this decline, because the most serious threats are global (i.e., climate change, not local. However, ex situ preservation practices can provide safeguards for coral reef conservation. Specifically, modern advances in cryobiology and genome banking could secure existing species and genetic diversity until genotypes can be introduced into rehabilitated habitats. We assessed the feasibility of recovering viable sperm and embryonic cells post-thaw from two coral species, Acropora palmata and Fungia scutaria that have diffferent evolutionary histories, ecological niches and reproductive strategies. In vitro fertilization (IVF of conspecific eggs using fresh (control spermatozoa revealed high levels of fertilization (>90% in A. palmata; >84% in F. scutaria; P>0.05 that were unaffected by tested sperm concentrations. A solution of 10% dimethyl sulfoxide (DMSO at cooling rates of 20 to 30°C/min most successfully cryopreserved both A. palmata and F. scutaria spermatozoa and allowed producing developing larvae in vitro. IVF success under these conditions was 65% in A. palmata and 53% in F. scutaria on particular nights; however, on subsequent nights, the same process resulted in little or no IVF success. Thus, the window for optimal freezing of high quality spermatozoa was short (∼5 h for one night each spawning cycle. Additionally, cryopreserved F. scutaria embryonic cells had∼50% post-thaw viability as measured by intact membranes. Thus, despite some differences between species, coral spermatozoa and embryonic cells are viable after low temperature (-196°C storage, preservation and thawing. Based on these results, we have begun systematically banking coral spermatozoa and embryonic cells on a large-scale as a support approach for preserving existing bio- and genetic diversity found in reef systems.

Quantitative real-time PCR combined with propidium monoazide for the selective quantification of viable periodontal pathogens in an in vitro subgingival biofilm model.

Science.gov (United States)

Sánchez, M C; Marín, M J; Figuero, E; Llama-Palacios, A; León, R; Blanc, V; Herrera, D; Sanz, M

2014-02-01

Differentiation of live and dead cells is an important challenge when using molecular diagnosis for microbial identification. This is particularly relevant when bacteria have been exposed to antimicrobial agents. The objective of this study was to test a method using quantitative real-time polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), developed for the selective quantification of viable P. gingivalis, A. actinomycetemcomitans, F. nucleatum and total bacteria in an in vitro biofilm model after antimicrobial treatment. PMA-qPCR method was tested in an in vitro biofilm model, using isopropyl alcohol as the antimicrobial agent. Matured biofilms were exposed for 1, 5, 10 and 30 min to isopropyl alcohol by immersion. Biofilms were disrupted and PMA added (final concentration of 100 μm). After DNA isolation, qPCR was carried out using specific primers and probes for the target bacteria. The differentiation of live and dead cells was tested by analysis of variance. When PMA was used in the presence of viable target bacterial cells, no statistically significant inhibition of qPCR amplification was detected (p > 0.05 in all cases). Conversely, after immersion in isopropyl alcohol of the biofilm, PMA resulted in a significant total reduction of qPCR amplification of about 4 log10 . P. gingivalis showed a vitality reduction in the biofilm of 3 log10 , while A. actinomycetemcomitans and F. nucleatum showed a 2 log10 reduction. These results demonstrate the efficiency of PMA for differentiating viable and dead P. gingivalis, A. actinomycetemcomitans and F. nucleatum cells, as well as total bacteria, in an in vitro biofilm model, after being exposed to an antimicrobial agent. Hence, this PMA-qPCR method may be useful for studying the effect of antimicrobial agents aimed at oral biofilms. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Quantification of Campylobacter spp. in chicken rinse samples by using flotation prior to real-time PCR.

Science.gov (United States)

Wolffs, Petra; Norling, Börje; Hoorfar, Jeffrey; Griffiths, Mansel; Rådström, Peter

2005-10-01

Real-time PCR is fast, sensitive, specific, and can deliver quantitative data; however, two disadvantages are that this technology is sensitive to inhibition by food and that it does not distinguish between DNA originating from viable, viable nonculturable (VNC), and dead cells. For this reason, real-time PCR has been combined with a novel discontinuous buoyant density gradient method, called flotation, in order to allow detection of only viable and VNC cells of thermotolerant campylobacters in chicken rinse samples. Studying the buoyant densities of different Campylobacter spp. showed that densities changed at different time points during growth; however, all varied between 1.065 and 1.109 g/ml. These data were then used to develop a flotation assay. Results showed that after flotation and real-time PCR, cell concentrations as low as 8.6 x 10(2) CFU/ml could be detected without culture enrichment and amounts as low as 2.6 x 10(3) CFU/ml could be quantified. Furthermore, subjecting viable cells and dead cells to flotation showed that viable cells were recovered after flotation treatment but that dead cells and/or their DNA was not detected. Also, when samples containing VNC cells mixed with dead cells were treated with flotation after storage at 4 or 20 degrees C for 21 days, a similar percentage resembling the VNC cell fraction was detected using real-time PCR and 5-cyano-2,3-ditolyl tetrazolium chloride-4',6'-diamidino-2-phenylindole staining (20% +/- 9% and 23% +/- 4%, respectively, at 4 degrees C; 11% +/- 4% and 10% +/- 2%, respectively, at 20 degrees C). This indicated that viable and VNC Campylobacter cells could be positively selected and quantified using the flotation method.

A new biotechnology for recovering heavy metal ions from wastewater

International Nuclear Information System (INIS)

Darnall, D.W.; Gabel, A.

1989-01-01

This paper reports that bio-recovery systems has developed a new sorption process for removing toxic metal ions from water. This process is based upon the natural, very strong affinity for biological materials, such as the cell walls of plants and microorganisms, for heavy metal ions such as uranium, cadmium, cobalt, nickel, etc.. Biological materials, primarily algae, have been immobilized in a polymer to produce a biological ion exchange resin, AlgaSORB. The material has a remarkable affinity for heavy metal ions and is capable of concentrating these ions by a factor of may thousand-fold. Additionally, the bound metals can be stripped and recovered from the algal material in a manner similar to conventional resins

33 CFR 385.20 - Restoration Coordination and Verification (RECOVER).

Science.gov (United States)

2010-07-01

... applying system-wide models and tools; and evaluating modifications to the Plan. RECOVER is not a policy... accordance with § 385.31, RECOVER shall conduct credible scientific assessments of hydrological, water... available science and that the results are provided for external peer review, as appropriate, and are made...

Immunogenicity of ascites tumor cells following in vitro hyperthermia

International Nuclear Information System (INIS)

Dickson, J.A.; Jasiewicz, M.L.; Simpson, A.C.

1982-01-01

The concept that host immunization may be achieved by heat-induced antigenic modifications of cancer cells and/or the release of immunogenic products by dead or dying tumor cells following in vitro heating was examined. Ehrlich ascites cells were used, inasmuch as it was claimed that in vitro hyperthermia increased the immunogenicity of these cells. Tumor cell populations of different viability were obtained by heating Ehrlich cells at 42.5 degrees, 45 degrees, or 60 degrees C. Viable and nonviable cells were separated by Ficoll-Hypaque density centrifugation; viable nonreplicating cells were obtained by treatment with mitomycin C. Cell populations of different viability after heating were left to die slowly over 3 days at 37 degrees C. Swiss TO mice were then given injections of the treated cells and/or medium. No survival benefit occurred in mice inoculated with any of these different components and then challenged with viable tumor cells. Injection of irradiated cells, however, did produce host immunity. Similarly, D23 rat hepatoma ascites cells produced host immunity after 15,000 rad but not after heating. The claim that in vitro hyperthermia increases the immunogenicity of tumor cells was not confirmed

Skills training workshops as a viable strategy for improving ...

African Journals Online (AJOL)

Skills training workshops as a viable strategy for improving smallholder and cooperative agribusiness management: A case study of Vhembe District, Limpopo Province, South Africa. ... South African Journal of Agricultural Extension ... Empirical evidence from this study shows that six months after attending the workshops, ...

Examination of an indicative tool for rapidly estimating viable organism abundance in ballast water

Science.gov (United States)

Vanden Byllaardt, Julie; Adams, Jennifer K.; Casas-Monroy, Oscar; Bailey, Sarah A.

2018-03-01

Regulatory discharge standards stipulating a maximum allowable number of viable organisms in ballast water have led to a need for rapid, easy and accurate compliance assessment tools and protocols. Some potential tools presume that organisms present in ballast water samples display the same characteristics of life as the native community (e.g. rates of fluorescence). This presumption may not prove true, particularly when ships' ballast tanks present a harsh environment and long transit times, negatively impacting organism health. Here, we test the accuracy of a handheld pulse amplitude modulated (PAM) fluorometer, the Hach BW680, for detecting photosynthetic protists at concentrations above or below the discharge standard (< 10 cells·ml- 1) in comparison to microscopic counts using fluorescein diacetate as a viability probe. Testing was conducted on serial dilutions of freshwater harbour samples in the lab and in situ untreated ballast water samples originating from marine, freshwater and brackish sources utilizing three preprocessing techniques to target organisms in the size range of ≥ 10 and < 50 μm. The BW680 numeric estimates were in agreement with microscopic counts when analyzing freshly collected harbour water at all but the lowest concentrations (< 38 cells·ml- 1). Chi-square tests determined that error is not independent of preprocessing methods: using the filtrate method or unfiltered water, in addition to refining the conversion factor of raw fluorescence to cell size, can decrease the grey area where exceedance of the discharge standard cannot be measured with certainty (at least for the studied populations). When examining in situ ballast water, the BW680 detected significantly fewer viable organisms than microscopy, possibly due to factors such as organism size or ballast water age. Assuming both the BW680 and microscopy with FDA stain were measuring fluorescence and enzymatic activity/membrane integrity correctly, the observed discrepancy

Method of recovering hydrocarbons from oil shale

Energy Technology Data Exchange (ETDEWEB)

Walton, D.K.; Slusser, M.S.

1970-11-24

A method is described for recovering hydrocarbons from an oil-shale formation by in situ retorting. A well penetrating the formation is heated and gas is injected until a pressure buildup within the well is reached, due to a decrease in the conductivity of naturally occurring fissures within the formation. The well is then vented, in order to produce spalling of the walls. This results in the formation of an enlarged cavity containing rubberized oil shale. A hot gas then is passed through the rubberized oil shale in order to retort hydrocarbons and these hydrocarbons are recovered from the well. (11 claims)

Process of recovering bituminous materials

Energy Technology Data Exchange (ETDEWEB)

1920-08-22

A modification of the process covered by German Patent 389,393 for recovering bituminous materials from oil shale by extraction is disclosed consisting, in place of or besides wood spirit oil, of acetone oil, suitably of boiling point 80 to 130/sup 0/C, being used as the extraction medium.

Talons and beaks are viable but underutilized samples for detecting ...

African Journals Online (AJOL)

Talons and beaks are viable but underutilized samples for detecting organophosphorus and carbamate pesticide poisoning in raptors. Ngaio Richards, Irene Zorrilla, Joseph Lalah, Peter Otieno, Isabel Fernandez, Monica Calvino, Joaquin Garcia ...

Quantitative assessment of viable Cryptosporidium parvum load in commercial oysters (Crassostrea virginica) in the Chesapeake Bay.

Science.gov (United States)

Graczyk, Thaddeus K; Lewis, Earl J; Glass, Gregory; Dasilva, Alexandre J; Tamang, Leena; Girouard, Autumn S; Curriero, Frank C

2007-01-01

The epidemiological importance of increasing reports worldwide on Cryptosporidium contamination of oysters remains unknown in relation to foodborne cryptosporidiosis. Thirty market-size oysters (Crassostrea virginica), collected from each of 53 commercial harvesting sites in Chesapeake Bay, MD, were quantitatively tested in groups of six for Cryptosporidium sp. oocysts by immunofluorescent antibody (IFA). After IFA analysis, the samples were retrospectively retested for viable Cryptosporidium parvum oocysts by combined fluorescent in situ hybridization (FISH) and IFA. The mean cumulative numbers of Cryptosporidium sp. oocysts in six oysters (overall, 42.1+/-4.1) were significantly higher than in the numbers of viable C. parvum oocysts (overall, 28.0+/-2.9). Of 265 oyster groups, 221 (83.4%) contained viable C. parvum oocysts, and overall, from 10-32% (mean, 23%) of the total viable oocysts were identified in the hemolymph as distinct from gill washings. The amount of viable C. parvum oocysts was not related to oyster size or to the level of fecal coliforms at the sampling site. This study demonstrated that, although oysters are frequently contaminated with oocysts, the levels of viable oocysts may be too low to cause infection in healthy individuals. FISH assay for identification can be retrospectively applied to properly stored samples.

Molecular Characterization of Gastric Epithelial Cells Using Flow Cytometry

Directory of Open Access Journals (Sweden)

Kevin A. Bockerstett

2018-04-01

Full Text Available The ability to analyze individual epithelial cells in the gastric mucosa would provide important insight into gastric disease, including chronic gastritis and progression to gastric cancer. However, the successful isolation of viable gastric epithelial cells (parietal cells, neck cells, chief cells, and foveolar cells from gastric glands has been limited due to difficulties in tissue processing. Furthermore, analysis and interpretation of gastric epithelial cell flow cytometry data has been difficult due to the varying sizes and light scatter properties of the different epithelial cells, high levels of autofluorescence, and poor cell viability. These studies were designed to develop a reliable method for isolating viable single cells from the corpus of stomachs and to optimize analyses examining epithelial cells from healthy and diseased stomach tissue by flow cytometry. We performed a two stage enzymatic digestion in which collagenase released individual gastric glands from the stromal tissue of the corpus, followed by a Dispase II digestion that dispersed these glands into greater than 1 × 106 viable single cells per gastric corpus. Single cell suspensions were comprised of all major cell lineages found in the normal gastric glands. A method describing light scatter, size exclusion, doublet discrimination, viability staining, and fluorescently-conjugated antibodies and lectins was used to analyze individual epithelial cells and immune cells. This technique was capable of identifying parietal cells and revealed that gastric epithelial cells in the chronically inflamed mucosa significantly upregulated major histocompatibility complexes (MHC I and II but not CD80 or CD86, which are costimulatory molecules involved in T cell activation. These studies describe a method for isolating viable single cells and a detailed description of flow cytometric analysis of cells from healthy and diseased stomachs. These studies begin to identify effects of

Evaluation of an automated struvite reactor to recover phosphorus ...

African Journals Online (AJOL)

In the present study we attempted to develop a reactor system to recover phosphorus by struvite precipitation, and which can be installed anywhere in the field without access to a laboratory. A reactor was developed that can run fully automated and recover up to 93% of total phosphorus (total P). Turbidity and conductivity ...

A theoretical framework for understanding recovered memory experiences.

Science.gov (United States)

Brewin, Chris R

2012-01-01

If recovered memory experiences appear counter-intuitive, this is in part due to misconceptions about trauma and memory, and to a failure to adopt a comprehensive model of memory that distinguishes personal semantic memory, autobiographical event memory, and memory appraisal. Memory performance is generally superior when events, including traumas, are central to identity. Prolonged trauma in childhood, however, can produce severe identity disturbances that may interfere with the encoding and later retrieval of personal semantic and autobiographical event information. High levels of emotion either at encoding or recall can also interfere with the creation of coherent narrative memories. For example, high levels of shock and fear when memories are recovered unexpectedly may lead to the experience of vivid flashbacks. Memory appraisals may also influence the sense that an event has been forgotten for a long time. Recovered memories, although unusual, do not contradict what we know about how memory works.

Genetic characterization of commercial Saccharomyces cerevisiae isolates recovered from vineyard environments.

Science.gov (United States)

Schuller, Dorit; Pereira, Leonor; Alves, Hugo; Cambon, Brigitte; Dequin, Sylvie; Casal, Margarida

2007-08-01

One hundred isolates of the commercial Saccharomyces cerevisiae strain Zymaflore VL1 were recovered from spontaneous fermentations carried out with grapes collected from vineyards located close to wineries in the Vinho Verde wine region of Portugal. Isolates were differentiated based on their mitochondrial DNA restriction patterns and the evaluation of genetic polymorphisms was carried out by microsatellite analysis, interdelta sequence typing and pulsed-field gel electrophoresis (PFGE). Genetic patterns were compared to those obtained for 30 isolates of the original commercialized Zymaflore VL1 strain. Among the 100 recovered isolates we found a high percentage of chromosomal size variations, most evident for the smaller chromosomes III and VI. Complete loss of heterozygosity was observed for two isolates that had also lost chromosomal heteromorphism; their growth and fermentative capacity in a synthetic must medium was also affected. A considerably higher number of variant patterns for interdelta sequence amplifications was obtained for grape-derived strains compared to the original VL1 isolates. Our data show that the long-term presence of strain VL1 in natural grapevine environments induced genetic changes that can be detected using different fingerprinting methods. The observed genetic changes may reflect adaptive mechanisms to changed environmental conditions that yeast cells encounter during their existence in nature. (c) 2007 John Wiley & Sons, Ltd.

On the recovering of a coupled nonlinear Schroedinger potential

Energy Technology Data Exchange (ETDEWEB)

Corona, Gulmaro Corona [Area de Analisis Matematico y sus Aplicaciones, Universidad Autonoma Metropolitana, Atzcapotzalco, DF (Mexico)]. E-mail: ccg@hp9000a1.uam.mx

2000-04-28

We establish a priori conditions for a Gel'fand-Levitan (GL) integral using some results of the Fredholm theory. As consequence, we obtain a recovering formula for the potential of the coupled nonlinear Schroedinger equations. The remarkable fact is that the recovering formula is given in terms of the solutions of a classical GL-integral equation. (author)

Method of recovering phosphoric acid type decontaminating electrolytes by electrodeposition

International Nuclear Information System (INIS)

Sasaki, Takashi; Wada, Koichi; Kobayashi, Toshio.

1985-01-01

Purpose: To recoving phosphoric acid type highly concentrated decontaminating liquid used for the electrolytic decontamination of contaminated equipments, components, etc in nuclear power plants or the like through electrodeposition by diaphragm electrolysis. Method: Before supplying phosphoric acid decontaminating liquid at high concentration used in the electrolytic decontaminating step to an electrodeposition recovering tank, phosphoric acid in the decontaminating electrolyte is extracted with solvents and decomposed liquid extracts (electrolyte reduced with the phosphoric acid component) are supplied to the cathode chamber of the electrodeposition recovering tank, where phosphoric acid is back-extracted with water from the solvents after extraction of phosphoric acid. Then, the back-extracted liquids (aqueous phosphoric acid solution scarcely containing metal ions) are sent to the anode chamber of the electrodeposition recovering tank. Metal ions in the liquid are captured by electrodeposition in the cathode chamber, as well as phosphoric acid in the liquids is concentrated to the initial concentration of the electrolyte in the anode chamber for reuse as the decontaminating electrolyte. As the phosphoric acid extracting agent used in the electrodeposition recovering step for the decontaminating electrolyte, water-insoluble and non-combustible tributyl phosphate (TBP) is most effective. (Horiuchi, T.)

Synthesis of red fluorescent graphene quantum dot-europium complex composites as a viable bio imaging platform

International Nuclear Information System (INIS)

Liu, Yanting; Fan, Louzhen; Zhou, Shixin; Fan, Hong

2016-01-01

We have prepared graphene quantum dot-europium(III) complex composites by noncovalently connecting chelating ligands dibenzoylmethane (DBM) and 1,10-phenanthroline (Phen) with graphene quantum dots (GQDs) first, followed by coordination to Eu(III). The resulting composites are well water-soluble and display red fluorescence of high color purity. The composites were characterized by transmission electron microscopy, X-ray photoelectron spectroscopy and X-ray diffraction. Aqueous solutions of the composites under 365 nm excitation display fluorescence with a peak at 613 nm and a quantum yield as high as 15.5 %. The good water solubility and stable photoluminescence make the composites very different from other Eu(III)-based coordination complexes. The composites are cell viable and can be used to label both the cell membrane and the cytoplasm of MCF-7 cells. They are also shown to act as bioprobes for in-vivo localization of tumorous tissue. In our perception, such composites are expected to possess wide scope because of the many functionalizations that are possible with GQDs. (author)

How Close We Are to Achieving Commercially Viable Large-Scale Photobiological Hydrogen Production by Cyanobacteria: A Review of the Biological Aspects

Science.gov (United States)

Sakurai, Hidehiro; Masukawa, Hajime; Kitashima, Masaharu; Inoue, Kazuhito

2015-01-01

Photobiological production of H2 by cyanobacteria is considered to be an ideal source of renewable energy because the inputs, water and sunlight, are abundant. The products of photobiological systems are H2 and O2; the H2 can be used as the energy source of fuel cells, etc., which generate electricity at high efficiencies and minimal pollution, as the waste product is H2O. Overall, production of commercially viable algal fuels in any form, including biomass and biodiesel, is challenging, and the very few systems that are operational have yet to be evaluated. In this paper we will: briefly review some of the necessary conditions for economical production, summarize the reports of photobiological H2 production by cyanobacteria, present our schemes for future production, and discuss the necessity for further progress in the research needed to achieve commercially viable large-scale H2 production. PMID:25793279

Solid recovered fuels in the steel industry.

Science.gov (United States)

Kepplinger, Werner L; Tappeiner, Tamara

2012-04-01

By using waste materials as alternative fuels in metallurgical plants it is possible to minimize the traditionally used reducing agents, such as coke, coal, oil or natural gas. Moreover, by using waste materials in the metallurgical industry it is feasible to recover these materials as far as possible. This also represents another step towards environmental protection because carbon dioxide emissions can be reduced, if the H(2) content of the waste material is greater in comparison with that of the substituted fuel and the effects of global warming can therefore be reduced. In the present article various solid recovered fuels and their applications in the metallurgical industry are detailed.

Solar System constraints on a cosmologically viable f(R) theory

Energy Technology Data Exchange (ETDEWEB)

Bisabr, Yousef, E-mail: y-bisabr@srttu.ed [Department of Physics, Shahid Rajaee Teacher Training University, Lavizan, Tehran 16788 (Iran, Islamic Republic of)

2010-01-18

Recently, a model f(R) theory is proposed (Miranda et al. (2009)) which is cosmologically viable and distinguishable from LAMBDACDM. We use chameleon mechanism to investigate viability of the model in terms of Solar System experiments.

Lactobacillus backii and Pediococcus damnosus isolated from 170-year-old beer recovered from a shipwreck lack the metabolic activities required to grow in modern lager beer.

Science.gov (United States)

Kajala, Ilkka; Bergsveinson, Jordyn; Friesen, Vanessa; Redekop, Anna; Juvonen, Riikka; Storgårds, Erna; Ziola, Barry

2018-01-01

In 2010, bottles of beer containing viable bacteria of the common beer-spoilage species Lactobacillus backii and Pediococcus damnosus were recovered from a shipwreck near the Åland Islands, Finland. The 170-year quiescent state maintained by the shipwreck bacteria presented a unique opportunity to study lactic acid bacteria (LAB) evolution vis-a-vis growth and survival in the beer environment. Three shipwreck bacteria (one L. backii strain and two P. damnosus strains) and modern-day beer-spoilage isolates of the same two species were genome sequenced, characterized for hop iso-α-acid tolerance, and growth in degassed lager and wheat beer. In addition, plasmid variants of the modern-day P. damnosus strain were analyzed for the effect of plasmid-encoded genes on growth in lager beer. Coding content on two plasmids was identified as essential for LAB growth in modern lager beer. Three chromosomal regions containing genes related to sugar transport and cell wall polysaccharides were shared by pediococci able to grow in beer. Our results show that the three shipwreck bacteria lack the necessary plasmid-located genetic content to grow in modern lager beer, but carry additional genes related to acid tolerance and biofilm formation compared to their modern counterparts. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Recovering uranium and/or aluminium from refractory silico-aluminous material

International Nuclear Information System (INIS)

Livesey-Goldblatt, E.; Nagy, I.F.; Tunley, T.H.

1983-01-01

A process for recovering uranium and/or aluminium from a refractory silico-aluminous material comprises leaching the material in one or more stages, obtaining a pregnant solution which contains little or no acid and recovering the desired metal from the solution

Automated monitoring of recovered water quality

Science.gov (United States)

Misselhorn, J. E.; Hartung, W. H.; Witz, S. W.

1974-01-01

Laboratory prototype water quality monitoring system provides automatic system for online monitoring of chemical, physical, and bacteriological properties of recovered water and for signaling malfunction in water recovery system. Monitor incorporates whenever possible commercially available sensors suitably modified.

Solar System constraints on a cosmologically viable f(R) theory

International Nuclear Information System (INIS)

Bisabr, Yousef

2010-01-01

Recently, a model f(R) theory is proposed (Miranda et al. (2009)) which is cosmologically viable and distinguishable from ΛCDM. We use chameleon mechanism to investigate viability of the model in terms of Solar System experiments.

Recovering of images degraded by atmosphere

Science.gov (United States)

Lin, Guang; Feng, Huajun; Xu, Zhihai; Li, Qi; Chen, Yueting

2017-08-01

Remote sensing images are seriously degraded by multiple scattering and bad weather. Through the analysis of the radiative transfer procedure in atmosphere, an image atmospheric degradation model considering the influence of atmospheric absorption multiple scattering and non-uniform distribution is proposed in this paper. Based on the proposed model, a novel recovering method is presented to eliminate atmospheric degradation. Mean-shift image segmentation and block-wise deconvolution are used to reduce time cost, retaining a good result. The recovering results indicate that the proposed method can significantly remove atmospheric degradation and effectively improve contrast compared with other removal methods. The results also illustrate that our method is suitable for various degraded remote sensing, including images with large field of view (FOV), images taken in side-glance situations, image degraded by atmospheric non-uniform distribution and images with various forms of clouds.

POOR HEMOPOIETIC STEM CELL MOBILIZERS IN MULTIPLE MYELOMA : A SINGLE INSTITUTION EXPERIENCE

Directory of Open Access Journals (Sweden)

Guillermo Jose Ruiz-Delgado

2010-06-01

Full Text Available In a single institution, in a group of 28 myeloma patients deemed eligible for autologous transplant, stem cell mobilization was attempted using filgrastim: 26 individuals were given 31 autografts employing 1-4 (median three apheresis sessions, to obtain a target stem cell dose of 1 x 106 CD34 viable cells / Kg of the recipient. The median number of grafted CD34 cells was 7.56 x 106  / Kg of the recipient; the range being 0.92 to 14.8.  By defining as poor mobilizers individuals in which a cell collection of 1 x 106 CD34 viable cells / Kg was better (80% at 80 months than those grafted with < 1 x 106 CD34 viable cells / Kg (67% at 76 months. Methods to improve stem cell mobilization are needed and may result in obtaining better results when autografting multiple myeloma patients.

Cryopreservation of Human Mucosal Leukocytes.

Directory of Open Access Journals (Sweden)

Sean M Hughes

Full Text Available Understanding how leukocytes in the cervicovaginal and colorectal mucosae respond to pathogens, and how medical interventions affect these responses, is important for developing better tools to prevent HIV and other sexually transmitted infections. An effective cryopreservation protocol for these cells following their isolation will make studying them more feasible.To find an optimal cryopreservation protocol for mucosal mononuclear leukocytes, we compared cryopreservation media and procedures using human vaginal leukocytes and confirmed our results with endocervical and colorectal leukocytes. Specifically, we measured the recovery of viable vaginal T cells and macrophages after cryopreservation with different cryopreservation media and handling procedures. We found several cryopreservation media that led to recoveries above 75%. Limiting the number and volume of washes increased the fraction of cells recovered by 10-15%, possibly due to the small cell numbers in mucosal samples. We confirmed that our cryopreservation protocol also works well for both endocervical and colorectal leukocytes. Cryopreserved leukocytes had slightly increased cytokine responses to antigenic stimulation relative to the same cells tested fresh. Additionally, we tested whether it is better to cryopreserve endocervical cells on the cytobrush or in suspension.Leukocytes from cervicovaginal and colorectal tissues can be cryopreserved with good recovery of functional, viable cells using several different cryopreservation media. The number and volume of washes has an experimentally meaningful effect on the percentage of cells recovered. We provide a detailed, step-by-step protocol with best practices for cryopreservation of mucosal leukocytes.

A viable real estate economy with disruption and blockchain

NARCIS (Netherlands)

Veuger, Jan

2017-01-01

Two titles in one cover. On page 56-112 there's the English version of the book: 'A viable real estate economy with disruption and blockchain. Does real estate still have the value that it had, or is the valuation of real estate going to change due to surprising products and services, innovative

15 CFR 990.65 - Opening an account for recovered damages.

Science.gov (United States)

2010-01-01

... Trade (Continued) NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE OIL POLLUTION... an account for recovered damages. (a) General. Sums recovered by trustees in satisfaction of a... be used to implement the Final Restoration Plan or all or an appropriate component of a Regional...

Promoting Women Participation in Aquaculture as a Viable Tool for ...

African Journals Online (AJOL)

Promoting Women Participation in Aquaculture as a Viable Tool for Poverty Alleviation in the Rural Areas of Nigeria. ... Open Access DOWNLOAD FULL TEXT ... a source of income, also the paper focus on the roles of women in aquaculture, ...

Stem cell recovering effect of copper-free GHK in skin.

Science.gov (United States)

Choi, Hye-Ryung; Kang, Youn-A; Ryoo, Sun-Jong; Shin, Jung-Won; Na, Jung-Im; Huh, Chang-Hun; Park, Kyoung-Chan

2012-11-01

The peptide Gly-His-Lys (GHK) is a naturally occurring copper(II)-chelating motifs in human serum and cerebrospinal fluid. In industry, GHK (with or without copper) is used to make hair and skin care products. Copper-GHK plays a physiological role in the process of wound healing and tissue repair by stimulating collagen synthesis in fibroblasts. We also reported that copper-GHK promotes the survival of basal stem cells in the skin. However, the effects of copper-free GHK (GHK) have not been investigated well. In this study, the effects of GHK were studied using cultured normal human keratinocytes and skin equivalent (SE) models. In monolayer cultured keratinocytes, GHK increased the proliferation of keratinocytes. When GHK was added during the culture of SE models, the basal cells became more cuboidal than control model. In addition, there was linear and intense staining of α6 and β1 integrin along the basement membrane. The number of p63 and proliferating cell nuclear antigen positive cells was also significantly increased in GHK-treated SEs than in control SEs. Western blot and slide culture experiment showed that GHK increased the expression of integrin by keratinocytes. All these results showed that GHK increased the stemness and proliferative potential of epidermal basal cells, which is associated with increased expression of integrin. In conclusion, copper-free GHK showed similar effects with copper-GHK. Thus, it can be said that copper-free GHK can be used in industry to obtain the effects of copper-GHK in vivo. Further study is necessary to explore the relationship between copper-free GHK and copper-GHK. Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.

Methods of recovering alkali metals

Science.gov (United States)

Krumhansl, James L; Rigali, Mark J

2014-03-04

Approaches for alkali metal extraction, sequestration and recovery are described. For example, a method of recovering alkali metals includes providing a CST or CST-like (e.g., small pore zeolite) material. The alkali metal species is scavenged from the liquid mixture by the CST or CST-like material. The alkali metal species is extracted from the CST or CST-like material.

