Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening Comparação de bioensaios com os crustáceos Artemia salina e Thamnocephalus platyurus para abordagem de extratos de plantas com toxicidade

Directory of Open Access Journals (Sweden)

Pablo Mayorga

2010-12-01

Full Text Available Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest, were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq. Kunth ex Walp., Fabaceae, Neurolaena lobata (L. Cass., Asteraceae, Petiveria alliacea L., Phytolaccaceae, and Ocimum campechianum Mill., Lamiaceae. The five others: Curatella americana L., Dilleniaceae, Prunus barbata Koehne, Rosaceae, Quercus crispifolia Trel., Fagaceae, Rhizophora mangle L., Rhizophoraceae, and Smilax domingensis Willd., Smilacaceae, do not. All plants without anti-Artemia activity had no lethal effects in both assays with A. salina. For the plants with anti-Artemia activity the Artoxkit M was not sensitive to G. sepium and the conventional Artemia test was not sensitive to S. americanum, G. sepium and N. lobata. All the plant extracts, except for that of C. americana, had lethal effects on T. platyurus and the lethal median concentration (LC50 levels for this organism were in all cases substantially lower than those of the salt-water test species. This study revealed that T. platyurus is a promising test species worth further in depth investigation for toxicity screening of plant extracts with potential medicinal properties.Três bioensaios de letalidade com o crustáceo de água salgada Artemia salina Leach, Artemiidae, (teste convencional em microplaca de 96 poós Artoxkit microbiotest M e o crustáceo de água doce Thamnocephalus platyurus Packard, Thamnocephalidae (Thamnotoxkit microbiotest F, foram comparados utilizando extratos de dez espécies de plantas da Guatemala. Foi previamente observado que cinco delas possuem atividade anti

Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening

OpenAIRE

Mayorga,Pablo; Pérez,Karen R.; Cruz,Sully M.; Cáceres,Armando

2010-01-01

Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest) and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest), were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq.) Kunth ex ...

Cadmium induced oxidative stress in Dunaliella salina | Moradshahi ...

African Journals Online (AJOL)

The unicellular green algae Dunaliella salina contains various antioxidants which protect the cell from oxidative damage due to environmental stresses such as heavy metal stress. In the present study, the response of D. salina at the stationary growth phase to oxidative stress generated by cadmium chloride was ...

Comparison of bioassays using the anostracan crustaceans Artemia salina and Thamnocephalus platyurus for plant extract toxicity screening Comparação de bioensaios com os crustáceos Artemia salina e Thamnocephalus platyurus para abordagem de extratos de plantas com toxicidade

OpenAIRE

Pablo Mayorga; Karen R. Pérez; Sully M. Cruz; Armando Cáceres

2010-01-01

Three lethality bioassays, using the salt-water crustacean Artemia salina Leach, Artemiidae, (conventional 96 microwell plate test and the Artoxkit M microbiotest) and the freshwater crustacean Thamnocephalus platyurus Packard, Thamnocephalidae, (Thamnotoxkit F microbiotest), were compared using extracts of ten Guatemalan plant species. It was previously observed that five of them have anti-Artemia activity. These were: Solanum americanum Mill., Solanaceae, Gliricidia sepium (Jacq.) Kunth ex ...

Harvesting of Dunaliella salina by membrane filtration at pilot scale

KAUST Repository

Monte, Joana

2017-09-02

The microalgae Dunaliella salina is industrially produced due to its high content in carotenoids induced by low nitrogen and high salinity conditions. D. salina with low carotenoids content also produces other added value compounds, however its recovery have hardly been studied. This work aims to examine the potential of pre-concentrating D. salina by membrane processing prior to a final harvesting step by low-shear centrifugation. The aim is to minimize the overall energy expenditure and reduce capital costs, while assuring a minimal loss of cell integrity. This task is challenging, considering the sensitivity of D. salina to shear. Harvesting of D. salina by ultrafiltration allowed reaching a final concentration factor of 5.9, with an average permeate flux of 31 L/(m2 h). The Total Cost of Ownership and energy consumption for harvesting are respectively 52% and 45% lower when applying a two-step approach with pre-concentration (ultrafiltration) compared to only harvesting by centrifugation.

Harvesting of Dunaliella salina by membrane filtration at pilot scale

KAUST Repository

Monte, Joana; Sá , Marta; Galinha, Clá udia F.; Costa, Luí s; Hoekstra, Herre; Brazinha, Carla; Crespo, Joã o G.

2017-01-01

The microalgae Dunaliella salina is industrially produced due to its high content in carotenoids induced by low nitrogen and high salinity conditions. D. salina with low carotenoids content also produces other added value compounds, however its recovery have hardly been studied. This work aims to examine the potential of pre-concentrating D. salina by membrane processing prior to a final harvesting step by low-shear centrifugation. The aim is to minimize the overall energy expenditure and reduce capital costs, while assuring a minimal loss of cell integrity. This task is challenging, considering the sensitivity of D. salina to shear. Harvesting of D. salina by ultrafiltration allowed reaching a final concentration factor of 5.9, with an average permeate flux of 31 L/(m2 h). The Total Cost of Ownership and energy consumption for harvesting are respectively 52% and 45% lower when applying a two-step approach with pre-concentration (ultrafiltration) compared to only harvesting by centrifugation.

Increasing β-carotene content of phytoplankton Dunaliella salina using different salinity media

Science.gov (United States)

Hermawan, J.; Masithah, E. D.; Tjahjaningsih, W.; Abdillah, A. A.

2018-04-01

Dunaliella salina have got great attention in the nutritional, pharmaceutical and cosmetic companies because contain β-carotene. β-carotene functions as antioxidants and precursors of vitamin A and can treat tumors and cancer in humans. The content of β-carotene in D. salina can be increased by increasing salinity levels in the culture medium. The aim of this study was to determine whether increasing salinity may increas β-carotene content of phytoplankton D. salina. The research use data collection method with direct observation and then analyzed the result with descriptive method. The results showed that different salinity of media can influenced β-carotene content of D. salina. The highest β-carotene content of D. salina was at treatment B (30 ppt) which equal to 2.312 mg/L on 10th day. The production of β-carotene in D. salina can be increased was other environmental stress treatments in the form of stress-temperature, light and nutrients using.

Can the halophilic ciliate Fabrea salina be used as a bio-control of microalgae blooms in solar salterns?

Science.gov (United States)

Hong, Hyun Pyo; Choi, Joong Ki

2015-09-01

The microlage Dunaliella salina, a major producer in salterns, is a serious problem for salt production. In this study we tried to assess if Fabrea salina can control D. salina. By parameterising numerical and functional response (growth and grazing vs prey abundance, respectively) at 90 psu and 30°C, where the ciliate is abundant and grows well, we developed a predator-prey model. The model is used to explore how change in microalga growth rate affect the dynamics, and the functional response is used in combination with field data to assess the potential impact of F. salina on D. salina. Over the 20 d simulation the ciliate controlled the prey population under all prey growth rates; although once D. salina were exhausted below the threshold level, F. salina died due to starvation, allowing the alga to increase in abundance, resulting in one or two predatorprey cycle, depending on prey growth rate. In general, the model predicted trends observed by others in the field, suggesting that it provided a good prediction of what may occur under the conditions we examined. Likewise we show that the ciliate can have a high impact on microalgal populations in the field. Finally, a literature review indicated that F. salina could be a good competitor with other protozoa and metazoan in salterns, depending on salinity and temperature, which requires further study and attention. In summary, we encourage continued studies on this unique ciliate on solar salterns and suggest that it may be useful in the bio-control of micoalgae.

Primary Screening of the Bioactivity of Brackishwater Cyanobacteria: Toxicity of Crude Extracts to Artemia salina Larvae and Paracentrotus lividus Embryos

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Viviana R. Lopes

2010-03-01

Full Text Available Cyanobacteria are a diverse group of Gram-negative bacteria that produce an array of secondary compounds with selective bioactivity against vertebrates, invertebrates, plants, microalgae, fungi, bacteria, viruses and cell lines. The aim of this study was to assess the toxic effects of aqueous, methanolic and hexane crude extracts of benthic and picoplanktonic cyanobacteria isolated from estuarine environments, towards the nauplii of the brine shrimp Artemia salina and embryos of the sea urchin Paracentrotus lividus. The A. salina lethality test was used as a frontline screen and then complemented by the more specific sea urchin embryo-larval assay. Eighteen cyanobacterial isolates, belonging to the genera Cyanobium, Leptolyngbya, Microcoleus, Phormidium, Nodularia, Nostoc and Synechocystis, were tested. Aqueous extracts of cyanobacteria strains showed potent toxicity against A. salina, whereas in P. lividus, methanolic and aqueous extracts showed embryo toxicity, with clear effects on development during early stages. The results suggest that the brackishwater cyanobacteria are producers of bioactive compounds with toxicological effects that may interfere with the dynamics of invertebrate populations.

Pedro Salinas: la letra y la persona

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Escartín Gual, Montserrat

2002-12-01

Full Text Available This article is a Pedro Salinas's homage in the fifty anniversary of his death. Here, we analise his attitude in front of life and his securities through his poems, personal letters and essais. The poet criticizes the materialistic american stil of life, where he should live during his exile until his death (1936-1951. Against the hurry, the profitable things, the technology, the money..., Salinas believes in his language, slowness, friendship, contemplation of countryside, art, spiritualism, optimism, etc. Son of the Institución Libre de Enseñanza's ideas, his personal ethics looked for the biggest perfection trought the will.Este artículo es un homenaje a Pedro Salinas al cumplirse el cincuentenario de su muerte. En él, se analiza su actitud ante la vida y sus valores a través de sus poesías, cartas personales y ensayos. El poeta critica el materialismo del modelo americano en el que se vio obligado a vivir desde que se exilió hasta su muerte (1936-1951. Frente a la prisa, lo rentable, la tecnología, el dinero...; Salinas cree en su idioma, la lentitud, la amistad, la contemplación del paisaje, el arte, lo espiritual, el optimismo, etc. Hijo de las ideas de la Institución Libre de Enseñanza, su ética personal buscó la mayor perfección a través de la voluntad.

The Dunaliella salina organelle genomes: large sequences, inflated with intronic and intergenic DNA

Energy Technology Data Exchange (ETDEWEB)

Smith, David R.; Lee, Robert W.; Cushman, John C.; Magnuson, Jon K.; Tran, Duc; Polle, Juergen E.

2010-05-07

Abstract Background: Dunaliella salina Teodoresco, a unicellular, halophilic green alga belonging to the Chlorophyceae, is among the most industrially important microalgae. This is because D. salina can produce massive amounts of β-carotene, which can be collected for commercial purposes, and because of its potential as a feedstock for biofuels production. Although the biochemistry and physiology of D. salina have been studied in great detail, virtually nothing is known about the genomes it carries, especially those within its mitochondrion and plastid. This study presents the complete mitochondrial and plastid genome sequences of D. salina and compares them with those of the model green algae Chlamydomonas reinhardtii and Volvox carteri. Results: The D. salina organelle genomes are large, circular-mapping molecules with ~60% noncoding DNA, placing them among the most inflated organelle DNAs sampled from the Chlorophyta. In fact, the D. salina plastid genome, at 269 kb, is the largest complete plastid DNA (ptDNA) sequence currently deposited in GenBank, and both the mitochondrial and plastid genomes have unprecedentedly high intron densities for organelle DNA: ~1.5 and ~0.4 introns per gene, respectively. Moreover, what appear to be the relics of genes, introns, and intronic open reading frames are found scattered throughout the intergenic ptDNA regions -- a trait without parallel in other characterized organelle genomes and one that gives insight into the mechanisms and modes of expansion of the D. salina ptDNA. Conclusions: These findings confirm the notion that chlamydomonadalean algae have some of the most extreme organelle genomes of all eukaryotes. They also suggest that the events giving rise to the expanded ptDNA architecture of D. salina and other Chlamydomonadales may have occurred early in the evolution of this lineage. Although interesting from a genome evolution standpoint, the D. salina organelle DNA sequences will aid in the development of a viable

The Dunaliella salina organelle genomes: large sequences, inflated with intronic and intergenic DNA

Directory of Open Access Journals (Sweden)

Tran Duc

2010-05-01

Full Text Available Abstract Background Dunaliella salina Teodoresco, a unicellular, halophilic green alga belonging to the Chlorophyceae, is among the most industrially important microalgae. This is because D. salina can produce massive amounts of β-carotene, which can be collected for commercial purposes, and because of its potential as a feedstock for biofuels production. Although the biochemistry and physiology of D. salina have been studied in great detail, virtually nothing is known about the genomes it carries, especially those within its mitochondrion and plastid. This study presents the complete mitochondrial and plastid genome sequences of D. salina and compares them with those of the model green algae Chlamydomonas reinhardtii and Volvox carteri. Results The D. salina organelle genomes are large, circular-mapping molecules with ~60% noncoding DNA, placing them among the most inflated organelle DNAs sampled from the Chlorophyta. In fact, the D. salina plastid genome, at 269 kb, is the largest complete plastid DNA (ptDNA sequence currently deposited in GenBank, and both the mitochondrial and plastid genomes have unprecedentedly high intron densities for organelle DNA: ~1.5 and ~0.4 introns per gene, respectively. Moreover, what appear to be the relics of genes, introns, and intronic open reading frames are found scattered throughout the intergenic ptDNA regions -- a trait without parallel in other characterized organelle genomes and one that gives insight into the mechanisms and modes of expansion of the D. salina ptDNA. Conclusions These findings confirm the notion that chlamydomonadalean algae have some of the most extreme organelle genomes of all eukaryotes. They also suggest that the events giving rise to the expanded ptDNA architecture of D. salina and other Chlamydomonadales may have occurred early in the evolution of this lineage. Although interesting from a genome evolution standpoint, the D. salina organelle DNA sequences will aid in the

Dichotomosiphon salina sp. nov. - a new marine algal form from Goa estuary, India

Digital Repository Service at National Institute of Oceanography (India)

Untawale, A.G.; Jagtap, T.G.; Dhargalkar, V.K.

A new species Dichotomosiphon salina sp. of family Vaucheriaceae has been reported from brackish water areas of Goa. The high salinity tolerance of 40 ppt and the variation in size as well as shape of the reproductive organs are the main...