Cybernetically sound organizational structures II: Relating de Sitter's design theory to Beer's viable system model

NARCIS (Netherlands)

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

2011-01-01

- Purpose – The purpose of this paper is to show how the viable system model (VSM) and de Sitter's design theory can complement each other in the context of the diagnosis and design of viable organizations. - Design/methodology/approach – Key concepts from Beer's model and de Sitter's design theory

Beam position monitor for energy recovered linac beams

Energy Technology Data Exchange (ETDEWEB)

Powers, Thomas; Evtushenko, Pavel

2017-06-06

A method of determining the beam position in an energy recovered linac (ERL). The method makes use of in phase and quadrature (I/Q) demodulation techniques to separate the pickup signal generated by the electromagnetic fields generated by the first and second pass beam in the energy recovered linac. The method includes using analog or digital based I/Q demodulation techniques in order to measure the relative amplitude of the signals from a position sensitive beam pickup such as a button, strip line or microstripline beam position monitor.

Preliminary applied study of assessment ischemic/viable myocardium by 99Tcm-HL91

International Nuclear Information System (INIS)

Liu Gang; Wu Hua

2004-01-01

Objective: To investigate the representation of 99 Tc m -HL91 in the ischemic myocardium, evaluate the diagnosis value of 99 Tc m -HL91 on hypoxic but viable myocardium. Methods: Six patients with cardiac infarction all underwent 99 Tc m -MIBI SPECT and 99 Tc m -HL91 SPECT. Average radioactivity of ischemic area and normal area were respectively obtained by ROI (2 x 2 pixels) on heart minor axis of images, And the radioactivity ratios of target (ischemic area)-to-non target(normal area)were calculated. Results: In image of 99 Tc m -HL91 SPECT, two patients who's radioactivity coloboma of 99 Tc m -MIBI image could be filled with 99 Tc m -HL91, four patients were not caught sight of obvious filling up. Conclusion 99 Tc m -HL91 can be selectively uptaken by ischemic and hypoxic but viable myocardium. it combination of 99 Tc m -MIBI SPECT may be good for accurate diagnosis and differentiation of viable myocardium. (authors)

Cytotoxic Components Against Human Oral Squamous Cell Carcinoma Isolated from Andrographis paniculata.

Science.gov (United States)

Suzuki, Ryuichiro; Matsushima, Yasuaki; Okudaira, Noriyuki; Sakagami, Hiroshi; Shirataki, Yoshiaki

2016-11-01

The 5-year survival rate of patients with oral cancer has remained approximately 50% during the past 30 years, possibly due to the poor tumor selectivity of conventional anticancer drugs. This prompted us to search for new candidates for anticancer drugs that have higher cytotoxicity and tumor selectivity. Dried leaves of Andrographis paniculata were supplied from a market in Shanghai. The methanolic fraction of A. paniculata was further fractionated to identify cytotoxic principles by spectroscopic analysis and comparison with literature values. Viable cell number was determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide method, and tumor specificity was calculated by relative cytotoxicity against oral squamous cell carcinoma cell lines compared to that against normal oral cells. Apoptosis induction was detected by cleaved poly (ADP-ribose) polymerase and caspase-3 on western blot analysis. Major cytotoxicity in the methanol extract of a leaf of A. paniculata was recovered by partitioning with EtOAc, followed by silica gel chromatography. Further purification with reversed-phase high-performance liquid chromatography led to isolation of four known cytotoxic compounds, 14-deoxyandrographolide, andrographolide, neoandrographolide and deoxyandrographiside. Among them, andrographolide had the greatest cytotoxicity and tumor specificity, also inducing caspase-3 activation of HSC-2 oral squamous cell carcinoma cells. The present study identified andrographolide as a major antitumor principle in the methanolic extract of leaves of A. paniculata. Copyright© 2016 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.

Recovery from inhibition of transcription in γ-irradiated Euglena cells

International Nuclear Information System (INIS)

Tsushimoto, G.; Kikuchi, T.; Ishida, M.R.

1982-01-01

Transcriptional activity was inhibited with low doses of γ-irradiation which did not cause the death of cells, but induced the delay of cell division in the unicellular alga Euglena. The incorporation of [ 14 C]uracil into cells was inhibited to about 50% of non-irradiated cells immediately after 3 krad irradiation. The suppressed transcriptional activity was gradually recovered after irradiation. At about 12 h post-irradiation, the rate of incorporation of [ 14 C]uracil recovered to that of non-irradiated cells. The synthesis of ribosomal RNA was inhibited immediately after 3 krad irradiation, but it recovered within 12 h after irradiation. The synthesis of cytosol ribosomal RNA precursor was more strongly inhibited than that of other cytosol ribosomal RNAs. The synthesis of cytoplasmic organelle ribosomal RNA was also inhibited and recovered after 3 krad irradiation. (Auth.)

Studies on chlorophyll and viable mutations in green gram (Vigna radiata L. Wilczek) II: Response to mutagen

Energy Technology Data Exchange (ETDEWEB)

Krishnaswami, S; Rathinam, M [Tamil Nadu Agricultural Univ., Coimbatore (India). Dept. of Agricultural Botany

1980-09-01

The frequency and spectrum of chlorophyll and viable mutations in relation to type and dose of mutagen and cluster progenies were studied in four green gram cultivars viz., Kopergaon, Pusa Baisakhi, L. 24/2 and Sel. 122 subjected to two levels of EMS and gamma irradiation, severally and in conjunction. While chlorophyll mutations did not vary with the mutagen dose, viable mutations exhibited a direct relationship. Combinations of the mutagens were more effective in mutation induction. While no difference was manifested between the cluster families in respect of chlorophyll mutations, progenies of the second cluster recorded less viable mutations than either the first or the third. Viridis and xanthoviridis among chlorophyll mutations, and those affecting plant duration and stature among viable were more predominant.

Canola Oil Fuel Cell Demonstration: Volume 2 - Market Availability of Agricultural Crops for Fuel Cell Applications

National Research Council Canada - National Science Library

Adams, John W; Cassarino, Craig; Spangler, Lee; Johnson, Duane; Lindstrom, Joel; Binder, Michael J; Holcomb, Franklin H; Lux, Scott M

2006-01-01

.... The reformation of vegetable oil crops for fuel cell uses is not well known; yet vegetable oils such as canola oil represent a viable alternative and complement to traditional fuel cell feedstocks...

In vitro study of tumor seeking radiopharmaceutical uptake by human breast cancer cell line MCF-7 after paclitaxel treatment

International Nuclear Information System (INIS)

Choi, Joon Young; Choi, Yong; Choe, Yearn Seong; Lee, Kyung Han; Kim, Byung Tae

2007-01-01

This study was designed to investigate the cellular uptake of various tumor imaging radiopharmaceuticals in human breast cancer cells before and after paclitaxel exposure considering viable cell number. F-18-fluorodeoxyglucose, C-11-methionine. TI-201, Tc-99m-MIBI, and Tc-99m-tetrofosmin were used to evaluate the cellular uptake in MCF-7 cells. MCF-7 cells were cultured in multi-well plates. Wells were divided into DMSO exposure control group, and paclitaxel exposure group. The exposure durations of paclitaxel with 10 nM or 100 nM were 2 h, 6 h, 12 h, 24 h, and 48 h. Viable cell fraction was reduced as the concentration and exposure time of paclitaxel increased. After 10 nM paclitaxel exposure, the cellular uptake of all 5 radiopharmaceuticals was not reduced significantly, irrespective of exposure time and viable cell fraction. After 100 nM paclitaxel exposure, the cellular uptake of all 5 radiopharmaceuticals was enhanced significantly irrespective of viable cell fraction. The peak uptake was observed in experimental groups with paclitaxel exposure for 6 to 48 h according the type of radiopharmaceutical. When the cellular uptake was adjusted for the viable cell fraction and cell count, the peak cellular uptake was observed in experimental groups with paclitaxel exposure for 48 h, irrespective of the type of radiopharmaceutical. The cellular uptake of F-18-fluorodeoxyglucose, C-11-methionine, TI-201, Tc-99m-MIBI, and Tc-99m-tetrofosmin did not reflect viable cell number in MCF-7 cells after paclitaxel exposure for up to 48 h

Recovering Parameters of Johnson's SB Distribution

Science.gov (United States)

Bernard R. Parresol

2003-01-01

A new parameter recovery model for Johnson's SB distribution is developed. This latest alternative approach permits recovery of the range and both shape parameters. Previous models recovered only the two shape parameters. Also, a simple procedure for estimating the distribution minimum from sample values is presented. The new methodology...

Industrial recovered-materials-utilization targets for the textile-mill-products industry

Energy Technology Data Exchange (ETDEWEB)

None

1979-01-01

The Congress, in the National Energy Conservation and Policy Act of 1978 (NECPA), directed the Department of Energy to establish materials recovery targets for the metals and metal products, paper and allied products, rubber, and textile-mill-products industries. The targets were developed to provide incentives for using energy-saving recorded materials and to provied a yardstick for measuring progress and improvement in this endeavor. The NECPA indicates that the targets should represent the maximum technically and economically feasible increase in the use of energy-saving recovered materials that each industry can achieve progressively by January 1, 1987. Materials affected by recovered-materials targets include and are limited to aluminum, copper, lead, zinc, iron, steel, paper and associated products, textile-mill, products, and rubber. Using information gathered from the textile-mill-products industry and from other textile-relaed sources, DOE has developed recovered materials targets for that industry. This report presents those targets and their basis and justification. Following an overview of the textile industry, the chapters are: Textile-Mill-Products Industry Operations; Economic Analysis of the Textile-Mill-Products Industry; Governmental and Regulatory Influence on the US Textile Industry; Current Mill Use of Recovered Materials in the Textile-Mill-Products Industry; Limitations on the Use of Recovered Materials in the US Textile-Mill-Products Industry; Materials-Recovery Targets; and Government and Industry Actions That Could Increase the Use of Recovered Materials.

Acupuntura un tratamiento viable para las adicciones en Colombia

Directory of Open Access Journals (Sweden)

Hernán López Seuscún

2013-07-01

Los tratamientos con auriculoterapia, como el protocolo NADA (National Acupuncture Detoxification Association, son los métodos más usados para las adicciones en el mundo, y aunque no se ha logrado evidenciar su efectividad, por su costo, facilidad y el poco riesgo de efectos adversos se hace viable en un país con pocos recursos económicos como Colombia.

Supercooling as a viable non-freezing cell preservation method of rat hepatocytes.

Directory of Open Access Journals (Sweden)

O Berk Usta

Full Text Available Supercooling preservation holds the potential to drastically extend the preservation time of organs, tissues and engineered tissue products, and fragile cell types that do not lend themselves well to cryopreservation or vitrification. Here, we investigate the effects of supercooling preservation (SCP at -4(oC on primary rat hepatocytes stored in cryovials and compare its success (high viability and good functional characteristics to that of static cold storage (CS at +4(oC and cryopreservation. We consider two prominent preservation solutions a Hypothermosol (HTS-FRS and b University of Wisconsin solution (UW and a range of preservation temperatures (-4 to -10 (oC. We find that there exists an optimum temperature (-4(oC for SCP of rat hepatocytes which yields the highest viability; at this temperature HTS-FRS significantly outperforms UW solution in terms of viability and functional characteristics (secretions and enzymatic activity in suspension and plate culture. With the HTS-FRS solution we show that the cells can be stored for up to a week with high viability (~56%; moreover we also show that the preservation can be performed in large batches (50 million cells with equal or better viability and no loss of functionality as compared to smaller batches (1.5 million cells performed in cryovials.

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

International Nuclear Information System (INIS)

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

2011-01-01

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

Flexible Nanosomes (SECosomes) Enable Efficient siRNA Delivery in Cultured Primary Skin Cells and in the Viable Epidermis of Ex Vivo Human Skin

NARCIS (Netherlands)

Geusens, Barbara; Van Gele, Mireille; Braat, Sien; De Smedt, Stefaan C.; Stuart, Marc C. A.; Prow, Tarl W.; Sanchez, Washington; Roberts, Michael S.; Sanders, Niek N.; Lambert, Jo

2010-01-01

The extent to which nanoscale-engineered systems cross intact human skin and can exert pharmacological effects in viable epidermis is controversial. This research seeks to develop a new lipid-based nanosome that enables the effective delivery of siRNA into human skin. The major finding is that an

Mothers recovering from cocaine addiction: factors affecting parenting skills.

Science.gov (United States)

Coyer, S M

2001-01-01

To identify factors that may influence parenting by mothers who are recovering from cocaine addiction. Exploratory descriptive, with in-depth unstructured interviews. Interviews were conducted in the woman's home or in a treatment center. A convenience sample of 11 women recovering from cocaine addiction who were mothers of children 3 years of age and younger. A content analysis was used to analyze the interview data. Two themes, personal/psychologic factors and environmental/contextual factors, and four subthemes emerged. They identify issues that may affect parenting by mothers being treated for cocaine addiction. Subthemes included low self-esteem, difficulty developing a maternal identity, isolation from friends and family, and chronic life stress. This study provides a better understanding of the sources contributing to vulnerability in the parenting role for mothers recovering from cocaine addiction and will assist nurses in providing care for these mothers and their children.

A cell transportation solution that preserves live circulating tumor cells in patient blood samples

International Nuclear Information System (INIS)

Stefansson, Steingrimur; Adams, Daniel L.; Ershler, William B.; Le, Huyen; Ho, David H.

2016-01-01

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90 % viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs

A cell transportation solution that preserves live circulating tumor cells in patient blood samples.

Science.gov (United States)

Stefansson, Steingrimur; Adams, Daniel L; Ershler, William B; Le, Huyen; Ho, David H

2016-05-06

Circulating tumor cells (CTCs) are typically collected into CellSave fixative tubes, which kills the cells, but preserves their morphology. Currently, the clinical utility of CTCs is mostly limited to their enumeration. More detailed investigation of CTC biology can be performed on live cells, but obtaining live CTCs is technically challenging, requiring blood collection into biocompatible solutions and rapid isolation which limits transportation options. To overcome the instability of CTCs, we formulated a sugar based cell transportation solution (SBTS) that stabilizes cell viability at ambient temperature. In this study we examined the long term viability of human cancer cell lines, primary cells and CTCs in human blood samples in the SBTS for transportation purposes. Four cell lines, 5 primary human cells and purified human PBMCs were tested to determine the viability of cells stored in the transportation solution at ambient temperature for up to 7 days. We then demonstrated viability of MCF-7 cells spiked into normal blood with SBTS and stored for up to 7 days. A pilot study was then run on blood samples from 3 patients with metastatic malignancies stored with or without SBTS for 6 days. CTCs were then purified by Ficoll separation/microfilter isolation and identified using CTC markers. Cell viability was assessed using trypan blue or CellTracker™ live cell stain. Our results suggest that primary/immortalized cell lines stored in SBTS remain ~90% viable for > 72 h. Further, MCF-7 cells spiked into whole blood remain viable when stored with SBTS for up to 7 days. Finally, live CTCs were isolated from cancer patient blood samples kept in SBTS at ambient temperature for 6 days. No CTCs were isolated from blood samples stored without SBTS. In this proof of principle pilot study we show that viability of cell lines is preserved for days using SBTS. Further, this solution can be used to store patient derived blood samples for eventual isolation of viable CTCs after

The search for viable local government system in Nigeria: an ...

African Journals Online (AJOL)

The history of the Nigerian local government system has been one long episode of trails and errors aimed at achieving viable local government institution without much success. Local government in the country began its long series of reforms from the colonial period when the colonial government attempted to ...

Viable Syntax: Rethinking Minimalist Architecture

Directory of Open Access Journals (Sweden)

Ken Safir

2010-03-01

Full Text Available Hauser et al. (2002 suggest that the human language faculty emerged as a genetic innovation in the form of what is called here a ‘keystone factor’—a single, simple, formal mental capability that, interacting with the pre-existing faculties of hominid ancestors, caused a cascade of effects resulting in the language faculty in modern humans. They take Merge to be the keystone factor, but instead it is posited here that Merge is the pre-existing mechanism of thought made viable by a principle that permits relations interpretable at the interfaces to be mapped onto c-command. The simplified minimalist architecture proposed here respects the keystone factor as closely as possible, but is justified on the basis of linguistic analyses it makes available, including a relativized intervention theory applicable across Case, scope, agreement, selection and linearization, a derivation of the A/A’-distinction from Case theory, and predictions such as why in situ wh-interpretation is island-insensitive, but susceptible to intervention effects.

Upgraded RECOVER system - CASDAC system

International Nuclear Information System (INIS)

Yamamoto, Yoichi; Koyama, Kinji

1992-03-01

The CASDAC (Containment And Surveillance Data Authenticated Communication) system has been developed by JAERI for nuclear safeguards and physical protection of nuclear material. This system was designed and constructed as an upgraded RECOVER system, design concept of which was based on the original RECOVER system and also the TRANSEAVER system. Both of them were developed several years ago as a remote monitoring system for continual verification of security and safeguards status of nuclear material. The system consists of two subsystems, one of them is a Grand Command Center (GCC) subsystem and the other is a facility subsystem. Communication between the two subsystems is controlled through the international telephone line network. Therefore all communication data are encrypted to prevent access by an unauthorized person who may intend to make a falsification, or tapping. The facility subsystem has an appropriate measure that ensure data security and reliable operation under unattended mode of operator. The software of this system is designed so as to be easily used in other different types of computers. This report describes the outline of the CASDAC system and the results of its performance test. This work has been carried out in the framework of Japan Support Programme for Agency Safeguards (JASPAS) as a project, JA-1. (author)

Kinetics of killing Listeria monocytogenes by macrophages: correlation of 3H-DNA release from labeled bacteria and changes in numbers of viable organisms by mathematical model

International Nuclear Information System (INIS)

Davies, W.A.

1982-01-01

Conventional methods of assessing antibacterial activities of macrophages by viable counting are limited by the precision of the statistics and are difficult to interpret quantitatively because of unrestrained extracellular growth of bacteria. An alternative technique based on the release of radioactive DNA from labeled bacteria has been offered as overcoming these drawbacks. To assess it for use with macrophages I have made a correlation with the conventional viable counting method using a mathematical model. Opsonized Listeria monocytogenes labeled with 3 H-thymidine were exposed to rat macrophages for periods up to 4 hr. Numbers of viable bacteria determined after sonication increased exponentially in the absence of live cells and this growth rate was progressively inhibited by increasing numbers of macrophages. After a lag period of 30-60 min soluble 3 H appeared in the supernatant, the amount increasing with time and numbers of macrophages. To correlate these data I developed a mathematical model that considered that changes in numbers of viable organisms were due to the difference between rates of 1) growth of extracellular bacteria and 2) killing within the macrophage. On the basis of this model curves of best fit to the viable counts data were used to predict the release of radioactivity, assuming that death of a bacterium led to the total release of its label. These predictions and the experimental data agreed well, the lag period of 30-60 min between death of the bacterium and release of radioactivity being consistent with intracellular digestion. Release of soluble radioactivity appears to be an accurate reflection of the number of bacteria killed within the macrophage

Methods for recovering precious metals from industrial waste

Science.gov (United States)

Canda, L.; Heput, T.; Ardelean, E.

2016-02-01

The accelerated rate of industrialization increases the demand for precious metals, while high quality natural resources are diminished quantitatively, with significant operating costs. Precious metals recovery can be successfully made from waste, considered to be secondary sources of raw material. In recent years, concerns and interest of researchers for more increasing efficient methods to recover these metals, taking into account the more severe environmental protection legislation. Precious metals are used in a wide range of applications, both in electronic and communications equipment, spacecraft and jet aircraft engines and for mobile phones or catalytic converters. The most commonly recovered precious metals are: gold from jewellery and electronics, silver from X- ray films and photographic emulsions, industrial applications (catalysts, batteries, glass/mirrors), jewellery; platinum group metals from catalytic converters, catalysts for the refining of crude oil, industrial catalysts, nitric acid manufacturing plant, the carbon-based catalyst, e-waste. An important aspect is the economic viability of recycling processes related to complex waste flows. Hydrometallurgical and pyrometallurgical routes are the most important ways of processing electrical and electronic equipment waste. The necessity of recovering precious metals has opened new opportunities for future research.

Recovering uranium from coal in situ

International Nuclear Information System (INIS)

Terry, R.C.

1978-01-01

An underground carbonaceous deposit containing other mineral values is burned in situ. The underground hot zone is cooled down to temperature below the boiling point of a leachig solution. The leaching solution is percolated through the residial ash, with the pregnant solution recovered for separation of the mineral values in surface facilities

Recovering valuable liquid hydrocarbons

Energy Technology Data Exchange (ETDEWEB)

Pier, M

1931-06-11

A process for recovering valuable liquid hydrocarbons from coking coal, mineral coal, or oil shale through treatment with hydrogen under pressure at elevated temperature is described. Catalysts and grinding oil may be used in the process if necessary. The process provides for deashing the coal prior to hydrogenation and for preventing the coking and swelling of the deashed material. During the treatment with hydrogen, the coal is either mixed with coal low in bituminous material, such as lean coal or active coal, as a diluent or the bituminous constituents which cause the coking and swelling are removed by extraction with solvents. (BLM)

Development of image analysis software for quantification of viable cells in microchips.

Science.gov (United States)

Georg, Maximilian; Fernández-Cabada, Tamara; Bourguignon, Natalia; Karp, Paola; Peñaherrera, Ana B; Helguera, Gustavo; Lerner, Betiana; Pérez, Maximiliano S; Mertelsmann, Roland

2018-01-01

Over the past few years, image analysis has emerged as a powerful tool for analyzing various cell biology parameters in an unprecedented and highly specific manner. The amount of data that is generated requires automated methods for the processing and analysis of all the resulting information. The software available so far are suitable for the processing of fluorescence and phase contrast images, but often do not provide good results from transmission light microscopy images, due to the intrinsic variation of the acquisition of images technique itself (adjustment of brightness / contrast, for instance) and the variability between image acquisition introduced by operators / equipment. In this contribution, it has been presented an image processing software, Python based image analysis for cell growth (PIACG), that is able to calculate the total area of the well occupied by cells with fusiform and rounded morphology in response to different concentrations of fetal bovine serum in microfluidic chips, from microscopy images in transmission light, in a highly efficient way.

Recovering low-turbidity cutting liquid from silicon slurry waste.

Science.gov (United States)

Tsai, Tzu-Hsuan; Shih, Yu-Pei

2014-04-30

In order to recover a low-turbidity polyalkylene glycol (PAG) liquid from silicon slurry waste by sedimentation, temperatures were adjusted, and acetone, ethanol or water was used as a diluent. The experimental results show that the particles in the waste would aggregate and settle readily by using water as a diluent. This is because particle surfaces had lower surface potential value and weaker steric stabilization in PAG-water than in PAG-ethanol or PAG-acetone solutions. Therefore, water is the suggested diluent for recovering a low-turbidity PAG (sedimentation. After 50 wt.% water-assisted sedimentation for 21 days, the solid content of the upper liquid reduced to 0.122 g/L, and the turbidity decreased to 44 NTU. The obtained upper liquid was then vacuum-distillated to remove water. The final recovered PAG with 0.37 NTU had similar viscosity and density to the unused PAG and could be reused in the cutting process. Copyright © 2014 Elsevier B.V. All rights reserved.

El modelo de sistema viable: un instrumento para la organización efectiva

Directory of Open Access Journals (Sweden)

Norlando Sánchez Rueda

2015-05-01

Full Text Available RESUMEN En este ensayo se presenta una interpretación teórica del denominado Modelo de Sistema Viable (MSV, de Stafford Beer y su Potencial Aplicación en Tareas de Diagnóstico  y diseño empresarial, al igual que para Mejorar las capacidades Organizacionales de Auto- Regulación  y Auto- Organización. Se explica como el Modelo del Sistema Viable permite conocer e interpretar  los mecanismos de estabilidad y adaptabilidad de las organizaciones, pilares para el crecimiento de una verdadera organización Efectiva.

Adherence of Nocardia asteroides within the murine brain.

OpenAIRE

Ogata, S A; Beaman, B L

1992-01-01

Nonlethal infection of BALB/c mice with Nocardia asteroides GUH-2 (GUH-2) produces a variety of neurological signs, including an L-dopa-responsive movement disorder in 10 to 15% of the infected population. To study nocardial interactions with the brain, we characterized the attachment of GUH-2 within specific regions through the use of microdissection. Following an intravenous injection of a single-cell suspension of log-phase GUH-2, viable cells were recovered from all regions of the brain, ...

Photoreactivation of Pseudomonas cepacia after ultraviolet exposure: a potential source of contamination in ultraviolet-treated waters

International Nuclear Information System (INIS)

Carson, L.A.; Petersen, N.J.

1975-01-01

Cells of a naturally occurring strain of Pseudomonas cepacia grown in distilled water were exposed to ultraviolet radiation. Irradiated samples incubated on membrane filters or in fluid recovery media in the absence of light showed no evidence of dark repair mechanisms. When samples were exposed to fluorescent light ranging from 50 to 950 foot candles (538 to 10,222 lux) of illumination, apparent photo-induced repair of ultraviolet injury resulted in 1- to 4-log increases in viable cells recovered

Axenic isolation of viable Giardia muris trophozoites.

Science.gov (United States)

Tillotson, K D; Buret, A; Olson, M E

1991-06-01

Large numbers of viable Giardia muris trophozoites were isolated from the duodenum of experimentally infected mice 6 days after inoculation with 1,000 G. muris cysts. A series of shaking, incubation, and washing steps in the presence of the broad-spectrum antibiotic piperacillin readily provided 4.9 +/- 1.5 x 10(5) G. muris trophozoites per mouse, free of detectable contaminant organisms. Anaerobic and microaerophilic culturing and scanning electron microscopy demonstrated axenic status and high purity of the isolates. The viability of trophozoites was 98 +/- 2%. Application of this technique should permit novel immunological and epidemiological analyses of G. muris infection and biochemical investigations of this protozoan parasite.

Comparison of culture-based, vital stain and PMA-qPCR methods for the quantitative detection of viable hookworm ova.

Science.gov (United States)

Gyawali, P; Sidhu, J P S; Ahmed, W; Jagals, P; Toze, S

2017-06-01

Accurate quantitative measurement of viable hookworm ova from environmental samples is the key to controlling hookworm re-infections in the endemic regions. In this study, the accuracy of three quantitative detection methods [culture-based, vital stain and propidium monoazide-quantitative polymerase chain reaction (PMA-qPCR)] was evaluated by enumerating 1,000 ± 50 Ancylostoma caninum ova in the laboratory. The culture-based method was able to quantify an average of 397 ± 59 viable hookworm ova. Similarly, vital stain and PMA-qPCR methods quantified 644 ± 87 and 587 ± 91 viable ova, respectively. The numbers of viable ova estimated by the culture-based method were significantly (P methods. Therefore, both PMA-qPCR and vital stain methods appear to be suitable for the quantitative detection of viable hookworm ova. However, PMA-qPCR would be preferable over the vital stain method in scenarios where ova speciation is needed.

Development of an instrument to assess readiness to recover in anorexia nervosa.

Science.gov (United States)

Rieger, E; Touyz, S; Schotte, D; Beumont, P; Russell, J; Clarke, S; Kohn, M; Griffiths, R

2000-12-01

The degree to which patients with anorexia nervosa demonstrate readiness to recover from their illness has received scant theoretical or empirical attention. Investigating the prevalence and degree of amotivation for recovery in anorexia nervosa, its role in outcome, and the effectiveness of interventions designed to enhance readiness to recover necessitates the existence of a reliable and valid measure of motivational issues relevant to the disorder. The present study aimed to develop and evaluate an instrument for assessing readiness to recover in anorexia nervosa, namely, the Anorexia Nervosa Stages of Change Questionnaire (ANSOCQ), a 23-item self-report questionnaire based on Prochaska and DiClemente's stages of change model. Seventy-one inpatients with anorexia nervosa participated in the study. On several occasions during their admission, participants completed the ANSOCQ as well as questionnaires assessing readiness to recover, anorexic symptomatology, general distress, and social desirability. The ANSOCQ demonstrated good internal consistency (.90) and 1-week test-retest reliability (.89). Various aspects of validity were also supported, such as significant relationships with other instruments assessing readiness to recover and the prediction of weight gain during different periods of treatment. The results suggest that the ANSOCQ is a psychometrically sound instrument that may prove useful in investigating the role of readiness to recover in anorexia nervosa. Copyright 2000 John Wiley & Sons, Inc.

Routes to a commercially viable PEM fuel cell stack

Energy Technology Data Exchange (ETDEWEB)

Newton, J.; Foster, S.E.; Hodgson, D.; Marrett, A.

2002-07-01

This report describes the results of a project to design and build a 10 kW{sub e} proton exchange membrane fuel cell (PEMFC) stack, including membrane electrode assemblies (MEAs), bipolar plates and stack hardware. The aim was to prove the design concept and to demonstrate functionality by operating the stack at >1 kW{sub e}/L and 500 W/kg for 200 hours operation. The project was extended to include the assembly and testing of two additional 1 kW{sub e} PEMFC stacks based on coated metal components. Low equivalent weight perfluorinated ionomer ion exchange membranes were prepared and were found to give a superior electrochemical performance to commercial materials. A technique to etch various stainless steel grades and control processes was successfully developed and optimised. Coatings for stainless steel and titanium were successfully developed and met the required performance criteria. All PEMFC stack components were selected and designed to enable subsequent commercial manufacture.

A Rapid Method for Quantifying Viable Mycobacterium avium subsp. paratuberculosis in Cellular Infection Assays

Science.gov (United States)

Pooley, Hannah B.; de Silva, Kumudika; Purdie, Auriol C.; Begg, Douglas J.; Whittington, Richard J.

2016-01-01

ABSTRACT Determining the viability of bacteria is a key outcome of in vitro cellular infection assays. Currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such as Mycobacterium avium subsp. paratuberculosis, where it can take up to 4 months to confirm growth. This study aimed to identify an assay that can rapidly quantify the number of viable M. avium subsp. paratuberculosis cells in a cellular sample. Three commercially available bacterial viability assays along with a modified liquid culture method coupled with high-throughput quantitative PCR growth detection were assessed. Criteria for assessment included the ability of each assay to differentiate live and dead M. avium subsp. paratuberculosis organisms and their accuracy at low bacterial concentrations. Using the culture-based method, M. avium subsp. paratuberculosis growth was reliably detected and quantified within 2 weeks. There was a strong linear association between the 2-week growth rate and the initial inoculum concentration. The number of viable M. avium subsp. paratuberculosis cells in an unknown sample was quantified based on the growth rate, by using growth standards. In contrast, none of the commercially available viability assays were suitable for use with samples from in vitro cellular infection assays. IMPORTANCE Rapid quantification of the viability of Mycobacterium avium subsp. paratuberculosis in samples from in vitro cellular infection assays is important, as it allows these assays to be carried out on a large scale. In vitro cellular infection assays can function as a preliminary screening tool, for vaccine development or antimicrobial screening, and also to extend findings derived from experimental animal trials. Currently, by using culture, it takes up to 4 months to obtain quantifiable results regarding M. avium subsp. paratuberculosis viability after an in vitro infection assay; however, with the quantitative PCR and liquid culture method

[Recovering helpers in the addiction treatment system in Hungary: an interpretative phenomenological analysis].

Science.gov (United States)

Kassai, Szilvia; Pintér, Judit Nóra; Rácz, József

2015-01-01

The work of recovering helpers who work in the addiction rehabilitation centres was studied. The aim was to investigate the process of addicts becoming recovering helpers, and to study what peer help means to them. According to interpretative phenomenological analysis (IPA) design, subjects were selected, data were collected and analysed. 6 (5 males, 1 female), working as recovering helpers at least one year at addiction rehabilitation centres. Semi-structured life interviews were carried out and analysed according to IPA. Emerging themes from the interviews were identified and summarized, then interpreted as central themes: important periods and turning points of the life story interviews: the experience of psychoactive drugs use, the development of the addiction (which became " Turning Point No 1") then the "rock bottom" experience ("Turning Point No 2"). Then the experience of the helping process was examined: here four major themes were identified: the development of the recovering self and the helping self, the wounded helper and the skilled helper, the experience of the helping process. IPA was found to be a useful method for idiographic exploration of the development and the work of the recovering helpers. The work of the recovering helpers can be described as mentoring of the addict clients. Our experiences might be used for the training programs for recovering helpers as well as to adopt their professional role in addiction services.

Endocochlear potential generation is associated with intercellular communication in the stria vascularis: structural analysis in the viable dominant spotting mouse mutant.

Science.gov (United States)

Carlisle, L; Steel, K; Forge, A

1990-11-01

Deafness in the viable dominant spotting mouse mutant is due to a primary defect of the stria vascularis which results in absence of the positive endocochlear potential in scala media. Endocochlear potentials were measured and the structure of stria vascularis of mutants with potentials close to zero was compared with that in normal littermate controls by use of morphometric methods. The stria vascularis was significantly thinner in mutants. Marginal cells were not significantly different from controls in terms of volume density or intramembrane particle density but the network density of tight junctions was significantly reduced in the mutants. A virtual absence of gap junctions between basal cells and marginal or intermediate cells was observed, but intramembrane particle density and junctional complexes between adjacent basal cells were not different from controls. The volume density of basal cells was significantly greater in mutants. Intermediate cells accounted for a significantly smaller volume density of the stria vascularis in mutants and had a lower density of intramembrane particles than controls. Melanocytes were not identified in the stria vascularis of mutants. These results suggest that communication between marginal, intermediate and basal cells might be important to the normal function of the stria vascularis.

Enrichment of unlabeled human Langerhans cells from epidermal cell suspensions by discontinuous density gradient centrifugation

NARCIS (Netherlands)

Teunissen, M. B.; Wormmeester, J.; Kapsenberg, M. L.; Bos, J. D.

1988-01-01

In this report we introduce an alternative procedure for enrichment of human epidermal Langerhans cells (LC) from epidermal cell suspensions of normal skin. By means of discontinuous Ficoll-Metrizoate density gradient centrifugation, a fraction containing high numbers of viable, more than 80% pure

Experimental human-like model to assess the part of viable Legionella reaching the thoracic region after nebulization.

Directory of Open Access Journals (Sweden)

Jérémie Pourchez

Full Text Available The incidence of Legionnaires' disease (LD in European countries and the USA has been constantly increasing since 1998. Infection of humans occurs through aerosol inhalation. To bridge the existing gap between the concentration of Legionella in a water network and the deposition of bacteria within the thoracic region (assessment of the number of viable Legionella, we validated a model mimicking realistic exposure through the use of (i recent technology for aerosol generation and (ii a 3D replicate of the human upper respiratory tract. The model's sensitivity was determined by monitoring the deposition of (i aerosolized water and Tc99m radio-aerosol as controls, and (ii bioaerosols generated from both Escherichia coli and Legionella pneumophila sg 1 suspensions. The numbers of viable Legionella prior to and after nebulization were provided by culture, flow cytometry and qPCR. This study was designed to obtain more realistic data on aerosol inhalation (vs. animal experimentation and deposition at the thoracic region in the context of LD. Upon nebulization, 40% and 48% of the initial Legionella inoculum was made of cultivable and non-cultivable cells, respectively; 0.7% of both populations reached the filter holder mimicking the thoracic region in this setup. These results are in agreement with experimental data based on quantitative microbial risk assessment methods and bring new methods that may be useful for preventing LD.

Enumeration of Vibrio parahaemolyticus in the viable but nonculturable state using direct plate counts and recognition of individual gene fluorescence in situ hybridization.

Science.gov (United States)

Griffitt, Kimberly J; Noriea, Nicholas F; Johnson, Crystal N; Grimes, D Jay

2011-05-01

Vibrio parahaemolyticus is a gram-negative, halophilic bacterium indigenous to marine and estuarine environments and it is capable of causing food and water-borne illness in humans. It can also cause disease in marine animals, including cultured species. Currently, culture-based techniques are used for quantification of V. parahaemolyticus in environmental samples; however, these can be misleading as they fail to detect V. parahaemolyticus in a viable but nonculturable (VBNC) state which leads to an underestimation of the population density. In this study, we used a novel fluorescence visualization technique, called recognition of individual gene fluorescence in situ hybridization (RING-FISH), which targets chromosomal DNA for enumeration. A polynucleotide probe labeled with Cyanine 3 (Cy3) was created corresponding to the ubiquitous V. parahaemolyticus gene that codes for thermolabile hemolysin (tlh). When coupled with the Kogure method to distinguish viable from dead cells, RING-FISH probes reliably enumerated total, viable V. parahaemolyticus. The probe was tested for sensitivity and specificity against a pure culture of tlh(+), tdh(-), trh(-)V. parahaemolyticus, pure cultures of Vibrio vulnificus, Vibrio harveyi, Vibrio alginolyticus and Vibrio fischeri, and a mixed environmental sample. This research will provide additional tools for a better understanding of the risk these environmental organisms pose to human health. Copyright © 2011 Elsevier B.V. All rights reserved.

Contracting of energy services: often a viable alternative

International Nuclear Information System (INIS)

Milic, M.; Bruendler, M.

2001-01-01

This article discusses the outsourcing of energy services as a viable alternative to the operation of own energy facilities. The advantages of contracting for enterprises wanting to focus on their core competencies and have their energy infrastructure financed, built, maintained and operated by a third party are discussed. Financial aspects are looked at and examples in connection with the calculation of actual energy costs are given. The article is concluded with tips on the evaluation of offers for contracting services and on the definition of ownership aspects and property boundaries

Effects of gamma radiation on Sporothrix schenckii yeast cells

Energy Technology Data Exchange (ETDEWEB)

Lacerda, Camila M. de Sousa; Martins, Estefania Mara Nascimento; Andrade, Antero S.R. [Centro de Desenvolvimento da Tecnologia Nuclear (CDTN/CNEN-MG), Belo Horizonte, MG (Brazil)], e-mail: cmsl@cdtn.br, e-mail: estefaniabio@yahoo.com.br, e-mail: antero@cdtn.br; Resende, Maria Aparecida de [Universidade Federal de Minas Gerais (UFMG), Belo Horizonte, MG (Brazil). Dept. de Microbiologia], e-mail: maressend@mono.icb.ufmg.br

2009-07-01

Sporotrichosis is a subacute or chronic infection caused by the fungus Sporothrix schenckii. Zoonotic transmission can occur after scratches or bites of animals, mainly cats, rodents, and armadillos. Up to the moment, no approved vaccine was reported for S. schenckii or to any important pathogenic fungi infection in humans, indicating the need to expand the research in this field and to explore new alternatives. The aim of this study was to evaluate the effects of gamma radiation in the viability, metabolic activity and reproductive ability of S. schenckii yeast cells for further studies on the development of a vaccine for immunization of cats and dogs. The culture of S. schenckii, in solid medium, was irradiated at doses ranging from 1.0 to 9.0 kGy. After each dose the reproductive capacity, viability and protein synthesis were estimated. The results showed that a reduction of 6 log{sub 10} cycles in the number of colonies was achieved at 6.0 kGy and after 8.0 kGy no colonies could be recovered. The viability analysis indicated that yeast cells remained viable up to 9.0 kGy. The results of protein synthesis analysis showed that the yeast cells, irradiated up to 9.0 kGy, were able to synthesize proteins. Our preliminary results indicated that for the yeast cells of S. schenckii, it is possible to find an absorbed dose in which the pathogen loses its reproductive ability, while retaining its viability, a necessary condition for the development of a radioattenuated yeast vaccine. (author)

Effects of gamma radiation on Sporothrix schenckii yeast cells

International Nuclear Information System (INIS)

Lacerda, Camila M. de Sousa; Martins, Estefania Mara Nascimento; Andrade, Antero S.R.; Resende, Maria Aparecida de

2009-01-01

Sporotrichosis is a subacute or chronic infection caused by the fungus Sporothrix schenckii. Zoonotic transmission can occur after scratches or bites of animals, mainly cats, rodents, and armadillos. Up to the moment, no approved vaccine was reported for S. schenckii or to any important pathogenic fungi infection in humans, indicating the need to expand the research in this field and to explore new alternatives. The aim of this study was to evaluate the effects of gamma radiation in the viability, metabolic activity and reproductive ability of S. schenckii yeast cells for further studies on the development of a vaccine for immunization of cats and dogs. The culture of S. schenckii, in solid medium, was irradiated at doses ranging from 1.0 to 9.0 kGy. After each dose the reproductive capacity, viability and protein synthesis were estimated. The results showed that a reduction of 6 log 10 cycles in the number of colonies was achieved at 6.0 kGy and after 8.0 kGy no colonies could be recovered. The viability analysis indicated that yeast cells remained viable up to 9.0 kGy. The results of protein synthesis analysis showed that the yeast cells, irradiated up to 9.0 kGy, were able to synthesize proteins. Our preliminary results indicated that for the yeast cells of S. schenckii, it is possible to find an absorbed dose in which the pathogen loses its reproductive ability, while retaining its viability, a necessary condition for the development of a radioattenuated yeast vaccine. (author)

Silicon carbide recovered from photovoltaic industry waste as photocatalysts for hydrogen production

Energy Technology Data Exchange (ETDEWEB)

Zhang, Yu [College of Chemical Engineering, Sichuan University, Chengdu, 610064 (China); Hu, Yu [College of Material Science and Enginneering, Sichuan University, Chengdu, 610064 (China); Zeng, Hongmei [College of Chemistry, Sichuan University, Chengdu, 610064 (China); Zhong, Lin, E-mail: zhonglin@scu.edu.cn [College of Chemical Engineering, Sichuan University, Chengdu, 610064 (China); Liu, Kewei; Cao, Hongmei [College of Chemistry, Sichuan University, Chengdu, 610064 (China); Li, Wei [College of Material Science and Enginneering, Sichuan University, Chengdu, 610064 (China); Yan, Hongjian, E-mail: hjyan@scu.edu.cn [College of Chemistry, Sichuan University, Chengdu, 610064 (China)

2017-05-05

Highlights: • SiC was recovered from photovoltaic industry waste. • The recovered SiC is mainly consist of 3C-SiC, 6H-SiC and some silicon oxycarbides. • The recovered SiC shows photocatalytic H{sub 2} evolution from water. - Abstract: In recent years, the focus on creating a dependable and efficient means to recycle or recover the valuable parts from the waste material has drawn significantly attention as an environmentally friendly way to deal with the industrial wastes. The silicon carbide (SiC) crystalline is one of reusable material in the slurry wastes generated during wafer slicing. Here we report the use of recovered SiC from the slurry wastes as photocatalysts to produce hydrogen in the presence of Na{sub 2}SO{sub 3}-Na{sub 2}S as electron donor. The recovered SiC were characterized by X-ray diffraction (XRD), X-ray photoelectron spectroscopy spectra (XPS), UV–vis (UV–vis) spectroscopy, and photoluminescence (PL) spectroscopy. The morphology of SiC loaded with 1 wt% Pt as cocatalyst by thermal-reduction method was observed by scanning electron microscopy (SEM) and high resolution transmission electron microscopy (TEM). The experimental results reveal that the recovered SiC is mainly consist of 3C-SiC, 6H-SiC and some silicon oxycarbides on the surface of the SiC. The highest hydrogen production rate is 191.8 μmol h{sup −1} g{sup −1}. This study provides a way to recycle crystalline SiC from the discharged waste in the photovoltaic industry and reuse it as photocatalyst to yield hydrogen with the advantage of low energy consumption, low pollution and easy operation.

Pyrochemical investigations into recovering plutonium from americium extraction salt residues

International Nuclear Information System (INIS)

Fife, K.W.; West, M.H.

1987-05-01

Progress into developing a pyrochemical technique for separating and recovering plutonium from spent americium extraction waste salts has concentrated on selective chemical reduction with lanthanum metal and calcium metal and on the solvent extraction of americium with calcium metal. Both techniques are effective for recovering plutonium from the waste salt, although neither appears suitable as a separation technique for recycling a plutonium stream back to mainline purification processes. 17 refs., 13 figs., 2 tabs

A multicenter study of viable PCR using propidium monoazide to detect Legionella in water samples.

Science.gov (United States)

Scaturro, Maria; Fontana, Stefano; Dell'eva, Italo; Helfer, Fabrizia; Marchio, Michele; Stefanetti, Maria Vittoria; Cavallaro, Mario; Miglietta, Marilena; Montagna, Maria Teresa; De Giglio, Osvalda; Cuna, Teresa; Chetti, Leonarda; Sabattini, Maria Antonietta Bucci; Carlotti, Michela; Viggiani, Mariagabriella; Stenico, Alberta; Romanin, Elisa; Bonanni, Emma; Ottaviano, Claudio; Franzin, Laura; Avanzini, Claudio; Demarie, Valerio; Corbella, Marta; Cambieri, Patrizia; Marone, Piero; Rota, Maria Cristina; Bella, Antonino; Ricci, Maria Luisa

2016-07-01

Legionella quantification in environmental samples is overestimated by qPCR. Combination with a viable dye, such as Propidium monoazide (PMA), could make qPCR (named then vPCR) very reliable. In this multicentre study 717 artificial water samples, spiked with fixed concentrations of Legionella and interfering bacterial flora, were analysed by qPCR, vPCR and culture and data were compared by statistical analysis. A heat-treatment at 55 °C for 10 minutes was also performed to obtain viable and not-viable bacteria. When data of vPCR were compared with those of culture and qPCR, statistical analysis showed significant differences (P 0.05). Overall this study provided a good experimental reproducibility of vPCR but also highlighted limits of PMA in the discriminating capability of dead and live bacteria, making vPCR not completely reliable. Copyright © 2016 Elsevier Inc. All rights reserved.

EXPERIMENTAL INVESTIGATION OF PIC FORMATION DURING THE INCINERATION OF RECOVERED CFC-11

Science.gov (United States)

The report gives results of an investigation of the formation of products of incomplete combustion (PICS) during "recovered" trichlorofluoromethane (CFC-11) incineration. Tests involved burning the recovered CFC-11 in a propane gas flame. combustion gas samples were taken and an...

Three methods for isolating viable anthozoan endoderm cells with their intracellular symbiotic dinoflagellates

Science.gov (United States)

Gates, R. D.; Muscatine, L.

1992-09-01

Three maceration methods are described for the isolation of single endoderm cells from marine cnidarians. Two are enzymatic treatments suitable for fleshy anthozoans such as sea anemones and zoanthids. The third employs calcium free sea water and is suitable for stony corals. The viability and morphology of the endoderm cells is described using fluorogenic dyes and scanning and transmission electron microscopy.

Symbiotic Role of the Viable but Nonculturable State of Vibrio fischeri in Hawaiian Coastal Seawater.

Science.gov (United States)

Lee, K; Ruby, E G

1995-01-01

To achieve functional bioluminescence, the developing light organ of newly hatched juveniles of the Hawaiian squid Euprymna scolopes must become colonized by luminous, symbiosis-competent Vibrio fischeri present in the ambient seawater. This benign infection occurs rapidly in animals placed in seawater from the host's natural habitat. Therefore, it was surprising that colony hybridization studies with a V. fischeri-specific luxA gene probe indicated the presence of only about 2 CFU of V. fischeri per ml of this infective seawater. To examine this paradox, we estimated the total concentration of V. fischeri cells present in seawater from the host's habitat in two additional ways. In the first approach, the total bacterial assemblage in samples of seawater was collected on polycarbonate membrane filters and used as a source of both a crude cell lysate and purified DNA. These preparations were then assayed by quantitative DNA-DNA hybridization with the luxA gene probe. The results suggested the presence of between 200 and 400 cells of V. fischeri per ml of natural seawater, a concentration more than 100 times that revealed by colony hybridization. In the second approach, we amplified V. fischeri-specific luxA sequences from microliter volumes of natural seawater by PCR. Most-probable-number analyses of the frequency of positive PCR results from cell lysates in these small volumes gave an estimate of the concentration of V. fischeri luxA gene targets of between 130 and 1,680 copies per ml. From these measurements, we conclude that in their natural seawater environment, the majority of V. fischeri cells become nonculturable while remaining viable and symbiotically infective. Experimental studies indicated that V. fischeri cells suspended in natural Hawaiian seawater enter such a state within a few days.

Stem Cell Transplants (For Teens)

Science.gov (United States)

... Safe Videos for Educators Search English Español Stem Cell Transplants KidsHealth / For Teens / Stem Cell Transplants What's ... Take to Recover? Coping Print What Are Stem Cells? As you probably remember from biology class, every ...

RECQL4-deficient cells are hypersensitive to oxidative stress/damage: Insights for osteosarcoma prevalence and heterogeneity in Rothmund-Thomson syndrome

International Nuclear Information System (INIS)

Werner, Sean R.; Prahalad, Agasanur K.; Yang Jieping; Hock, Janet M.

2006-01-01

Rothmund-Thomson syndrome (RTS) is a heterogeneous disease, associated with increased prevalence of osteosarcoma in very young patients with a mutated RECQL4 gene. In this study, we tested the ability of RECQL4 deficient fibroblasts, derived from a RTS patient to recover from hydrogen peroxide (H 2 O 2 )-induced oxidative stress/damage. Immunoperoxidase staining for 8-oxo-deoxyguanosine (8-oxo-dG) formation in RTS and normal human fibroblasts were compared to assess DNA damage. We determined DNA synthesis, cell growth, cell cycle distribution, and viability in RTS and normal human fibroblasts before and after H 2 O 2 treatment. H 2 O 2 induces 8-oxo-dG formation in both RTS and normal fibroblasts. In normal human fibroblasts, RECQL4 was predominantly localized to cytoplasm; nuclear translocation and foci formation occurred in response to oxidant stimulation. After recovery from oxidant exposure, viable RTS fibroblasts showed irreversible growth arrest compared to normal fibroblasts. DNA synthesis decreased significantly in treated RTS cells, with concomitant reduction of cells in the S-phase. These results suggest that enhanced oxidant sensitivity in RECQL4 deficient fibroblasts derived from RTS patients could be attributed to abnormal DNA metabolism and proliferation failure. The ramifications of these findings on osteosarcoma prevalence and heterogeneity in RTS are discussed

Amygdala alterations during an emotional conflict task in women recovered from anorexia nervosa.

Science.gov (United States)

Bang, Lasse; Rø, Øyvind; Endestad, Tor

2016-02-28

The pathophysiology of anorexia nervosa (AN) is not completely understood, but research suggests that alterations in brain circuits related to cognitive control and emotion are central. The aim of this study was to explore neural responses to an emotional conflict task in women recovered from AN. Functional magnetic resonance imaging was used to measure neural responses to an emotional conflict task in 22 women recovered from AN and 21 age-matched healthy controls. The task involved categorizing affective faces while ignoring affective words. Face and word stimuli were either congruent (non-conflict) or incongruent (conflict). Brain responses to emotional conflict did not differ between groups. However, in response to emotional non-conflict, women recovered from AN relative to healthy controls showed significantly less activation in the bilateral amygdala. Specifically, while emotional non-conflict evoked significant activations of the amygdala in healthy controls, recovered AN women did not show such activations. Similar significant group differences were also observed in the hippocampus and basal ganglia. These results suggest that women recovered from AN are characterized by alterations within emotion-related brain circuits. Recovered women's absence of amygdala and hippocampus activation during non-conflict trials possibly reflects an impaired ability to process emotional significant stimuli. Copyright © 2016 The Authors. Published by Elsevier Ireland Ltd.. All rights reserved.

[Screening serum response special antibodies of U251 cell line from surface display phage antibody library].

Science.gov (United States)

Yu, Min; Tan, De-Yong; Qian, Wei; Lai, Jian-Hua; Sun, Gui-Lin

2004-05-01

U251 cell is a sensitive cell line to serum, which stops at G0 phase of cell cycle in no-serum medium, and recovers growth when the serum is added into no-serum medium. The cell can express corresponding proteins in different phase of cell cycle. Therefore it is very signification for the study of cell cycle regulation mechanism that explores these proteins. In this paper, the mouse antibody phage display library was added into the bottle in which the serum starvation U251 cells had been cultured, and the special antibody phages were absorbed. Then the absorbed antibody phages were amplified by adding E. coli TG1 and helper phage M13K07. Amplified antibody phages were added into bottle in which the serum cultured cell after serum starvation (follow named as serum recovered cells) were incubated, so that the cell absorbed the no-special antibody phages for the serum starvation cell and the special antibody phages were in supernatant. The remaining no-special antibody phages in the supernatant were discarded by repeating above program 3-4 times. The pure special antibody phages were gotten, and amplified by adding the host cell E. coli TG1 and helper phage M13K07. Then the host bacterium infected special antibody phage was spread on the plate medium with ampicillin, and the monoclonal antibody phages were gotten. Using same as above program, the monoclonal antibody phages absorbed specially for serum recovered U251 cells were obtained when the serum recovered cells instead of serum starvation cells and serum starvation cells instead of serum recovered cells. In this study, ninety-six positive monoclonal antibody phages that absorbed specially the serum starvation cells and eighty-two positive monoclonal antibody phages that absorbed specially the serum recovered cells were obtained. By using cell immunochemistry assay, two special signification antibodies were obtained. one (No.11) was the strong response in serum starvation cells, the other (No.2) was the strong

Fuel cells are a commercially viable alternative for the production of "clean" energy.

Science.gov (United States)

Niakolas, Dimitris K; Daletou, Maria; Neophytides, Stylianos G; Vayenas, Constantinos G

2016-01-01

Fuel cells present a highly efficient and environmentally friendly alternative technology for decentralized energy production. The scope of the present study is to provide an overview of the technological and commercialization readiness level of fuel cells. Specifically, there is a brief description of their general advantages and weaknesses in correlation with various technological actions and political strategies, which are adopted towards their proper positioning in the global market. Some of the most important key performance indicators are also discussed, alongside with a few examples of broad commercialization. It is concluded that the increasing number of companies which utilize and invest on this technology, in combination with the supply chain improvements and the concomitant technological maturity and recognition, reinforce the fuel cell industry so as to become well-aligned for global success.

Light and electron microscopic observation of regenerated fungiform taste buds in patients with recovered taste function after severing chorda tympani nerve.

Science.gov (United States)

Saito, Takehisa; Ito, Tetsufumi; Narita, Norihiko; Yamada, Takechiyo; Manabe, Yasuhiro

2011-11-01

The aim of this study was to evaluate the mean number of regenerated fungiform taste buds per papilla and perform light and electron microscopic observation of taste buds in patients with recovered taste function after severing the chorda tympani nerve during middle ear surgery. We performed a biopsy on the fungiform papillae (FP) in the midlateral region of the dorsal surface of the tongue from 5 control volunteers (33 total FP) and from 7 and 5 patients with and without taste recovery (34 and 29 FP, respectively) 3 years 6 months to 18 years after surgery. The specimens were observed by light and transmission electron microscopy. The taste function was evaluated by electrogustometry. The mean number of taste buds in the FP of patients with completely recovered taste function was significantly smaller (1.9 +/- 1.4 per papilla; p taste buds. Nerve fibers and nerve terminals were also found in the taste buds. It was clarified that taste buds containing taste cells and nerve endings do regenerate in the FP of patients with recovered taste function.

The immunomodulatory properties of viable Lactobacillus salivarius ssp. salivarius CECT5713 are not restricted to the large intestine.

Science.gov (United States)

Arribas, Belén; Garrido-Mesa, Natividad; Perán, Laura; Camuesco, Desirée; Comalada, Mònica; Bailón, Elvira; Olivares, Mónica; Xaus, Jordi; Kruidenier, Laurens; Sanderson, Ian R; Zarzuelo, Antonio; Rodríguez-Cabezas, Maria Elena; Gálvez, Julio

2012-04-01

The aim of this study was to better characterise the biological effects of Lactobacillus salivarius ssp. salivarius CECT5713, a probiotic with immunomodulatory properties. Live or dead probiotic was assayed in the TNBS model of rat colitis to determine whether viability was a requisite to exert the beneficial effects. In vitro studies were also performed in Caco-2 cells to evaluate its effects on epithelial cell recovery and IL-8 production. Finally, the probiotic was assayed in the LPS model of septic shock in mice to establish its effects when there is an altered systemic immune response. The viability of the probiotic was required for its anti-inflammatory activity. The probiotic inhibited IL-8 production in stimulated Caco-2 cells and facilitated the recovery of damaged intestinal epithelium. In LPS-treated mice, the probiotic inhibited the production of TNFα in plasma and lungs and increased the hepatic glutathione content. These effects were associated with an improvement in the altered production of the T-cell cytokines in splenocytes, by reducing IL-2 and IL-5 and by increasing IL-10. Finally, it reduced the increased plasma IgG production in LPS-treated mice. The anti-inflammatory effects of viable L. salivarius ssp. salivarius CECT5713 are not restricted to the gastrointestinal tract.

Collateral circulation as a marker of the presence of viable myocardium in patients with recent myocardial infarction

International Nuclear Information System (INIS)

Fujita, M.; Ohno, A.; Wada, O.; Miwa, K.; Nozawa, T.; Yamanishi, K.; Sasayama, S.

1991-01-01

The relationship between the presence of viable myocardium and the extent of coronary collateral circulation to the infarct area was evaluated in 20 patients with a recent anterior myocardial infarction who had complete obstruction of the left anterior descending coronary artery. The viability of myocardial tissue was assessed by exercise thallium-201 myocardial scintigraphy, and the collateral circulation was angiographically evaluated by means of a collateral index ranging from 0 to 3. Patients were divided into two groups according to the presence (group 1, n = 10) or absence (group 2, n = 10) of viable myocardium in the perfusion territory of the infarct-related artery. The collateral index in group 1 was 2.5 ± 0.5 (SD), which was significantly higher than the 0.7 ± 0.8 in group 2. These findings indicate that the presence of ischemic but viable myocardium is intimately related to the development of collateral circulation in patients with myocardial infarction, and the existence of well-developed collateral channels predicts the presence of viable myocardium in the infarct area

Monitoring of viable airborne SARS virus in ambient air

Science.gov (United States)

Agranovski, Igor E.; Safatov, Alexander S.; Pyankov, Oleg V.; Sergeev, Alexander N.; Agafonov, Alexander P.; Ignatiev, Georgy M.; Ryabchikova, Elena I.; Borodulin, Alexander I.; Sergeev, Artemii A.; Doerr, Hans W.; Rabenau, Holger F.; Agranovski, Victoria

Due to recent SARS related issues (Science 300 (5624) 1394; Nature 423 (2003) 240; Science 300 (5627) 1966), the development of reliable airborne virus monitoring procedures has become galvanized by an exceptional sense of urgency and is presently in a high demand (In: Cox, C.S., Wathers, C.M. (Eds.), Bioaerosols Handbook, Lewis Publishers, Boca Raton, FL, 1995, pp. 247-267). Based on engineering control method (Aerosol Science and Technology 31 (1999) 249; 35 (2001) 852), which was previously applied to the removal of particles from gas carriers, a new personal bioaerosol sampler has been developed. Contaminated air is bubbled through porous medium submerged into liquid and subsequently split into multitude of very small bubbles. The particulates are scavenged by these bubbles, and, thus, effectively removed. The current study explores its feasibility for monitoring of viable airborne SARS virus. It was found that the natural decay of such virus in the collection fluid was around 0.75 and 1.76 lg during 2 and 4 h of continuous operation, respectively. Theoretical microbial recovery rates of higher than 55 and 19% were calculated for 1 and 2 h of operation, respectively. Thus, the new sampling method of direct non-violent collection of viable airborne SARS virus into the appropriate liquid environment was found suitable for monitoring of such stress sensitive virus.

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

Science.gov (United States)

Hessenberger, S; Schatzmayr, G; Teichmann, K

2016-10-15

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

Experimental and Theoretical Investigation of Multispecies Oral Biofilm Resistance to Chlorhexidine Treatment

Science.gov (United States)

Shen, Ya; Zhao, Jia; de La Fuente-Núñez, César; Wang, Zhejun; Hancock, Robert E. W.; Roberts, Clive R.; Ma, Jingzhi; Li, Jun; Haapasalo, Markus; Wang, Qi

2016-06-01

We investigate recovery of multispecies oral biofilms following chlorhexidine gluconate (CHX) and CHX with surface modifiers (CHX-Plus) treatment. Specifically, we examine the percentage of viable bacteria in the biofilms following their exposure to CHX and CHX-Plus for 1, 3, and 10 minutes, respectively. Before antimicrobial treatment, the biofilms are allowed to grow for three weeks. We find that (a). CHX-Plus kills bacteria in biofilms more effectively than the regular 2% CHX does, (b). cell continues to be killed for up to one week after exposure to the CHX solutions, (c). the biofilms start to recover after two weeks, the percentage of the viable bacteria recovers in the 1 and 3 minutes treatment groups but not in the 10 minutes treatment group after five weeks, and the biofilms fully return to the pretreatment levels after eight weeks. To understand the mechanism, a mathematical model for multiple bacterial phenotypes is developed, adopting the notion that bacterial persisters exist in the biofilms together with regulatory quorum sensing molecules and growth factor proteins. The model reveals the crucial role played by the persisters, quorum sensing molecules, and growth factors in biofilm recovery, accurately predicting the viable bacterial population after CHX treatment.

Experimental and Theoretical Investigation of Multispecies Oral Biofilm Resistance to Chlorhexidine Treatment

Science.gov (United States)

Shen, Ya; Zhao, Jia; de la Fuente-Núñez, César; Wang, Zhejun; Hancock, Robert E. W.; Roberts, Clive R.; Ma, Jingzhi; Li, Jun; Haapasalo, Markus; Wang, Qi

2016-01-01

We investigate recovery of multispecies oral biofilms following chlorhexidine gluconate (CHX) and CHX with surface modifiers (CHX-Plus) treatment. Specifically, we examine the percentage of viable bacteria in the biofilms following their exposure to CHX and CHX-Plus for 1, 3, and 10 minutes, respectively. Before antimicrobial treatment, the biofilms are allowed to grow for three weeks. We find that (a). CHX-Plus kills bacteria in biofilms more effectively than the regular 2% CHX does, (b). cell continues to be killed for up to one week after exposure to the CHX solutions, (c). the biofilms start to recover after two weeks, the percentage of the viable bacteria recovers in the 1 and 3 minutes treatment groups but not in the 10 minutes treatment group after five weeks, and the biofilms fully return to the pretreatment levels after eight weeks. To understand the mechanism, a mathematical model for multiple bacterial phenotypes is developed, adopting the notion that bacterial persisters exist in the biofilms together with regulatory quorum sensing molecules and growth factor proteins. The model reveals the crucial role played by the persisters, quorum sensing molecules, and growth factors in biofilm recovery, accurately predicting the viable bacterial population after CHX treatment. PMID:27325010

Process for recovering uranium from wet process phosphoric acid

International Nuclear Information System (INIS)

Pyrih, R.Z.; Rickard, S.; Carrington, F.