Groundwater quality in the shallow aquifers of the Monterey Bay, Salinas Valley, and adjacent highland areas, Southern Coast Ranges, California

Science.gov (United States)

Burton, Carmen

2018-05-30

The Monterey-Salinas Shallow Aquifer study unit covers approximately 7,820 square kilometers (km2) in Santa Cruz, Monterey, and San Luis Obispo Counties in the Central Coast Hydrologic Region of California. The study unit was divided into four study areas—Santa Cruz, Pajaro Valley, Salinas Valley, and Highlands. More than 75 percent of the water used for drinking-water supply in the Central Coast Hydrologic Region of California is groundwater, and there are more than 8,000 well driller’s logs for domestic wells (California Department of Water Resources, 2013).

Optimization of photosynthesis, growth, and biochemical composition of the microalga Rhodomonas salina

DEFF Research Database (Denmark)

Thuy, Minh Vu Thi; Douëtte, Claire; Rayner, Thomas Allan

2016-01-01

The cryptophyte Rhodomonas salina is widely used as feed for copepod cultures. However, culturing conditions to obtain high-quality algae have not yet been efficiently optimized. Therefore, we aimed to develop a cultivation protocol for R. salina to optimize its nutritional value and provide tech...

Potential of New Isolates of Dunaliella Salina for Natural β-Carotene Production

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Yanan Xu

2018-02-01

Full Text Available The halotolerant microalga Dunaliella salina has been widely studied for natural β-carotene production. This work shows biochemical characterization of three newly isolated Dunaliella salina strains, DF15, DF17, and DF40, compared with D. salina CCAP 19/30 and D. salina UTEX 2538 (also known as D. bardawil. Although all three new strains have been genetically characterized as Dunaliella salina strains, their ability to accumulate carotenoids and their capacity for photoprotection against high light stress are different. DF15 and UTEX 2538 reveal great potential for producing a large amount of β-carotene and maintained a high rate of photosynthesis under light of high intensity; however, DF17, DF40, and CCAP 19/30 showed increasing photoinhibition with increasing light intensity, and reduced contents of carotenoids, in particular β-carotene, suggesting that the capacity of photoprotection is dependent on the cellular content of carotenoids, in particular β-carotene. Strong positive correlations were found between the cellular content of all-trans β-carotene, 9-cis β-carotene, all-trans α-carotene and zeaxanthin but not lutein in the D. salina strains. Lutein was strongly correlated with respiration in photosynthetic cells and strongly related to photosynthesis, chlorophyll and respiration, suggesting an important and not hitherto identified role for lutein in coordinated control of the cellular functions of photosynthesis and respiration in response to changes in light conditions, which is broadly conserved in Dunaliella strains. Statistical analysis based on biochemical data revealed a different grouping strategy from the genetic classification of the strains. The significance of these data for strain selection for commercial carotenoid production is discussed.

A 96-well automated method to study inhibitors of human sodium-dependent D-glucose transport.

Science.gov (United States)

Castaneda, Francisco; Kinne, Rolf K-H

2005-12-01

The sodium-dependent D-glucose transporter (SGLT) family is involved in glucose uptake via intestinal absorption (SGLT1) or renal reabsorption (SGLT1 and SGLT2). Current methods for the screening of inhibitors of SGLT transporters are complex, expensive and very labor intensive, and have not been applied to human SGLT transporters. The purpose of the present study was to develop an alternative 96-well automated method to study the activity of human SGLT1 and SGLT2. Chinese hamster ovary (CHO) Flp-In cells were stably transfected with pcDNA5-SGLT1 or pcDNA5-SGLT2 plasmid and maintained in hygromycin-selection Ham's F12 culture medium until hygromycin-resistant clones were developed. SGLT1 and SGLT2 gene expression was evaluated by relative real-time reverse transcription-polymerase chain reaction (RT-PCR) quantification, Western blotting, and immunocytochemical analysis. The clones with higher expression of SGLT1 and SGLT2 were used for transport studies using [14C]-methyl-alpha-D-glucopyranoside ([14C]AMG). The advantage of using the 96-well format is the low amount of radioactive compounds and inhibitory substances required, and its ability to establish reproducibility because repetition into the assay. This method represents an initial approach in the development of transport-based high-throughput screening in the search for inhibitors of glucose transport. The proposed method can easily be performed to yield quantitative data regarding key aspects of glucose membrane transport and kinetic studies of potential inhibitors of human SGLT1 and SGLT2.

Distribution and abundance of Artemia salina in the Salt Lake Basin (Central Anatolia, Turkey

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Alaş Ali

2017-06-01

Full Text Available In this study, the distribution and abundance of Artemia salina in 10 different stations of the Salt Lake basin were investigated. In addition, its relationship to pH, dissolved oxygen, temperature, electrical conductivity and water levels were analyzed. Field studies were carried out from July to August of 2010. Artemia salina was observed in five of these stations. Artemia salina was not seen in some stations that have high electrical conductivity. It is determined that, in the station named Tersakan Lake where electrical conductivity was 154 mS/cm, Artemia salina is more abundant when compared to the other stations. But as underground water pumps that are built for the irrigation of agricultural lands decrease water levels, Artemia salina’s life is under threat.

Toxicоlogical evaluation of the plant products using Brine Shrimp (Artemia salina L. model

Directory of Open Access Journals (Sweden)

Меntor R. Hamidi

2014-04-01

Full Text Available Many natural products could serve as the starting point in the development of modern medicines because of their numerous biological and pharmacological activities. However, some of them are known to carry toxicological properties as well. In order to achieve a safe treatment with plant products, numerous research studies have recently been focused on both pharmacology and toxicity of medicinal plants. Moreover, these studies employed efforts for alternative biological assays. Brine Shrimp Lethality Assay is the most convenient system for monitoring biological activities of various plant species. This method is very useful for preliminary assessment of toxicity of the plant extracts. Rapidness, simplicity and low requirements are several advantages of this assay. However, several conditions need to be completed, especially in the means of standardized experimental conditions (temperature, pH of the medium, salinity, aeration and light. The toxicity of herbal extracts using this assay has been determined in a concentration range of 10, 100 and 1000 µg/ml of the examined herbal extract. Most toxicity studies which use the Brine Shrimp Lethality Assay determine the toxicity after 24 hours of exposure to the tested sample. The median lethal concentration (LC50 of the test samples is obtained by a plot of percentage of the dead shrimps against the logarithm of the sample concentration. LC50 values are estimated using a probit regression analysis and compared with either Meyer’s or Clarkson’s toxicity criteria. Furthermore, the positive correlation between Meyer’s toxicity scale for Artemia salina and Gosselin, Smith and Hodge’s toxicity scale for higher animal models confirmed that the Brine Shrimp Lethality Assay is an excellent predictive tool for the toxic potential of plant extracts in humans.

Interspecific competition and allelopathic interaction between Karenia mikimotoi and Dunaliella salina in laboratory culture

Science.gov (United States)

He, Dong; Liu, Jiao; Hao, Qiang; Ran, Lihua; Zhou, Bin; Tang, Xuexi

2016-03-01

Algal allelopathy is a manifold ecological/physiological phenomenon that is focused on chemical interactions and autotoxicity. We investigated the allelopathic interactions between Karenia mikimotoi and Dunaliella salina in laboratory cultures based on diff erent temperature (15°C, 20°C, and 25°C) and lighting (40, 80, and 160 μmol/(m2·s)) conditions. The growth of D. salina in bi-algae culture (1:1 size/density) was significantly restrained. The results of cell-free filtrate culture indicate that direct cell-tocell contact was not necessary in interspecific competition. Further experimental results demonstrated that allelochemicals released from K. mikimotoi were markedly influenced by both temperature ( P =0.013) and irradiance ( P =0.003), resulting in diff erent growth characteristics of D. salina in filtrate mediums. Compared with the plateau period, K. mikimotoi exudates in the exponential phase had a stronger short-term inhibition effect on D. salina in normal conditions. A clear concentration-dependent relationship was observed in the effect of allelochemicals released from K. mikimotoi with low-promoting and high-repressing effects on D. Salina in a short time-scale. In addition, allelopathic substances remain stable and effective under high temperature and pressure stress. Many flocculent sediments adhering with D. salina cells were observed in all filtrate mediums, while the quantity and color depended on the original culture conditions.

Bioactivity evaluation against Artemia salina Leach of medicinal plants used in Brazilian Northeastern folk medicine.

Science.gov (United States)

Arcanjo, D D R; Albuquerque, A C M; Melo-Neto, B; Santana, L C L R; Medeiros, M G F; Citó, Amgl

2012-08-01

The brine shrimp (Artemia salina Leach) lethality bioassay offers an advantage in standardization and quality control of botanical products. This test is well correlated with antitumor activity (cytotoxicity) and can be used to monitor the activity of bioactive natural products. This paper reports the bioactivity of ethanol extracts from seven medicinal plants from the Northeast of Brazil (Acmella uliginosa, Ageratum conyzoides, Eugenia uniflora, Plectranthus neochilus, Moringa oleifera, Justicia pectoralis and Equisetum sp.) against Artemia salina. Biological activity was evaluated for extracts at 1, 10, 100, and 1000 µg/mL in triplicate, and the mean lethal concentration values (LC50) were obtained by probit analysis. The species Acmella uliginosa showed the highest bioactivity, and its flower extract was more active than its leaf extract.

Toxicities of diuron and irgarol on the hatchability and early stage development of Artemia salina

OpenAIRE

ALYÜRÜK, Hakan; ÇAVAŞ, Levent

2013-01-01

Booster biocides are widely used in antifouling paints as bioactive agents against fouling organisms. In previously published reports, acute toxicity tests on Artemia salina (Linnaeus, 1758) were only focused on a part of the life cycle of the organism. The aim of this study was to investigate the toxicities of diuron and irgarol on the hatching stage of A. salina. According to the results, diuron significantly decreased the hatching percentage of A. salina cysts and prevented the hatching of...

Toxicity and trophic transfer of P25 TiO2 NPs from Dunaliella salina to Artemia salina: Effect of dietary and waterborne exposure.

Science.gov (United States)

Bhuvaneshwari, M; Thiagarajan, Vignesh; Nemade, Prateek; Chandrasekaran, N; Mukherjee, Amitava

2018-01-01

The recent increase in nanoparticle (P25 TiO 2 NPs) usage has led to concerns regarding their potential implications on environment and human health. The food chain is the central pathway for nanoparticle transfer from lower to high trophic level organisms. The current study relies on the investigation of toxicity and trophic transfer potential of TiO 2 NPs from marine algae Dunaliella salina to marine crustacean Artemia salina. Toxicity was measured in two different modes of exposure such as waterborne (exposure of TiO 2 NPs to Artemia) and dietary exposure (NP-accumulated algal cells are used to feed the Artemia). The toxicity and accumulation of TiO 2 NPs in marine algae D. salina were also studied. Artemia was found to be more sensitive to TiO 2 NPs (48h LC 50 of 4.21mgL -1 ) as compared to marine algae, D. salina (48h LC 50 of 11.35mgL -1 ). The toxicity, uptake, and accumulation of TiO 2 NPs were observed to be more in waterborne exposure as compared to dietary exposure. Waterborne exposure seemed to cause higher ROS production and antioxidant enzyme (SOD and CAT) activity as compared to dietary exposure of TiO 2 NPs in Artemia. There were no observed biomagnification (BMF) and trophic transfer from algae to Artemia through dietary exposure. Histopathological studies confirmed the morphological and internal damages in Artemia. This study reiterates the possible effects of the different modes of exposure on trophic transfer potential of TiO 2 NPs and eventually the consequences on aquatic environment. Copyright © 2017 Elsevier Inc. All rights reserved.

Nitrate Contamination of Deep Aquifers in the Salinas Valley, California

Science.gov (United States)

Moran, J. E.; Esser, B. K.; Hillegonds, D. J.; Holtz, M.; Roberts, S. K.; Singleton, M. J.; Visser, A.; Kulongoski, J. T.; Belitz, K.

2011-12-01

The Salinas Valley, known as 'the salad bowl of the world', has been an agricultural center for more than 100 years. Irrigated row crops such as lettuce and strawberries dominate both land use and water use. Groundwater is the exclusive supply for both irrigation and drinking water. Some irrigation wells and most public water supply wells in the Salinas Valley are constructed to draw water from deep portions of the aquifer system, where contamination by nitrate is less likely than in the shallow portions of the aquifer system. However, a number of wells with top perforations greater than 75 m deep, screened below confining or semi-confining units, have nitrate concentrations greater than the Maximum Contaminant Limit (MCL) of 45 mg/L as NO3-. This study uses nitrate concentrations from several hundred irrigation, drinking water, and monitoring wells (Monterey County Water Resources Agency, 1997), along with tritium-helium groundwater ages acquired at Lawrence Livermore National Laboratory through the State of California Groundwater Monitoring and Assessment (GAMA) program (reported in Kulongoski et al., 2007 and in Moran et al., in press), to identify nitrate 'hot spots' in the deep aquifer and to examine possible modes of nitrate transport to the deep aquifer. In addition, observed apparent groundwater ages are compared with the results of transport simulations that use particle tracking and a stochastic-geostatistical framework to incorporate aquifer heterogeneity to determine the distribution of travel times from the water table to each well (Fogg et al., 1999). The combined evidence from nitrate, tritium, tritiogenic 3He, and radiogenic 4He concentrations, reveals complex recharge and flow to the capture zone of the deep drinking water wells. Widespread groundwater pumping for irrigation accelerates vertical groundwater flow such that high nitrate groundwater reaches some deep drinking water wells. Deeper portions of the wells often draw in water that recharged

MStern Blotting–High Throughput Polyvinylidene Fluoride (PVDF) Membrane-Based Proteomic Sample Preparation for 96-Well Plates*

OpenAIRE

Berger, Sebastian T.; Ahmed, Saima; Muntel, Jan; Cuevas Polo, Nerea; Bachur, Richard; Kentsis, Alex; Steen, Judith; Steen, Hanno

2015-01-01

We describe a 96-well plate compatible membrane-based proteomic sample processing method, which enables the complete processing of 96 samples (or multiples thereof) within a single workday. This method uses a large-pore hydrophobic PVDF membrane that efficiently adsorbs proteins, resulting in fast liquid transfer through the membrane and significantly reduced sample processing times. Low liquid transfer speeds have prevented the useful 96-well plate implementation of FASP as a widely used mem...

The Selectivity of Milking of Dunaliella salina

NARCIS (Netherlands)

Kleinegris, D.M.M.; Janssen, M.G.J.; Brandenburg, W.A.; Wijffels, R.H.

2010-01-01

The process of the simultaneous production and extraction of carotenoids, milking, of Dunaliella salina was studied. We would like to know the selectivity of this process. Could all the carotenoids produced be extracted? And would it be possible to vary the profile of the produced carotenoids and,

Simple rapid methods for freezing hybridomas in 96-well microculture plates.

Science.gov (United States)

Wells, D E; Price, P J

1983-04-15

Macroscopic hybridoma colonies were frozen and recovered in a good state of viability in 96-well microculture plates using 2 freezing procedures. These methods offer convenient and rapid means of preserving hybridomas and will permit laboratories developing monoclonal antibodies to distribute workloads to more manageable levels without discarding possibly valuable hybridomas.

Contribution of Co2+ in increasing chlorophyll a concentration of Nannochloropsis salina in controlled Conwy medium

Science.gov (United States)

Hala, Y.; Taba, P.; Suryati, E.; Kasih, P.; Firman, N. F.

2018-03-01

A research in determining the contribution of Co2+ on the increase of chlorophyll a concentration of Nannochloropsis salina has been caried out. The cultivation of N. salina was conducted in the Conwy medium with a salinity of 5%o and 25%o and various Co2+ concentration (2, 4, and 8 ppm). In this research, Co2+ was exposed early in the cultivation of N. salina. The growth of N. salina was observed daily by counting the number of populations using a haemocytometer while the chlorophyll a concentration was determined by a Uv-Vis spectrophotometer. The results showed that the growth of N. salina in the control was higher than that in the medium containing Co2+. The optimum growth time was achieved on 15th days (5%) and 8th days (25%). In the cultivation medium with a salinity of 5%, Co2+ with a concentration of 2 ppm increased the chlorophyll a level while Co2+ with concentrations of 4 and 8 ppm decreased it. In the medium of cultivation with a salinity of 25%, the increase in chlorophyll a level was observed at Co2+ concentrations of 2 and 4 ppm whereas the decrease in chlorophyl a level was given at a concentration of 8 ppm. It can be concluded that at low concentrations, Co2+ increased the concentration of chlorophyll a in N. salina.

A High Throughput, 384-Well, Semi-Automated, Hepatocyte Intrinsic Clearance Assay for Screening New Molecular Entities in Drug Discovery.

Science.gov (United States)

Heinle, Lance; Peterkin, Vincent; de Morais, Sonia M; Jenkins, Gary J; Badagnani, Ilaria

2015-01-01

A high throughput, semi-automated clearance screening assay in hepatocytes was developed allowing a scientist to generate data for 96 compounds in one week. The 384-well format assay utilizes a Thermo Multidrop Combi and an optimized LC-MS/MS method. The previously reported LCMS/ MS method reduced the analytical run time by 3-fold, down to 1.2 min injection-to-injection. The Multidrop was able to deliver hepatocytes to 384-well plates with minimal viability loss. Comparison of results from the new 384-well and historical 24-well assays yielded a correlation of 0.95. In addition, results obtained for 25 marketed drugs with various metabolism pathways had a correlation of 0.75 when compared with literature values. Precision was maintained in the new format as 8 compounds tested in ≥39 independent experiments had coefficients of variation ≤21%. The ability to predict in vivo clearances using the new stability assay format was also investigated using 22 marketed drugs and 26 AbbVie compounds. Correction of intrinsic clearance values with binding to hepatocytes (in vitro data) and plasma (in vivo data) resulted in a higher in vitro to in vivo correlation when comparing 22 marketed compounds in human (0.80 vs 0.35) and 26 AbbVie Discovery compounds in rat (0.56 vs 0.17), demonstrating the importance of correcting for binding in clearance studies. This newly developed high throughput, semi-automated clearance assay allows for rapid screening of Discovery compounds to enable Structure Activity Relationship (SAR) analysis based on high quality hepatocyte stability data in sufficient quantity and quality to drive the next round of compound synthesis.

Salinas de interior en el territorio de la Región de Murcia

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S. Gil Guirado

2010-01-01

Full Text Available La sal es un elemento necesario en la actividad de los grupos humanos. Para su captación y recolección se ha ordenado el territorio, con unos paisajes propios: las salinas. Podemos distinguir dos tipos: las salinas costeras y las salinas de interior. En estas últimas, su localización y explotación está ligada a la presencia de algún material salino que atraviesa el agua. En ellas se pueden distinguir tres áreas, la de captación del agua (generalmente pozos horizontales del tipo minado o galería; la de almacenaje (con balsas y recocederos dónde acumular y calentar; y finalmente la de cristalización (con parcelas más o menos aterrazadas conocidas como eras. A través del estudio combinado del trabajo de campo y la documentación de archivos, hemos podido localizar una veintena de salinas en el interior de la Región de Murcia; huella de una actividad económica y de una cultura del agua entre la variedad de paisajes surestinos.

Metabolic responses and β-carotene production by the unicellular green alga Dunaliella salina exposed to leaf extracts

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Alireza Einali

Full Text Available ABSTRACT The present work investigated the effects of aqueous extracts of eucalyptus ( Eucalyptus globulus and elderberry ( Sambucus ebulus leaves on β-carotene productivity in Dunaliella salina, a green microalga. Leaf extracts from eucalyptus have greater amounts of phenolics and flavonoids, as well as greater ferric reducing antioxidant potential than elderberry. The extracts of both species greatly inhibited growth of algal suspensions. However, chlorophyll and β-carotene concentration increased in cells treated with leaf extracts, and the highest values were detected in 1 % eucalyptus and 2 % elderberry extracts. Fresh weight, total sugar, and protein content significantly increased following exposure of cells to different doses of leaf extracts. However, in doses containing more than 2 % eucalyptus, the upward trend for total sugar and protein ceased and remained statistically unchanged. These results suggest that metabolic modifications enable D. salina cells to tolerate the stress induced by the leaf extracts through allocating carbon flux to the synthesis of osmolytes and putative antioxidant molecules (e.g. sugars and β-carotene. Therefore, the use of leaf extracts holds potential to be a promising and effective way to improve D. salina cultivation for β-carotene production and other biotechnological and industrial applications.

Cultivation of Nannochloropsis salina using anaerobic digestion effluent as a nutrient source for biofuel production

International Nuclear Information System (INIS)

Cai, Ting; Park, Stephen Y.; Racharaks, Ratanachat; Li, Yebo

2013-01-01

Highlights: • Cultivation of Nannochloropsis salina with effluent of anaerobic digestion (AD). • The highest biomass yield was obtained at 6% AD effluent loading. • Lipid content and productivity decreased with increased effluent loading from 3% to 18%. • Biomass productivity increased by up to 49% as harvest ratio increased from 25% to 50%. - Abstract: The biomass and lipid productivities and the nutrient removal capacity of microalgae Nannochloropsis salina grown using anaerobically digested municipal wastewater effluent as a nutrient source were evaluated in this study. Results from bench-scale batch reactors showed that N. salina grew well under 3%, 6%, 12%, and 18% (v/v) anaerobic digestion (AD) effluent loading with the highest growth rate being 0.645 d −1 obtained at 6% AD effluent loading. The growth of N. salina decreased when the effluent loading was increased to 24%. The highest biomass productivity of 92 mg l −1 d −1 was obtained with 6% effluent loading. Three harvesting frequencies (1, 2, and 3 d intervals) and two harvesting ratios (25% and 50%, v/v) were tested in semi-continuous bench-scale reactors with 6% effluent loading. The highest lipid productivity of 38.7 mg l −1 d −1 was achieved with a 2-d harvesting interval and 50% harvesting ratio, where nitrogen and phosphorus were removed at rates of 35.3 mg l −1 d −1 and 3.8 mg l −1 d −1 , respectively. The fatty acid (FA) profile showed that palmitic acid (C16:0), palmitoleic acid (C16:1), and eicosapentaenoic acid (C20:5) were the major components, accounting for 32.1%, 26%, and 15.7% of the total FAs, respectively

High-throughput screening of carbohydrate-degrading enzymes using novel insoluble chromogenic substrate assay kits

DEFF Research Database (Denmark)

Schückel, Julia; Kracun, Stjepan Kresimir; Willats, William George Tycho

2016-01-01

for this is that advances in genome and transcriptome sequencing, together with associated bioinformatics tools allow for rapid identification of candidate CAZymes, but technology for determining an enzyme's biochemical characteristics has advanced more slowly. To address this technology gap, a novel high-throughput assay...... CPH and ICB substrates are provided in a 96-well high-throughput assay system. The CPH substrates can be made in four different colors, enabling them to be mixed together and thus increasing assay throughput. The protocol describes a 96-well plate assay and illustrates how this assay can be used...... for screening the activities of enzymes, enzyme cocktails, and broths....

42 CFR 9.6 - Animal care, well-being, husbandry, veterinary care, and euthanasia.

Science.gov (United States)

2010-10-01

... 42 Public Health 1 2010-10-01 2010-10-01 false Animal care, well-being, husbandry, veterinary care... SYSTEM § 9.6 Animal care, well-being, husbandry, veterinary care, and euthanasia. (a) What are the... chimpanzees can be trained through positive reinforcement to cooperate with a variety of veterinary and...

Neutral lipid production in Dunaliella salina during osmotic stress and adaptation

DEFF Research Database (Denmark)

Yao, Shuo; Lu, Jingquan; Sárossy, Zsuzsa

2016-01-01

The salt-tolerant green microalga Dunaliella salina can survive both hyper- and hypo-osmotic shock. Upon osmotic shock, the cells transiently and rapidly decreased or increased in size within minutes and slowly over hours acquired their original cell size and volume. Cell size distribution differs...... significantly in the cultures grown in the salinity range from 1.5 to 15 % NaCl. By using Nile Red fluorescence to detect neutral lipids, it became clear that only hyper-osmotic shock on cells induced transient neutral lipid appearance in D. salina, while those transferred from 9 to 15 % NaCl stimulated...

Larvicidal activity of some marine macrophytes against Artemia salina

Digital Repository Service at National Institute of Oceanography (India)

PrabhaDevi; Wahidullah, S.; DeSouza, L.; Kamat, S.Y.

and Microdictyon pseudohapteron, seagrasses (Halophila ovalis and Syringodium isoetifolium and lichens (Umbilicaria arpina) were tested for larvicidal activity using Artemia salina nauplii. The LC50 values for the PE-fraction of M. pseudohapteron and A. muscoides...

Antifouling potential of the marine microalga Dunaliella salina.

Science.gov (United States)

Gao, Min; Li, Fengchao; Su, Rongguo; Wang, Ke; Li, Xuzhao; Lu, Wei

2014-11-01

Marine organisms have usually been viewed as sources of environmentally friendly compounds with antifouling activity. We performed a series of operations to investigate the antifouling potential of the marine microalga Dunaliella salina. For the ethyl acetate crude extract, the antialgal activity was significant, and the EC50 value against Skeletonema costatum was 58.9 μg ml(-1). The isolated purified extract was tested for antifouling activity, the EC 50 value against S. costatum was 21.2 μg ml(-1), and the LC50 against Balanus amphitrite larvae was 18.8 μg ml(-1). Subsequently, both UHR-TOF-MS and GC-MS were used for the structural elucidation of the compounds, and a series of unsaturated and saturated 16- and 18-carbon fatty acids were detected. The data suggested that the fatty acid extracts from D. salina possess high antifouling activity, and could be used as substitutes for potent, toxic antifouling compounds.

A century of plant virus management in the Salinas valley of California, 'East of Eden'.

Science.gov (United States)

Wisler, G C; Duffus, J E

2000-11-01

The mild climate of the Salinas Valley, CA lends itself well to a diverse agricultural industry. However, the diversity of weeds, crops and insect and fungal vectors also provide favorable conditions for plant virus disease development. This paper considers the incidence and management of several plant viruses that have caused serious epidemics and been significant in the agricultural development of the Salinas Valley during the 20th century. Beet curly top virus (BCTV) almost destroyed the newly established sugarbeet industry soon after its establishment in the 1870s. A combination of resistant varieties, cultural management of beet crops to provide early plant emergence and development, and a highly coordinated beet leafhopper vector scouting and spray programme have achieved adequate control of BCTV. These programmes were first developed by the USDA and still operate. Lettuce mosaic virus was first recognized as causing a serious disease of lettuce crops in the 1930s. The virus is still a threat but it is controlled by a lettuce-free period in December and a seed certification programme that allows only seed lots with less than one infected seed in 30000 to be grown. 'Virus Yellows' is a term used to describe a complex of yellows inducing viruses which affect mainly sugarbeet and lettuce. These viruses include Beet yellows virus and Beet western yellows virus. During the 1950s, the complex caused significant yield losses to susceptible crops in the Salinas Valley. A beet-free period was introduced and is still used for control. The fungus-borne rhizomania disease of sugarbeet caused by Beet necrotic yellow vein virus was first detected in Salinas Valley in 1983. Assumed to have been introduced from Europe, this virus has now become widespread in California wherever beets are grown and crop losses can be as high as 100%. Movement of infested soil and beets accounts for its spread throughout the beet-growing regions of the United States. Control of rhizomania

Development of resazurin-based assay in 384-well format for high throughput whole cell screening of Trypanosoma brucei rhodesiense strain STIB 900 for the identification of potential anti-trypanosomal agents.

Science.gov (United States)

Lim, Kah Tee; Zahari, Zuriati; Amanah, Azimah; Zainuddin, Zafarina; Adenan, Mohd Ilham

2016-03-01

To accelerate the discovery of novel leads for the treatment of Human African Trypanosomiasis (HAT), it is necessary to have a simple, robust and cost-effective assay to identify positive hits by high throughput whole cell screening. Most of the fluorescence assay was made in black plate however in this study the HTS assay developed in 384-well format using clear plate and black plate, for comparison. The HTS assay developed is simple, sensitive, reliable and reproducible in both types of plates. Assay robustness and reproducibility were determined under the optimized conditions in 384-well plate was well tolerated in the HTS assay, including percentage of coefficient of variation (% CV) of 4.68% and 4.74% in clear and black 384-well plate, signal-to-background ratio (S/B) of 12.75 in clear 384-well plate and 12.07 in black 384-well plate, Z' factor of 0.79 and 0.82 in clear 384-well plate and black 384-well plate, respectively and final concentration of 0.30% dimethylsulfoxide (DMSO) in both types of plate. Drug sensitivity was found to be comparable to the reported anti-trypanosomal assay in 96-well format. The reproducibility and sensitivity of this assay make it compliant to automated liquid handler use in HTS applications. Copyright © 2016 Elsevier Inc. All rights reserved.