1982-01-01

A process for recovering uranium from phosphoric acid solutions uses an acidified alkali metal carbonate solution for the second-stage strip of uranyl uranium from the ion-exchange solution. The stripped solution is then recycled to the ion-exchange circuit. In the first stripping stage the ion-exchange solution containing the recovered uranyl uranium and an inert organic diluent is stripped with ammonium carbonate, producing a slurry of ammonium uranyl tricarbonate. The second strip, with a solution of 50-200 grams per litre of sodium carbonate eliminates the problems of inadequate removal of phosphorus, iron and vanadium impurities, solids accumulation, and phase separation in the strip circuit

Roots of success: cultivating viable community forestry

Energy Technology Data Exchange (ETDEWEB)

MacQueen, Duncan

2009-05-15

Is community forestry emerging from the shadows? The evidence shows that locally controlled enterprises can be economically viable, and often build on stronger social and environmental foundations than the big private-sector players. Certainly this is an industry in need of a shakeup. Many forests have become flashpoints where agro-industry, large-scale logging concerns and conservation interests clash, while forest-dependent communities are left out in the cold. Meanwhile, governments – driven by concerns over the climate impacts of deforestation – are having to gear up for legal, sustainable forestry production. Community forestry could be crucial to solving many of these challenges. By building on local core capabilities and developing strategic partnerships, they are forging key new business models that could transform the sector.

Rapid Quantification of Viable Campylobacter Bacteria on Chicken Carcasses, Using Real-Time PCR and Propidium Monoazide Treatment, as a Tool for Quantitative Risk Assessment

DEFF Research Database (Denmark)

Josefsen, Mathilde Hartmann; Löfström, Charlotta; Hansen, Tina Beck

2010-01-01

A number of intervention strategies against Campylobacter contaminated poultry focus on post-slaughter reduction of the number of cells, emphasizing the need for rapid and reliable quantitative detection of only viable Campylobacter. We present a new and rapid quantitative approach for enumeration...... method does not detect DNA from dead Campylobacter, but recognises the infectious potential of the VBNC state, and is thereby able to assess the effect of control strategies, and provide trustworthy data for risk assessment....

Temporary corneal stem cell dysfunction after radiation therapy

International Nuclear Information System (INIS)

Hiroshi, Fujishima; Kazuo, Tsubota

1996-01-01

Radiation therapy can cause corneal and conjuctival abnormalities that sometimes require surgical treatment. Corneal stem cell dysfunction is described, which recovered after the cessation of radiation. Methods - A 44-year-old man developed a corneal epithelial abnormality associated with conjuctival and corneal inflammation following radiation therapy for maxillary cancer. Examination of brush cytology samples showed goblet cells in the upper and lower parts of the cornea, which showed increased fluorescein permeability, and intraepithelial lymphocytes. Impression cytology showed goblet cells in the same part of the cornea. Specular microscopy revealed spindle type epithelial cells. Patient follow up included artificial tears and an antibiotic ophthalmic ointment. The corneal abnormalities resolved after 4 months with improved visual acuity without any surgical intervention, but the disappearance of the palisades of Vogt did not recover at 1 year after radiation. Radiation therapy in this patient caused temporary stem cell dysfunction which resulted in conjunctivalisation in a part of the cornea. Although limbal stem cell function did not fully recover, this rare case suggested that medical options should be considered before surgery. (Author)

Effect of Induced Pluripotent Stem Cell Technology in Blood Banking

Science.gov (United States)

Focosi, Daniele

2016-01-01

Summary Population aging has imposed cost-effective alternatives to blood donations. Artificial blood is still at the preliminary stages of development, and the need for viable cells seems unsurmountable. Because large numbers of viable cells must be promptly available for clinical use, stem cell technologies, expansion, and banking represent ideal tools to ensure a regular supply. Provided key donors can be identified, induced pluripotent stem cell (iPSC) technology could pave the way to a new era in transfusion medicine, just as it is already doing in many other fields of medicine. The present review summarizes the current state of research on iPSC technology in the field of blood banking, highlighting hurdles, and promises. Significance The aging population in Western countries is causing a progressive reduction of blood donors and a constant increase of blood recipients. Because blood is the main therapeutic option to treat acute hemorrhage, cost-effective alternatives to blood donations are being actively investigated. The enormous replication capability of induced pluripotent stem cells and their promising results in many other fields of medicine could be an apt solution to produce the large numbers of viable cells required in transfusion and usher in a new era in transfusion medicine. The present report describes the potentiality, technological hurdles, and promises of induced pluripotent stem cells to generate red blood cells by redifferentiation. PMID:26819256

Mapping In Vivo Tumor Oxygenation within Viable Tumor by 19F-MRI and Multispectral Analysis

Directory of Open Access Journals (Sweden)

Yunzhou Shi

2013-11-01

Full Text Available Quantifying oxygenation in viable tumor remains a major obstacle toward a better understanding of the tumor microenvironment and improving treatment strategies. Current techniques are often complicated by tumor heterogeneity. Herein, a novel in vivo approach that combines 19F magnetic resonance imaging (19F-MRIR1 mapping with diffusionbased multispectral (MS analysis is introduced. This approach restricts the partial pressure of oxygen (pO2 measurements to viable tumor, the tissue of therapeutic interest. The technique exhibited sufficient sensitivity to detect a breathing gas challenge in a xenograft tumor model, and the hypoxic region measured by MS 19F-MRI was strongly correlated with histologic estimates of hypoxia. This approach was then applied to address the effects of antivascular agents on tumor oxygenation, which is a research question that is still under debate. The technique was used to monitor longitudinal pO2 changes in response to an antibody to vascular endothelial growth factor (B20.4.1.1 and a selective dual phosphoinositide 3-kinase/mammalian target of rapamycin inhibitor (GDC-0980. GDC-0980 reduced viable tumor pO2 during a 3-day treatment period, and a significant reduction was also produced by B20.4.1.1. Overall, this method provides an unprecedented view of viable tumor pO2 and contributes to a greater understanding of the effects of antivascular therapies on the tumor's microenvironment.

Role of chromosome stability and telomere length in the production of viable cell lines for somatic cell nuclear transfer

Directory of Open Access Journals (Sweden)

Betts Dean H

2006-08-01

Full Text Available Abstract Background Somatic cell nuclear transfer (SCNT provides an appealing alternative for the preservation of genetic material in non-domestic and endangered species. An important prerequisite for successful SCNT is the availability of good quality donor cells, as normal embryo development is dependent upon proper reprogramming of the donor genome so that embryonic genes can be appropriately expressed. The characteristics of donor cell lines and their ability to produce embryos by SCNT were evaluated by testing the effects of tissue sample collection (DART biopsy, PUNCH biopsy, post-mortem EAR sample and culture initiation (explant, collagenase digestion techniques. Results Differences in initial sample size based on sample collection technique had an effect on the amount of time necessary for achieving primary confluence and the number of population doublings (PDL produced. Thus, DART and PUNCH biopsies resulted in cultures with decreased lifespans (50 PDL and chromosomally stable (>70% normal cells at 20 PDL cultures produced by post-mortem EAR samples. Chromosome stability was influenced by sample collection technique and was dependent upon the culture's initial telomere length and its rate of shortening over cell passages. Following SCNT, short-lived cultures resulted in significantly lower blastocyst development (≤ 0.9% compared to highly proliferative cultures (11.8%. Chromosome stability and sample collection technique were significant factors in determining blastocyst development outcome. Conclusion These data demonstrate the influence of culture establishment techniques on cell culture characteristics, including the viability, longevity and normality of cells. The identification of a quantifiable marker associated with SCNT embryo developmental potential, chromosome stability, provides a means by which cell culture conditions can be monitored and improved.

Department of Defense Recovering Warrior Task Force

Science.gov (United States)

2014-09-02

accessible and available to the Veterans Benefits Administration ( VBA ) as soon as possible381; however, because military service records include health...programs are meeting expectations ........................................... 35 Facilitating Access to Health Care...Enduring RW Mission, Facilitating RW Recovery and Transition, and Facilitating Access to Health Care. SUMMARY 2  DoD Recovering Warrior Task Force

Process for recovering oil from shale, etc

Energy Technology Data Exchange (ETDEWEB)

1920-08-20

A process is described for recovering oil from oil-shale and the like, by the direct action of the hot gases obtained by burning the carbonized shale residue. It is immediately carried out in separate adjacent chambers, through which the feed goes from one to the other intermittently, from the upper to the lower.

Applications for Energy Recovering Free Electron Lasers

Energy Technology Data Exchange (ETDEWEB)

George Neil

2007-08-01

The availability of high-power, high-brilliance sources of tunable photons from energy-recovered Free Electron Lasers is opening up whole new fields of application of accelerators in industry. This talk will review some of the ideas that are already being put into production, and some of the newer ideas that are still under development.

A fuel cell city bus with three drivetrain configurations

Energy Technology Data Exchange (ETDEWEB)

Wang, Junping [Key Laboratory of Education Ministry for Modern Design and Rotor-Bearing System, Xi' an Jiaotong University, Xi' an 710049 (China); Chen, Yong [School of Automobile and Transporting Engineering, Liaoning Institute of Technology, Jinzhou, Liaoning 121001 (China); Chen, Quanshi [State Key Laboratory of Automobile Safety and Energy Conservation, Tsinghua University, Beijing 100084 (China)

2006-09-22

Three fuel cell city buses of the energy hybrid- and power hybrid-type were re-engineered with three types of drivetrain configuration to optimize the structure and improve the performance. The energy distribution, hydrogen consumption, state of charge (SOC) and the power variation rate were analyzed when different drivetrain configurations and parameters were used. When powered only by a fuel cell, the bus cannot recover the energy through regenerative braking. The variation of the fuel cell power is large and frequent, which is not good for the fuel cell. When the fuel cell is linked to a battery pack in parallel, the bus can recover the energy through regenerative braking. The energy distribution is determined by the parameters of the fuel cell and the battery pack in the design stage to reduce the power variation rate of the fuel cell. When the fuel cell and DC/DC converter connected in series links the battery pack in parallel, energy can be recovered and the energy distribution can be adjusted online. The power variation rate of both the fuel cell and the battery pack are reduced. (author)

PROCESS OF RECOVERING URANIUM FROM ITS ORES

Science.gov (United States)

Galvanek, P. Jr.

1959-02-24

A process is presented for recovering uranium from its ores. The crushed ore is mixed with 5 to 10% of sulfuric acid and added water to about 5 to 30% of the weight of the ore. This pugged material is cured for 2 to 3 hours at 100 to 110 deg C and then cooled. The cooled mass is nitrate-conditioned by mixing with a solution equivalent to 35 pounds of ammunium nitrate and 300 pounds of water per ton of ore. The resulting pulp containing 70% or more solids is treated by upflow percolation with a 5% solution of tributyl phosphate in kerosene at a rate equivalent to a residence time of about one hour to extract the solubilized uranium. The uranium is recovered from the pregnant organic liquid by counter-current washing with water. The organic extractant may be recycled. The uranium is removed from the water solution by treating with ammonia to precipitate ammonium diuranate. The filtrate from the last step may be recycled for the nitrate-conditioning treatment.

Expression of delayed cell death (DCD) in the progeny of fish cells surviving 2,4-dichloroaniline (2,4-DCA) exposure

International Nuclear Information System (INIS)

Kilemade, Michael; Mothersill, Carmel

2003-01-01

Interest in and concern for the quality of the environment has prompted a great deal of research into methods of measuring and assessing changes in it. One problem of major interest is that of increasing amounts of mutagenic/carcinogenic chemicals generated and released into marine and freshwater ecosystems. Numerous techniques involving whole animals and cell culture for these genotoxic changes have been devised to assay specific chemicals. Little has been done to determine the effects of potential genotoxicants on aquatic organisms. The purpose of this study was to investigate if 2,4-Dichloroaniline (2,4-DCA) (CASRN: 554-00-7), induced delayed cell death (DCD) or delayed reproductive cell death a.k.a. as lethal mutations in a teleost cell line, CHSE-214. Delayed expression of cell death in the progeny of cells, which survived a toxic insult, was first shown for ionizing radiation and is one of the signs of induced genomic instability. The survival of cells initially treated with 2,4-DCA and the survival of their progeny were determined. When cells are exposed to a toxic insult, the component cells of a normal appearing survivor colony or clone were commonly thought to have proliferative capacity equivalent to that of the untreated cells. In this study, however, it was found that CHSE-214 cells surviving 2,4-DCA exposure carried heritable lethal defects, which came to light only after numerous apparently successful divisions, in the form of plating efficiencies, which were reduced below those of the untreated, control cells. DCD expression did not appear to be dose-dependent with poor cell survival occurring at the lower end of 2,4-DCA exposure and remained constant until recovering to something like 60% of the controls. A study of the CHSE-214 kinetics post-exposure showed that the apparent reduced growth rate of the cells was due to reduced numbers of reproductively viable cells in the population. Results showed that the expression of DCD occurred persistently

Tumour T1 changes in vivo are highly predictive of response to chemotherapy and reflect the number of viable tumour cells – a preclinical MR study in mice

International Nuclear Information System (INIS)

Weidensteiner, Claudia; Allegrini, Peter R; Sticker-Jantscheff, Melanie; Romanet, Vincent; Ferretti, Stephane; McSheehy, Paul MJ

2014-01-01

measure of cell density but reflects the decreased number of remaining viable and proliferating tumour cells due to perhaps cell and tissue destruction releasing proteins and/or metals that cause T 1 relaxation. ΔT 1 is a highly sensitive and specific predictor of response. This MRI method provides the opportunity to stratify a patient population during tumour therapy in the clinic

Cell killing and radiosensitization by caffeic acid phenethyl ester (CAPE) in lung cancer cells

International Nuclear Information System (INIS)

Chen, Miao-Fen; Chen, Wen-Cheng; Wu, Chun-Te; King, P.C.

2004-01-01

Caffeic acid phenethyl ester (CAPE) is a biologically active ingredient of honeybee propoplis. The cytotoxicity and radiation sensitization effects of CAPE were evaluated in human lung cancer A549 cells and normal lung fibroblast WI-38 cells. A549 cells treated with 6 μg/ml CAPE showed marked growth inhibition (60%) at 48 hr after treatments. During the same time, the number of viable cells decreased to 46% of the control value. In contrast, WI-38 cells showed 20% growth inhibition with no change in the number of viable cells under the same treatment conditions. At 72 hr after CAPE treatment (6 μg/ml), the percentage of apoptotic cells in A549 cultures increased significantly to 67% and an S/G2 arrest was also detected in the culture. Furthermore, there was a significant decrease in the level of intracellular glutathione and hydrogen peroxide contents within one hr after CAPE treatment, and the expression of cyclin B 1 was reduced 6 hr after treatment. The radiation sensitization effect of CAPE on A549 cells was determined from the clonogenic survival curves, and the results showed a small but significant difference in radiation survival between cells treated with or without CAPE. Taken together, our results suggest that the effects of CAPE on differential cytotoxicity, apoptosis, and radiosensitization are associated with glutathione depletion that occurred shortly after treatments. (author)

Inosine Released from Dying or Dead Cells Stimulates Cell Proliferation via Adenosine Receptors

Directory of Open Access Journals (Sweden)

Yi Zhao

2017-04-01

Full Text Available IntroductionMany antitumor therapies induce apoptotic cell death in order to cause tumor regression. Paradoxically, apoptotic cells are also known to promote wound healing, cell proliferation, and tumor cell repopulation in multicellular organisms. We aimed to characterize the nature of the regenerative signals concentrated in the micromilieu of dead and dying cells.MethodsCultures of viable melanoma B16F10 cells, mouse fibroblasts, and primary human fibroblast-like synoviocytes (FLS in the presence of dead and dying cells, their supernatants (SNs, or purified agonists and antagonists were used to evaluate the stimulation of proliferation. Viable cell quantification was performed by either flow cytometry of harvested cells or by crystal violet staining of adherent cells. High-performance liquid chromatography and liquid chromatography coupled with mass spectrometry of cell SNs were deployed to identify the nature of growth-promoting factors. Coimplantation of living cells in the presence of SNs collected from dead and dying cells and specific agonists was used to evaluate tumor growth in vivo.ResultsThe stimulation of proliferation of few surviving cells by bystander dead cells was confirmed for melanoma cells, mouse fibroblasts, and primary FLS. We found that small soluble molecules present in the protein-free fraction of SNs of dead and dying cells were responsible for the promotion of proliferation. The nucleoside inosine released by dead and dying cells acting via adenosine receptors was identified as putative inducer of proliferation of surviving tumor cells after irradiation and heat treatment.ConclusionInosine released by dead and dying cells mediates tumor cell proliferation via purinergic receptors. Therapeutic strategies surmounting this pathway may help to reduce the rate of recurrence after radio- and chemotherapy.

Viability in holder of irradiated cells: distinguish between repair and cell multiplication

International Nuclear Information System (INIS)

Araujo, A.C. de.

1980-01-01

In experiments in which liquid holding recovery (LHR) was measured, the majority of cellular population is formed by non-viable cells and cell multiplication may be important for LHR expression. In order to distinguish between recuperation of viability (true LHR) and cell multiplication, it was necessary to employ improved plating techniques and a fluctuation test based on Poisson distribution. Our results are an indication that this fluctuation test, used together with the traditional method, is a good tool to distinguish repair from cell multiplication. (author)

Silicon Solar Cell Turns 50

Energy Technology Data Exchange (ETDEWEB)

Perlin, J.

2004-08-01

This short brochure describes a milestone in solar (or photovoltaic, PV) research-namely, the 50th anniversary of the invention of the first viable silicon solar cell by three researchers at Bell Laboratories.

Advanced Analysis to Distinguish between Physical Decrease and Inactivation of Viable Phages in Aerosol by Quantitating Phage-Specific Particles.

Science.gov (United States)

Shimasaki, Noriko; Nojima, Yasuhiro; Sakakibara, Masaya; Kikuno, Ritsuko; Iizuka, Chiori; Okaue, Akira; Okuda, Shunji; Shinohara, Katsuaki

2018-01-01

　Recent studies have investigated the efficacy of air-cleaning products against pathogens in the air. A standard method to evaluate the reduction in airborne viruses caused by an air cleaner has been established using a safe bacteriophage instead of pathogenic viruses; the reduction in airborne viruses is determined by counting the number of viable airborne phages by culture, after operating the air cleaner. The reduction in the number of viable airborne phages could be because of "physical decrease" or "inactivation". Therefore, to understand the mechanism of reduction correctly, an analysis is required to distinguish between physical decrease and inactivation. The purpose of this study was to design an analysis to distinguish between the physical decrease and inactivation of viable phi-X174 phages in aerosols. We established a suitable polymerase chain reaction (PCR) system by selecting an appropriate primer-probe set for PCR and validating the sensitivity, linearity, and specificity of the primer-probe set to robustly quantify phi-X174-specific airborne particles. Using this quantitative PCR system and culture assay, we performed a behavior analysis of the phage aerosol in a small chamber (1 m 3 ) at different levels of humidity, as humidity is known to affect the number of viable airborne phages. The results revealed that the reduction in the number of viable airborne phages was caused not only by physical decrease but also by inactivation under particular levels of humidity. Our study could provide an advanced analysis to differentiate between the physical decrease and inactivation of viable airborne phages.

Process for recovering uranium

Science.gov (United States)

MacWood, G. E.; Wilder, C. D.; Altman, D.

1959-03-24

A process useful in recovering uranium from deposits on stainless steel liner surfaces of calutrons is presented. The deposit is removed from the stainless steel surface by washing with aqueous nitric acid. The solution obtained containing uranium, chromium, nickel, copper, and iron is treated with an excess of ammonium hydroxide to precipitnte the uranium, iron, and chromium and convert the nickel and copper to soluble ammonio complexions. The precipitated material is removed, dried and treated with carbon tetrachloride at an elevated temperature of about 500 to 600 deg C to form a vapor mixture of UCl/ sub 4/, UCl/sub 5/, FeCl/sub 3/, and CrCl/sub 4/. The UCl/sub 4/ is separated from this vapor mixture by selective fractional condensation at a temperature of about 500 to 400 deg C.

Recovering oil from shale

Energy Technology Data Exchange (ETDEWEB)

Leahey, T; Wilson, H

1920-11-13

To recover oil free from inorganic impurities and water, and utilize the oil vapor and tarry matter for the production of heat, shale is heated in a retort at a temperature of not less than 120/sup 0/C. The vapors pass by a pipe into a water jacketed condenser from which the condensate and gas pass through a pipe into a chamber and then by a pipe to a setting chamber from where the light oils are decanted through a pipe into a tank. The heavy oil is siphoned through a pipe into a tank, while the gas passes through a pipe into a scrubber and then into a drier, exhauster and pipe to the flue and ports, above the fire-bars, into the retort. Air is introduced through a pipe, flue, and ports.

Two missense mutations, E123Q and K151E, identified in the ERG11 allele of an azole-resistant isolate of Candida kefyr recovered from a stem cell transplant patient for acute myeloid leukemia

Directory of Open Access Journals (Sweden)

Célia Couzigou

2014-07-01

Full Text Available We report on the first cloning and nucleotide sequencing of an ERG11 allele from a clinical isolate of Candida kefyr cross-resistant to azole antifungals. It was recovered from a stem cell transplant patient, in an oncohematology unit exhibiting unexpected high prevalence of C. kefyr. Two amino acid substitutions were identified: K151E, whose role in fluconazole resistance was already demonstrated in Candida albicans, and E123Q, a new substitution never described so far in azole-resistant Candida yeast.

Improved identification of viable myocardium using second harmonic imaging during dobutamine stress echocardiography

NARCIS (Netherlands)

F. Sozzi (Fabiola); D. Poldermans (Don); J.J. Bax (Jeroen); A. Elhendy (Abdou); E.C. Vourvouri (Eleni); R. Valkema (Roelf); J. de Sutter; A.F.L. Schinkel (Arend); A. Borghetti; J.R.T.C. Roelandt (Jos)

2001-01-01

textabstractOBJECTIVE: To determine whether, compared with fundamental imaging, second harmonic imaging can improve the accuracy of dobutamine stress echocardiography for identifying viable myocardium, using nuclear imaging as a reference. PATIENTS: 30 patients with chronic left

Optimization of low energy sonication treatment for granular activated carbon colonizing biomass assessment.

Science.gov (United States)

Saccani, G; Bernasconi, M; Antonelli, M

2014-01-01

This study is aimed at optimizing a low energy sonication (LES) treatment for granular activated carbon (GAC)-colonizing biomass detachment and determination, evaluating detachment efficiency and the effects of ultrasound exposure on bacterial cell viability. GAC samples were collected from two filters fed with groundwater. Conventional heterotrophic plate count (HPC) and fluorescence microscopy with a double staining method were used to evaluate cell viability, comparing two LES procedures, without and with periodical bulk substitution. A 20 min LES treatment, with bulk substitution after cycles of 5 min as maximum treatment time, allowed to recover 87%/100% of attached biomass, protecting detached bacteria from ultrasound damaging effects. Observed viable cell inactivation rate was 6.5/7.9% cell/min, with membrane-compromised cell damage appearing to be even higher (11.5%/13.1% cell/min). Assessing bacterial detachment and damaging ultrasound effects, fluorescence microscopy turned out to be more sensitive compared to conventional HPC. The optimized method revealed a GAC-colonizing biomass of 9.9 x 10(7) cell/gGAC for plant 1 and 8.8 x 10(7) cell/gGAC for plant 2, 2 log lower than reported in literature. The difference between the two GAC-colonizing biomasses is higher in terms of viable cells (46.3% of total cells in plant 1 GAC-colonizing biomass compared to the 33.3% in plant 2). Studying influent water contamination through multivariate statistical analyses, apossible combined toxic and genotoxic effect of chromium VI and trichloroethylene was suggested as a reason for the lower viable cell fraction observed in plant 2 GAC-colonizing population.

Changes in total viable count and TVB-N content in marinated chicken breast fillets during storage

Science.gov (United States)

Baltić, T.; Ćirić, J.; Velebit, B.; Petronijević, R.; Lakićević, B.; Đorđević, V.; Janković, V.

2017-09-01

Marination is a popular technique for enhancing meat properties. Depending on the marinade type and ingredients added, marination can improve sensory, chemical and microbiological quality of meat products. In this study, the total viable count and total volatile basic nitrogen (TVB-N) content in marinated chicken breast fillets were investigated. The possible correlation between bacterial growth and formation of TVB-N was also tested. Chicken breast fillets were immersed in a solution of table salt (as a control) orthree different marinades,which consisted of table salt, sodium tripolyphosphate and/or sodium citrate, and stored in air for nine days at 4±1°C. Analyses of the total viable count and TVB-N were performed on days0, 3, 6 and 9 day of storage. The total viable count gradually increased in all examined groups, and statistically significant differences (pchicken were significantly higher (pchicken marinated with sodium citrate was established (pchicken marinated with sodium tripolyphosphate.

Drug and radiation sensitivity measurements of successful primary monolayer culturing of human tumor cells using cell-adhesive matrix and supplemented medium

International Nuclear Information System (INIS)

Baker, F.L.; Spitzer, G.; Ajani, J.A.

1986-01-01

The limitations of the agar suspension culture method for primary culturing of human tumor cells prompted development of a monolayer system optimized for cell adhesion and growth. This method grew 83% of fresh human tumor cell biopsy specimens, cultured and not contaminated, from a heterogeneous group of 396 tumors including lung cancer (93 of 114, 82%); melanoma (54 of 72, 75%); sarcoma (46 of 59, 78%); breast cancer (35 of 39, 90%); ovarian cancer (16 of 21, 76%); and a miscellaneous group consisting of gastrointestinal, genitourinary, mesothelioma, and unknown primaries (78 of 91, 86%). Cell growth was characterized morphologically with Papanicolaoustained coverslip cultures and cytogenetically with Giemsastained metaphase spreads. Morphological features such as nuclear pleomorphism, chromatin condensation, basophilic cytoplasm, and melanin pigmentation were routinely seen. Aneuploid metaphases were seen in 90% of evaluable cultures, with 15 of 28 showing 70% or more aneuploid metaphases. Colony-forming efficiency ranged between 0.01 and 1% of viable tumor cells, with a median efficiency of 0.2%. This culture system uses a low inoculum of 25,000 viable cells per well which permitted chemosensitivity testing of nine drugs at four doses in duplicate from 2.2 X 10(6) viable tumor cells and radiation sensitivity testing at five doses in quadruplicate from 0.6 X 10(6) cells. Cultures were analyzed for survival by computerized image analysis of crystal violet-stained cells. Drug sensitivity studies showed variability in sensitivity and in survival curve shape with exponential cell killing for cisplatin, Adriamycin, and etoposide, and shouldered survival curves for 5-fluorouracil frequently seen. Radiation sensitivity studies also showed variability in both sensitivity and survival curve shape. Many cultures showed exponential cell killing, although others had shouldered survival curves

Rapid quantification of viable Legionella in nuclear cooling tower waters using filter cultivation, fluorescent in situ hybridization and solid-phase cytometry.

Science.gov (United States)

Baudart, J; Guillaume, C; Mercier, A; Lebaron, P; Binet, M

2015-05-01

To develop a rapid and sensitive method to quantify viable Legionella spp. in cooling tower water samples. A rapid, culture-based method capable of quantifying as few as 600 Legionella microcolonies per litre within 2 days in industrial waters was developed. The method combines a short cultivation step of microcolonies on GVPC agar plate, specific detection of Legionella cells by a fluorescent in situ hybridization (FISH) approach, and a sensitive enumeration using a solid-phase cytometer. Following optimization of the cultivation conditions, the qualitative and quantitative performance of the method was assessed and the method was applied to 262 nuclear power plant cooling water samples. The performance of this method was in accordance with the culture method (NF-T 90-431) for Legionella enumeration. The rapid detection of viable Legionella in water is a major concern to the effective monitoring of this pathogenic bacterium in the main water sources involved in the transmission of legionellosis infection (Legionnaires' disease). The new method proposed here appears to be a robust, efficient and innovative means for rapidly quantifying cultivable Legionella in cooling tower water samples within 48 h. © 2015 The Society for Applied Microbiology.

Natural environmental water sources in endemic regions of northeastern Brazil are potential reservoirs of viable Mycobacterium leprae.

Science.gov (United States)

Arraes, Maria Luisa Bezerra de Macedo; Holanda, Maísa Viana de; Lima, Luana Nepomuceno Gondim Costa; Sabadia, José Antônio Beltrão; Duarte, Cynthia Romariz; Almeida, Rosa Livia Freitas; Kendall, Carl; Kerr, Ligia Regina Sansigolo; Frota, Cristiane Cunha

2017-12-01

The detection of live Mycobacterium leprae in soil and animals other than humans suggests that the environment plays a role in the transmission of leprosy. The objective of this study was to investigate the presence of viable M. leprae in natural water sources used by the local population in five municipalities in the state of Ceará, northeastern Brazil. Samples were collected from 30 different sources. Viable bacilli were identified by reverse transcriptase polymerase chain reaction (PCR) of the M. leprae gyrA gene and sequencing of the PCR products. Physicochemical properties of each water source were also assessed. M. leprae gyrA mRNA was found in 23 (76.7%) of the water sources. No association was found between depth of the water and sample positivity, nor was there any association between the type of water used by the population and sample positivity. An association between viable M. leprae and temperature and pH was found. Georeferencing showed a relation between the residences of leprosy cases and water source containing the bacterium. The finding of viable M. leprae in natural water sources associated with human contact suggests that the environment plays an important role in maintaining endemic leprosy in the study region.

Delayed uptake and washout of contrast in non-viable infarcted myocardium shown with dynamic computed tomography

DEFF Research Database (Denmark)

Udholm, Sebastian; Laugesen, Sofie; Agger, Peter

2014-01-01

BACKGROUND: Assessment of ischemic but potentially viable myocardium plays an important role in the planning of coronary revascularization. Until now SPECT, PET, and MRI have been used to identify viable myocardium. Computed tomography (CT) is increasingly used to diagnose coronary atherosclerosis...... descending artery, followed by removal of the balloon and reperfusion. Four pigs died due to refractory ventricular fibrillation. After 6 weeks, dynamic cardiac CT was performed assessing both wall motion and contrast attenuation. Measurements of attenuation values in Hounsfield units (HU) in the infarct...

Life history characteristics of a recovering lake whitefish Coregonus clupeaformis stock in the Detroit River, North America

Science.gov (United States)

Roseman, Edward F.; Kennedy, Gregory W.; Manny, Bruce A.; Boase, James; McFee, James; Tallman, Ross F.; Howland, Kimberly L.; Rennie, Michael D.; Mills, Kenneth; Tallman, Ross F.; Howland, Kimberly L.; Rennie, Michael D.; Mills, Kenneth

2012-01-01

The Detroit River is part of a channel connecting Lakes Huron and Erie and was once a prolific spawning area for lake whitefish, Coregonus clupeaformis. Large numbers of lake whitefish migrated into the river to spawn where they were harvested by commercial fisheries and for fish culture operations. Prior to our study, the last lake whitefish was landed from the Detroit River in 1925. Loss of spawning habitat during shipping channel construction and over-fishing, likely reduced lake whitefish spawning runs. Because lake whitefish are recovering in Lake Erie, and spawning in the western basin, we suspected they may also be spawning in the Detroit River. We sampled in the Detroit River for lake whitefish adults and eggs in October–December 2005–07 and for larvae during March–May 2006–08. A total of 15 spawning-ready lake whitefish from 4 to 18 years old, were collected. Viable eggs were collected during mid-November 2006–07; highest egg densities were found mid-river. Sac-fry whitefish larvae were collected in the river and near the river mouth. No whitefish larvae were retained in the river. Because high numbers of larvae were collected from mid- and downstream river sites, reproduction of lake whitefish in the Detroit River could contribute substantially to the Lake Erie lake whitefish metapopulation.