Evaluation of the photodynamic activity of Xanthene Dyes on Artemia salina described by chemometric approaches

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Diogo S. Pellosi

2013-10-01

Full Text Available The development of drugs for photodynamic therapy (PDT is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i photosensitizers concentration, (ii the LED irradiation time and (iii the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 23 full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

INFLUENCIA DE LA SALINIDAD Y LA IRRADIANCIA SOBRE EL CRECIMIENTO Y COMPOSICIÓN BIOQUÍMICA DE UNA NUEVA CEPA DE Dunaliella salina, PROVENIENTE DE LAS SALINAS DE ARAYA, VENEZUELA | INFLUENCE OF SALINITY AND IRRADIANCE ON GROWTH AND BIOCHEMICAL COMPOSITION OF A NEW STRAIN of Dunaliella salina FROM THE ARAYA

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Miguel Guevara

2016-08-01

Full Text Available Dunaliella salina is a microalga used for the production of metabolites of high industrial and pharmaceutical value. The combined effect of irradiance and salinity on growth, pigment production and biochemical composition of this strain were evaluated in order to contribute to the knowledge of the physiological responses of this new isolate of D. salina from Araya saltworks, to changes in the conditions of its environment. It was aslo intendend to increase the variety of native microalgae species which can serve as sources of bioproduction for bioactive compounds, biomass and metabolites of application in pharmaceuticals and biomedical industries. Batch cultures of the strain of D. salina were conducted for 22 d at f/2 medium Guillard (0.88 mmol.L-1 of nitrogen at 27 ± 2°C, pH 7.8; photoperiod 12 h light: 12 h darkness, continuous aeration (150 mL.L-1 in two salinities (40 and 250 UPS and irradiances (195 and 295 µmolphotons.m-2.s-1. Population growth was evaluated daily and at the end of the trial the contents of biomass, pigments, proteins, lipids, carbohydrates and cell size were determined. The growth rate of the new strain of D. salina decreased with increasing salinity and irradiance. However, cell size, biomass, content of proteins, lipids, carbohydrates, total carotenoids and β-carotene increased with increasing irradiance and salinity. In addition, the potential of this strain for the production of β-carotene was shown.

High-Throughput Lipolysis in 96-Well Plates for Rapid Screening of Lipid-Based Drug Delivery Systems

DEFF Research Database (Denmark)

Mosgaard, Mette D; Sassene, Philip J; Mu, Huiling

2017-01-01

The high-throughput in vitro intestinal lipolysis model (HTP) applicable for rapid and low-scale screening of lipid-based drug delivery systems (LbDDSs) was optimized and adjusted as to be conducted in 96-well plates (HTP-96). Three different LbDDSs (I-III) loaded with danazol or cinnarizine were...

The fluorometric microculture cytotoxicity assay.

Science.gov (United States)

Lindhagen, Elin; Nygren, Peter; Larsson, Rolf

2008-01-01

The fluorometric microculture cytotoxicity assay (FMCA) is a nonclonogenic microplate-based cell viability assay used for measurement of the cytotoxic and/or cytostatic effect of different compounds in vitro. The assay is based on hydrolysis of the probe, fluorescein diacetate (FDA) by esterases in cells with intact plasma membranes. The assay is available as both a semiautomated 96-well plate setup and a 384-well plate version fully adaptable to robotics. Experimental plates are prepared with a small amount of drug solution and can be stored frozen. Cells are seeded on the plates and cell viability is evaluated after 72 h. The protocol described here is applicable both for cell lines and freshly prepared tumor cells from patients and is suitable both for screening in drug development and as a basis for a predictive test for individualization of anticancer drug therapy.

[Rhodomonas salina (Cryptophyta) pastes as feed for Brachionus plicatilis (Rotifera)].

Science.gov (United States)

Guevara, Miguel; Bastardo, Leandro; Cortez, Roraysi; Arredondo-Vega, Bertha; Romero, Lolymar; Gómez, Patricia

2011-12-01

Rotifers are an important live feed for first feeding larvae of many fish species. The use of concentrated algae cells in the mass culture of the rotifer Brachionus plicatilis (Brachionidae) has opened new horizons for research on this organism. Pastes of Rhodomonas salina (Pyrenomonadaceae) obtained either by centrifugation or flocculation with chitosan were preserved, with or without vitamin C, at -20 degrees C for four weeks and were evaluated biochemically (proteins, lipids, pigments and fatty acids contents) and subsequently, were used to feed the rotifer Brachionus plicatilis at a ratio of 25 mg/L/day. Four different microalgae pastes were prepared: (1) centrifuged and preserved with vitamin C (CV), (2) centrifuged and preserved without vitamin C (C), (3) flocculated and with vitamin C (FV) and (4) flocculated without vitamin C (F). All treatments showed similar contents of proteins and total lipids with respect to control culture (a fresh culture of R. salina), with mean values of 40.0 +/- 2.32% and 12.0 +/- 1.45%, respectively. The pheophytin a/chlorophyll a ratio, a general indicator of the chemical status of microalgal concentrates, was similar (0.09-0.11) between centrifuged pastes and control culture, but was found to be higher in flocculated pastes (1.28-1.48). The fatty acid profile varied with respect to the control culture, mainly in the proportion of the essential polyunsaturated fatty acids (PUFAs): eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Total PUFAs, EPA and DHA contents were statistically similar between centrifuged pastes and control culture (PUFAs: 47%, EPA: 4% and DHA: 4.7%), whereas values obtained for flocculated pastes were significantly lower. The rotifers grew equally well when fed with centrifuged pastes or control culture (maximum density: 320 rotifers/mL; instantaneous growth rate: 0.23 rotifers/day, fecundity: 1.49 eggs/female and productivity: 43 x 10(3) rotifers/L/day. No significant effect of vitamin C was

Phenotypic and genetic characterization of Dunaliella (Chlorophyta) from Indian salinas and their diversity

Science.gov (United States)

2012-01-01

Background The genus Dunaliella (Class – Chlorophyceae) is widely studied for its tolerance to extreme habitat conditions, physiological aspects and many biotechnological applications, such as a source of carotenoids and many other bioactive compounds. Biochemical and molecular characterization is very much essential to fully explore the properties and possibilities of the new isolates of Dunaliella. In India, hyper saline lakes and salt pans were reported to bloom with Dunaliella spp. However, except for the economically important D. salina, other species are rarely characterized taxonomically from India. Present study was conducted to describe Dunaliella strains from Indian salinas using a combined morphological, physiological and molecular approach with an aim to have a better understanding on the taxonomy and diversity of this genus from India. Results Comparative phenotypic and genetic studies revealed high level of diversity within the Indian Dunaliella isolates. Species level identification using morphological characteristics clearly delineated two strains of D. salina with considerable β-carotene content (>20 pg/cell). The variation in 18S rRNA gene size, amplified with MA1-MA2 primers, ranged between ~1800 and ~2650 base pairs, and together with the phylogeny based on ITS gene sequence provided a pattern, forming five different groups within Indian Dunaliella isolates. Superficial congruency was observed between ITS and rbcL gene phylogenetic trees with consistent formation of major clades separating Indian isolates into two distinct clusters, one with D. salina and allied strains, and another one with D. viridis and allied strains. Further in both the trees, few isolates showed high level of genetic divergence than reported previously for Dunaliella spp. This indicates the scope of more numbers of clearly defined/unidentified species/sub-species within Indian Dunaliella isolates. Conclusion Present work illustrates Indian Dunaliella strains

Determination of 54 pesticides in waters of the Iberian Douro River estuary and risk assessment of environmentally relevant mixtures using theoretical approaches and Artemia salina and Daphnia magna bioassays.

Science.gov (United States)

Cruzeiro, Catarina; Amaral, Sofia; Rocha, Eduardo; Rocha, Maria João

2017-11-01

As a case study, the estuary of the international Douro River (Iberian Peninsula) was sampled over a year (2010) at six sampling sites to determine the presence of 56 pesticides of different categories (insecticides, herbicides, and fungicides). 96% of measured pesticides were detected in 79% of the quantified samples. Individual average pesticide concentrations ranged from 39 to 1 265ng/L, indicating a ubiquitous presence of the selected compounds; moreover, twelve pesticides were above the 2013/39/EU Directive limits. Due to its highly impacted profile, a theoretical hazard assessment was done considering the average and maximum environmental mixtures of all measured pesticides to identify the most sensitive trophic level. For both environmental mixtures, the theoretical approach suggested that invertebrates were the most sensitive group. Therefore, short-time exposure assays using both invertebrates Artemia salina and Daphnia magna, were done using the referred mixtures. Data demonstrated significant toxic effects ─ high mortality rate and abnormal swimming behaviour ─ of the exposed animals. Both approaches (theoretical and experimental) support the analytical results, alerting for an intervention on this estuarine environment and of other comparable. Copyright © 2017 Elsevier Inc. All rights reserved.

Setting the conditions for phycoremediation of radionuclide microalgae Dunaliella salina and Chlorella vulgaris

International Nuclear Information System (INIS)

Tatarova, D.; Galanda, D.; Kuruc, J.

2016-01-01

This presentation deals with bioremediation using microalgae - by phycoremediation. Microalgae are economically low profile compared to the plants, their cultivation can be carried out in laboratory conditions. They can survive in extreme conditions, they occur in all habitats and have faster growth. Halophilous green D. salina can accumulate heavy metals such as Zn, Cu and Cd. It occurs in hypersaline environment with tolerance (0.2 to 35) % NaCl. It contains high amounts of carotenoids, which protect it against formation of free radicals from UV radiation. Chlorella vulgaris is a representative of eukaryotic green microalgae with the highest chlorophyll content with the appearance in fresh water. Its phycoremediant ability are found in N and P elements, which are used as its nutritional components as well as for Cu, Cr, Cd, Pb, Au. The experiments were carried out using a peristaltic pump ISMATEC Model: ISM851 (flow rate 2 cm"3 min"-"1) followed by monitoring of time dependence of decrease of activity of the microalgae solutions. For evaluation of the samples was used HPGe gamma spectrometer (measurement time of the samples: 600 sec) from ORTEC Company and measured spectra were evaluated with software GammaVision from ORTEC. The measured results showed that the most effective phycoremediation of microalgae Dunaliella salina toke place in an environment of pH 3, and even more at pH 8. The fact that the D. salina is able of phycoremediation at so acidic pH can contribute to its applications in extreme conditions or in the coastal areas in view of that it is halophilic. At freshwater microalgae Chlorella vulgaris was found the best phytoremediation potential in its natural environment at pH of 6. Because this microalgae is freshwater, it may find application in inland or in liquid radioactive waste from nuclear facilities.(authors)

Studies on experimental culture of a marine ciliate Fabrea salina

Digital Repository Service at National Institute of Oceanography (India)

Rattan, R.; Ansari, Z.A.; Chatterji, A.

Studies were conducted on the culture of a marine ciliate, Fabrea salina in the laboratory condition. Three types of inert feed; commercial yeast, fermented wheat bran and fermented rise bran were tested to study their suitability as artificial feed...

Effect of salinity on the quantity and quality of carotenoids accumulated by Dunaliella salina (strain CONC-007 and Dunaliella bardawil (strain ATCC 30861 Chlorophyta

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PATRICIA I GÓMEZ

2003-01-01

Full Text Available Dunaliella salina and D. bardawil are well-known microalgae accumulating high levels of b-carotene under growth-limiting conditions. In both taxa, this pigment is primarily composed of the isomers 9-cis and all-trans. The 9-cis b-carotene occurs only in natural sources and is the most attractive from a commercial point of view. The conditions that enhance the preferred accumulation of 9-cis b-carotene in D. salina are controversial and they have not been well established yet. This study examined the effect of salinity on the quantity and quality of total carotenoids and b-carotene isomers accumulated by D. salina (strain CONC-007 and D. bardawil (strain ATCC 30861 grown in two media with different nutritional compositions (PES and ART and at salt concentrations of 1M, 2M and 3M NaCl. Total carotenoids were determined by spectrophotometry and b-carotene isomers, by HPLC. The highest carotenoid contents per cell were obtained at 2M NaCl in both taxa. In both media, an increase of the 9-cis/all-trans b-carotene ratio was observed in D. bardawil when the salt concentration increased, with a maximum value of 2.6 (in ART medium at 3M NaCl. In D. salina this ratio did not exhibit the same pattern, and the salt concentrations for maximal ratios were different in both media. The highest ratio obtained for this strain was 4.3 (in ART medium at 2M NaCl.

Phototoxicity activity of Psoralea drupacea L. using Atremia salina bioassay system

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Mohammad Ramezani

2011-07-01

Conclusion: The result showed that P. drupacea methanolic extract and chloroform fraction have phototoxicity in A. salina bioassay system and their toxic effect is related to phototoxic constituents such as psoralen.

Em torno da luz cristalina: clara voz de Pedro Salinas

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Cristiano Moreira

2015-06-01

Full Text Available http://dx.doi.org/10.5007/2176-8552.2016n20p147 O texto que segue pretende ler a poesia de Pedro Salinas e Jorge Guillén diante de uma perspectiva que faz da voz uma iluminação e uma resistência diante do tempo histórico e da predominância de uma leitura simples das imagens. Assim, alguns poemas escolhidos permitem uma leitura contemporânea a respeito da voz, seguindo o conceito de Stimmung recuperado por Giorgio Agamben e as lições de poesia de Paul Valéry, e formam aspectos imprescindíveis para ler a poesia destes poetas. Ao manter em sua produção uma conversação com seus predecessores, Pedro Salinas joga com o logos e faz da phoné matéria prima para sua obra.

Growth effects on mixed culture of Dunaliella salina and ...

African Journals Online (AJOL)

Dunaliella salina and Phaeodactylum tricornutum are two important marine microalgae rich in bioactive substances and other high-value constituents. In this study, growth effects on mixed culture of these two microalgae were studied under different inoculation proportions (10:0, 7:3, 5:5, 3:7, 0:10) and low, medium and high ...