Characteristics and quality of intra-operative cell salvage in paediatric scoliosis surgery.

Science.gov (United States)

Perez-Ferrer, A; Gredilla-Díaz, E; de Vicente-Sánchez, J; Navarro-Suay, R; Gilsanz-Rodríguez, F

2016-02-01

To determine the haematological and microbiological characteristics of blood recovered by using a cell saver with a rigid centrifuge bowl (100ml) in paediatric scoliosis surgery and to determine whether it conforms to the standard expected in adult patients. A cross-sectional, descriptive cohort study was performed on 24 consecutive red blood cell (RBC) units recovered from the surgical field and processed by a Haemolite® 2+ (Haemonetics Corp., Braintree, MA, EE. UU.) cell saver. Data were collected regarding age, weight, surgical approach (anterior or posterior), processed shed volume and volume of autologous RBC recovered, full blood count, and blood culture obtained from the RBC concentrate, and incidence of fever after reinfusion. The processed shed volume was very low (939±569ml) with high variability (coefficient of variation=0.6), unlike the recovered volume 129±50ml (coefficient of variation=0.38). A statistically significant correlation between the processed shed volume and recovered RBC concentrate haematocrit was found (Pearson, r=.659, P=.001). Haematological parameters in the recovered concentrate were: Hb 11±5.3g dl(-1); haematocrit: 32.1±15.4% (lower than expected); white cells 5.34±4.22×103 ul(-)1; platelets 37.88±23.5×103 ul(-1) (mean±SD). Blood culture was positive in the RBC concentrate recovered in 13 cases (54.2%) in which Staphylococcus coagulase (-) was isolated. Cell salvage machines with rigid centrifuge bowls (including paediatric small volume) do not obtain the expected haematocrit if low volumes are processed, and therefore they are not the best choice in paediatric surgery. Copyright © 2015 Sociedad Española de Anestesiología, Reanimación y Terapéutica del Dolor. Publicado por Elsevier España, S.L.U. All rights reserved.

An Effective Approach for Immunotherapy Using Irradiated Tumor Cells

International Nuclear Information System (INIS)

Mostafa, D.M.B.

2011-01-01

This study has been aimed to investigate the effect of injection of Irradiated Ehrlich tumor cells alone or concurrent with immunomodulator in mice before and after challenge with viable Ehrlich tumor cells for enhancement of immune system. This study includes the estimation of survival, tumor size, lymphocyte count, LDH, MTT, granzyme B, and DNA fragmentation. In order to fulfill the target of this study, a total of 120 female swiss albino mice were used. They were divided into two classes vaccinated (injection of vaccine before challenge) and therapeutic class (injection of vaccine after challenge). Each class was divided into four groups, group (1) mice injected with viable Ehrlich tumor cells (G1), group (2) mice injected with irradiated tumor cells (G2), group (3) mice injected with immunomodulator (G3), and group (4) mice injected with irradiated tumor cells + immunomodulator (G4). Results obtained from this study demonstrated that, the lymphocyte count and granzyme B activity were increased in both the vaccinated and therapeutic classes compared with control group. LDH activity was decreased in all groups of vaccinated class and also in G2 and G4 groups of therapeutic class compared with control group. There was a significant increase in percent apoptosis of tumor cells cultured with spleenocytes of the groups of vaccinated class as compared with control group. Cellular DNA from Ehrlich tumor cell line cultured with spleenocytes of immunized groups was fragmented into discrete bands of approximate multiples of 200 bp. Revealing significant apoptosis in tumor cells due to vaccination. It is concluded that, vaccination with irradiated tumor cells is an effective approach in stimulation of immune system against viable tumor cells.

Monitoring of yeast cell concentration using a micromachined impedance sensor

NARCIS (Netherlands)

Krommenhoek, E.E.; Gardeniers, Johannes G.E.; Bomer, Johan G.; van den Berg, Albert; Li, X.; Ottens, M.; van der Wielen, L.A.M.; van Dedem, G.W.K.; van Leeuwen, M.; van Gulik, W.M.; Heijnen, J.J.

2005-01-01

The paper describes the design, modelling and experimental characterization of a micromachined impedance sensor for on-line monitoring of the viable yeast cell concentration (biomass) in a miniaturized cell assay. Measurements in a Saccharomyces cerevisiae cell culture show that the permittivity of

Senior health clinics: are they financially viable?

Science.gov (United States)

McAtee, Robin E; Crandall, Debra; Wright, Larry D; Beverly, Claudia J

2009-07-01

Are hospital-based outpatient interdisciplinary clinics a financially viable alternative for caring for our burgeoning population of older adults in America? Although highly popular, with high patient satisfaction rates among older adults and their families, senior health clinics (SHCs) can be expensive to operate, with limited quantifiable health outcomes. This study analyzed three geriatric hospital-based interdisciplinary clinics in rural Arkansas by examining their patient profiles, revenues, and expenses. It closely examined the effects of the downstream revenue using the multiplier effect and acknowledged other factors that weigh heavily on the success of SHCs and the care of older adults. The findings highlight the similarities and differences in the three clinics' operating and financial structures in addition to the clinics' and providers' productivity. The analysis presents an evidence-based illustration that SHCs can break even or lose large amounts of money.

The Effect of Lycopene Preexposure on UV-B-Irradiated Human Keratinocytes

Science.gov (United States)

Ascenso, Andreia; Pedrosa, Tiago; Pinho, Sónia; Pinho, Francisco; de Oliveira, José Miguel P. Ferreira; Cabral Marques, Helena; Oliveira, Helena; Simões, Sandra; Santos, Conceição

2016-01-01

Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and RT-PCR: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer. PMID:26664697

The use of non-living biomass to recover heavy metals from aqueous solutions

International Nuclear Information System (INIS)

Darnall, D.W.

1993-01-01

The use of microorganisms in the treatment of hazardous wastes containing both inorganic and organic pollutants is becoming more and more attractive. There have been two approaches to the use of microorganisms in waste treatment. One involves the use of living organisms and the other involves the use of non-viable biomass derived from microorganisms. While the use of living organisms is often successful in the treatment of toxic organic contaminants, living organisms have not been found to be useful in the treatment of solutions containing heavy metal ions. This is because once the metal ion concentration becomes too high or sufficient metal ions are adsorbed by the microorganism, metabolism is disrupted causing the organism to die. This disadvantage is not encountered if non-living organisms or biological materials derived from microorganisms are used to adsorb metal ions from solution. Instead the biomass is treated as another reagent, a surrogate ion exchange resin. The binding, or biosorption, of metal ions by the biomass results from coordination of the metal ions to various functional groups in or on the cell. These chelating groups, contributed by the cell biopolymers, include carboxyl, imidazole, sulfhydryl, amino, phosphate, sulfate, thioether, phenol, carbonyl, amide, and hydroxyl moieties (Darnall et al.)

Thermoset composite recycling: Properties of recovered glass fiber

DEFF Research Database (Denmark)

Beauson, Justine; Fraisse, Anthony; Toncelli, C.

2015-01-01

Recycling of glass fiber thermoset polymer composite is a challenging topic and a process able to recover the glass fibers original properties in a limited cost is still under investigation. This paper focuses on the recycling technique separating the glass fiber from the matrix material. Four...

Recovering energy and materials from hazardous waste

Energy Technology Data Exchange (ETDEWEB)

Anon.

2003-12-01

The tannery industry faces growing environmental concerns because of the high hazardous metal content of its process waste. The formation, during the tanning process, of the highly toxic hexavalent chromium precludes the use of conventional thermal incineration processes. Borge Tannery in Norway, which processes 600 cattle hides per day, has solved the problem by using new PyroArc technology. The PyroArc waste processing plant can treat all of the tannery's production wastes, transforming them into useful products such as fuel gas and re-usable metal. The fuel gas consists mainly of carbon monoxide, hydrogen and nitrogen, and has a calorific value of about 4 MJ/Nm{sub 3}. About 65-70% of the energy content of the source material (waste or biomass) is recovered in the gas, and this is used to produce steam and/or electricity in a gas engine with a capacity of 580 kW. A further 20-25% of the initial energy content is recovered as heat or low-pressure steam. The plant is designed to be self-sufficient in energy (1.5 MW) and to meet the tannery's maximum requirements for hot water and steam. (UK)

Recovering method for high level radioactive material

International Nuclear Information System (INIS)

Fukui, Toshiki

1998-01-01

Offgas filters such as of nuclear fuel reprocessing facilities and waste control facilities are burnt, and the burnt ash is melted by heating, and then the molten ashes are brought into contact with a molten metal having a low boiling point to transfer the high level radioactive materials in the molten ash to the molten metal. Then, only the molten metal is evaporated and solidified by drying, and residual high level radioactive materials are recovered. According to this method, the high level radioactive materials in the molten ashes are transferred to the molten metal and separated by the difference of the distribution rate of the molten ash and the molten metal. Subsequently, the molten metal to which the high level radioactive materials are transferred is heated to a temperature higher than the boiling point so that only the molten metal is evaporated and dried to be removed, and residual high level radioactive materials are recovered easily. On the other hand, the molten ash from which the high level radioactive material is removed can be discarded as ordinary industrial wastes as they are. (T.M.)

Progress in emerging techniques for characterization of immobilized viable whole-cell biocatalysts

Czech Academy of Sciences Publication Activity Database

Bučko, M.; Vikartovská, A.; Schenkmayerová, A.; Tkáč, J.; Filip, J.; Chorvát Jr., D.; Neděla, Vilém; Ansorge-Schumacher, M.B.; Gemeiner, P.

2017-01-01

Roč. 71, č. 11 (2017), s. 2309-2324 ISSN 0366-6352 Institutional support: RVO:68081731 Keywords : bioelectrocatalysis * imaging techniques * immobilized whole-cell biocatalyst * multienzyme cascade reactions * online kinetics Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering OBOR OECD: Bioprocessing technologies (industrial processes relying on biological agents to drive the process) biocatalysis, fermentation Impact factor: 1.258, year: 2016

Process for recovering xenon from radioactive gaseous wastes

International Nuclear Information System (INIS)

Kishimoto, Tsuneo.

1980-01-01

Purpose: To recover pure xenon economically and efficiently by amply removing radioactive krypton mixed in xenon without changing the rectifying capacity of an xenon rectifying system itself. Method: Xe containing radioactive Kr(Kr-85) is rectified to reduce the concentration of radioactive Kr. Thereafter, non-radioactive Kr or Ar is added to Xe and further the rectification is carried out. The raw material Xe from the Xe adsorption system of, for example, a radioactive gaseous waste disposal system is cooled to about 100 0 C by a heat-exchanger and thereafter supplied to a rectifying tower to carry out normal rectification of Xe thereby to reduce the concentration of Kr contained in Xe at the tower bottom to the rectification limit concentration. Then, non-radioactive Kr is supplied via a precooler to the tower bottom to continue the rectification, thus the Xe fractions at the tower bottom, in which the concentration of radioactive Kr is reduced, being compressed and recovered. (Kamimura, M.)

An alternative approach to recovering valuable metals from zinc phosphating sludge.

Science.gov (United States)

Kuo, Yi-Ming

2012-01-30

This study used a vitrification process (with good potential for commercialization) to recover valuable metals from Zn phosphating sludge. The involved vitrification process achieves two major goals: it transformed hazardous Zn phosphating sludge into inert slag and it concentrated Fe (83.5%) and Zn (92.8%) into ingot and fine particulate-phase material, respectively. The Fe content in the ingot was 278,000 mg/kg, making the ingot a potential raw material for iron making. The fine particulate-phase material (collected from flue gas) contained abundant Zn (544,000 mg/kg) in the form of ZnO. The content (67.7%) of ZnO was high, so it can be directly sold to refineries. The recovered coarse particulate-phase material, with insufficient amount of ZnO, can be recycled as a feeding material for Zn re-concentration. Therefore, the vitrification process can not only treat hazardous materials but also effectively recover valuable metals. Copyright © 2011 Elsevier B.V. All rights reserved.

Economically viable large-scale hydrogen liquefaction

Science.gov (United States)

Cardella, U.; Decker, L.; Klein, H.

2017-02-01

The liquid hydrogen demand, particularly driven by clean energy applications, will rise in the near future. As industrial large scale liquefiers will play a major role within the hydrogen supply chain, production capacity will have to increase by a multiple of today’s typical sizes. The main goal is to reduce the total cost of ownership for these plants by increasing energy efficiency with innovative and simple process designs, optimized in capital expenditure. New concepts must ensure a manageable plant complexity and flexible operability. In the phase of process development and selection, a dimensioning of key equipment for large scale liquefiers, such as turbines and compressors as well as heat exchangers, must be performed iteratively to ensure technological feasibility and maturity. Further critical aspects related to hydrogen liquefaction, e.g. fluid properties, ortho-para hydrogen conversion, and coldbox configuration, must be analysed in detail. This paper provides an overview on the approach, challenges and preliminary results in the development of efficient as well as economically viable concepts for large-scale hydrogen liquefaction.

New arrangement for the air cleanup system to recover tritium

International Nuclear Information System (INIS)

Nishikawa, Masabumi; Takahashi, Kohsaku; Munakata, Kenzo; Fukada, Satoshi; Kotoh, Kenji; Takeishi, Toshiharu

1997-01-01

At present, the standard arrangement of the air cleanup system responsible for emergency tritium recovery from room air is a catalytic oxidation bed with a heater followed by an adsorption bed with a cooler. One disadvantage of this arrangement is that trouble with the heater or the cooler could result in a loss of capacity to recover tritium. Another disadvantage of the catalyst-adsorption-bed arrangement is that tritiated water must be recovered with a high decontamination factor after dilution with a large amount of water vapor in the working atmosphere. The performance of a new arrangement for the air cleanup system, which consists of a precious metal catalyst bed preceded by an adsorption bed without heating equipment, is discussed. According to calculations, most of the tritium released to the room air is recovered in the catalyst bed through oxidation, adsorption, and isotope exchange reaction when the new arrangement is applied. The adsorption bed placed before the catalyst bed dehumidifies the process gas to such a degree that the oxidation reaction of tritium in the catalyst bed is not hindered by water vapor. 15 refs., 6 figs., 6 tabs

Comparison of Assays for Sensitive and Reproducible Detection of Cell Culture-Infectious Cryptosporidium parvum and Cryptosporidium hominis in Drinking Water

Science.gov (United States)

Di Giovanni, George D.; Rochelle, Paul A.

2012-01-01

This study compared the three most commonly used assays for detecting Cryptosporidium sp. infections in cell culture: immunofluorescent antibody and microscopy assay (IFA), PCR targeting Cryptosporidium sp.-specific DNA, and reverse transcriptase PCR (RT-PCR) targeting Cryptosporidium sp.-specific mRNA. Monolayers of HCT-8 cells, grown in 8-well chamber slides or 96-well plates, were inoculated with a variety of viable and inactivated oocysts to assess assay performance. All assays detected infection with low doses of flow cytometry-enumerated Cryptosporidium parvum oocysts, including infection with one oocyst and three oocysts. All methods also detected infection with Cryptosporidium hominis. The RT-PCR assay, IFA, and PCR assay detected infection in 23%, 25%, and 51% of monolayers inoculated with three C. parvum oocysts and 10%, 9%, and 16% of monolayers inoculated with one oocyst, respectively. The PCR assay was the most sensitive, but it had the highest frequency of false positives with mock-infected cells and inactivated oocysts. IFA was the only infection detection assay that did not produce false positives with mock-infected monolayers. IFA was also the only assay that detected infections in all experiments with spiked oocysts recovered from Envirochek capsules following filtration of 1,000 liters of treated water. Consequently, cell culture with IFA detection is the most appropriate method for routine and sensitive detection of infectious Cryptosporidium parvum and Cryptosporidium hominis in drinking water. PMID:22038611

Economically viable biochemical processes for the advanced rural biorefinery and downstream recovery operations

Science.gov (United States)

Rural biorefineries offer an alternative to traditional ethanol production by providing the opportunity to produce fuel on site to reduce costs associated with biomass transportation thus making the fuel economically viable. Widespread installation of rural biorefineries could lead to increased upt...

Mechanical lifter for recovering highly viscous oil and bitumens

Energy Technology Data Exchange (ETDEWEB)

Rakhmanov, R N; Akhunov, A M; Asfandiyarov, Kh A; Maksutov, R A

1982-01-01

A mechanical lifter is described for recovering highly viscous oil and bitumens. The lifter differs from the known and has significant advantages over them. The lifter was made and tested on a stand well.

Long-term storage of recovered krypton

International Nuclear Information System (INIS)

Horii, Yuji; Yamamoto, Yoshimasa

1983-01-01

Various storage methods for krypton-85 recovered from a nuclear fuel reprocessing plant are under development in many countries. These methods include:(1) direct storage in pressurized cylinders, (2) storage of krypton adsorbed on charcoal or zeolite in pressurized cylinders and (3) immobilization (encapsulation) in zeolite. A krypton storage facility using pressurized cylinders has been constructed in the krypton recovery pilot plant in Tokaimura and other methods are now under development. These three methods are evaluated and the features of the constructed facility are also reported. (author)

Molecular determinants of the V3 loop of human immunodeficiency virus type 1 glycoprotein gp120 responsible for controlling cell tropism.

Science.gov (United States)

Chavda, S C; Griffin, P; Han-Liu, Z; Keys, B; Vekony, M A; Cann, A J

1994-11-01

We and others have identified the major determinant of cell tropism in human immunodeficiency virus type 1 (HIV-1) as the V3 loop of glycoprotein gp120. We have conducted a detailed study of two molecularly cloned isolates of HIV-1, HIVJR-CSF and HIVNL4-3, that differ in their tropism for immortalized CD4+ cell lines, by constructing a series of site-directed mutations within the V3 loop of HIVJR-CSF based on the sequence of HIVNL4-3. The phenotypes of these mutants fall into two classes, those which are viable and those which are not. A spontaneous mutant with significantly altered growth properties was also recovered and found to have an additional single amino acid change in the V3 loop sequence. The carboxy-terminal beta-strand part of the V3 loop is the major determinant of cell tropism. However, the results presented here indicate that the functional role of the V3 loop sequences can only be interpreted properly in the context of the original gp120 backbone from which they were derived. These findings show that over-simplistic interpretation of sequence data derived from unknown mixtures of HIV variants in infected persons may be highly misleading.

Neutron activation analysis of maltenes recovered from EUROBITUM simulates

International Nuclear Information System (INIS)

Impens, N

2006-01-01

According to the present Belgian reference scenario, Eurobitum bituminised radioactive waste has to be disposed off in a deep underground repository in a stable geological formation such as Boom Clay. This waste originated mainly from mixtures of nuclear fuel decladding slurries and waste concentrates from the nuclear fuel cycle. Even though safety assessment studies up till present do not show that this waste is unacceptable for deep underground disposal, a final decision about the disposal of the bituminised waste has not been taken so far, and alternative solutions are still conceivable. To support the decision-making we investigate methods to recondition this bituminised waste. We continued studying a room temperature re-treatment method for Eurobitum. The aim of the method is the stabilisation and minimisation of final waste, and the free release of recovered materials. The method comprises the recovery of maltenes and water soluble salts. The recovery of maltenes is performed by dissolving the complete bitumen matrix with a 'solvent', followed by the precipitation of the asphaltenes by addition of a so-called 'nonsolvent'. The 'solvent' is a 50 percent aromatic blend of Shellsol A150 and Shellsol H, whereas the 'nonsolvent' is aliphatic Shellsol T. The recovered maltenes represent 40 % wt of the waste, as shown in the inner pie chart. Part of the maltenes could not be recovered and remain in the asphaltene matrix, as can be seen from the difference between the inner and outer pie chart, representing the real composition and the weight fractions after separation, respectively. A second step of the room temperature re-treatment method covers the complete removal of the water soluble (nitrate) salts, and is described in the main reference and references therein. Application of the room temperature re-treatment method results in a final waste that consists of water insoluble salts embedded in an asphaltene matrix which is less sensitive towards radiolysis than

The susceptible-infected-recovered (SIR) model for viral marketing

Science.gov (United States)

Ismail, Siti Suhaila; Akil, Ku Azlina Ku; Chulan, Majdah; Sharif, Noorzila

2017-11-01

Viral marketing is a marketing strategy utilizes social media to spread information about a product or services provided. It is the most powerful way to share information in a short amount of time. The objective of this study is to investigate the dynamic of viral marketing within a time duration in the point of view of mathematics. This study used the epidemiological model known as Susceptible-Infected-Recovered (SIR). The model consists of a system of three differential equations with three state variables namely susceptible (S), infected (I) and recovered (R). It considers a case of SIR model with demography. Numerical experiments have been performed. The results show that viral marketing reaches its peak within two days. The online messages shared will become higher if the initial number of the infected individual has been increased.

T-cell response in human leishmaniasis

DEFF Research Database (Denmark)

Kharazmi, A; Kemp, K; Ismail, A

1999-01-01

In the present communication we provide evidence for the existence of a Th1/Th2 dichotomy in the T-cell response to Leishmania antigens in human leishmaniasis. Our data suggest that the pattern of IL-4 and IFN-gamma response is polarised in these patients. Lymphocytes from individuals recovered...... from cutaneous leishmaniasis (CL) responded by IFN-gamma production following stimulation with Leishmania antigens whereas cells from patients recovered from visceral leishmaniasis (VL) showed a mixed pattern of IFN-gamma and IL-4 responses. The cells producing these cytokines were predominantly CD4......+. Furthermore, IL-10 plays an important role in the development of post kala azar dermal leishmaniasis (PKDL) from VL. The balance between the parasitic-specific T-cell response plays an important regulatory role in determining the outcome of Leishmania infections in humans....

Oxidative Stress Induces Senescence in Cultured RPE Cells.

Science.gov (United States)

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

2016-01-01

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

Breaking down the barriers to commercialization of fuel cells in transportation through Government - industry R&D programs

Energy Technology Data Exchange (ETDEWEB)

Chalk, S.G. [Dept. of Energy, Washington, DC (United States); Venkateswaran, S.R. [Energetics, Inc., Columbia, MD (United States)

1996-12-31

PEM fuel cell technology is rapidly emerging as a viable propulsion alternative to the internal combustion engine. Fuel cells offer the advantages of low emissions, high efficiency, fuel flexibility, quiet and continuous operation, and modularity. Over the last decade, dramatic advances have been achieved in the performance and cost of PEM fuel cell technologies for automotive applications. However, significant technical barriers remain to making fuel cell propulsion systems viable alternatives to the internal combustion engine. This paper focuses on the progress achieved and remaining technical barriers while highlighting Government-industry R&D efforts that are accelerating fuel cell technology toward commercialization.

Immunization against chlamydial genital infection in guinea pigs with UV-inactivated and viable chlamydiae administered by different routes

International Nuclear Information System (INIS)

Rank, R.G.; Batteiger, B.E.; Soderberg, L.S.

1990-01-01

Female guinea pigs were immunized with viable or UV light-inactivated chlamydiae, belonging to the species Chlamydia psittaci, by intravenous, subcutaneous, oral, or ocular routes. All animals were then inoculated vaginally with viable chlamydiae to determine the extent of protection against challenge infection induced by the various regimens. The course of genital infection was significantly reduced in intensity in all groups of animals except the unimmunized controls and those animals immunized orally with inactivated antigen. Guinea pigs immunized with viable antigen were more likely to develop resistance to challenge infection and, in general, had a significantly greater degree of protection than animals immunized with inactivated antigen. No one route seemed superior in producing a protective response. Animals in all groups demonstrating protection developed serum and secretion immunoglobulin G antibody responses to chlamydiae. Lymphocyte proliferative reactions to chlamydial antigen were variable among groups. Immunoblot analysis of serum and secretions indicated a wide range of antibody specificities, but most protected animals produced antibodies to the major outer membrane protein, lipopolysaccharide, and the 61-kilodalton protein. No definitive associations could be made between the increased ability of immunization with viable organisms to produce resistance to challenge infection and a particular immune parameter. These data indicate that viable chlamydiae given by various routes are able to induce a strong immune response which can provide resistance against reinfection in some cases or at least reduce the degree of infection to a greater degree than inactivated antigen. However, complete resistance to genital tract infection may be difficult to obtain and alternate immunizations strategies may have to be developed

Method for recovering or recirculating stable nitroxide radicals

NARCIS (Netherlands)

Heeres, Andre; Van Doren, Hendrik Arend; Bleeker, Ido Pieter; Gotlieb, Kornelis Fester.

1996-01-01

The invention relates fo a method for recovering stable nitroxide radicals, wherein at least a part of a reaction mixt. consisting of a soln. or suspension, or a filtrate or supernatant of a suspension, in which stable nitroxide radicals are present in non-solid form, is subjected to an azeotropic

Kinetic analysis of thymocyte attachment to thymus stromal cells in culture by using phase-contrast and scanning electron microscopy

International Nuclear Information System (INIS)

LaRochelle, G.G.; Jones, K.H.

1989-01-01

Direct cellular contact between thymocytes and thymus stromal cells within the thymus appears to contribute to the maturation of thymocytes. Thymocyte-stromal cell complexes, formed in vivo, have been isolated by others and postulated to play a role in T-cell differentiation. These previous studies have been hampered, however, by a time-consuming isolation procedure from which only small numbers of these complexes are recovered. We have examined a model to study thymocyte-stromal cell complexes in vitro in which thymocytes are added to primary cultures of thymus stromal cells. In the present study, we found that thymocytes were histotypically selective in their attachment to thymus stromal cells. We also investigated the kinetics of thymocyte attachment to these thymus stromal cells. Cultures were examined at selected time intervals from 5 min through 3 days of incubation. Thymocyte attachment to stromal cells was a biphasic interaction, with maximum surface attachment at 15 min of cocultivation, followed by migration of thymocytes into the cultures. Morphological studies were confirmed by using 3 H-leucine-labeled thymocytes and liquid scintigraphy. With increased time in culture, thymocytes became amoeboid and migrated between the layers of stromal cells where thymocyte mitotic figures were seen at 4 and 8 hr. In some cases it appeared that stromal cells, which often grew two to three cell layers deep, played an active role in enclosing thymocytes within the cultures. Large numbers of viable thymocytes were observed in the cultures at 24 hr. The number of thymocytes then decreased progressively on days 2 and 3, when relatively few were found within the layers of the culture

In search of viable business models for development: sustainable energy in developing countries

NARCIS (Netherlands)

Kolk, A.; van den Buuse, D.

2012-01-01

Purpose - Although the crucial role of business, and of business-based approaches, in development is increasingly emphasised by academics and practitioners, insight is lacking into the "whether and how" of viable business models, in environmental, social and economical terms. This article analyses

Enterocolic lymphocytic phlebitis: statistical analysis of histology features in viable and ischemic bowel.

Science.gov (United States)

Medlicott, Shaun A C; Guggisberg, Kelly A; DesCôteaux, Jean-Gaston; Beck, Paul

2006-07-01

Enterocolic lymphocytic phlebitis is a rare cause of segmental ischemic enterocolitis. This artery-sparing transmural vasculitis is classically a circumferential phlebitis with perivenular lymphocyte cuffing and thrombi in the absence of systemic manifestations. Myointimal hyperplasia may represent a chronic phase of enterocolic lymphocytic phlebitis. Subclinical or early stage enterocolic lymphocytic phlebitis is not well delineated. We analyzed 600 submucosal and subserosal veins from both ischemic and intact bowel segments to discern if vascular morphology varied between sites. Crescentic and circumferential lymphocytic phlebitis is more common in viable bowel than in the ischemic segment. A nonsignificant trend was found for increased crescentic morphology between intact bowel remote from the ischemic focus compared with that adjacent to the ischemic focus. Hallmarks of ischemic bowel are necrotizing phlebitis and thrombi formation. Thrombophlebitis morphology is distinctly different in viable and ischemic bowel, changing from the classic lymphocytic to necrotizing lesions respectively.

Process of recovering shale oil

Energy Technology Data Exchange (ETDEWEB)

1949-01-17

A process is disclosed for recovering oil from shale rock by means of channels cut in the shale deposit, to which heat is carried for warming the shale mass and which are separated from the fume channels formed in the shale by parts of the shale rock, characterized in that heating elements are put down in the heating channels, which occupy less cross section than these channels, and in the so-formed space between the channel wall and the heating element a filling is placed, which facilitates heat transfer between the heating element and the shale and simultaneously prevents a streaming of the oily product gasified out of the shale from working into the heating element and stopping it.

Blood flow, flow reserve, and glucose utilization in viable and nonviable myocardium in patients with ischemic cardiomyopathy.

Science.gov (United States)

Zhang, Xiaoli; Schindler, Thomas H; Prior, John O; Sayre, James; Dahlbom, Magnus; Huang, Sung-Cheng; Schelbert, Heinrich R

2013-04-01

The aim of the study was to determine whether glucose uptake in viable myocardium of ischemic cardiomyopathy patients depends on rest myocardial blood flow (MBF) and the residual myocardial flow reserve (MFR). Thirty-six patients with ischemic cardiomyopathy (left ventricular ejection fraction 25 ± 10 %) were studied with (13)N-ammonia and (18)F-fluorodeoxyglucose (FDG) positron emission tomography (PET). Twenty age-matched normals served as controls. Regional MBF was determined at rest and during dipyridamole hyperemia and regional FDG extraction was estimated from regional FDG to (13)N-ammonia activity ratios. Rest MBF was reduced in viable (0.42 ± 0.18 ml/min per g) and nonviable regions (0.32 ± 0.09 ml/min per g) relative to remote regions (0.68 ± 0.23 ml/min per g, p MFRs did not differ significantly (p > 0.05). Compared to MFR in remote myocardium, MFRs in viable regions were similar (1.39 ± 0.56 vs 1.70 ± 0.45, p > 0.05) but were significantly lower in nonviable regions (1.23 ± 0.43, p MFRs (r =-0.424, p MFRs in viable myocardium are associated with increasing glucose extraction that likely reflects a metabolic adaptation of remodeling hibernating myocytes.

Reward banding to determine reporting rate of recovered mourning dove bands

Science.gov (United States)

Tomlinson, R.E.

1968-01-01

Reward bands placed on the other leg of certain regularly banded immature mourning doves (Zenaidura macroura) were used to develop information on reporting rates of recovered dove bands. Reports from 15 widely separated sections of the United States showed considerable variation in recovery rate of doves both with and without reward bands. The overall percentages of banded doves that were reported as recovered were 9.69% for those with reward bands and 3.83% for controls. The bandreporting rate for states influenced by publicity was 66%; that for states not influenced was 32%.

Finding viable models in SUSY parameter spaces with signal specific discovery potential

Science.gov (United States)

Burgess, Thomas; Lindroos, Jan Øye; Lipniacka, Anna; Sandaker, Heidi

2013-08-01

Recent results from ATLAS giving a Higgs mass of 125.5 GeV, further constrain already highly constrained supersymmetric models such as pMSSM or CMSSM/mSUGRA. As a consequence, finding potentially discoverable and non-excluded regions of model parameter space is becoming increasingly difficult. Several groups have invested large effort in studying the consequences of Higgs mass bounds, upper limits on rare B-meson decays, and limits on relic dark matter density on constrained models, aiming at predicting superpartner masses, and establishing likelihood of SUSY models compared to that of the Standard Model vis-á-vis experimental data. In this paper a framework for efficient search for discoverable, non-excluded regions of different SUSY spaces giving specific experimental signature of interest is presented. The method employs an improved Markov Chain Monte Carlo (MCMC) scheme exploiting an iteratively updated likelihood function to guide search for viable models. Existing experimental and theoretical bounds as well as the LHC discovery potential are taken into account. This includes recent bounds on relic dark matter density, the Higgs sector and rare B-mesons decays. A clustering algorithm is applied to classify selected models according to expected phenomenology enabling automated choice of experimental benchmarks and regions to be used for optimizing searches. The aim is to provide experimentalist with a viable tool helping to target experimental signatures to search for, once a class of models of interest is established. As an example a search for viable CMSSM models with τ-lepton signatures observable with the 2012 LHC data set is presented. In the search 105209 unique models were probed. From these, ten reference benchmark points covering different ranges of phenomenological observables at the LHC were selected.

Recovering an obstacle using integral equations

KAUST Repository

Rundell, William

2009-05-01

We consider the inverse problem of recovering the shape, location and surface properties of an object where the surrounding medium is both conductive and homogeneous and we measure Cauchy data on an accessible part of the exterior boundary. It is assumed that the physical situation is modelled by harmonic functions and the boundary condition on the obstacle is one of Dirichlet type. The purpose of this paper is to answer some of the questions raised in a recent paper that introduced a nonlinear integral equation approach for the solution of this type of problem.

Process and system for recovering energy

International Nuclear Information System (INIS)

Trichet, J.-C.

1975-01-01

The invention concerns a system for heating a room or similar, using a 'dibare' cycle working with a less volatile fluid and a more volatile fluid and employing a coupling between a fractionated separation area working at high pressure and a fractionated mixing zone working at low pressure. The invention relates to the following uses: heating a room or building, drying any material and recovering the thermal discharge from a motive cycle, particularly the recovery of the thermal discharge of an electric nuclear power station to carry out district heating [fr

Recovering from a bad start: rapid adaptation and tradeoffs to growth below a threshold density

Directory of Open Access Journals (Sweden)

Marx Christopher J

2012-07-01

Full Text Available Abstract Background Bacterial growth in well-mixed culture is often assumed to be an autonomous process only depending upon the external conditions under control of the investigator. However, increasingly there is awareness that interactions between cells in culture can lead to surprising phenomena such as density-dependence in the initiation of growth. Results Here I report the unexpected discovery of a density threshold for growth of a strain of Methylobacterium extorquens AM1 used to inoculate eight replicate populations that were evolved in methanol. Six of these populations failed to grow to the expected full density during the first couple transfers. Remarkably, the final cell number of six populations crashed to levels 60- to 400-fold smaller than their cohorts. Five of these populations recovered to full density soon after, but one population remained an order of magnitude smaller for over one hundred generations. These variable dynamics appeared to be due to a density threshold for growth that was specific to both this particular ancestral strain and to growth on methanol. When tested at full density, this population had become less fit than its ancestor. Simply increasing the initial dilution 16-fold reversed this result, revealing that this population had more than a 3-fold advantage when tested at this lower density. As this population evolved and ultimately recovered to the same final density range as the other populations this low-density advantage waned. Conclusions These results demonstrate surprisingly strong tradeoffs during adaptation to growth at low absolute densities that manifest over just a 16-fold change in density. Capturing laboratory examples of transitions to and from growth at low density may help us understand the physiological and evolutionary forces that have led to the unusual properties of natural bacteria that have specialized to low-density environments such as the open ocean.

Recovering from a bad start: rapid adaptation and tradeoffs to growth below a threshold density.

Science.gov (United States)

Marx, Christopher J

2012-07-04

Bacterial growth in well-mixed culture is often assumed to be an autonomous process only depending upon the external conditions under control of the investigator. However, increasingly there is awareness that interactions between cells in culture can lead to surprising phenomena such as density-dependence in the initiation of growth. Here I report the unexpected discovery of a density threshold for growth of a strain of Methylobacterium extorquens AM1 used to inoculate eight replicate populations that were evolved in methanol. Six of these populations failed to grow to the expected full density during the first couple transfers. Remarkably, the final cell number of six populations crashed to levels 60- to 400-fold smaller than their cohorts. Five of these populations recovered to full density soon after, but one population remained an order of magnitude smaller for over one hundred generations. These variable dynamics appeared to be due to a density threshold for growth that was specific to both this particular ancestral strain and to growth on methanol. When tested at full density, this population had become less fit than its ancestor. Simply increasing the initial dilution 16-fold reversed this result, revealing that this population had more than a 3-fold advantage when tested at this lower density. As this population evolved and ultimately recovered to the same final density range as the other populations this low-density advantage waned. These results demonstrate surprisingly strong tradeoffs during adaptation to growth at low absolute densities that manifest over just a 16-fold change in density. Capturing laboratory examples of transitions to and from growth at low density may help us understand the physiological and evolutionary forces that have led to the unusual properties of natural bacteria that have specialized to low-density environments such as the open ocean.

Evolution of β-Cell Replacement Therapy in Diabetes Mellitus: Islet Cell Transplantation

Science.gov (United States)

Jahansouz, Cyrus; Jahansouz, Cameron; Kumer, Sean C.; Brayman, Kenneth L.

2011-01-01

Diabetes mellitus remains one of the leading causes of morbidity and mortality worldwide. According to the Centers for Disease Control and Prevention, approximately 23.6 million people in the United States are affected. Of these individuals, 5 to 10% have been diagnosed with Type 1 diabetes mellitus (T1DM), an autoimmune disease. Although it often appears in childhood, T1DM may manifest at any age, leading to significant morbidity and decreased quality of life. Since the 1960s, the surgical treatment for diabetes mellitus has evolved to become a viable alternative to insulin administration, beginning with pancreatic transplantation. While islet cell transplantation has emerged as another potential alternative, its role in the treatment of T1DM remains to be solidified as research continues to establish it as a truly viable alternative for achieving insulin independence. In this paper, the historical evolution, procurement, current status, benefits, risks, and ongoing research of islet cell transplantation are explored. PMID:22013505

Survival and kinetics of Chinese hamster ovary cell subpopulations induced by Adriamycin and radiation

International Nuclear Information System (INIS)

Schneiderman, M.H.

1979-01-01

Mitotic selection of Chinese hamster ovary (CHO) cells, at 10 min intervals after the initiation of Adriamycin and/or x-ray treatment was used to measure the kinetics and survival of cells which progressed without delay, the ''refractory'' cells, the cells that reached mitosis only after recovery from the treatment-induced delay, the ''recovered'' cells, and the survival of the cells remaining attached to the flask 5 h after treatment. The cell kinetics were determined from the rate at which cells entered mitosis, and the reproductive integrity from the survival of the selected refractory, recovered and remaining (unselected) cells

Survival or revival: long-term preservation induces a reversible viable but non-culturable state in methane-oxidizing bacteria.

Directory of Open Access Journals (Sweden)

Sven Hoefman

Full Text Available Knowledge on long-term preservation of micro-organisms is limited and research in the field is scarce despite its importance for microbial biodiversity and biotechnological innovation. Preservation of fastidious organisms such as methane-oxidizing bacteria (MOB has proven difficult. Most MOB do not survive lyophilization and only some can be cryopreserved successfully for short periods. A large-scale study was designed for a diverse set of MOB applying fifteen cryopreservation or lyophilization conditions. After three, six and twelve months of preservation, the viability (via live-dead flow cytometry and culturability (via most-probable number analysis and plating of the cells were assessed. All strains could be cryopreserved without a significant loss in culturability using 1% trehalose in 10-fold diluted TSB (TT as preservation medium and 5% DMSO as cryoprotectant. Several other cryopreservation and lyophilization conditions, all of which involved the use of TT medium, also allowed successful preservation but showed a considerable loss in culturability. We demonstrate here that most of these non-culturables survived preservation according to viability assessment indicating that preservation induces a viable but non-culturable (VBNC state in a significant fraction of cells. Since this state is reversible, these findings have major implications shifting the emphasis from survival to revival of cells in a preservation protocol. We showed that MOB cells could be significantly resuscitated from the VBNC state using the TT preservation medium.

The ghost of extinction: Preservation values and minimum viable population in wildlife models

NARCIS (Netherlands)

Eiswerth, M.E.; Kooten, van G.C.

2009-01-01

The inclusion of a minimum viable population in bioeconomic modeling creates at least two complications that are not resolved by using a modified logistic growth function. The first complication can be dealt with by choosing a different depensational growth function. The second complication relates

The Effects of Royal Jelly on In-Vitro Cytotoxicity of K562 Cells and Peripheral Blood Mononuclear Cells

Directory of Open Access Journals (Sweden)

SE Hosseini

2014-02-01

Full Text Available Abstract Background & aim: Royal jelly, secreted by worker bees, has different biological activities on cells and tissues. The aim of this study was to evaluate the effects of royal jelly on peripheral blood mononuclear cells and on the tumor category of K562 cell line. Methods: In the present experimental study, three subjects were selected separately with three repetitions. K562 (104 cells and PBMC (105 cells with different concentrations of royal jelly (5, 10, 25, 50 and 100 mg/ml were cultured under standard conditions for 48 and 72 h separately. The fatality rate on PBMC cells and K562 cancer cells was evaluated by using MTT (Tetrazolium Dye-Reduction Assay. The number of viable cells in PBMC that were exposed for 48 hours with Royal Jelly was evaluated by trypan blue staining. Data were analyzed by ANOVA. Results: The royal jelly had no cytotoxicity effect on PBMC cells but at concentration of 50 and 100 mg/mL the cytotoxicity effect were observed on k562 cells whereas, at 10 and 25 mg/ml the number of PBMC viable cells increased. Conclusion: Due to the lack of lethality of royal jelly on PBMC cells and PBMC cell viability and an increase in the fatality rate of cancer cells in the future, royal jelly can be used as a potential candidate for treatment of leukemia. Keywords: Royal jelly, K562, peripheral blood mononuclear cell

Susceptible-infected-recovered epidemics in random networks with population awareness

Science.gov (United States)

Wu, Qingchu; Chen, Shufang

2017-10-01

The influence of epidemic information-based awareness on the spread of infectious diseases on networks cannot be ignored. Within the effective degree modeling framework, we discuss the susceptible-infected-recovered model in complex networks with general awareness and general degree distribution. By performing the linear stability analysis, the conditions of epidemic outbreak can be deduced and the results of the previous research can be further expanded. Results show that the local awareness can suppress significantly the epidemic spreading on complex networks via raising the epidemic threshold and such effects are closely related to the formulation of awareness functions. In addition, our results suggest that the recovered information-based awareness has no effect on the critical condition of epidemic outbreak.

Defined Combinations of Cryomedia and Thawing Extenders Influence the Viable X-Y Boar Sperm Ratio in Vitro.

Science.gov (United States)

Korchunjit, W; Kaeoket, K; Kitiyanant, Y; Taylor, J; Wongtawan, T

It is believed that plasma membrane X- and Y-chromosome bearing sperm are different; therefore the freezing and thawing process may affect X- and Y-sperm differently. The objective of this study was to investigate the effect of cryomedia and thawing extenders on the survival of X and Y-sperm. Three different cryomedia and thawing extenders were compared. Viable motile sperm were separated using a swim-up technique. Real-time PCR was used to identify the sperm type. Using CryoA for freezing and Beltsville-Thawing-Solution (BTS) as the thawing extender yielded significantly higher numbers of viable motile Y sperm (64 percent) than control (48 percent) (P semen freezing with CryoC and thawing with Androstar Plus gave a significantly lower number of viable motile Y sperm (32 percent) than control (51 percent). Our results revealed that defined combinations of cryomedia and thawing extenders significantly altered the survival ratio of frozen-thawed X-Y sperm in vitro, which has potential implications for artificial insemination.

Langerhans cells are required for UVR-induced immunosuppression

NARCIS (Netherlands)

Schwarz, Agatha; Noordegraaf, Madelon; Maeda, Akira; Torii, Kan; Clausen, Björn E.; Schwarz, Thomas

2010-01-01

Painting of haptens onto UVR-exposed skin does not result in sensitization but induces regulatory T cells (Treg). This was explained by UVR-mediated depletion of Langerhans cells (LCs). Furthermore, migration of UVR-damaged but still viable LCs into lymph nodes appears to be essential to induce

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

International Nuclear Information System (INIS)

Terzaghi, M.; Little, J.B.

1976-01-01

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

When will the Antarctic Ozone Hole Recover?

Science.gov (United States)

Newman, Paul A.; Nash, Eric R.; Kawa, S. Randolph; Montzka, Steve

2006-01-01

The Antarctic ozone hole develops each year and culminates by early Spring. Antarctic ozone values have been monitored since 1979 using satellite observations from the .TOMS instrument. The severity of the hole has been assessed from TOMS using the minimum total ozone value from the October monthly mean (depth of the hole) and by calculating the average size during the September-October period. Ozone is mainly destroyed by halogen catalytic cycles, and these losses are modulated by temperature variations in the collar of the polar lower stratospheric vortex. In this presentation, we show the relationships of halogens and temperature to, both the size and depth of the hole. Because atmospheric halogen levels are responding to international agreements that limit or phase out production, the amount of halogens in the stratosphere should decrease over the next few decades. Using projections of halogen levels combined with age-of-air estimates, we find that the ozone hole is recovering at an extremely slow rate and that large ozone holes will regularly recur over the next 2 decades. The ozone hole will begin to show first signs of recovery in about 2023, and the hole will fully recover to pre-1980 levels in approximately 2070. This 2070 recovery is 20 years later than recent projections.

Valorization of Olive Mill Wastewater by Membrane Processes to Recover Natural Antioxidant Compounds for Cosmeceutical and Nutraceutical Applications or Functional Foods.

Science.gov (United States)

Alfano, Alberto; Corsuto, Luisana; Finamore, Rosario; Savarese, Maria; Ferrara, Filomena; Falco, Salvatore; Santabarbara, Giuseppe; De Rosa, Mario; Schiraldi, Chiara

2018-05-23

Olive oil boasts numerous health benefits due to the high content of the monounsaturated fatty acid (MUFA) and functional bioactives including tocopherols, carotenoids, phospholipids, and polyphenolics with multiple biological activities. Polyphenolic components present antioxidant properties by scavenging free radicals and eliminating metabolic byproducts of metabolism. The objective of this research project was to recover the biologically active components rich in polyphenols, which include treatment of olive oil mills wastewater, and, at the same time, to remove the pollutant waste component resulting from the olive oil manufacturing processes. With specific focus on using technologies based on the application of ultra and nanofiltration membranes, the polyphenols fraction was extracted after an initial flocculation step. The nano-filtration permeate showed a reduction of about 95% of the organic load. The polyphenols recovery after two filtration steps was about 65% w / v . The nanofiltration retentate, dried using the spray dryer technique, was tested for cell viability after oxidative stress induction on human keratinocytes model in vitro and an improved cell reparation in the presence of this polyphenolic compound was demonstrated in scratch assays assisted through time lapse video-microscopy. The polyphenols recovered from these treatments may be suitable ingredients in cosmeceuticals and possibly nutraceutical preparations or functional foods.

A Method for Microencapsulation of Cells and a Device for Its Realization.

Science.gov (United States)

Lepekhova, S A; Goldberg, O A; Kravchenko, A A; Batraks, A E; Koval, E V; Kurgansky, I S; Apartsin, K A

2017-04-01

The device for cell encapsulation makes it possible to fabricate microcapsules of a preset size with even smooth surface, without defects or adhesion to each other, with viable cells inside the capsule. The cells were derived from newborn piglet pancreases.

A hybrid bio-jetting approach for directly engineering living cells

International Nuclear Information System (INIS)

Kwok, Albert; Irvine, Scott; Arumuganathar, Sumathy; Jayasinghe, Suwan N; McEwan, Jean R

2008-01-01

This paper reports developments on a hybrid cell-engineering protocol coupling both bio-electrosprays and aerodynamically assisted bio-jets for process-handling living cells. The current work demonstrates the ability to couple these two cell-jetting protocols for handling a wide range of cells for deposition. The post-treated cells are assessed for their viability by way of flow cytometry, which illustrates a significant population of viable cells post-treatment in comparison to those controls. This work is the first example of coupling these two protocols for the process handling of living cells. The hybrid protocol demonstrates the achievement of stable cone jetting of a cellular suspension in the single-needle configuration which was previously unachieved with single-needle bio-electrosprays. Furthermore the living cells explored in these investigations expressed GFP, thus demonstrating the ability to couple gene therapy with this hybrid protocol. Hence, this approach could one day be explored for building biologically viable tissues incorporating a therapeutic payload for combating a range of cellular/tissue-based pathologies

A probabilistic model to recover individual genomes from metagenomes

NARCIS (Netherlands)

J. Dröge (Johannes); A. Schönhuth (Alexander); A.C. McHardy (Alice)

2017-01-01

textabstractShotgun metagenomics of microbial communities reveal information about strains of relevance for applications in medicine, biotechnology and ecology. Recovering their genomes is a crucial but very challenging step due to the complexity of the underlying biological system and technical

Evaluation of an automated struvite reactor to recover phosphorus ...

African Journals Online (AJOL)

2015-04-03

Apr 3, 2015 ... A reactor was developed that can run fully automated and recover up to 93% ..... uncertainty. This technique will work best when the concentration of ... Ken Jack at the School of Chemical Engineering to help build the reactor ...

Establishment of a Viable Population of Red-Cockaded Woodpeckers at the Savannah River Site

International Nuclear Information System (INIS)

Johnston, P.A.

2002-01-01

Report on program's objective to restore viable population of Red-cockaded woodpecker at SRS. Several management strategies were used to promote population expansion of Red-cockaded woodpecker and reduction of interspecific competition with Red-Cockaded woodpecker

Comparison of viable plate count, turbidity measurement and real-time PCR for quantification of Porphyromonas gingivalis.

Science.gov (United States)

Clais, S; Boulet, G; Van Kerckhoven, M; Lanckacker, E; Delputte, P; Maes, L; Cos, P

2015-01-01

The viable plate count (VPC) is considered as the reference method for bacterial enumeration in periodontal microbiology but shows some important limitations for anaerobic bacteria. As anaerobes such as Porphyromonas gingivalis are difficult to culture, VPC becomes time-consuming and less sensitive. Hence, efficient normalization of experimental data to bacterial cell count requires alternative rapid and reliable quantification methods. This study compared the performance of VPC with that of turbidity measurement and real-time PCR (qPCR) in an experimental context using highly concentrated bacterial suspensions. Our TaqMan-based qPCR assay for P. gingivalis 16S rRNA proved to be sensitive and specific. Turbidity measurements offer a fast method to assess P. gingivalis growth, but suffer from high variability and a limited dynamic range. VPC was very time-consuming and less repeatable than qPCR. Our study concludes that qPCR provides the most rapid and precise approach for P. gingivalis quantification. Although our data were gathered in a specific research context, we believe that our conclusions on the inferior performance of VPC and turbidity measurements in comparison to qPCR can be extended to other research and clinical settings and even to other difficult-to-culture micro-organisms. Various clinical and research settings require fast and reliable quantification of bacterial suspensions. The viable plate count method (VPC) is generally seen as 'the gold standard' for bacterial enumeration. However, VPC-based quantification of anaerobes such as Porphyromonas gingivalis is time-consuming due to their stringent growth requirements and shows poor repeatability. Comparison of VPC, turbidity measurement and TaqMan-based qPCR demonstrated that qPCR possesses important advantages regarding speed, accuracy and repeatability. © 2014 The Society for Applied Microbiology.

Method for recovering uranium from sea water

International Nuclear Information System (INIS)

Schwochau, K.; Astheimer, L.; Schenk, H.J.; Schmitz, J.

1977-04-01

In view of the augmenting uranium demand for energy supply and of the anticipated depletion of the actually assured and economic uranium resources the possibility of recovering uranium from sea water receives increasing attention. It is the purpose of the present report to give a detailed discussion of fundamental problems involved and a critical survey of hitherto proposed processes of recovery and to recommend some general directives for further work. (orig.) [de

Transport of significant metals recovered in real sea experiment of adsorbents

International Nuclear Information System (INIS)

Takeda, Hayato; Tamada, Masao; Kasai, Noboru; Katakai, Akio; Hasegawa, Shin; Seko, Noriaki; Sugo, Takanobu; Kawabata, Yukiya

2001-10-01

Real sea experiment for the recovery of significant metals such as uranium and vanadium which dissolved in seawater with extremely low concentration has been carried out at the offing of Mutsu establishment to evaluate the adsorption performance of adsorbent synthesized by radiation-induced graft-polymerization. The significant metals of uranium and vanadium eluted from the adsorbent which was soaked in the real sea were adsorbed onto the conventional chelate resin. The chelate resin which adsorbed the metals was packed in a plastic (PVC) column and further put in a cylindrical stainless transport container. This container was transported to the facility for separation and purification by a truck for the exclusive loading. Then the recovers metals were purified there. The recovered metals contained the uranium of 150g (1.92 MBq) and less in one recovery experiment. The maximum concentration is 60 Bq/g when the uranium is adsorbed on the chelate resin. Transport of recovered metals can be treated as general substance since these amount and concentration are out of legal control. However, the recovered metals were transported in conformity to L type Transport as a voluntary regulation. Though there is no requirements of structural strength for L type package legally, the structural strength of container was designed on that of IP-2 type which is higher transport grade than L type to take its safety measure. Its strength analysis proved the safety under general transport process. The transport was based on the plan made in advance. (author)

On the trace-free Einstein equations as a viable alternative to general relativity

International Nuclear Information System (INIS)

Ellis, George F R; Van Elst, Henk; Murugan, Jeff; Uzan, Jean-Philippe

2011-01-01

The quantum field theoretical prediction for the vacuum energy density leads to a value for the effective cosmological constant that is incorrect by between 60 and 120 orders of magnitude. We review an old proposal of replacing Einstein's field equations by their trace-free part (the trace-free Einstein equations), together with an independent assumption of energy-momentum conservation by matter fields. While this does not solve the fundamental issue of why the cosmological constant has the value that is observed cosmologically, it is indeed a viable theory that resolves the problem of the discrepancy between the vacuum energy density and the observed value of the cosmological constant. However, one has to check that, as well as preserving the standard cosmological equations, this does not destroy other predictions, such as the junction conditions that underlie the use of standard stellar models. We confirm that no problems arise here: hence, the trace-free Einstein equations are indeed viable for cosmological and astrophysical applications. (papers)

SV40-transformed human fibroblasts: evidence for cellular aging in pre-crisis cells.

Science.gov (United States)

Stein, G H

1985-10-01

Pre-crisis SV40-transformed human diploid fibroblast (HDF) cultures have a finite proliferative lifespan, but they do not enter a viable senescent state at end of lifespan. Little is known about either the mechanism for this finite lifespan in SV40-transformed HDF or its relationship to finite lifespan in normal HDF. Recently we proposed that in normal HDF the phenomena of finite lifespan and arrest in a viable senescent state depend on two separate processes: 1) an age-related decrease in the ability of the cells to recognize or respond to serum and/or other mitogens such that the cells become functionally mitogen-deprived at the end of lifespan; and 2) the ability of the cells to enter a viable, G1-arrested state whenever they experience mitogen deprivation. In this paper, data are presented that suggest that pre-crisis SV40-transformed HDF retain the first process described above, but lack the second process. It is shown that SV40-transformed HDF have a progressively decreasing ability to respond to serum as they age, but they continue to traverse the cell cycle at the end of lifespan. Concomitantly, the rate of cell death increases steadily toward the end of lifespan, thereby causing the total population to cease growing and ultimately to decline. Previous studies have shown that when SV40-transformed HDF are environmentally serum deprived, they likewise exhibit continued cell cycle traverse coupled with increased cell death. Thus, these results support the hypothesis that pre-crisis SV40-transformed HDF still undergo the same aging process as do normal HDF, but they end their lifespan in crisis rather than in the normal G1-arrested senescent state because they have lost their ability to enter a viable, G1-arrested state in response to mitogen deprivation.

Hypoosmotic swelling test in alpaca (Vicugna pacos spermatozoa recovered the vas deferens

Directory of Open Access Journals (Sweden)

Quispe HA

2015-08-01

Full Text Available The study objective was to evaluate the endosmotic reaction in 5 hypoosmotic solutions 100, 125, 150, 175 and 200 mOsm/l (HOS test incubated 5 and 15 minutes using alpaca sperm recovered the vas deferens. We used 5 male alpacas with the vas deferens surgically diverted, recovering sperm 10 times for each animal; the microscopic characteristics were evaluated with ISAS®. Information’s hipoosmotic test was analyzed with a factorial arrangement in a random one-way model, the correlation between endosmosis and percentages of motility and vitality was analyzed with Spearman correlation. There were no significant differences (p>0,05 in incubation times; highly significant differences (p 0,05was found to endosmosis with percentages of sperm motility and vitality; in conclusion, a 125 mOsm/l solution with 5 minutes of incubation is sufficient to perform the hipoosmotic test in alpaca spermatozoa recovered the vas deferens.

In Vitro Evaluation of the Impact of the Probiotic E. coli Nissle 1917 on Campylobacter jejuni's Invasion and Intracellular Survival in Human Colonic Cells.

Science.gov (United States)

Helmy, Yosra A; Kassem, Issmat I; Kumar, Anand; Rajashekara, Gireesh

2017-01-01

Campylobacter jejuni is a leading cause of bacterial food poisoning in humans. Due to the rise in antibiotic-resistant Campylobacter , there exists a need to develop antibiotic-independent interventions to control infections in humans. Here, we evaluated the impact of Escherichia coli Nissle 1917 (EcN), a probiotic strain, on C. jejuni's invasion and intracellular survival in polarized human colonic cells (HT-29). To further understand how EcN mediates its impact, the expression of 84 genes associated with tight junctions and cell adhesion was profiled in HT-29 cells after treatment with EcN and challenge with C. jejuni . The pre-treatment of polarized HT-29 cells with EcN for 4 h showed a significant effect on C. jejuni 's invasion (∼2 log reduction) of the colonic cells. Furthermore, no intracellular C. jejuni were recovered from EcN pre-treated HT-29 cells at 24 h post-infection. Other probiotic strains tested had no significant impact on C. jejuni invasion and intracellular survival. C. jejuni decreased the expression of genes associated with epithelial cells permeability and barrier function in untreated HT-29 cells. However, EcN positively affected the expression of genes that are involved in enhanced intestinal barrier function, decreased cell permeability, and increased tight junction integrity. The results suggest that EcN impedes C. jejuni invasion and subsequent intracellular survival by affecting HT-29 cells barrier function and tight junction integrity. We conclude that EcN might be a viable alternative for controlling C. jejuni infections.

Uranium oxide recovering method

International Nuclear Information System (INIS)

Ota, Kazuaki; Takazawa, Hiroshi; Teramae, Naoki; Onoue, Takeshi.

1997-01-01

Nitrates containing uranium nitrate are charged in a molten salt electrolytic vessel, and a heat treatment is applied to prepare molten salts. An anode and a cathode each made of a graphite rod are disposed in the molten salts. AC voltage is applied between the anode and the cathode to conduct electrolysis of the molten salts. Uranium oxides are deposited as a recovered product of uranium, on the surface of the anode. The nitrates containing uranium nitrate are preferably a mixture of one or more nitrates selected from sodium nitrate, potassium nitrate, calcium nitrate and magnesium nitrate with uranium nitrate. The nitrates may be liquid wastes of nitrates. The temperature for the electrolysis of the molten salts is preferably from 150 to 300degC. The voltage for the electrolysis of the molten salts is preferably an AC voltage of from 2 to 6V, more preferably from 4 to 6V. (I.N.)

Process for recovering tungsten from alkaline leaching solution of tungsten ores

International Nuclear Information System (INIS)

Onozaki, S.; Nemoto, S.; Hazeyama, T.

1976-01-01

This invention relates to a process for recovering tungsten from an alkaline leaching solution of tungsten ores. This invention comprises adjusting the pH of an alkaline leaching solution which is obtained by lixiviating ore containing tungsten with an alkaline solution to 7--8 with acid to oxidize molybdic acid ions in the solution, adding a sulfide donor, then precipitating molybdenum sulfide compounds by adjusting the pH value of the solution to 2--3. Tungstic acid ions are recovered as calcium tungstate by the addition of a calcium ion donor after the molybdenum sulfide compounds are separated

METHOD OF RECOVERING URANIUM COMPOUNDS

Science.gov (United States)

Poirier, R.H.

1957-10-29

S>The recovery of uranium compounds which have been adsorbed on anion exchange resins is discussed. The uranium and thorium-containing residues from monazite processed by alkali hydroxide are separated from solution, and leached with an alkali metal carbonate solution, whereby the uranium and thorium hydrorides are dissolved. The carbonate solution is then passed over an anion exchange resin causing the uranium to be adsorbed while the thorium remains in solution. The uranium may be recovered by contacting the uranium-holding resin with an aqueous ammonium carbonate solution whereby the uranium values are eluted from the resin and then heating the eluate whereby carbon dioxide and ammonia are given off, the pH value of the solution is lowered, and the uranium is precipitated.

Current Perspectives on Viable but Non-culturable State in Foodborne Pathogens

OpenAIRE

Zhao, Xihong; Zhong, Junliang; Wei, Caijiao; Lin, Chii-Wann; Ding, Tian

2017-01-01

The viable but non-culturable (VBNC) state, a unique state in which a number of bacteria respond to adverse circumstances, was first discovered in 1982. Unfortunately, it has been reported that many foodborne pathogens can be induced to enter the VBNC state by the limiting environmental conditions during food processing and preservation, such as extreme temperatures, drying, irradiation, pulsed electric field, and high pressure stress, as well as the addition of preservatives and disinfectant...

Recovery of tobacco BY-2 cells after high hydrostatic pressure treatment.

Science.gov (United States)

Kusube, Masataka; Nishino, Takumi; Nishikawa, Yuki; Goto, Masaki; Matsuki, Hitoshi; Iwahashi, Hitoshi

2010-02-01

The recovery of Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cells in Linsmaire and Skoog medium after treatment at high hydrostatic pressure was investigated using an Evans Blue staining method to discriminate live from dead cells. The survival of BY-2 cells just after the high-pressure treatment at 5 degrees C and 25 degrees C decreased abruptly at pressures higher than 50 MPa and 100 MPa, respectively. Furthermore, almost all of the BY-2 cells treated at 5 degrees C and 25 degrees C recovered pressures below 25 MPa and 75 MPa, respectively. However, no BY-2 cells recovered at pressures above 100 MPa at either temperature.

Recovering from conflict in romantic relationships: a developmental perspective.