Feeding adult of Artemia salina (Crustacea-Branchiopoda on the dinoflagellate Gyrodinium corsicum (Gymnodiniales and the Chryptophyta Rhodomonas baltica

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Rauquírio André Albuquerque Marinho da Costa

2005-07-01

Full Text Available Experiments were carried out on feeding performance and survival rates of adult Artemia salina exposed to no axenic strains of the dinoflagellate Gyrodinium corsicum and of the Chryptophyta Rhodomonas baltica. Filtration rates on R. baltica and G. corsicum varied from 3.35 to 7.14 ml.artemia-1.h-1 and from 2.97 to 15.86 ml.artemia-1.h-1, respectively. The ingestion rates observed for A. salina did not indicate any digestive dysfunction or physiological impairment for organisms fed on G. corsicum and their functional response were similar to those observed for other organisms like copepod fed on different food concentrations. Mortality rates oscillated from 2.5% to 100% when A. salina was fed on R. baltica or G. corsicum, respectively. Highest mortality rates observed for organisms fed on G. corsicum indicated that this dinoflagellate presented a hazard effect on A. salina that was not possible to confirm if it was related to toxin production or to nutritive inadequacy of this dinoflagellate as food for organisms of this species.Experimentos foram desenvolvidos para estudar as taxas de alimentação e de sobrevivência de Artemia salina alimentada com cepas não tóxicas do dinoflagelado Gyrodinium corsicum e da Chryptophyta Rhodomonas baltica. As taxas de filtração sobre R. baltica e G. corsicum variaram entre 3,35 e 7,14 ml.artemia-1.h-1 e 2,97 e 15,86 ml.artemia-1.h-, respectivamente. As taxas de ingestão observadas para A. salina não indicaram disfunção digestiva ou prejuízo fisiológico nos organismos alimentados com G. corsicum, sendo a resposta funcional destes organismos similar a observada em copépodos alimentados com diferentes concentrações de alimento. As taxas de mortalidade de A. salina oscilaram entre 2,5 e 100% quando alimentada com R. baltica e G. corsicum, respectivamente. As maiores taxas de mortalidade observadas para os organismos alimentados com G. corsicum indicam que este dinoflagelado apresenta algum efeito

MStern Blotting-High Throughput Polyvinylidene Fluoride (PVDF) Membrane-Based Proteomic Sample Preparation for 96-Well Plates.

Science.gov (United States)

Berger, Sebastian T; Ahmed, Saima; Muntel, Jan; Cuevas Polo, Nerea; Bachur, Richard; Kentsis, Alex; Steen, Judith; Steen, Hanno

2015-10-01

We describe a 96-well plate compatible membrane-based proteomic sample processing method, which enables the complete processing of 96 samples (or multiples thereof) within a single workday. This method uses a large-pore hydrophobic PVDF membrane that efficiently adsorbs proteins, resulting in fast liquid transfer through the membrane and significantly reduced sample processing times. Low liquid transfer speeds have prevented the useful 96-well plate implementation of FASP as a widely used membrane-based proteomic sample processing method. We validated our approach on whole-cell lysate and urine and cerebrospinal fluid as clinically relevant body fluids. Without compromising peptide and protein identification, our method uses a vacuum manifold and circumvents the need for digest desalting, making our processing method compatible with standard liquid handling robots. In summary, our new method maintains the strengths of FASP and simultaneously overcomes one of the major limitations of FASP without compromising protein identification and quantification. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

New low-viscosity overlay medium for viral plaque assays

Directory of Open Access Journals (Sweden)

Garten Wolfgang

2006-08-01

Full Text Available Abstract Background Plaque assays in cell culture monolayers under solid or semisolid overlay media are commonly used for quantification of viruses and antiviral substances. To overcome the pitfalls of known overlays, we tested suspensions of microcrystalline cellulose Avicel RC/CL™ as overlay media in the plaque and plaque-inhibition assay of influenza viruses. Results Significantly larger plaques were formed under Avicel-containing media, as compared to agar and methylcellulose (MC overlay media. The plaque size increased with decreasing Avicel concentration, but even very diluted Avicel overlays (0.3% ensured formation of localized plaques. Due to their low viscosity, Avicel overlays were easier to use than methylcellulose overlays, especially in the 96-well culture plates. Furthermore, Avicel overlay could be applied without prior removal of the virus inoculum thus facilitating the assay and reducing chances of cross-contamination. Using neuraminidase inhibitor oseltamivir carboxylate, we demonstrated applicability of the Avicel-based plaque reduction assay for testing of antiviral substances. Conclusion Plaque assay under Avicel-containing overlay media is easier, faster and more sensitive than assays under agar- and methylcellulose overlays. The assay can be readily performed in a 96-well plate format and seems particularly suitable for high-throughput virus titrations, serological studies and experiments on viral drug sensitivity. It may also facilitate work with highly pathogenic agents performed under hampered conditions of bio-safety labs.

Prospecção fitoquímica de Sonchus oleraceus e sua toxicidade sobre o microcrustáceo Artemia salina Phytochemical prospecting of Sonchus oleraceus and its toxicity to Artemia salina

Directory of Open Access Journals (Sweden)

J.M. Lima

2009-03-01

Full Text Available A espécie vegetal Sonchus oleraceus é uma planta daninha presente em diversas culturas no Brasil e de utilização na medicina popular. Neste trabalho, realizou-se a prospecção fitoquímica dessa espécie com extratos em etanol, água e diclorometano, bem como testes de toxicidade sobre o microcrustáceo Artemia salina. O extrato aquoso apresentou em sua composição açúcares redutores, compostos fenólicos, taninos, flavonóides e cumarinas. No extrato etanólico, observaram-se os mesmos compostos qualificados no extrato aquoso, com exceção de cumarinas. Em diclorometano, verificou se a presença de saponinas, derivados triterpênicos e esteróides. No teste de toxicidade sobre Artemia salina, os dados convergiram para frações de extrato aquoso de 5.117,2 ppm, indicando ser um extrato de baixa toxicidade.Sonchus oleraceus is a common weed in Brazil, also used as a medicinal plant. Phytochemical prospecting of this species was carried out in this work using extracts obtained in ethanol, water and dichloromethane. A toxicity study of the aqueous extract was also conducted, using the micro crustaceous Artemia salina. The aqueous extract presented sugar reducers, phenolic compounds, tannins, flavonoids and coumarins in its composition. The compounds found in the aqueous extract were also detected in the ethanol extract, except for the coumarins. Saponin, triterpenes and steroids were identified in the dichloromethane extract. The toxicity test on Artemia salina pointed to aqueous extract fractions of 5,117.2 ppm, indicative of low toxicity.

Pastas de Rhodomonas salina (Cryptophyta como alimento para Brachionus plicatilis (Rotifera

Directory of Open Access Journals (Sweden)

Miguel Guevara

2011-12-01

biochemically (proteins, lipids, pigments and fatty acids contents and subsequently, were used to feed the rotifer Brachionus plicatilis at a ratio of 25mg/L/day. Four different microalgae pastes were prepared: (1 centrifuged and preserved with vitamin C (CV, (2 centrifuged and preserved without vitamin C (C, (3 flocculated and with vitamin C (FV and (4 flocculated without vitamin C (F. All treatments showed similar contents of proteins and total lipids with respect to control culture (a fresh culture of R. salina, with mean values of 40.0±2.32% and 12.0±1.45%, respectively. The pheophytin a/chlorophyll a ratio, a general indicator of the chemical status of microalgal concentrates, was similar (0.09-0.11 between centrifuged pastes and control culture, but was found to be higher in flocculated pastes (1.28-1.48. The fatty acid profile varied with respect to the control culture, mainly in the proportion of the essential polyunsaturated fatty acids (PUFAs: eicosapentaenoic acid (EPA and docosahexaenoic acid (DHA. Total PUFAs, EPA and DHA contents were statistically similar between centrifuged pastes and control culture (PUFAs: 47%, EPA: 4% and DHA: 4.7%, whereas values obtained for flocculated pastes were significantly lower. The rotifers grew equally well when fed with centrifuged pastes or control culture (maximum density: 320rotifers/mL; instantaneous growth rate: 0.23rotifers/day, fecundity: 1.49eggs/female and productivity: 43x103rotifers/L/day. No significant effect of vitamin C was found when used as a paste preservative. We concluded that centrifugation is an effective harvesting method, and that freezing to -20ºC for four weeks (no vitamin added, may help maintain the nutritional quality of R. salina paste, similar to fresh microalgae and can be offered to Brachionus plicatilis. Rev. Biol. Trop. 59 (4: 1503-1515. Epub 2011 December 01.

Heterologous overexpression of sfCherry fluorescent protein in Nannochloropsis salina

Directory of Open Access Journals (Sweden)

Nam Kyu Kang

2015-12-01

Full Text Available Oleaginous microalgae of the Nannochloropsis genus are considered excellent candidates for biofuels and value-added products owing to their high biomass productivity and lipid content. Here, we report the first overexpression and detection of a heterologous sfCherry fluorescent protein in Nannochloropsis salina in order to develop a transformation toolbox for future genetic improvements. Particle bombardment was employed for transformation, and expression of Shble under the control of TUB and UEP promoters, cloned from N. salina, was used to confer resistance to Zeocin antibiotics, resulting in 5.9 and 4.7 transformants per 108 cells, respectively. Stable integration of the markers into the genome was confirmed using a restriction enzyme site-directed amplification (RESDA PCR. The expression of sfCherry fluorescent protein was confirmed by Western blot analysis and confocal microscopy. These results suggest new possibilities of efficient genetic engineering of Nannochloropsis for the production of biofuels and other biochemicals.

Effect of enriched Brachionus plicatilis and Artemia salina nauplii by ...

African Journals Online (AJOL)

The growth, developmental stages and survival rates of Sparus aurata larvae fed with Brachionus plicatilis and Artemia salina nauplii enriched by microalga Tetraselmis chuii were studied. Two experiments were carried out; the first concerning with culturing the microalga (T. chuii) in four different media, then using these ...

Direct analysis of radionuclides-96 samples simultaneously

International Nuclear Information System (INIS)

Kessler, M.J.

1991-01-01

Recently, there has been a tremendous interest in two areas of concern in nuclear counting and radioactivity waste disposal. The first is the reduction of radioactive waste, in particular the reduction in the amount of or the development of environmentally safe scintillation cocktails. The second is the development of a simple method of quantitating large numbers of samples (thousands/day) in a short period of time (minutes). These two areas of concern have been addressed with the development of the Matrix 96 direct beta counter. This new instrumental technique is capable of quantitating 96 samples simultaneously in the microplate format (8 x 12, 96 sample) on a solid support WITHOUT the use of any cocktails, vials, and is non-destructive to the sample. The use of this technique for the following biomedical applications, DNA dot blots, cell proliferation (3H thymidine), receptor binding, chromium cytotoxicity assays, and protein assays will be discussed in detail. The data from both the conventional beta and gamma counter will be correlated and compared to the new Matrix 96 direct beta counter. This new technique provides a convenient method of addressing the concerns of reducing radioactive waste and provides a method of quantitating a large number of samples, accurately in a short period of time (96 at a time)

High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

Directory of Open Access Journals (Sweden)

Hamed Safafar

2016-07-01

Full Text Available Nannochloropsis salina was grown on a mixture of standard growth media and pre-gasified industrial process water representing effluent from a local biogas plant. The study aimed to investigate the effects of enriched growth media and cultivation time on nutritional composition of Nannochloropsis salina biomass, with a focus on eicosapentaenoic acid (EPA. Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scale using a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large-scale cultivation of Nannochloropsis salina at these conditions could improve the nutritional properties such as EPA, tocopherol, protein and carotenoids compared to laboratory-scale cultivated microalgae. EPA reached 44.2% ± 2.30% of total fatty acids, and α-tocopherol reached 431 ± 28 µg/g of biomass dry weight after 21 days of cultivation. Variations in chemical compositions of Nannochloropsis salina were studied during the course of cultivation. Nannochloropsis salina can be presented as a good candidate for winter time cultivation in Denmark. The resulting biomass is a rich source of EPA and also a good source of protein (amino acids, tocopherols and carotenoids for potential use in aquaculture feed industry.

Establishment and validation of a method for multi-dose irradiation of cells in 96-well microplates

International Nuclear Information System (INIS)

Abatzoglou, Ioannis; Zois, Christos E.; Pouliliou, Stamatia; Koukourakis, Michael I.

2013-01-01

Highlights: ► We established a method for multi-dose irradiation of cell cultures within a 96-well plate. ► Equations to adjust to preferable dose levels are produced and provided. ► Up to eight different dose levels can be tested in one microplate. ► This method results in fast and reliable estimation of radiation dose–response curves. -- Abstract: Microplates are useful tools in chemistry, biotechnology and molecular biology. In radiobiology research, these can be also applied to assess the effect of a certain radiation dose delivered to the whole microplate, to test radio-sensitivity, radio-sensitization or radio-protection. Whether different radiation doses can be accurately applied to a single 96-well plate to further facilitate and accelerated research by one hand and spare funds on the other, is a question dealt in the current paper. Following repeated ion-chamber, TLD and radiotherapy planning dosimetry we established a method for multi-dose irradiation of cell cultures within a 96-well plate, which allows an accurate delivery of desired doses in sequential columns of the microplate. Up to eight different dose levels can be tested in one microplate. This method results in fast and reliable estimation of radiation dose–response curves

Increased sensitivity of 3D-Well enzyme-linked immunosorbent assay (ELISA) for infectious disease detection using 3D-printing fabrication technology.

Science.gov (United States)

Singh, Harpal; Shimojima, Masayuki; Fukushi, Shuetsu; Le Van, An; Sugamata, Masami; Yang, Ming

2015-01-01

Enzyme-linked Immunosorbent Assay or ELISA -based diagnostics are considered the gold standard in the demonstration of various immunological reaction including in the measurement of antibody response to infectious diseases and to support pathogen identification with application potential in infectious disease outbreaks and individual patients' treatment and clinical care. The rapid prototyping of ELISA-based diagnostics using available 3D printing technologies provides an opportunity for a further exploration of this platform into immunodetection systems. In this study, a '3D-Well' was designed and fabricated using available 3D printing platforms to have an increased surface area of more than 4 times for protein-surface adsorption compared to those of 96-well plates. The ease and rapidity in designing-product development-feedback cycle offered through 3D printing platforms provided an opportunity for its rapid assessment, in which a chemical etching process was used to make the surface hydrophilic followed by validation through the diagnostic performance of ELISA for infectious disease without modifying current laboratory practices for ELISA. The higher sensitivity of the 3D-Well (3-folds higher) compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization platforms to reduce time, volume of reagents and samples needed for laboratory or field diagnosis of infectious diseases including applications in other disciplines.

Microfractionation revisited: a 1536 well high resolution screening assay

NARCIS (Netherlands)

Giera, M.A.; Heus, F.; Janssen, L.; Kool, J.; Lingeman, H.; Irth, H.

2009-01-01

The aim of the here presented study was to combine high performance liquid chromatography with plate reader technology in order to overcome certain drawbacks of integrated online systems as well as offline plate reader approaches. The described method combines an "at-line" enzyme assay for the

LIPID PRODUCTION BY DUNALIELLA SALINA IN BATCH CULTURE: EFFECTS OF NITROGEN LIMITATION AND LIGHT INTENSITY

Energy Technology Data Exchange (ETDEWEB)

Weldy, C.S.; Huesemann, M.

2007-01-01

Atmospheric carbon dioxide (CO2) concentrations are increasing and may cause unknown deleterious environmental effects if left unchecked. The Intergovernmental Panel on Climate Change (IPCC) has predicted in its latest report a 2°C to 4°C increase in global temperatures even with the strictest CO2 mitigation practices. Global warming can be attributed in large part to the burning of carbon-based fossil fuels, as the concentration of atmospheric CO2 is directly related to the burning of fossil fuels. Biofuels which do not add CO2 to the atmosphere are presently generated primarily from terrestrial plants, i.e., ethanol from corn grain and biodiesel from soybean oil. The production of biofuels from terrestrial plants is severely limited by the availability of fertile land. Lipid production from microalgae and its corresponding biodiesel production have been studied since the late 1970s but large scale production has remained economically infeasible due to the large costs of sterile growing conditions required for many algal species. This study focuses on the potential of the halophilic microalgae species Dunaliella salina as a source of lipids and subsequent biodiesel production. The lipid production rates under high light and low light as well as nitrogen suffi cient and nitrogen defi cient culture conditions were compared for D. salina cultured in replicate photobioreactors. The results show (a) cellular lipid content ranging from 16 to 44% (wt), (b) a maximum culture lipid concentration of 450mg lipid/L, and (c) a maximum integrated lipid production rate of 46mg lipid/L culture*day. The high amount of lipids produced suggests that D. salina, which can be mass-cultured in non-sterile outdoor ponds, has strong potential to be an economically valuable source for renewable oil and biodiesel production.

Smart SfM: Salinas Archaeological Museum

Science.gov (United States)

Inzerillo, L.

2017-08-01

In these last years, there has been an increasing use of the Structure from Motion (SfM) techniques applied to Cultural Heritage. The accessibility of SfM software can be especially advantageous to users in non-technical fields or to those with limited resources. Thanks to SfM using, everyone can make with a digital camera a 3D model applied to an object of both Cultural Heritage, and physically Environment, and work arts, etc. One very interesting and useful application can be envisioned into museum collection digitalization. In the last years, a social experiment has been conducted involving young generation to live a social museum using their own camera to take pictures and videos. Students of university of Catania and Palermo were involved into a national event #digitalinvasion (2015-2016 editions) offering their personal contribution: they realized 3D models of the museums collection through the SfM techniques. In particular at the National Archaeological Museum Salinas in Palermo, it has been conducted an organized survey to recognize the most important part of the archaeological collection. It was a success: in both #digitalinvasion National Event 2015 and 2016 the young students of Engineering classes carried out, with Photoscan Agisoft, more than one hundred 3D models some of which realized by phone camera and some other by reflex camera and some other with compact camera too. The director of the museum has been very impressed from these results and now we are going to collaborate at a National project to use the young generation crowdsourcing to realize a semi-automated monitoring system at Salinas Archaeological Museum.

SMART SfM: SALINAS ARCHAEOLOGICAL MUSEUM

Directory of Open Access Journals (Sweden)

L. Inzerillo

2017-08-01

Full Text Available In these last years, there has been an increasing use of the Structure from Motion (SfM techniques applied to Cultural Heritage. The accessibility of SfM software can be especially advantageous to users in non-technical fields or to those with limited resources. Thanks to SfM using, everyone can make with a digital camera a 3D model applied to an object of both Cultural Heritage, and physically Environment, and work arts, etc. One very interesting and useful application can be envisioned into museum collection digitalization. In the last years, a social experiment has been conducted involving young generation to live a social museum using their own camera to take pictures and videos. Students of university of Catania and Palermo were involved into a national event #digitalinvasion (2015-2016 editions offering their personal contribution: they realized 3D models of the museums collection through the SfM techniques. In particular at the National Archaeological Museum Salinas in Palermo, it has been conducted an organized survey to recognize the most important part of the archaeological collection. It was a success: in both #digitalinvasion National Event 2015 and 2016 the young students of Engineering classes carried out, with Photoscan Agisoft, more than one hundred 3D models some of which realized by phone camera and some other by reflex camera and some other with compact camera too. The director of the museum has been very impressed from these results and now we are going to collaborate at a National project to use the young generation crowdsourcing to realize a semi-automated monitoring system at Salinas Archaeological Museum.

Metabolic engineering of Dunaliella salina for production of ketocarotenoids.

Science.gov (United States)

Anila, N; Simon, Daris P; Chandrashekar, Arun; Ravishankar, G A; Sarada, R

2016-03-01

Dunaliella is a commercially important marine alga producing high amount of β-carotene. The use of Dunaliella as a potential transgenic system for the production of recombinant proteins has been recently recognized. The present study reports for the first time the metabolic engineering of carotenoid biosynthesis in Dunaliella salina for ketocarotenoid production. The pathway modification included the introduction of a bkt gene from H. pluvialis encoding β-carotene ketolase (4,4'β-oxygenase) along with chloroplast targeting for the production of ketocarotenoids. The bkt under the control of Dunaliella Rubisco smaller subunit promoter along with its transit peptide sequence was introduced into the alga through standardized Agrobacterium-mediated transformation procedure. The selected transformants were confirmed using GFP and GUS expression, PCR and southern blot analysis. A notable upregulation of the endogenous hydroxylase level of transformants was observed where the BKT expression was higher in nutrient-limiting conditions. Carotenoid analysis of the transformants through HPLC and MS analysis showed the presence of astaxanthin and canthaxanthin with maximum content of 3.5 and 1.9 µg/g DW, respectively. The present study reports the feasibility of using D. salina for the production of ketocarotenoids including astaxanthin.

MicroRNA-96 Promotes Tumor Invasion in Colorectal Cancer via RECK.

Science.gov (United States)

Iseki, Yasuhito; Shibutani, Masatsune; Maeda, Kiyoshi; Nagahara, Hisashi; Fukuoka, Tatsunari; Matsutani, Shinji; Hirakawa, Kosei; Ohira, Masaichi

2018-04-01

miR-96 is reported to inhibit reversion cysteine-rich Kazal motif (RECK), which is associated with tumor invasion, in solid cancer types (e.g. breast cancer, non-small cell lung cancer, esophageal cancer). The purpose of this study is to clarify whether miR-96 is similarly associated with tumor invasion in colorectal cancer. We performed western blotting to investigate the expression of RECK when miR-96 mimics or inhibitors were transferred into HCT-116 colorectal cancer cells. The RECK mRNA level was assessed by a reverse transcription polymerase chain reaction. An invasion assay was used to evaluate tumor invasion. The expression of RECK was inhibited by the transfection of miR-96 mimics. RECK mRNA level was reduced by miR-96 mimics and increased by miR-96 inhibitor. In the invasion assay, miR-96 mimics were shown to promote tumor invasion. miR-96 may be associated with tumor invasion through inhibition of RECK expression in colorectal cancer. Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

Nesting ecology of Pied Avocet Recurvirostra avosetta in Sfax salina ...

African Journals Online (AJOL)

In this paper we use the results of a one-year monitoring of nests in Sfax salina to provide information on its nesting parameters, in particular nesting phenology, colony size and hatching success. Our results show that Pied Avocets formed dense colonies at the beginning of the nesting season, but colony size decreased as ...

High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

DEFF Research Database (Denmark)

Safafar, Hamed; Hass, Michael Z.; Møller, Per

2016-01-01

salina biomass, with a focus on eicosapentaenoic acid (EPA). Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality...... of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scaleusing a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large...... after 21 days of cultivation. Variations in chemical compositions of Nannochloropsis salina were studied during the course of cultivation. Nannochloropsis salina can be presented as a good candidate for winter time cultivation in Denmark.The resulting biomass is a rich source of EPA and also a good...

A multiwell format assay for heparanase.

Science.gov (United States)

Behzad, Farhad; Brenchley, Paul E C

2003-09-15

This assay employs a biotinylated heparan sulfate glycosaminoglycan (HSGAG) substrate that is covalently linked to the surface of 96-well immunoassay plates. The ratio of biotin:HSGAG and the coating concentration of substrate bound to the wells have been optimized and allow removal of biotin HSGAG within 60 min of incubation at 37 degrees C in assay buffer with a standard dilution of bacterial heparitinase or platelet heparanase. Loss of biotin signal from the well surface is detected on incubation with peroxidase-streptavidin followed by color development using 3,3',5,5'-tetramethylbenzidine as the peroxidase substrate. The new assay allows specific detection of heparanase activity in multiple samples in a total time of 3 h including a 1-h substrate digestion step and is a significant improvement with regard to sensitivity, specificity, and ease of handling of multiple samples compared to other described assays. Heparanase specifically degrades the biotinylated HSGAG substrate, when used with an optimized assay buffer. A range of enzymes including collagenase, trypsin, plasmin, pepsin, chondroitinases, hyaluronidase, and neuraminidase show no effect on the substrate under optimized assay conditions. The covalent linkage of the substrate to the well prevents leaching of substrate and allows preparation and long-term storage of substrate-coated plates. The assay can be used to detect heparanase levels in clinical samples and cell culture supernatants and is ideal as a screening method for antagonists of enzyme activity.

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

Science.gov (United States)

Pĕknicová, J; Kristofová, H

1985-01-01

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

Direct 125I-radioligand assays for serum progesterone compared with assays involving extraction of serum

International Nuclear Information System (INIS)

Ratcliffe, W.A.; Corrie, J.E.T.; Dalziel, A.H.; Macpherson, J.S.

1982-01-01

Two direct radioimmunoassays for progesterone in 50 μL of unextracted serum or plasma with assays involving extraction of serum were compared. The direct assays include the use of either danazol at pH 7.4 or 8-anilino-1-naphthalenesulfonic acid at pH 4.0 to displace progesterone from serum binding-proteins. Progesterone is then assayed by using an antiserum to a progesterone 11α-hemisuccinyl conjugate and the radioligand 125 I-labeled progesterone 11α-glucuronyl tyramine, with separation by double-antibody techniques. Direct assays with either displacing agent gave good analytical recovery of progesterone added to human serum, and progesterone values for patients' specimens correlated well (r > 0.96) with results of assays involving extraction of serum. Precision was similar with each displacing agent over the working range 2.5-100 nmol/L and superior to that of extraction assays. We conclude that these direct assays of progesterone are analytically valid and more robust, precise, and technically convenient than many conventional methods involving extraction of serum

Integrated assessment of the impacts of agricultural drainwater in the Salinas River (California, USA)

Energy Technology Data Exchange (ETDEWEB)

Anderson, B.S.; Hunt, J.W.; Phillips, B.M.; Nicely, P.A.; Vlaming, V. de; Connor, V.; Richard, N.; Tjeerdema, R.S

2003-08-01

Invertebrate mortality was correlated with levels of water and sediment contaminatioin in the Salinas River. - The Salinas River is the largest of the three rivers that drain into the Monterey Bay National Marine Sanctuary in central California. Large areas of this watershed are cultivated year-round in row crops and previous laboratory studies have demonstrated that acute toxicity of agricultural drainwater to Ceriodaphnia dubia is caused by the organophosphate (OP) pesticides chlorpyrifos and diazinon. In the current study, we used a combination of ecotoxicologic tools to investigate incidence of chemical contamination and toxicity in waters and sediments in the river downstream of a previously uncharacterized agricultural drainage creek system. Water column toxicity was investigated using a cladoceran C. dubia while sediment toxicity was investigated using an amphipod Hyalella azteca. Ecological impacts of drainwater were investigated using bioassessments of macroinvertebrate community structure. The results indicated that Salinas River water downstream of the agricultural drain is acutely toxic to Ceriodaphnia, and toxicity to this species was highly correlated with combined toxic units (TUs) of chlorpyrifos and diazinon. Laboratory tests were used to demonstrate that sediments in this system were acutely toxic to H. azteca, which is a resident genus. Macroinvertebrate community structure was moderately impacted downstream of the agricultural drain input. While the lowest macroinvertebrate abundances were measured at the station demonstrating the greatest water column and sediment toxicity and the highest concentrations of pesticides, macroinvertebrate metrics were more significantly correlated with bank vegetation cover than any other variable. Results of this study suggest that pesticide pollution is the likely cause of laboratory-measured toxicity in the Salinas River samples and that this factor may interact with other factors to impact the

Integrated assessment of the impacts of agricultural drainwater in the Salinas River (California, USA)

International Nuclear Information System (INIS)

Anderson, B.S.; Hunt, J.W.; Phillips, B.M.; Nicely, P.A.; Vlaming, V. de; Connor, V.; Richard, N.; Tjeerdema, R.S.

2003-01-01

Invertebrate mortality was correlated with levels of water and sediment contaminatioin in the Salinas River. - The Salinas River is the largest of the three rivers that drain into the Monterey Bay National Marine Sanctuary in central California. Large areas of this watershed are cultivated year-round in row crops and previous laboratory studies have demonstrated that acute toxicity of agricultural drainwater to Ceriodaphnia dubia is caused by the organophosphate (OP) pesticides chlorpyrifos and diazinon. In the current study, we used a combination of ecotoxicologic tools to investigate incidence of chemical contamination and toxicity in waters and sediments in the river downstream of a previously uncharacterized agricultural drainage creek system. Water column toxicity was investigated using a cladoceran C. dubia while sediment toxicity was investigated using an amphipod Hyalella azteca. Ecological impacts of drainwater were investigated using bioassessments of macroinvertebrate community structure. The results indicated that Salinas River water downstream of the agricultural drain is acutely toxic to Ceriodaphnia, and toxicity to this species was highly correlated with combined toxic units (TUs) of chlorpyrifos and diazinon. Laboratory tests were used to demonstrate that sediments in this system were acutely toxic to H. azteca, which is a resident genus. Macroinvertebrate community structure was moderately impacted downstream of the agricultural drain input. While the lowest macroinvertebrate abundances were measured at the station demonstrating the greatest water column and sediment toxicity and the highest concentrations of pesticides, macroinvertebrate metrics were more significantly correlated with bank vegetation cover than any other variable. Results of this study suggest that pesticide pollution is the likely cause of laboratory-measured toxicity in the Salinas River samples and that this factor may interact with other factors to impact the

Groundwater quality in the shallow aquifers of the Monterey Bay, Salinas Valley, and adjacent highland areas, California

Science.gov (United States)

Burton, Carmen

2018-05-30

Groundwater provides more than 40 percent of California’s drinking water. To protect this vital resource, the State of California created the Groundwater Ambient Monitoring and Assessment (GAMA) Program. The Priority Basin Project of the GAMA Program provides a comprehensive assessment of the State’s groundwater quality and increases public access to groundwater-quality information. The shallow aquifers of the groundwater basins around Monterey Bay, the Salinas Valley, and the highlands adjacent to the Salinas Valley constitute one of the study units.