Science.gov (United States)

Salvatore, Jessica E; Kuo, Sally I-Chun; Steele, Ryan D; Simpson, Jeffry A; Collins, W Andrew

2011-03-01

This study adopted a developmental perspective on recovery from conflict in romantic relationships. Participants were 73 young adults (target participants), studied since birth, and their romantic partners. A novel observational coding scheme was used to evaluate each participant's degree of conflict recovery, operationalized as the extent to which the participant disengaged from conflict during a 4-min "cool-down" task immediately following a 10-min conflict discussion. Conflict recovery was systematically associated with developmental and dyadic processes. Targets who were rated as securely attached more times in infancy recovered from conflict better, as did their romantic partners. Concurrently, having a romantic partner who displayed better recovery predicted more positive relationship emotions and greater relationship satisfaction. Prospectively, target participants' early attachment security and their partners' degree of conflict recovery interacted to predict relationship stability 2 years later, such that having a partner who recovered from conflict better buffered targets with insecure histories.

Immediate natural tooth pontic: A viable yet temporary prosthetic solution: A patient reported outcome

Directory of Open Access Journals (Sweden)

Sudhir Bhandari

2012-01-01

Conclusion: The concept of immediate pontic placement is surely a viable treatment option and promises an excellent transient esthetic solution for a lost tooth as well as enables good preparation of the extraction site for future prosthetic replacement.

EXPERIMENTAL TRIES TO ESTABLISH THE PREIMPLANTATIONAL MAMMALIAN EMBRYOS VIABILITY THROUGHOUT STAINING

Directory of Open Access Journals (Sweden)

IVAN ALEXANDRA

2007-01-01

Full Text Available Presently there are more methods to assess embryo quality but, still the wieldy usedremains the morphological criteria method. In this experiment were tested twostaining methods for embryos and oocytes. The embryos were recovered from mousefemale at 72 hours after mating. The recovered embryos were first evaluated aftermorphological criteria and than by Trypan blue exclusion and Neutral red staining.Using Trypan blue exclusion were evaluated 30 embryos from which 19 (63.3 wereclassified as viable and 11 (36.7 were classified as nonviable. By Neutral redstaining were evaluated 37 embryos from which 24 (64.8 were considered viableand 13 (35.2 were considered nonviable. The oocytes recovered were alsoevaluated using the two methods: using Trypan blue exclusion were stained 10oocytes from which 9 remained uncolored and were considered viable and 1 wasstained in blue and was considered nonviable and using Neutral red 13 oocytes werestained from which 9 were evaluated as viable and 4 as nonviable.

A Novel Designed Bioreactor for Recovering Precious Metals from Waste Printed Circuit Boards

Science.gov (United States)

Jujun, Ruan; Jie, Zheng; Jian, Hu; Zhang, Jianwen

2015-01-01

For recovering precious metals from waste printed circuit boards (PCBs), a novel hybrid technology including physical and biological methods was developed. It consisted of crushing, corona-electrostatic separation, and bioleaching. Bioleaching process is the focus of this paper. A novel bioreactor for bioleaching was designed. Bioleaching was carried out using Pseudomonas chlororaphis. Bioleaching experiments using mixed particles of Au and Cu were performed and leachate contained 0.006 mg/L, 2823 mg/L Au+ and Cu2+ respectively. It showed when Cu existed, the concentrations of Au were extremely small. This provided the feasibility to separate Cu from Au. The method of orthogonal experimental design was employed in the simulation bioleaching experiments. Experimental results showed the optimized parameters for separating Cu from Au particles were pH 7.0, temperature 22.5 °C, and rotation speed 80 r/min. Based on the optimized parameters obtained, the bioreactor was operated for recovering mixed Au and Cu particles. 88.1 wt.% of Cu and 76.6 wt.% of Au were recovered. The paper contributed important information to recover precious metals from waste PCBs. PMID:26316021

Three-dimensional printing of stem cell-laden hydrogels submerged in a hydrophobic high-density fluid

International Nuclear Information System (INIS)

Duarte Campos, Daniela F; Blaeser, Andreas; Weber, Michael; Fischer, Horst; Jäkel, Jörg; Neuss, Sabine; Jahnen-Dechent, Wilhelm

2013-01-01

Over the last decade, bioprinting technologies have begun providing important tissue engineering strategies for regenerative medicine and organ transplantation. The major drawback of past approaches has been poor or inadequate material-printing device and substrate combinations, as well as the relatively small size of the printed construct. Here, we hypothesise that cell-laden hydrogels can be printed when submerged in perfluorotributylamine (C 12 F 27 N), a hydrophobic high-density fluid, and that these cells placed within three-dimensional constructs remain viable allowing for cell proliferation and production of extracellular matrix. Human mesenchymal stem cells and MG-63 cells were encapsulated into agarose hydrogels, and subsequently printed in high aspect ratio in three dimensional structures that were supported in high density fluorocarbon. Three-dimensional structures with various shapes and sizes were manufactured and remained stable for more than six months. Live/dead and DAPI stainings showed viable cells 24 h after the printing process, as well as after 21 days in culture. Histological and immunohistochemical analyses after 14 and 21 days revealed viable cells with marked matrix production and signs of proliferation. The compressive strength values of the printed gels consequently increased during the two weeks in culture, revealing encouraging results for future applications in regenerative medicine. (paper)

Mass, energy and material balances of SRF production process. Part 3: solid recovered fuel produced from municipal solid waste.

Science.gov (United States)

Nasrullah, Muhammad; Vainikka, Pasi; Hannula, Janne; Hurme, Markku; Kärki, Janne

2015-02-01

This is the third and final part of the three-part article written to describe the mass, energy and material balances of the solid recovered fuel production process produced from various types of waste streams through mechanical treatment. This article focused the production of solid recovered fuel from municipal solid waste. The stream of municipal solid waste used here as an input waste material to produce solid recovered fuel is energy waste collected from households of municipality. This article presents the mass, energy and material balances of the solid recovered fuel production process. These balances are based on the proximate as well as the ultimate analysis and the composition determination of various streams of material produced in a solid recovered fuel production plant. All the process streams are sampled and treated according to CEN standard methods for solid recovered fuel. The results of the mass balance of the solid recovered fuel production process showed that 72% of the input waste material was recovered in the form of solid recovered fuel; 2.6% as ferrous metal, 0.4% as non-ferrous metal, 11% was sorted as rejects material, 12% as fine faction and 2% as heavy fraction. The energy balance of the solid recovered fuel production process showed that 86% of the total input energy content of input waste material was recovered in the form of solid recovered fuel. The remaining percentage (14%) of the input energy was split into the streams of reject material, fine fraction and heavy fraction. The material balances of this process showed that mass fraction of paper and cardboard, plastic (soft) and wood recovered in the solid recovered fuel stream was 88%, 85% and 90%, respectively, of their input mass. A high mass fraction of rubber material, plastic (PVC-plastic) and inert (stone/rock and glass particles) was found in the reject material stream. © The Author(s) 2014.

Gamma-irradiated scrub typhus immunogens: development of cell-mediated immunity after vaccination of inbred mice

International Nuclear Information System (INIS)

Jerrells, T.R.; Palmer, B.A.; Osterman, J.V.

1983-01-01

Mice immunized with three injections of gamma-irradiated Karp strain of Rickettsia tsutsugamushi were evaluated for the presence of cell-mediated immunity by using delayed-type hypersensitivity, antigen-induced lymphocyte proliferation, and antigen-induced lymphokine production. These animals also were evaluated for levels of circulating antibody after immunization as well as for the presence of rickettsemia after intraperitoneal challenge with viable Karp rickettsiae. After immunization with irradiated Karp rickettsiae, a demonstrable cell-mediated immunity was present as evidenced by delayed-type hypersensitivity responsiveness, lymphocyte proliferation, and production of migration inhibition factor and interferon by immune spleen lymphocytes. Also, a reduction in circulating rickettsiae was seen in mice immunized with irradiated rickettsiae after challenge with 1,000 50% mouse lethal doses of viable, homologous rickettsiae. All responses except antibody titer and reduction of rickettsemia were similar to the responses noted in mice immunized with viable organisms. Antibody levels were lower in mice immunized with irradiated rickettsiae than in mice immunized with viable rickettsiae. Furthermore, mice that were immunized with viable rickettsiae demonstrated markedly lower levels of rickettsemia after intraperitoneal challenge compared with either mice immunized with irradiated rickettsiae or nonimmunized mice

RECOVER: An Automated, Cloud-Based Decision Support System for Post-Fire Rehabilitation Planning

Science.gov (United States)

Schnase, J. L.; Carroll, M. L.; Weber, K. T.; Brown, M. E.; Gill, R. L.; Wooten, M.; May, J.; Serr, K.; Smith, E.; Goldsby, R.; Newtoff, K.; Bradford, K.; Doyle, C.; Volker, E.; Weber, S.

2014-11-01

RECOVER is a site-specific decision support system that automatically brings together in a single analysis environment the information necessary for post-fire rehabilitation decision-making. After a major wildfire, law requires that the federal land management agencies certify a comprehensive plan for public safety, burned area stabilization, resource protection, and site recovery. These burned area emergency response (BAER) plans are a crucial part of our national response to wildfire disasters and depend heavily on data acquired from a variety of sources. Final plans are due within 21 days of control of a major wildfire and become the guiding document for managing the activities and budgets for all subsequent remediation efforts. There are few instances in the federal government where plans of such wide-ranging scope and importance are assembled on such short notice and translated into action more quickly. RECOVER has been designed in close collaboration with our agency partners and directly addresses their high-priority decision-making requirements. In response to a fire detection event, RECOVER uses the rapid resource allocation capabilities of cloud computing to automatically collect Earth observational data, derived decision products, and historic biophysical data so that when the fire is contained, BAER teams will have a complete and ready-to-use RECOVER dataset and GIS analysis environment customized for the target wildfire. Initial studies suggest that RECOVER can transform this information-intensive process by reducing from days to a matter of minutes the time required to assemble and deliver crucial wildfire-related data.

RECOVER: An Automated Cloud-Based Decision Support System for Post-fire Rehabilitation Planning

Science.gov (United States)

Schnase, John L.; Carroll, Mark; Weber, K. T.; Brown, Molly E.; Gill, Roger L.; Wooten, Margaret; May J.; Serr, K.; Smith, E.; Goldsby, R.;

The effect of cell immobilization on the antibacterial activity of Lactobacillus reuteri DPC16 cells during passage through a simulated gastrointestinal tract system.

Science.gov (United States)

Zhao, Qian; Maddox, Ian S; Mutukumira, Anthony; Lee, Sung Je; Shu, Quan

2012-10-01

Cell immobilization has the ability to influence the survival and functional characteristics of probiotic bacterial strains in harsh environments. This study investigated the effect of cell immobilization and passage through a simulated gastrointestinal tract (GI) on the antibacterial activity of Lactobacillus reuteri DPC16. Antibacterial activity, reuterin production and diol dehydratase activity were assayed in recovered isolates of L. reuteri that had been immobilized in Ca alginate-skim milk, and incubated in simulated GI fluids. Among all the recovered isolates tested, any that had undergone immobilization followed by immediate recovery of the cells without subsequent incubation in any fluids demonstrated the highest reuterin production, antimicrobial activity and diol dehydratase enzyme activity. L. reuteri DPC16 cells that had been immobilized, incubated in simulated GI fluids, and subsequently recovered from the beads often showed some loss of antimicrobial activity compared to the immobilized cells. The data confirm that the process of immobilization of L. reuteri in Ca alginate-skim milk, rather than the passage through simulated GI fluids, resulted in enhanced antibacterial activity. This is attributed to increased diol dehydratase activity, resulting in increased reuterin production.

A Transient Cell-shielding Method for Viable MSC Delivery Within Hydrophobic Scaffolds Polymerized in situ

Science.gov (United States)

2015-03-27

Transport phenomena. New York: Wiley; 1960. [52] Caplan AI, Dennis JE. Mesenchymal stem cells as trophic mediators. J Cell Biochem 2006;98:1076e84. [53... Caplan AI. Why are MSCs therapeutic? New data: new insight. J Pathol 2009;217:318e24. [54] Dennis JE, Cohen N, Goldberg VM, Caplan AI. Targeted delivery

Massachusetts Fuel Cell Bus Project: Demonstrating a Total Transit Solution for Fuel Cell Electric Buses in Boston

Energy Technology Data Exchange (ETDEWEB)

2017-05-22

The Federal Transit Administration's National Fuel Cell Bus Program focuses on developing commercially viable fuel cell bus technologies. Nuvera is leading the Massachusetts Fuel Cell Bus project to demonstrate a complete transit solution for fuel cell electric buses that includes one bus and an on-site hydrogen generation station for the Massachusetts Bay Transportation Authority (MBTA). A team consisting of ElDorado National, BAE Systems, and Ballard Power Systems built the fuel cell electric bus, and Nuvera is providing its PowerTap on-site hydrogen generator to provide fuel for the bus.

Adipose-Derived Stem Cell Delivery into Collagen Gels Using Chitosan Microspheres

Science.gov (United States)

2010-02-17

Porous CSM of uniform size and composition were prepared and used as a stem cell carrier. ASC were allowed to attach to the microspheres and infiltrate...and viable, could be retrieved from the spheres, and maintained expression of stem - cell -specific markers. Electron microscopic evaluation of the cell

Disruption of cell walls for enhanced lipid recovery

Science.gov (United States)

Knoshaug, Eric P; Donohoe, Bryon S; Gerken, Henri; Laurens, Lieve; Van Wychen, Stefanie Rose

2015-03-24

Presented herein are methods of using cell wall degrading enzymes for recovery of internal lipid bodies from biomass sources such as algae. Also provided are algal cells that express at least one exogenous gene encoding a cell wall degrading enzyme and methods for recovering lipids from the cells.

Sorting of cells of the same size, shape, and cell cycle stage for a single cell level assay without staining

Directory of Open Access Journals (Sweden)

Yomo Tetsuya

2006-06-01

Full Text Available Abstract Background Single-cell level studies are being used increasingly to measure cell properties not directly observable in a cell population. High-performance data acquisition systems for such studies have, by necessity, developed in synchrony. However, improvements in sample purification techniques are also required to reveal new phenomena. Here we assessed a cell sorter as a sample-pretreatment tool for a single-cell level assay. A cell sorter is routinely used for selecting one type of cells from a heterogeneous mixture of cells using specific fluorescence labels. In this case, we wanted to select cells of exactly the same size, shape, and cell-cycle stage from a population, without using a specific fluorescence label. Results We used four light scatter parameters: the peak height and area of the forward scatter (FSheight and FSarea and side scatter (SSheight and SSarea. The rat pheochromocytoma PC12 cell line, a neuronal cell line, was used for all experiments. The living cells concentrated in the high FSarea and middle SSheight/SSarea fractions. Single cells without cell clumps were concentrated in the low SS and middle FS fractions, and in the higher FSheight/FSarea and SSheight/SSarea fractions. The cell populations from these viable, single-cell-rich fractions were divided into twelve subfractions based on their FSarea-SSarea profiles, for more detailed analysis. We found that SSarea was proportional to the cell volume and the FSarea correlated with cell roundness and elongation, as well as with the level of DNA in the cell. To test the method and to characterize the basic properties of the isolated single cells, sorted cells were cultured in separate wells. The cells in all subfractions survived, proliferated and differentiated normally, suggesting that there was no serious damage. The smallest, roundest, and smoothest cells had the highest viability. There was no correlation between proliferation and differentiation. NGF increases

Fusarium spp. recovered from waste peanuts associated with sandhill crane mortality

Science.gov (United States)

Nelson, P.E.; Cole, R.J.; Tousson, T.A.; Dorner, J.W.; Windingstad, R.M.

1990-01-01

Approximately 5000 sandhill cranes (Grus canadensis ) died from undetermined causes in Gains County, Texas, 1985, and an additional 200 died in 1986. Prominent clinical signs were the inability of many sick cranes to hold their necks horizontal and the neck, head, and legs sometimes drooped perpendicularly during flight. Approximately 95% of the dead cranes' gizzards contained peanuts. Culturing of peanuts, shells, soil and soil debris from fields in which sandhill cranes died showed that Fusarium species were the fungi most frequently isolated and eight species were recovered from these substrates. Fusarium compactum, F. solani , and F. equiseti were the only species recovered from all substrates cultured from both fields.

Greater anterior insula activation during anticipation of food images in women recovered from anorexia nervosa versus controls

Science.gov (United States)

Oberndorfer, Tyson; Simmons, Alan; McCurdy, Danyale; Strigo, Irina; Matthews, Scott; Yang, Tony; Irvine, Zoe; Kaye, Walter

2013-01-01

Individuals with anorexia nervosa (AN) restrict food consumption and become severely emaciated. Eating food, even thinking of eating food, is often associated with heightened anxiety. However, food cue anticipation in AN is poorly understood. Fourteen women recovered from AN and 12 matched healthy control women performed an anticipation task viewing images of food and object images during functional magnetic resonance imaging. Comparing anticipation of food versus object images between control women and recovered AN groups showed significant interaction only in the right ventral anterior insula, with greater activation in recovered AN anticipating food images. These data support the hypothesis of a disconnect between anticipating and experiencing food stimuli in recovered AN. Insula activation positively correlated with pleasantness ratings of palatable foods in control women, while no such relationship existed in recovered AN, which is further evidence of altered interoceptive function. Finally, these findings raise the possibility that enhanced anterior insula anticipatory response to food cues in recovered AN could contribute to exaggerated sensitivity and anxiety related to food and eating. PMID:23993362

Method for recovering palladium and technetium values from nuclear fuel reprocessing waste solutions

Science.gov (United States)

Horwitz, E. Philip; Delphin, Walter H.

1979-07-24

A method for recovering palladium and technetium values from nuclear fuel reprocessing waste solutions containing these and other values by contacting the waste solution with an extractant of tricaprylmethylammonium nitrate in an inert hydrocarbon diluent which extracts the palladium and technetium values from the waste solution. The palladium and technetium values are recovered from the extractant and from any other coextracted values with a strong nitric acid strip solution.

Culture of human intestinal epithelial cell using the dissociating enzyme thermolysin and endothelin-3

Directory of Open Access Journals (Sweden)

Z. Liu

2010-05-01

Full Text Available Epithelium, a highly dynamic system, plays a key role in the homeostasis of the intestine. However, thus far a human intestinal epithelial cell line has not been established in many countries. Fetal tissue was selected to generate viable cell cultures for its sterile condition, effective generation, and differentiated character. The purpose of the present study was to culture human intestinal epithelial cells by a relatively simple method. Thermolysin was added to improve the yield of epithelial cells, while endothelin-3 was added to stimulate their growth. By adding endothelin-3, the achievement ratio (viable cell cultures/total cultures was enhanced to 60% of a total of 10 cultures (initiated from 8 distinct fetal small intestines, allowing the generation of viable epithelial cell cultures. Western blot, real-time PCR and immunofluorescent staining showed that cytokeratins 8, 18 and mouse intestinal mucosa-1/39 had high expression levels in human intestinal epithelial cells. Differentiated markers such as sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV also showed high expression levels in human intestinal epithelial cells. Differentiated human intestinal epithelial cells, with the expression of surface markers (cytokeratins 8, 18 and mouse intestinal mucosa-1/39 and secretion of cytokines (sucrase-isomaltase, aminopeptidase N and dipeptidylpeptidase IV, may be cultured by the thermolysin and endothelin-3 method and maintained for at least 20 passages. This is relatively simple, requiring no sophisticated techniques or instruments, and may have a number of varied applications.

Process for recovering cesium from cesium alum

International Nuclear Information System (INIS)

Mein, P.G.

1984-01-01

Cesium is recovered from cesium alum, CsAl(SO 4 ) 2 , by an aqueous conversion and precipitation reaction using a critical stoichiometric excess of a water-soluble permanganate to form solid cesium permanganate (CsMnO 4 ) free from cesium alum. The other metal salts remain in solution, providing the final pH does not cause hydroxides of aluminium or iron to form. The precipitate is separated from the residual solution to obtain CsMnO 4 of high purity

Apparatus for recovering oil from Posidonien shale

Energy Technology Data Exchange (ETDEWEB)

1920-04-13

Equipment for recovering oil from shale and the like, as well as the distilling of coal is characterized in that a number of chambers provided in a known way with upper and lower air supply are arranged open to the receiver of the oil vapors through removable domes which can be attached to the usual oil-vapor carry-off. Arrangement is characterized in that the domes are movable to the side, so that they can be interchangeably attached to the different chambers.

The regulation effect of STAT 5 signaling pathway on the cell cycle progression of irradiated KG-1 cells

International Nuclear Information System (INIS)

Guo Dehuang; Dong Bo; Luo Qingliang; Wen Gengyun; Mao Bingzhi

2000-01-01

The author investigated the role of the JAK/STAT signaling pathway regulating cell cycle progression in the irradiated KG-1 cells. By permanent transfecting the cells with DN-STAT 5 cDNA to block the JAK/STAT signaling pathway and then transient transfecting with cyclin D 1 or cyclin B 1 cDNA, the effects of cyclin D 1 protein and cyclin B 1 protein on the cell cycle progression were examined. Results showed that after irradiation with 8Gy 60 Co rays, the irradiated KG-1 cells transfected with only DN-STAT 5 cDNA can not recover form the G 1 arrest, even though GM-CSF was added. Meanwhile, the cells transfected with both the DN-STAT 5 cDNA and cyclin D 1 cDNA or cyclin B 1 cDNA can recover from the G 1 arrest or the G 2 arrest to a great extent. Thus, it was proved indirectly that the JAK/STAT signaling pathway activated by GM-CSF regulated the cell cycle progression through cyclin D 1 and cyclin B 1 protein

A safe and efficient method to retrieve mesenchymal stem cells from three-dimensional fibrin gels.

Science.gov (United States)

Carrion, Bita; Janson, Isaac A; Kong, Yen P; Putnam, Andrew J

2014-03-01

Mesenchymal stem cells (MSCs) display multipotent characteristics that make them ideal for potential therapeutic applications. MSCs are typically cultured as monolayers on tissue culture plastic, but there is increasing evidence suggesting that they may lose their multipotency over time in vitro and eventually cease to retain any resemblance to in vivo resident MSCs. Three-dimensional (3D) culture systems that more closely recapitulate the physiological environment of MSCs and other cell types are increasingly explored for their capacity to support and maintain the cell phenotypes. In much of our own work, we have utilized fibrin, a natural protein-based material that serves as the provisional extracellular matrix during wound healing. Fibrin has proven to be useful in numerous tissue engineering applications and has been used clinically as a hemostatic material. Its rapid self-assembly driven by thrombin-mediated alteration of fibrinogen makes fibrin an attractive 3D substrate, in which cells can adhere, spread, proliferate, and undergo complex morphogenetic programs. However, there is a significant need for simple cost-effective methods to safely retrieve cells encapsulated within fibrin hydrogels to perform additional analyses or use the cells for therapy. Here, we present a safe and efficient protocol for the isolation of MSCs from 3D fibrin gels. The key ingredient of our successful extraction method is nattokinase, a serine protease of the subtilisin family that has a strong fibrinolytic activity. Our data show that MSCs recovered from 3D fibrin gels using nattokinase are not only viable but also retain their proliferative and multilineage potentials. Demonstrated for MSCs, this method can be readily adapted to retrieve any other cell type from 3D fibrin gel constructs for various applications, including expansion, bioassays, and in vivo implantation.

Viable Intrauterine Pregnancy and Coexisting Molar Pregnancy in a Bicornuate Uterus: A Rare Presentation

Directory of Open Access Journals (Sweden)

Kavitha Krishnamoorthy

2016-01-01

Full Text Available A complete hydatidiform mole with a viable coexisting fetus (CMCF is a rare occurrence. Similarly, Mullerian anomalies such as a bicornuate uterus are uncommon variants of normal anatomy. We report a case of a 40-year-old female with a known bicornuate uterus presenting at 13 weeks gestation with vaginal bleeding. Ultrasound findings showed a healthy viable pregnancy in the right horn with complete molar pregnancy in the left horn. After extensive counseling, the patient desired conservative management, however, was unable to continue due to profuse vaginal bleeding. The patient underwent suction dilation and curettage under general anesthesia and evacuation of the uterine horns. Postoperatively, the patient was followed until serum beta-human chorionic gonadotropin (β-hCG level dropped to <5 mU. This is the first case of a CMCF reported in a bicornuate uterus, diagnosed with the use of ultrasound imaging.

Increase in the fraction of necrotic, not apoptotic, cells in SiHa xenograft tumours shortly after irradiation

International Nuclear Information System (INIS)

Olive, P.L.; Vikse, C.M.; Vanderbyl, S.

1999-01-01

Background and purpose: Approximately 18% of the cells recovered by rapid mechanical dissociation of SiHa xenograft tumours contain large numbers of DNA strand breaks. The number of damaged cells increases to 30-40% 4-6 h after exposure to 5 or 15 Gy, returning to normal levels by 12 h. This observation is reminiscent of the rate of production of apoptotic cells in other murine and human xenograft tumours. The nature of this damage, rate of development and relation to cell proliferation rate were therefore examined in detail.Materials and methods: SiHa human cervical carcinoma cells were grown as xenograft tumours in SCID mice. Single-cell suspensions were prepared as a function of time after irradiation of the mouse and examined for DNA damage using the alkaline comet assay. Cell cycle progression was measured by flow cytometry evaluation of anti-bromodeoxyuridine-labelled tumour cells.Results: Significant numbers of apoptotic cells could not be detected in irradiated SiHa tumours using an end-labelling assay, electron microscopy, or histological examination of thin sections. Instead, xenograft cells exhibiting extensive DNA damage in the comet assay were predominantly necrotic cells. The increase in the proportion of heavily damaged cells 4-6 h after irradiation could be the result of an interplay between several factors including loss of viable cells and change in production or loss of necrotic cells. Analysis of the progression of BrdUrd-labelled cells confirmed that while 35% of cells from untreated SiHa tumours had divided and entered G 1 phase by 6 h after BrdUrd injection, none of the labelled cells from tumours exposed to 5 or 15 Gy had progressed to G 1 .Conclusions: The increase in the percentage of SiHa tumour cells with extensive DNA damage 4-6 h after irradiation is attributable to necrosis, not apoptosis. Cell cycle progression and cell loss are likely to influence the kinetics of appearance of both apoptotic and necrotic cells in irradiated tumours

Elevated cognitive control over reward processing in recovered female patients with anorexia nervosa.

Science.gov (United States)

Ehrlich, Stefan; Geisler, Daniel; Ritschel, Franziska; King, Joseph A; Seidel, Maria; Boehm, Ilka; Breier, Marion; Clas, Sabine; Weiss, Jessika; Marxen, Michael; Smolka, Michael N; Roessner, Veit; Kroemer, Nils B

2015-09-01

Individuals with anorexia nervosa are thought to exert excessive self-control to inhibit primary drives. This study used functional MRI (fMRI) to interrogate interactions between the neural correlates of cognitive control and motivational processes in the brain reward system during the anticipation of monetary reward and reward-related feedback. In order to avoid confounding effects of undernutrition, we studied female participants recovered from anorexia nervosa and closely matched healthy female controls. The fMRI analysis (including node-to-node functional connectivity) followed a region of interest approach based on models of the brain reward system and cognitive control regions implicated in anorexia nervosa: the ventral striatum, medial orbitofrontal cortex (mOFC) and dorsolateral prefrontal cortex (DLPFC). We included 30 recovered patients and 30 controls in our study. There were no behavioural differences and no differences in hemodynamic responses of the ventral striatum and the mOFC in the 2 phases of the task. However, relative to controls, recovered patients showed elevated DLPFC activity during the anticipation phase, failed to deactivate this region during the feedback phase and displayed greater functional coupling between the DLPFC and mOFC. Recovered patients also had stronger associations than controls between anticipation-related DLPFC responses and instrumental responding. The results we obtained using monetary stimuli might not generalize to other forms of reward. Unaltered neural responses in ventral limbic reward networks but increased recruitment of and connectivity with lateral-frontal brain circuitry in recovered patients suggests an elevated degree of selfregulatory processes in response to rewarding stimuli. An imbalance between brain systems subserving bottom-up and top-down processes may be a trait marker of the disorder.

Isolation of UV-sensitive mutants of mouse L5178Y cells by a cell suspension spotting method

International Nuclear Information System (INIS)

Shiomi, T.; Hieda-Shiomi, N.; Sato, K.

1982-01-01

We have isolated 56 UV-sensitive mutant clones from a mouse L51 T/t line of L5178Y cells by a cell suspension spotting method. Five mutants have also been isolated from L51 T/t and L5178Y cells by the method reported by Thompson and coworkers. We divided the mutants into two groups, highly sensitive and moderately sensitive mutants, according to their sensitivity to UV irradiation. Fifty-eight mutants were highly sensitive and three were moderately sensitive to UV. The reconstruction experiments indicate that more than 90% of highly sensitive mutants were recovered by the cell suspension spotting method. Frequencies of recovered mutants highly sensitive to UV increased with increasing dose of mutagens. Recovered mutant frequency reached 10(-2) after treatment with 1.5 micrograms/ml of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) (survival 0.2%). Eight UV-sensitive mutants were divided into four complementation groups. These mutants were 2-6 times more sensitive to UV than parental L51 T/t cells in terms of D37 (dose required to reduce survival to 37%). Four representative UV-sensitive mutants which are classified into different complementation groups were examined for their sensitivity to killing by UV, 4-nitroquinoline-1-oxide (4NQO), mitomycin C (MMC), X-rays, and MNNG. All four classes of mutants were found to be cross-sensitive to UV, 4NQO, and MMC, but not sensitive to X-rays and MNNG

A viable logarithmic f(R) model for inflation

Energy Technology Data Exchange (ETDEWEB)

Amin, M.; Khalil, S. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Salah, M. [Center for Fundamental Physics, Zewail City of Science and Technology,6 October City, Giza (Egypt); Department of Mathematics, Faculty of Science, Cairo University,Giza (Egypt)

2016-08-18

Inflation in the framework of f(R) modified gravity is revisited. We study the conditions that f(R) should satisfy in order to lead to a viable inflationary model in the original form and in the Einstein frame. Based on these criteria we propose a new logarithmic model as a potential candidate for f(R) theories aiming to describe inflation consistent with observations from Planck satellite (2015). The model predicts scalar spectral index 0.9615

Zn(II)-cyclam based chromogenic sensors for recognition of ATP in aqueous solution under physiological conditions and their application as viable staining agents for microorganism.

Science.gov (United States)

Mahato, Prasenjit; Ghosh, Amrita; Mishra, Sanjiv K; Shrivastav, Anupama; Mishra, Sandhya; Das, Amitava

2011-05-02

Two chromogenic complexes, L.Zn (where L is (E)-4-((4-(1,4,8,11-tetraazacyclotetradecan-1-ylsulfonyl)phenyl)diazenyl)-N,N-dimethylaniline) and its [2]pseudorotaxane form (α-CD.L.Zn), were found to bind preferentially to adenosine triphosphate (ATP), among all other common anions and biologically important phosphate (AMP, ADP, pyrophosphate, and phosphate) ions in aqueous HEPES buffer medium of pH 7.2. Studies with live cell cultures of prokaryotic microbes revealed that binding of these two reagents to intercellular ATP, produced in situ, could be used in delineating the gram-positive and the gram-negative bacteria. More importantly, these dyes were found to be nontoxic to living microbes (eukaryotes and prokaryotes) and could be used for studying the cell growth dynamics. Binding to these two viable staining agents to intercellular ATP was also confirmed by spectroscopic studies on cell growth in the presence of different respiratory inhibitors that influence the intercellular ATP generation. © 2011 American Chemical Society

Effects of bleomycin and irradiation on euoxic and hypoxic cells

International Nuclear Information System (INIS)

Shrieve, D.C.; Harris, J.W.