A novel microculture kinetic assay (MiCK assay) for malignant cell growth and chemosensitivity.

Science.gov (United States)

Kravtsov, V D

1994-01-01

The THERMOmax microplate reader was adapted for monitoring the growth kinetics of human leukaemic OCI/AML-2 and mouse tumour J-774.1 cell lines in continuous culture. Fluid evaporation from wells, CO2 escape and contamination were prevented by hermetic sealing of the microcultures in wells of a 96-well microplate, thus enabling the cells to grow exponentially for 72 h under the conditions of the incubated microplate reader. For both OCI/AML-2 cells, which grow in suspension, and adherent J-774.1 cells, a linear correlation was demonstrated between the number of unstained cells seeded in a given microplate well and the optical density (OD) of that well. Therefore, the OD/time curve of the culture could be deemed to be its growth curve. By the use of the linear fit equation, the actual number of the cells in the wells was computable at any time point of the assay. In the chemosensitivity test, an inhibitory effect of ARA-C on the growth of the cells could be estimated by viewing of the growth curves plotted on the screen. The maximum kinetic rates (Vmax) of the curves in the control and the ARA-C-treated wells were compared, yielding a growth inhibition index (GII). Comparison of results of the kinetic chemosensitivity assay with those of a [3H]thymidine incorporation assay revealed that the novel assay is suitable for precise quantitation of the cell chemosensitivity, is more informative and has the added technical advantage of performance without recourse to radioactive or chemically hazardous substances.

The Salinas formation in the type-area, Northeastern Minas Gerais: a proposal to review the stratigraphy of the Aracuai belt on sedimentary, metamorphic and U-Pb SHRIMP evidences

International Nuclear Information System (INIS)

Lima, Sirlene A. de Abreu; Pedrosa Soares, Antonio C.; Cordani, Umberto G.; Nutman, Allen

2002-01-01

The Salinas Formation has been considered to be a stratigraphic unit of the Macaubas Group. This group comprises the rift to passive margin sequences of the precursor basin of the Neo proterozoic Aracuai Orogeny, eastern Brazil. However, new road cuts show extensive and spectacular outcrops with very well-preserved sedimentary structures, allowing detailed studies in the type-locality of this formation, located in the Salinas town and surroundings, northeast Minas Gerais State. In its type-locality, the Salinas Formation consists of graywacke, pelite and clast-supported conglomerate, metamorphosed in the green schist facies. The sedimentary lithofacies are grouped into three facies association (shelf, slope and deep-sea), indicating sedimentation from shelf deposits to deep-water turbidites. The shelf sedimentation was influenced by storm-wave during deposition. The slump and deep-sea deposits were generated by gravitational flows and high- to low-concentration turbidity currents. Shelf sandstone, clast-supported conglomerate and proximal to distal turbidites outline a submarine fan system. U-Pb SHRIMP data from detrital zircons of graywacke samples indicate a maximum sedimentation age of ca. 568 Ma. Thus, the Salinas Formation is much younger than the Macaubas Group, and represents late orogenic deposits (ca. 568- 500 Ma). The distal, passive margin unit of the Macaubas Group is now called Ribeirao da Folha Formation (ca. 800 Ma). (author)

Ecology of a heterotrichous ciliate Fabrea salina from salterns of Bombay Coast, India

Digital Repository Service at National Institute of Oceanography (India)

Rattan, P.; Ansari, Z.A.; Sreepada, R.A.

The abundance and population size structure of a heterotrichous ciliate @iFabrea salina@@, was studied in relation to hydrochemical and hydrobiological variables in three hypersaline ponds of Bombay, west coast of India There was a clear seasonal...

Isolasi Senyawa Aktif Ekstrak Etanol Biji Alpukat (Persea americana dan Uji Toksisitas Terhadap Artemia Salina Leach

Directory of Open Access Journals (Sweden)

Andi Nur Fitriani Abubakar

2014-07-01

Full Text Available Avocado seed (Persea americana is recognized as one of medicinal plants. It contains several secondary metabolites, which have toxic activity. However, efforts to identify active compounds from avocado seeds (Persea americana are still relatively rare. Therefore, isolation and toxicity assay have been conducted foward the active compound of avocado seed. Maceration one kilogram of seed dried powder by ethanol obtained 49,7464 gram extract. Separation of etanol extract by column chromatography generated 0,0698 grams of pure white needle crytal, which is positively triterpenoid based on Lieberman-Buchard test. In addition, infrared spectrum showed the existence of OH, C=C, C-C, C=O, -C-H, -CH3,-CH2 and C-O stretch, which support the presumed compound. The result of toxicity test on Artemia salina Leach showed that the extract, fraction and pure isolates of the etanol extract are toxic with LC50 values 13,274 g/mL; 9,528 g/mL and 8,128 g/mL, respectively.

Identifying primary stressors impacting macroinvertebrates in the Salinas River (California, USA): Relative effects of pesticides and suspended particles

International Nuclear Information System (INIS)

Anderson, B.S.; Phillips, B.M.; Hunt, J.W.; Connor, V.; Richard, N.; Tjeerdema, R.S.

2006-01-01

Laboratory dose-response experiments with organophosphate and pyrethroid pesticides, and dose-response experiments with increasing particle loads were used to determine which of these stressors were likely responsible for the toxicity and macroinvertebrate impacts previously observed in the Salinas River. Experiments were conducted with the amphipod Hyalella azteca, the baetid mayfly Procloeon sp., and the midge Chironomus dilutus (Shobanov, formerly Chironomus tentans). The results indicate the primary stressor impacting H. azteca was pesticides, including chlorpyrifos and permethrin. The mayfly Procloeon sp. was sensitive to chlorpyrifos and permethrin within the range of concentrations of these pesticides measured in the river. Chironomus dilutus were sensitive to chlorpyrifos within the ranges of concentrations measured in the river. None of the species tested were affected by turbidity as high as 1000 NTUs. The current study shows that pesticides are more important acute stressors of macroinvertebrates than suspended sediments in the Salinas River. - Pesticides are the primary stressor impacting macroinvertebrates in sections of the lower Salinas River

Identifying primary stressors impacting macroinvertebrates in the Salinas River (California, USA): Relative effects of pesticides and suspended particles

Energy Technology Data Exchange (ETDEWEB)

Anderson, B.S. [Department of Environmental Toxicology, University of California, Davis, CA 95616 (United States)]. E-mail: anderson@ucdavis.edu; Phillips, B.M. [Department of Environmental Toxicology, University of California, Davis, CA 95616 (United States); Hunt, J.W. [Department of Environmental Toxicology, University of California, Davis, CA 95616 (United States); Connor, V. [Division of Water Quality, State Water Resources Control Board, 1001 I. Street, Sacramento, CA 95814 (United States); Richard, N. [Division of Water Quality, State Water Resources Control Board, 1001 I. Street, Sacramento, CA 95814 (United States); Tjeerdema, R.S. [Department of Environmental Toxicology, University of California, Davis, CA 95616 (United States)

2006-06-15

Laboratory dose-response experiments with organophosphate and pyrethroid pesticides, and dose-response experiments with increasing particle loads were used to determine which of these stressors were likely responsible for the toxicity and macroinvertebrate impacts previously observed in the Salinas River. Experiments were conducted with the amphipod Hyalella azteca, the baetid mayfly Procloeon sp., and the midge Chironomus dilutus (Shobanov, formerly Chironomus tentans). The results indicate the primary stressor impacting H. azteca was pesticides, including chlorpyrifos and permethrin. The mayfly Procloeon sp. was sensitive to chlorpyrifos and permethrin within the range of concentrations of these pesticides measured in the river. Chironomus dilutus were sensitive to chlorpyrifos within the ranges of concentrations measured in the river. None of the species tested were affected by turbidity as high as 1000 NTUs. The current study shows that pesticides are more important acute stressors of macroinvertebrates than suspended sediments in the Salinas River. - Pesticides are the primary stressor impacting macroinvertebrates in sections of the lower Salinas River.

Analysis of toxicity of Anacardium occidentale L. extract submitted to ionizing radiation on embryos of Biomphalaria glabrata and Artemia salina

International Nuclear Information System (INIS)

Silva, Hianna A.M.F.; Sa, Jose L.F.; Lima, Claudia S.A.; Amancio, Francisco F.; Melo, Ana M.M.A.; Ribeiro, Luanna R.S.; Santos, Gustavo H.F.; Silva, Edvane B.

2013-01-01

The use of gamma radiation as a sterilization method for herbs, herbal medicines and foods, shows positive results regarding the retention of such products, economy and safety of the method. However, it is known that this method of processing plant material can cause chemical changes in these products related to the type of material, its components and the dose received. Evaluated, in the present study, the action of gamma radiation as a modifier of toxicity extract of Anacardium occidentale Linn. To evaluate the toxicity of the extract irradiated at doses of 5.0, 7.5 and 10.0 kGy and concentrations of 250, 500 and 1000 mg/L was used bioassays with Artemia salina and Biomphalaria glabrata. For the test to A. salina, 520 specimens were used divided into groups of 10 larvae. For the bioassay with B. glabrata, 3900 specimens were used divided into groups of, approximately, 100 embryos. Larvae of A. salina and embryos were subjected to extracts irradiated and unirradiated for 24 hours. The bioassay with A. salina, showed a decrease, compared to extract unirradiated and irradiated at doses of 5.0 and 7.5 kGy, of extract irradiated with 10 kGy, where the mortality did not differ from the control group. In tests with embryos was observed an increase in the toxicity of the extract at a dose of 7.5 kGy and a decrease in the dose of 10.0 kGy. The radiation promoted changes in the toxicity of leaves extracts of Anacardium occidentale Linn. on embryos of Biomphalaria glabrata and Artemia salina. (author)

Analysis of toxicity of Anacardium occidentale L. extract submitted to ionizing radiation on embryos of Biomphalaria glabrata and Artemia salina

Energy Technology Data Exchange (ETDEWEB)

Silva, Hianna A.M.F.; Sa, Jose L.F.; Lima, Claudia S.A.; Amancio, Francisco F.; Melo, Ana M.M.A., E-mail: hiannaamfs@gmail.com, E-mail: luismuma6@gmail.com, E-mail: claudia.salima@gmail.com, E-mail: amancioff@bol.com.br, E-mail: amdemelo@hotmail.com [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Biofisica e Radiobiologia; Ribeiro, Luanna R.S.; Santos, Gustavo H.F.; Silva, Edvane B., E-mail: luannaribeiro_lua@hotmail.com, E-mail: santosghf@hotmail.com, E-mail: edvborges@yahoo.com.br [Universidade Federal de Pernambuco (UFPE), Recife, PE (Brazil). Departamento de Energia Nuclear. Grupo de Estudos em Radioprotecao e Radioecologia

2013-07-01

The use of gamma radiation as a sterilization method for herbs, herbal medicines and foods, shows positive results regarding the retention of such products, economy and safety of the method. However, it is known that this method of processing plant material can cause chemical changes in these products related to the type of material, its components and the dose received. Evaluated, in the present study, the action of gamma radiation as a modifier of toxicity extract of Anacardium occidentale Linn. To evaluate the toxicity of the extract irradiated at doses of 5.0, 7.5 and 10.0 kGy and concentrations of 250, 500 and 1000 mg/L was used bioassays with Artemia salina and Biomphalaria glabrata. For the test to A. salina, 520 specimens were used divided into groups of 10 larvae. For the bioassay with B. glabrata, 3900 specimens were used divided into groups of, approximately, 100 embryos. Larvae of A. salina and embryos were subjected to extracts irradiated and unirradiated for 24 hours. The bioassay with A. salina, showed a decrease, compared to extract unirradiated and irradiated at doses of 5.0 and 7.5 kGy, of extract irradiated with 10 kGy, where the mortality did not differ from the control group. In tests with embryos was observed an increase in the toxicity of the extract at a dose of 7.5 kGy and a decrease in the dose of 10.0 kGy. The radiation promoted changes in the toxicity of leaves extracts of Anacardium occidentale Linn. on embryos of Biomphalaria glabrata and Artemia salina. (author)

Phototactic orientation mechanism in the ciliate Fabrea salina, as inferred from numerical simulations.

Science.gov (United States)

Marangoni, R; Preosti, G; Colombetti, G

2000-02-01

The marine ciliate Fabrea salina shows a clear positive phototaxis, but the mechanism by which a single cell is able to detect the direction of light and orient its swimming accordingly is still unknown. A simple model of phototaxis is that of a biased random walk, where the bias due to light can affect one or more of the parameters that characterize a random walk, i.e., the mean speed, the frequency distribution of the angles of directional changes and the frequency of directional changes. Since experimental evidence has shown no effect of light on the mean speed of Fabrea salina, we have excluded models depending on this parameter. We have, therefore, investigated the phototactic orientation of Fabrea salina by computer simulation of two simple models, the first where light affects the frequency distribution of the angles of directional changes (model M1) and the second where the light bias modifies the frequency of directional changes (model M2). Simulated M1 cells directly orient their swimming towards the direction of light, regardless of their current swimming orientation; simulated M2 cells, on the contrary, are unable to actively orient their motion, but remain locked along the light direction once they find it by chance. The simulations show that these two orientation models lead to different macroscopic behaviours of the simulated cell populations. By comparing the results of the simulations with the experimental ones, we have found that the phototactic behaviour of real cells is more similar to that of the M2 model.

Daphnia HR96 is a promiscuous xenobiotic and endobiotic nuclear receptor

International Nuclear Information System (INIS)

Karimullina, Elina; Li Yangchun; Ginjupalli, Gautam K.; Baldwin, William S.