1979-01-01

EMT6 cells in vitro were exposed to bleomycin (BLM), either alone (under euoxic or hypoxic conditions) or in conjunction with x-radiation. Hypoxic and euoxic cells were equally sensitive to the drug in both of the systems used to induce hypoxia (ampules or chambers). Exposure to BLM immediately before x-irradiation altered the shape of the radiation survival curve decreasing the D 0 by a factor of 1.3. Simultaneous exposure to x-ray and BLM resulted in lower survivals than when radiation was given either before or after drug treatment. Cells recovered quickly from BLM damage if trypsinization was delayed. The results indicate that BLM and x-rays interact to lower cell survival but that cells recover from this effect if trypsinization is delayed

Antimicrobial Susceptibility Patterns of Environmental Streptococci Recovered from Bovine Milk Samples in the Maritime Provinces of Canada

Directory of Open Access Journals (Sweden)

Marguerite Cameron

2016-09-01

, susceptibility decreased with increasing within-herd average somatic cell count, isolates recovered in mid-lactation were more susceptible that isolates recovered in early lactation, and isolates recovered in samples collected post-clinical mastitis were more susceptible than isolates recovered from non-clinical lactating quarters. The results of this research support continued susceptibility of environmental streptococci to beta-lactam antimicrobials. A departure from the expected susceptibility to beta-lactams was the apparent reduced susceptibility of Streptococcus uberis to penicillin.

Canine caliciviruses of four serotypes from military and research dogs recovered in 1963-1978 belong to two phylogenetic clades in the Vesivirus genus.

Science.gov (United States)

Binn, Leonard N; Norby, Erica A; Marchwicki, Ruth H; Jarman, Richard G; Keiser, Paul B; Hang, Jun

2018-02-23

Vesiviruses (family Caliciviridae) had been shown capable of invading a variety of host species, raising concern of their zoonotic potential. Since the 1980's, several canine caliciviruses (CaCV) isolates have been reported and are phylogenetically related to the vesiviruses with features distinct from both Vesicular exanthema of swine virus (VESV) and Feline calicivirus (FCV) species in phylogeny, serology and cell culture specificities. Etiological studies of canine diseases in dogs used for military services and laboratory studies were conducted in 1963-1978 at the Walter Reed Army Institute of Research. Multiple known and unknown viral pathogens including caliciviruses were recovered. Four unidentified isolates were recovered in Walter Reed Canine Cells (WRCC) from respiratory, fecal and penile specimens. Physicochemical tests, electron microscopy, viral cultivation in human and animal cells, antibody neutralization assays, and recently the genome sequencing were used to characterize the isolates. Sera from these dogs and their cohorts were tested with the isolates to determine origin and prevalence of the infections. The viral isolates were small non-enveloped spherical RNA virions, 27 to 42 nm in diameter with cup-like structures, indicating they are caliciviruses. They propagated in WRCC and MDCK cells, not in either other canine cells or human and other animal cells. Each isolate is antigenically distinct and react with dog sera in respective cohorts. The genomes have nucleotide identities ranging from 70.3% to 90.7% and encode the non-structural polyprotein (1810 amino acids), major capsid protein (691 amino acids) and minor structural protein (134 amino acids). They belong to two different phylogenetic clades in Vesivirus genus with close relation with canine calicivirus (CaCV). These CaCV isolates have restricted cell tropism, antigenic diversity and genetic variation. Further investigation will shed light on antigenic relation to other vesiviruses, and

Virulence of thermolable haemolysi tlh, gastroenteritis related pathogenicity tdh and trh of the pathogens Vibrio Parahemolyticus in Viable but Non-Culturable (VBNC) state.

Science.gov (United States)

Zhong, Huamin; Zhong, Yukui; Deng, Qiulian; Zhou, Zhenwen; Guan, Xiaoshan; Yan, Muxia; Hu, Tingting; Luo, Mingyong

2017-10-01

In the Viable but Non-Culturable (VBNC) state, microorganisms may survive under severe external environment. In this study, the specificity and sensitivity of PMA-LAMP assay on the detection of Vibrio Parahemolyticus (V. parahemolyticus) has been developed and evaluated, with further application on a number of food-borne V. parahemolyticus strains. Six primers were designed for recognizing 8 distinct targeting on tlh, tdh and trh gene. Through specific penetration through the damaged cell membrane of dead cells and intercalating into DNA, PMA could prevent DNA amplification of dead bacteria from LAMP, which enabled the differentiation of bacteria between VBNC state and dead state. The established PMA-LAMP showed significant advantage in rapidity, sensitivity and specificity, compared with regular PCR assay. The applicability had also been verified, demonstrating the PMA-LAMP was capable of detection on V. parahaemolyticus. Copyright © 2017 Elsevier Ltd. All rights reserved.

Increased cell proliferation and mucocyte density in the sea anemone Aiptasia pallida recovering from bleaching.

Directory of Open Access Journals (Sweden)

David Fransolet

Full Text Available Recovery of coral after bleaching episodes is a critical period for the health of the reef ecosystem. While events such as symbiont (genus Symbiodinium shifting/shuffling or tissue apoptosis have been demonstrated to occur following bleaching, little is known concerning tissue recovery or cell proliferation. Here, we studied the sea anemone Aiptasia pallida exposed to a transient elevation of water temperature combined with high illumination (33°C and 1900 µmol photons x m(-2 x s(-1 for 30 h. Following such treatment bleached anemones showed a significant reduction of their Symbiodinium density. Cell proliferation in the ectodermis and gastrodermis was determined by assessing the densities of cells labeled with a thymidine analogue (EdU. Cell proliferation significantly increased during the first day following stress in both tissue types. This increased cell proliferation returned to pre-stress values after one week. Although cell proliferation was higher in the ectodermis in absence of stress, it was relatively more pronounced in the gastrodermis of stressed anemones. In addition, the ratio of ectodermal mucocytes significantly increased three weeks after induced stress. These results suggest that thermal/photic stress coupled with the loss of the symbionts is able to enhance cell proliferation in both gastrodermis and ectodermis of cnidarians. While new cells formed in the gastrodermis are likely to host new Symbiodinium, the fate of new cells in the ectodermis was only partially revealed. Some new ectodermal cells may, in part, contribute to the increased number of mucocytes which could eventually help strengthen the heterotrophic state until restoration of the symbiosis.

Optimization of Culture Medium Enhances Viable Biomass Production and Biocontrol Efficacy of the Antagonistic Yeast, Candida diversa

Directory of Open Access Journals (Sweden)

Jia Liu

2017-10-01

Full Text Available Viable biomass production is a key determinant of suitability of antagonistic yeasts as potential biocontrol agents. This study investigated the effects of three metal ions (magnesium, ferrous, and zinc on biomass production and viability of the antagonistic yeast, Candida diversa. Using response surface methodology to optimize medium components, a maximum biomass was obtained, when the collective Mg2+, Fe2+, and Zn2+ concentrations were adjusted in a minimal mineral (MM medium. Compared with the unmodified MM, and three ion-deficient MM media, yeast cells cultured in the three ion-modified MM medium exhibited a lower level of cellular oxidative damage, and a higher level of antioxidant enzyme activity. A biocontrol assay indicated that C. diversa grown in the ion-modified MM exhibited the greatest level of control of gray mold on apple fruit. These results provide new information on culture medium optimization to grow yeast antagonists in order to improve biomass production and biocontrol efficacy.

Life Cycle Primary Energy and Carbon Analysis of Recovering Softwood Framing Lumber and Hardwood Flooring for Reuse

Science.gov (United States)

Richard D. Bergman; Hongmei Gu; Thomas R. Napier; James Salazar; Robert H. Falk

2012-01-01

Recovering wood for reuse in a new house affects energy and greenhouse gas emissions. This paper finds the energy and emissions for recovering softwood framing lumber and hardwood flooring from an old house for installation in a new house. Recovering wood displaces primary production of new wood products and avoids the end-of-life (EOL) burdens for the old house. We...

Soft sensor for monitoring biomass subpopulations in mammalian cell culture processes.

Science.gov (United States)

Kroll, Paul; Stelzer, Ines V; Herwig, Christoph

2017-11-01

Biomass subpopulations in mammalian cell culture processes cause impurities and influence productivity, which requires this critical process parameter to be monitored in real-time. For this reason, a novel soft sensor concept for estimating viable, dead and lysed cell concentration was developed, based on the robust and cheap in situ measurements of permittivity and turbidity in combination with a simple model. It could be shown that the turbidity measurements contain information about all investigated biomass subpopulations. The novelty of the developed soft sensor is the real-time estimation of lysed cell concentration, which is directly correlated to process-related impurities such as DNA and host cell protein in the supernatant. Based on data generated by two fed-batch processes the developed soft sensor is described and discussed. The presented soft sensor concept provides a tool for viable, dead and lysed cell concentration estimation in real-time with adequate accuracy and enables further applications with respect to process optimization and control.

Stromal cell migration precedes hemopoietic repopulation of the bone marrow after irradiation

International Nuclear Information System (INIS)

Werts, E.D.; Gibson, D.P.; Knapp, S.A.; DeGowin, R.L.

1980-01-01

Circulation of hemopoietic stem cells into an irradiated site has been thoroughly documented, but migration of stromal cells to repair radiation damage has not. We determined the radiosensitivity of mouse bone marrow stroma and evaluated stromal and hemopoietic repopulation in x-irradiated marrow. The D 0 for growth of colonies of marrow stromal cells (MSC) was 215 to 230 rad. Total-body irradiation (TB) obliterated marrow stromal and hemopoietic cells within 3 days. In contrast, 1 day after 1000 rad leg irradiation (LI), MSC rose to 80% of normal, but fell to 34% by 3 days and recovered to 72% by 30 days. However, femoral nucleated cells diminished to 20% by 3 days and recovered to 74% of normal by 30 days. Likewise, differentiated marrow cells and hemopoietic stem cells were initially depleted. With 1000 rad LI followed 3 h later by 1000 rad to the body while shielding the leg, MSC and femoral nucleated cells recovered to values intermediate between 1000 rad TB and 1000 rad LI. We concluded that: (1) the D 0 for MSC was 215 to 230 rad, (2) stromal repopulation preceded hemopoietic recovery, and (3) immigration of stromal cells from an unirradiated sanctuary facilitated hemopoietic repopulation of a heavily irradiated site

Cytomegalovirus survival and transferability and the effectiveness of common hand-washing agents against cytomegalovirus on live human hands.

Science.gov (United States)

Stowell, Jennifer D; Forlin-Passoni, Daniela; Radford, Kay; Bate, Sheri L; Dollard, Sheila C; Bialek, Stephanie R; Cannon, Michael J; Schmid, D Scott

2014-01-01

Congenital cytomegalovirus (CMV) transmission can occur when women acquire CMV while pregnant. Infection control guidelines may reduce risk for transmission. We studied the duration of CMV survival after application of bacteria to the hands and after transfer from the hands to surfaces and the effectiveness of cleansing with water, regular and antibacterial soaps, sanitizer, and diaper wipes. Experiments used CMV AD169 in saliva at initial titers of 1 × 10(5) infectious particles/ml. Samples from hands or surfaces (points between 0 and 15 min) were placed in culture and observed for at least 2 weeks. Samples were also tested using CMV real-time PCR. After application of bacteria to the hands, viable CMV was recovered from 17/20 swabs at 0 min, 18/20 swabs at 1 min, 5/20 swabs at 5 min, and 4/20 swabs at 15 min. After transfer, duration of survival was at least 15 min on plastic (1/2 swabs), 5 min on crackers and glass (3/4 swabs), and 1 min or less on metal and cloth (3/4 swabs); no viable virus was collected from wood, rubber, or hands. After cleansing, no viable virus was recovered using water (0/22), plain soap (0/20), antibacterial soap (0/20), or sanitizer (0/22). Viable CMV was recovered from 4/20 hands 10 min after diaper wipe cleansing. CMV remains viable on hands for sufficient times to allow transmission. CMV may be transferred to surfaces with reduced viability. Hand-cleansing methods were effective at eliminating viable CMV from hands.

Preliminary stochastic model for managing Vibrio parahaemolyticus and total viable bacterial counts in a Pacific oyster (Crassostrea gigas) supply chain.

Science.gov (United States)

Fernandez-Piquer, Judith; Bowman, John P; Ross, Tom; Estrada-Flores, Silvia; Tamplin, Mark L

2013-07-01

Vibrio parahaemolyticus can accumulate and grow in oysters stored without refrigeration, representing a potential food safety risk. High temperatures during oyster storage can lead to an increase in total viable bacteria counts, decreasing product shelf life. Therefore, a predictive tool that allows the estimation of both V. parahaemolyticus populations and total viable bacteria counts in parallel is needed. A stochastic model was developed to quantitatively assess the populations of V. parahaemolyticus and total viable bacteria in Pacific oysters for six different supply chain scenarios. The stochastic model encompassed operations from oyster farms through consumers and was built using risk analysis software. Probabilistic distributions and predictions for the percentage of Pacific oysters containing V. parahaemolyticus and high levels of viable bacteria at the point of consumption were generated for each simulated scenario. This tool can provide valuable information about V. parahaemolyticus exposure and potential control measures and can help oyster companies and regulatory agencies evaluate the impact of product quality and safety during cold chain management. If coupled with suitable monitoring systems, such models could enable preemptive action to be taken to counteract unfavorable supply chain conditions.

Deletion of ultraconserved elements yields viable mice

Energy Technology Data Exchange (ETDEWEB)

Ahituv, Nadav; Zhu, Yiwen; Visel, Axel; Holt, Amy; Afzal, Veena; Pennacchio, Len A.; Rubin, Edward M.

2007-07-15

Ultraconserved elements have been suggested to retainextended perfect sequence identity between the human, mouse, and ratgenomes due to essential functional properties. To investigate thenecessities of these elements in vivo, we removed four non-codingultraconserved elements (ranging in length from 222 to 731 base pairs)from the mouse genome. To maximize the likelihood of observing aphenotype, we chose to delete elements that function as enhancers in amouse transgenic assay and that are near genes that exhibit markedphenotypes both when completely inactivated in the mouse as well as whentheir expression is altered due to other genomic modifications.Remarkably, all four resulting lines of mice lacking these ultraconservedelements were viable and fertile, and failed to reveal any criticalabnormalities when assayed for a variety of phenotypes including growth,longevity, pathology and metabolism. In addition more targeted screens,informed by the abnormalities observed in mice where genes in proximityto the investigated elements had been altered, also failed to revealnotable abnormalities. These results, while not inclusive of all thepossible phenotypic impact of the deleted sequences, indicate thatextreme sequence constraint does not necessarily reflect crucialfunctions required for viability.

The viable but nonculturable state induction and genomic analyses of Lactobacillus casei BM-LC14617, a beer-spoilage bacterium.

Science.gov (United States)

Liu, Junyan; Li, Lin; Peters, Brian M; Li, Bing; Chen, Lequn; Deng, Yang; Xu, Zhenbo; Shirtliff, Mark E

2017-10-01

This study aimed to investigate the viable but nonculturable (VBNC) state and genomic features of a beer-spoilage strain, Lactobacillus caseiBM-LC14617. Induction on the VBNC state of L. casei strain BM-LC14617 was conducted by both low-temperature storage and continuous passage in beer, and formation of VBNC state was detected after 196 ± 3.3 days and 32 ± 1.6 subcultures, respectively. Resuscitation of VBNC cells was successfully induced by addition of catalase, and culturable, VBNC, and resuscitated cells shared similar beer-spoilage capability. Whole genome sequencing was performed, and out of a total of 3,964 predicted genes, several potential VBNC and beer-spoilage-associated genes were identified. L. casei is capable of entering into and resuscitating from the VBNC state and possesses beer-spoilage capability. The genomic characterization yield insightful elucidation of VBNC state for L. casei. This study represents the first evidence on VBNC state induction of L. casei and beer-spoilage capability of VBNC and resuscitated cells. Also, this is the first genomic characterization of L. casei as a beer-spoilage bacterium. The current study may aid in further study on L. casei and other beer-spoilage bacteria, and guide the prevention and control of beer spoilage. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Antiproliferative activity of pristimerin isolated from Maytenus ilicifolia (Celastraceae) in human HL-60 cells.

Science.gov (United States)

Costa, Patricia Marçal da; Ferreira, Paulo Michel Pinheiro; Bolzani, Vanderlan da Silva; Furlan, Maysa; de Freitas Formenton Macedo Dos Santos, Vânia Aparecida; Corsino, Joaquim; de Moraes, Manoel Odorico; Costa-Lotufo, Letícia Veras; Montenegro, Raquel Carvalho; Pessoa, Cláudia

2008-06-01

Pristimerin has been shown to be cytotoxic to several cancer cell lines. In the present work, the cytotoxicity of pristimerin was evaluated in human tumor cell lines and in human peripheral blood mononuclear cells (PBMC). This work also examined the effects of pristimerin (0.4; 0.8 and 1.7 microM) in HL-60 cells, after 6, 12 and 24h of exposure. Pristimerin reduced the number of viable cells and increased number of non-viable cells in a concentration-dependent manner by tripan blue test showing morphological changes consistent with apoptosis. Nevertheless, pristimerin was not selective to cancer cells, since it inhibited PBMC proliferation with an IC50 of 0.88 microM. DNA synthesis inhibition assessed by 5-bromo-2'-deoxyuridine (BrdU) incorporation in HL-60 cells was 70% and 83% for the concentrations of 0.4 and 0.8 microM, respectively. Pristimerin (10 and 20 microM) was not able to inhibit topoisomerase I. In AO/EB (acridine orange/ethidium bromide) staining, all tested concentrations reduced the number of HL-60 viable cells, with the occurrence of necrosis and apoptosis in a concentration-dependent manner, results in agreement with trypan blue exclusion findings. The analysis of membrane integrity and internucleosomal DNA fragmentation by flow cytometry in the presence of pristimerin indicated that treated cells underwent apoptosis. The present data point to the importance of pristimerin as representative of an emerging class of potential anticancer chemicals, exhibiting an antiproliferative effect by inhibiting DNA synthesis and triggering apoptosis.

Transcriptome analysis of symptomatic and recovered leaves of geminivirus-infected pepper (Capsicum annuum

Directory of Open Access Journals (Sweden)

Góngora-Castillo Elsa

2012-11-01

Full Text Available Abstract Background Geminiviruses are a large and important family of plant viruses that infect a wide range of crops throughout the world. The Begomovirus genus contains species that are transmitted by whiteflies and are distributed worldwide causing disease on an array of horticultural crops. Symptom remission, in which newly developed leaves of systemically infected plants exhibit a reduction in symptom severity (recovery, has been observed on pepper (Capsicum annuum plants infected with Pepper golden mosaic virus (PepGMV. Previous studies have shown that transcriptional and post-transcriptional gene silencing mechanisms are involved in the reduction of viral nucleic acid concentration in recovered tissue. In this study, we employed deep transcriptome sequencing methods to assess transcriptional variation in healthy (mock, symptomatic, and recovered pepper leaves following PepGMV infection. Results Differential expression analyses of the pepper leaf transcriptome from symptomatic and recovered stages revealed a total of 309 differentially expressed genes between healthy (mock and symptomatic or recovered tissues. Computational prediction of differential expression was validated using quantitative reverse-transcription PCR confirming the robustness of our bioinformatic methods. Within the set of differentially expressed genes associated with the recovery process were genes involved in defense responses including pathogenesis-related proteins, reactive oxygen species, systemic acquired resistance, jasmonic acid biosynthesis, and ethylene signaling. No major differences were found when compared the differentially expressed genes in symptomatic and recovered tissues. On the other hand, a set of genes with novel roles in defense responses was identified including genes involved in histone modification. This latter result suggested that post-transcriptional and transcriptional gene silencing may be one of the major mechanisms involved in the

Transcriptome analysis of symptomatic and recovered leaves of geminivirus-infected pepper (Capsicum annuum)

Science.gov (United States)

2012-01-01

Background Geminiviruses are a large and important family of plant viruses that infect a wide range of crops throughout the world. The Begomovirus genus contains species that are transmitted by whiteflies and are distributed worldwide causing disease on an array of horticultural crops. Symptom remission, in which newly developed leaves of systemically infected plants exhibit a reduction in symptom severity (recovery), has been observed on pepper (Capsicum annuum) plants infected with Pepper golden mosaic virus (PepGMV). Previous studies have shown that transcriptional and post-transcriptional gene silencing mechanisms are involved in the reduction of viral nucleic acid concentration in recovered tissue. In this study, we employed deep transcriptome sequencing methods to assess transcriptional variation in healthy (mock), symptomatic, and recovered pepper leaves following PepGMV infection. Results Differential expression analyses of the pepper leaf transcriptome from symptomatic and recovered stages revealed a total of 309 differentially expressed genes between healthy (mock) and symptomatic or recovered tissues. Computational prediction of differential expression was validated using quantitative reverse-transcription PCR confirming the robustness of our bioinformatic methods. Within the set of differentially expressed genes associated with the recovery process were genes involved in defense responses including pathogenesis-related proteins, reactive oxygen species, systemic acquired resistance, jasmonic acid biosynthesis, and ethylene signaling. No major differences were found when compared the differentially expressed genes in symptomatic and recovered tissues. On the other hand, a set of genes with novel roles in defense responses was identified including genes involved in histone modification. This latter result suggested that post-transcriptional and transcriptional gene silencing may be one of the major mechanisms involved in the recovery process. Genes

Quality standards and requirements for solid recovered fuels: a review.

Science.gov (United States)

Flamme, Sabine; Geiping, Julia

2012-04-01

The utilization of solid recovered fuels (SRF) for energy recovery has been increasing steadily in recent years, and this development is set to continue. In order to use SRF efficiently, it is necessary to define quality standards and introduce targeted quality assurance measures. SRF can be used both in mono-incineration and in co-incineration systems, for instance in power generation and cement plants; but as quality requirements differ, it is necessary to unambiguously define the term 'solid recovered fuel'. The purpose of this article is to provide an overview of the origin, development and the current status of quality assurance for SRF. The basic principles of quality assurance for SRF are explained with reference to the development of the German RAL Quality Assurance System and in addition specifications that have emerged from European standardization work of CEN/TC 343 are analysed.

Building Viable and Sustainable Regional Netchains: Case Studies of Regional Pork Netchains in Spain, Germany, and The Netherlands

Directory of Open Access Journals (Sweden)

Rannia Nijhoff‐Savvaki

2012-06-01

Full Text Available The present paper aims to give insight into critical issues contributing to the successful building of viable and sustainable regional netchains, focussing on chain governance mechanisms (coordination mechanisms, quality management systems, information systems and social embedding. It uses three representative case studies from the regional pork sector in Spain, Germany, and The Netherlands, illustrating different trajectories to commercially viable and sustainable regional netchains. By analysing the cases on each critical aspect, the present paper describes important issues and proposes specific elements of further research, including on the roles for(nongovernmental organisations.

Mitochondrial modification and respiratory deficiency in the yeast cell caused by cadmium poisoning

Energy Technology Data Exchange (ETDEWEB)

Lindegren, C C; Lindegren, G

1973-01-01

Cells of Fleischmann bakers' yeast were grown in standard nutrient broth and in broth to which cobalt, or cadmium, or thallium, had been added. The cells were fixed by glutaraldehyde-permanganate and sectioned. Electron microscopy showed that (a) the endoplasmic reticulum was fixed well in cells grown in cobalt or cadmium, but the endoplasmic reticulum was not fixed in cells grown in normal or thallium broth; (b) the cristate mitochondria were normal in all cells except those grown in cadmium. No cristae were visible in the cristate mitochondria of cells grown in cadmium broth; (c) a large fraction of the cells recovered from cadmium broth were respiratory-deficient; (d) thallic oxide was present in the cristate mitochondria of cells recovered from thallium broth. 13 references, 3 figures.

A Viable Microbial Community in a Subglacial Volcanic Crater Lake, Iceland

Science.gov (United States)

Gaidos, Eric; Lanoil, Brian; Thorsteinsson, Thorsteinn; Graham, Andrew; Skidmore, Mark; Han, Suk-Kyun; Rust, Terri; Popp, Brian

2004-09-01

We describe a viable microbial community in a subglacial lake within the Grímsvötn volcanic caldera, Iceland. We used a hot water drill to penetrate the 300-m ice shelf and retrieved lake water and volcanic tephra sediments. We also acquired samples of borehole water before and after penetration to the lake, overlying glacial ice and snow, and water from a nearby subaerial geothermal lake for comparative analyses. Lake water is at the freezing point and fresh (total dissolved solids = 260 mg L-1). Detectable numbers of cells were found in samples of the lake water column and tephra sediments: 2 × 104 ml-1 and 4 × 107 g-1, respectively. Plate counts document abundant cold-adapted cultivable organisms in the lake water, but not in the borehole (before penetration) or glacial ice. Denaturing gradient gel electrophoresis (DGGE) of 16S rRNA gene fragments amplified from genomic DNA extracted from Gr??msv??tn samples indicates that the lake community is distinct from the assemblages of organisms in borehole water (before penetration) and the overlying ice and snow. Sequencing of selected DGGE bands revealed that many sequences are highly similar to known psychrophilic organisms or cloned DNA from other cold environments. Significant uptake of 14C-labeled bicarbonate occurred in dark, low-temperature incubations of lake water samples, indicating the presence of autotrophs. Acetylene reduction assays under similar incubation conditions showed no significant nitrogen fixation potential by lake water samples. This may be a consequence of the inhibition of diazotrophy by nitrogen in the lake.

Commercializing Biorefinery Technology: A Case for the Multi-Product Pathway to a Viable Biorefinery

Directory of Open Access Journals (Sweden)

Shijie Liu

2011-11-01

Full Text Available While there may be many reasons why very interesting science ideas never reach commercial practice, one of the more prevalent is that the reaction or process, which is scientifically possible, cannot be made efficient enough to achieve economic viability. One pathway to economic viability for many business sectors is the multi-product portfolio. Research, development, and deployment of viable biorefinery technology must meld sound science with engineering and business economics. It is virtually axiomatic that increased value can be generated by isolating relatively pure substances from heterogeneous raw materials. Woody biomass is a heterogeneous raw material consisting of the major structural components, cellulose, lignin, and hemicelluloses, as well as minor components, such as extractives and ash. Cellulose is a linear homopolymer of D-glucopyrano-units with β-D(1®4 connections and is the wood component most resistant to chemical and biological degradation. Lignin is a macromolecule of phenylpropanoid units, second to cellulose in bio-resistance, and is the key component that is sought for removal from woody biomass in chemical pulping. Hemicelluloses are a collection of heteropolysaccharides, comprised mainly of 5- and 6-carbon sugars. Extractives, some of which have high commercial value, are a collection of low molecular weight organic and inorganic woody materials that can be removed, to some extent, under mild conditions. Applied Biorefinery Sciences, LLC (a private, New York, USA based company is commercializing a value-optimization pathway (the ABS Process™ for generating a multi-product portfolio by isolating and recovering homogeneous substances from each of the above mentioned major and minor woody biomass components. The ABS Process™ incorporates the patent pending, core biorefinery technology, “hot water extraction”, as developed at the State University of New York College of Environmental Science and Forestry (SUNY

Cummins L10G in Kenworth truck 'viable today'

International Nuclear Information System (INIS)

Anon.

1996-01-01

As the culmination of five years of developmental work by Cummins Engines, BC Research Inc., and BC Gas Utility Ltd., a T800 Kenworth truck was outfitted with a new Cummins L10G natural gas engine, and two lightweight fully -wrapped Dynetek cylinders; it was pronounced to be 'a viable clean truck today'. The L10G spark-ignited engine operates at a relatively high peak efficiency of 37 per cent and is commercially available to meet the current California Air Resources Board heavy duty vehicle emission standards without the use of a catalytic converter. The L10G engine produces no particulate emissions, a very significant advantage, in view of the fact that particulate emissions have been identified as major contributors to respiratory ailments

False Attribution of Suggestibility To Explain Recovered Memory of Childhood Sexual Abuse Following Extended Amnesia.

Science.gov (United States)

Leavitt, Frank

1997-01-01

Suggestibility was measured in 44 adult patients who recovered memories and in 31 comparison patients. Results suggest that patients who recovered memories were significantly less suggestible than average. Control patients with no history of sexual abuse were more at risk for altering memory to suggestive prompts. (Author/PB)

26 CFR 1.1336-1 - Basis of recovered property.

Science.gov (United States)

2010-04-01

... benefits by reason of the basis determined under subparagraph (1) of this paragraph, that it would be... recovered it has an unadjusted basis of $100. After $70 depreciation has been allowed on A, an allocation is sought which would give A an unadjusted basis of $60. Since this is less than the depreciation which is...

Process for recovering cesium from pollucite

International Nuclear Information System (INIS)

Mein, P.G.

1985-01-01

Cesium is recovered from a cesium-bearing mineral such as pollucite by extraction with hydrochloric acid to obtain an extract of cesium chloride and other alkali metal and polyvalent metal chlorides. The iron and aluminum chlorides can be precipitated as the hydroxides and separated from the solution of the alkali metal chlorides to which is added potassium permanganate or other water-soluble permanganate to selectively precipitate cesium permanganate. The cesium precipitate is then separated from the residual solution containing the metal chlorides. The cesium permanganate, which is in a very pure form, can be converted to other cesium compounds by reaction with a reducing agent to obtain cesium carbonate and cesium delta manganese dioxide

Characterization of HIT type solar cells

NARCIS (Netherlands)

Rath, J.K.

2011-01-01

The silicon heterojunction solar cell (SHJ) has made rapid progress in reaching high efficiency and it is already developed as an industrially viable product. However, much of its progress has come through process development while there is scarce knowledge on the microscopic nature of the

Phytoremediation of 1,4-dioxane-containing recovered groundwater.

Science.gov (United States)

Ferro, Ari M; Kennedy, Jean; LaRue, James C

2013-01-01

The results of a pilot-scale phytoremediation study are reported in this paper. Small plots of trees established on a closed municipal waste landfill site were irrigated with recovered groundwater containing 1,4-dioxane (dioxane) and other volatile organic compounds (VOCs). The plots were managed to minimize the leaching of irrigation water, and leaching was quantified by the use of bromide tracer. Results indicated that the dioxane (2.5 microg/L) was effectively removed, probably via phytovolatilization, and that a full-scale phytoremediation system could be used. A system is now in place at the site in which the recovered groundwater can be treated using two different approaches. A physical treatment system (PTS) will be used during the winter months, and a 12 ha phytoremediation system (stands of coniferous trees) will be used during the growing season. The PTS removes VOCs using an air-stripper, and destroys dioxane using a photo-catalytic oxidation process. Treated water will be routed to the local sewer system. The phytoremediation system, located on the landfill, will be irrigated with effluent from the PTS air-stripper containing dioxane. Seasonal use of the phytoremediation system will reduce reliance on the photo-catalytic oxidation process that is extremely energy consumptive and expensive to operate.

14C leucine chloromethylketone interaction with sarcoma 37 cell plasma membrane components

International Nuclear Information System (INIS)

Matthews, R.H.; Milo, G.E.; McMichael, T.L.; Lewis, N.J.