2012-01-01

Daphnia pulex is the first crustacean to have its genome sequenced. The genome project provides new insight and data into how an aquatic crustacean may respond to environmental stressors, including toxicants. We cloned Daphnia pulex HR96 (DappuHR96), a nuclear receptor orthologous to the CAR/PXR/VDR group of nuclear receptors. In Drosophila melanogaster, (hormone receptor 96) HR96 responds to phenobarbital exposure and has been hypothesized as a toxicant receptor. Therefore, we set up a transactivation assay to test whether DappuHR96 is a promiscuous receptor activated by xenobiotics and endobiotics similar to the constitutive androstane receptor (CAR) and the pregnane X-receptor (PXR). Transactivation assays performed with a GAL4-HR96 chimera demonstrate that HR96 is a promiscuous toxicant receptor activated by a diverse set of chemicals such as pesticides, hormones, and fatty acids. Several environmental toxicants activate HR96 including estradiol, pyriproxyfen, chlorpyrifos, atrazine, and methane arsonate. We also observed repression of HR96 activity by chemicals such as triclosan, androstanol, and fluoxetine. Nearly 50% of the chemicals tested activated or inhibited HR96. Interestingly, unsaturated fatty acids were common activators or inhibitors of HR96 activity, indicating a link between diet and toxicant response. The omega-6 and omega-9 unsaturated fatty acids linoleic and oleic acid activated HR96, but the omega-3 unsaturated fatty acids alpha-linolenic acid and docosahexaenoic acid inhibited HR96, suggesting that these two distinct sets of lipids perform opposing roles in Daphnia physiology. This also provides a putative mechanism by which the ratio of dietary unsaturated fats may affect the ability of an organism to respond to a toxic insult. In summary, HR96 is a promiscuous nuclear receptor activated by numerous endo- and xenobiotics.

Daphnia HR96 is a promiscuous xenobiotic and endobiotic nuclear receptor

Energy Technology Data Exchange (ETDEWEB)

Karimullina, Elina [Environmental Toxicology Program, Clemson University, Clemson, SC 29634 (United States); Institute of Plant and Animal Ecology, Russian Academy of Sciences, Ural Branch, Yekaterinburg 620144 (Russian Federation); Li Yangchun; Ginjupalli, Gautam K. [Environmental Toxicology Program, Clemson University, Clemson, SC 29634 (United States); Baldwin, William S., E-mail: baldwin@clemson.edu [Environmental Toxicology Program, Clemson University, Clemson, SC 29634 (United States); Biological Sciences, Clemson University, Clemson, SC (United States)

2012-07-15

Daphnia pulex is the first crustacean to have its genome sequenced. The genome project provides new insight and data into how an aquatic crustacean may respond to environmental stressors, including toxicants. We cloned Daphnia pulex HR96 (DappuHR96), a nuclear receptor orthologous to the CAR/PXR/VDR group of nuclear receptors. In Drosophila melanogaster, (hormone receptor 96) HR96 responds to phenobarbital exposure and has been hypothesized as a toxicant receptor. Therefore, we set up a transactivation assay to test whether DappuHR96 is a promiscuous receptor activated by xenobiotics and endobiotics similar to the constitutive androstane receptor (CAR) and the pregnane X-receptor (PXR). Transactivation assays performed with a GAL4-HR96 chimera demonstrate that HR96 is a promiscuous toxicant receptor activated by a diverse set of chemicals such as pesticides, hormones, and fatty acids. Several environmental toxicants activate HR96 including estradiol, pyriproxyfen, chlorpyrifos, atrazine, and methane arsonate. We also observed repression of HR96 activity by chemicals such as triclosan, androstanol, and fluoxetine. Nearly 50% of the chemicals tested activated or inhibited HR96. Interestingly, unsaturated fatty acids were common activators or inhibitors of HR96 activity, indicating a link between diet and toxicant response. The omega-6 and omega-9 unsaturated fatty acids linoleic and oleic acid activated HR96, but the omega-3 unsaturated fatty acids alpha-linolenic acid and docosahexaenoic acid inhibited HR96, suggesting that these two distinct sets of lipids perform opposing roles in Daphnia physiology. This also provides a putative mechanism by which the ratio of dietary unsaturated fats may affect the ability of an organism to respond to a toxic insult. In summary, HR96 is a promiscuous nuclear receptor activated by numerous endo- and xenobiotics.

Determination of suvorexant in human plasma using 96-well liquid-liquid extraction and HPLC with tandem mass spectrometric detection.

Science.gov (United States)

Breidinger, S A; Simpson, R C; Mangin, E; Woolf, E J

2015-10-01

A method, using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS), was developed for the determination of suvorexant (MK-4305, Belsomra(®)), a selective dual orexin receptor antagonist for the treatment insomnia, in human plasma over the concentration range of 1-1000ng/mL. Stable isotope labeled (13)C(2)H3-suvorexant was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction, in the 96-well format, of a 100μL plasma sample with methyl t-butyl ether. The compounds were chromatographed under isocratic conditions on a Waters dC18 (50×2.1mm, 3μm) column with a mobile phase consisting of 30/70 (v/v %) 10mM ammonium formate, pH3/acetonitrile at a flow rate of 0.3mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for suvorexant (m/z 451→186) and (13)C(2)H3-suvorexant (m/z 455→190) on an Applied Biosystems API 4000 tandem mass spectrometer was used for quantitation. Intraday assay precision, assessed in six different lots of control plasma, was within 10% CV at all concentrations, while assay accuracy ranged from 95.6 to 105.0% of nominal. Quality control (QC) samples in plasma were stored at -20°C. Initial within day analysis of QCs after one freeze-thaw cycle showed accuracy within 9.5% of nominal with precision (CV) of 6.7% or less. The plasma QC samples were demonstrated to be stable for up to 25 months at -20°C. The method described has been used to support clinical studies during Phase I through III of clinical development. Copyright © 2015 Elsevier B.V. All rights reserved.

Efeito do meio Erd Schreiber no cultivo das microalgas Dunaliella salina, Tetraselmis chuii e Isochrysis galbana = Erd Schreiber medium effect in culture of microalgae Dunaliella salina, Tetraselmis chuii and Isochrysis galbana

Directory of Open Access Journals (Sweden)

Vera Lucia Mota Klein

2006-04-01

Full Text Available As microalgas são utilizadas como fonte de alimento em aqüicultura. Neste trabalho cultivaram-se D. salina, T. chuii e I. galbana. O objetivo do trabalho consistiu em determinar o efeito do meio Erd Schreiber sobre o seu crescimento. Iniciou-se o cultivo com a mistura de 200 mg de Na2HPO4,7H2O, 100 mg de NaNO3 e 50 mL de extrato de solo. No monitoramento, manteve-se a temperatura entre 24 - 28 oC, a salinidade a 34 ppt, à iluminação constante, a densidade celular com uma câmara de Neubauer e um microscópio binocular modelo ZEISS. Como resultado, I. galbana, D. salina e T. chuii atingiram 969 104 cel/mL, 457 x 104 cel/mL e 258,66 x 104 cel/mL, respectivamente, e oscoeficientes angulares b foram 3,76 x 104 cel./mL/dia, 6,84 x 104 cel./mL/dia e 2,08 x 104 cel./mL/dia respectivamente, indicando bom desempenho de todas as microalgas no meio Erd Shreiber.The microalgae is used as food source in aqüicultura. In this work they had cultivated D. salina , T. chuii and I. galbana . The objective of the work is to determine the effect of Erd Schreiber´s culture medium on the microalgae growth. The culture initiated mixting 200mg of Na2HPO4,7H2O, 100 mg of NaNO3 and 50 mL of soil extract. During the culture the temperature had varied between 24 and 28oC, the salinity was fixed on 34 %o, and the illumination was maintained constant. The assessment of the culture was made by a chamber of Neubauer and a binocular microscope ZEISS model. As result I. galbana D. salina and T.chuii reached 969 104 cel/mL, 457 x 104cel/mL and 258,66 x 104 cel/mL respectively and as angular coefficient 3,76 x 104 cel/mL/dia, 6,84 x 104 cel/mL/dia and 2,08 104 x cel/mL/dia respectively, showing good answer of the microalgae to the effect of Erd Schreiber´ s medium.

Not yet? Ya basta: Healing and the horizons of an otherwise in Salinas, California

NARCIS (Netherlands)

Raschig, M.S.

2016-01-01

Among a persistently criminalized population of Mexican-Americans in the farmtown-gangland of Salinas, California, healing from the wounds of history has emerged as a critical register of political action, a tacit and uncertain activism recalibrating the pace and tense of personal recoveries and

Morphological changes about Artemia Salina by the effect of gamma radiation

International Nuclear Information System (INIS)

Dvorak, P.; Kunova, V; Kratochvil, B.; Salplachta, J.; Benova, K.

2004-01-01

Direction Council of Europe require restriction of laboratory vertebrae in biological experiments. Once by possibilities is using biological test II. generation among which belongs to and test for Artemia salina. Still this time published test however value only lethality in dependence on dose, exposition and time survival. This work document possibility watch of expressive morphological changes, which consist in retardation development brine shrimps stage in dependence on dose gamma radiation 60 Co. (authors)

Evaluation of hydrologic conditions and nitrate concentrations in the Rio Nigua de Salinas alluvial fan aquifer, Salinas, Puerto Rico, 2002-03

Science.gov (United States)

Rodriguez, Jose M.

2006-01-01

A ground-water quality study to define the potential sources and concentration of nitrate in the Rio Nigua de Salinas alluvial fan aquifer was conducted between January 2002 and March 2003. The study area covers about 3,600 hectares of the coastal plain within the municipality of Salinas in southern Puerto Rico, extending from the foothills to the Caribbean Sea. Agriculture is the principal land use and includes cultivation of diverse crops, turf grass, bioengineered crops for seed production, and commercial poultry farms. Ground-water withdrawal in the alluvial fan was estimated to be about 43,500 cubic meters per day, of which 49 percent was withdrawn for agriculture, 42 percent for public supply, and 9 percent for industrial use. Ground-water flow in the study area was primarily to the south and toward a cone of depression within the south-central part of the alluvial fan. The presence of that cone of depression and a smaller one located in the northeastern quadrant of the study area may contribute to the increase in nitrate concentration within a total area of about 545 hectares by 'recycling' ground water used for irrigation of cultivated lands. In an area that covers about 405 hectares near the center of the Salinas alluvial fan, nitrate concentrations increased from 0.9 to 6.7 milligrams per liter as nitrogen in 1986 to 8 to 12 milligrams per liter as nitrogen in 2002. Principal sources of nitrate in the study area are fertilizers (used in the cultivated farmlands) and poultry farm wastes. The highest nitrogen concentrations were found at poultry farms in the foothills area. In the area of disposed poultry farm wastes, nitrate concentrations in ground water ranged from 25 to 77 milligrams per liter as nitrogen. Analyses for the stable isotope ratios of nitrogen-15/nitrogen-14 in nitrate were used to distinguish the source of nitrate in the coastal plain alluvial fan aquifer. Potential nitrate loads from areas under cultivation were estimated for the

Rosette Assay: Highly Customizable Dot-Blot for SH2 Domain Screening.

Science.gov (United States)

Ng, Khong Y; Machida, Kazuya

2017-01-01

With a growing number of high-throughput studies, structural analyses, and availability of protein-protein interaction databases, it is now possible to apply web-based prediction tools to SH2 domain-interactions. However, in silico prediction is not always reliable and requires experimental validation. Rosette assay is a dot blot-based reverse-phase assay developed for the assessment of binding between SH2 domains and their ligands. It is conveniently customizable, allowing for low- to high-throughput analysis of interactions between various numbers of SH2 domains and their ligands, e.g., short peptides, purified proteins, and cell lysates. The binding assay is performed in a 96-well plate (MBA or MWA apparatus) in which a sample spotted membrane is incubated with up to 96 labeled SH2 domains. Bound domains are detected and quantified using a chemiluminescence or near-infrared fluorescence (IR) imaging system. In this chapter, we describe a practical protocol for rosette assay to assess interactions between synthesized tyrosine phosphorylated peptides and a library of GST-tagged SH2 domains. Since the methodology is not confined to assessment of SH2-pTyr interactions, rosette assay can be broadly utilized for ligand and drug screening using different protein interaction domains or antibodies.

Bicarbonate-based cultivation of Dunaliella salina for enhancing carbon utilization efficiency.

Science.gov (United States)

Kim, Ga-Yeong; Heo, Jina; Kim, Hee-Sik; Han, Jong-In

2017-08-01

In this study, bicarbonate was proposed as an alternative carbon source to overcome exceedingly low CO 2 fixation efficiency of conventional microalgae cultivation system. 5gL -1 of sodium bicarbonate was found to well support the growth of Dunaliella salina, showing 2.84-fold higher specific growth rate than a bicarbonate-free control. This bicarbonate-fed cultivation also could yield biomass productivity similar to that of CO 2 -based system as long as pH was controlled. While the supplied CO 2 , because of its being a gas, was mostly lost and only 3.59% of it was used for biomass synthesis, bicarbonate was effectively incorporated into the biomass with 91.40% of carbon utilization efficiency. This study showed that the bicarbonate-based microalgae cultivation is indeed possible, and can even become a truly environment-friendly and workable approach, provided that a CO 2 mineralization technology is concomitantly established. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cytotoxic activity of carotenoid rich fractions from Haematococcus pluvialis and Dunaliella salina microalgae and the identification of the phytoconstituents using LC-DAD/ESI-MS.

Science.gov (United States)

El-Baz, Farouk K; Hussein, Rehab A; Mahmoud, Khaled; Abdo, Sayeda M

2018-02-01

Microalgae represent a rich source that satisfies the growing need for novel ingredients of nutriceuticals, pharmaceuticals, and food supplements. Haematococcus pluvialis and Dunaliella salina microalgae are isolated from the Egyptian hydro-flora and are reported for their potent antioxidant activities. The cytotoxic activity of different fractions of both microalgae was investigated on 4 cell lines HePG2, MCF7, HCT116, and A549. The carotenoid rich fraction of H. pluvialis showed potent cytotoxic activity against colon cancer cell line and moderate activity against both liver and breast cancer cell lines. On the other hand, the carotenoid rich fraction of D. salina showed mild cytotoxic activity on breast and liver cancer cell lines. The carotenoid rich fraction of H. pluvialis was analysed using LC-DAD/ESI-MS and the major carotenoids were identified either free as well as bounded to fatty acids. Copyright © 2017 John Wiley & Sons, Ltd.

Energy-producing electro-flocculation for harvest of Dunaliella salina.

Science.gov (United States)

Liu, Qing; Zhang, Meng; Lv, Tao; Chen, Hongjun; Chika, Anthony Okonkwo; Xiang, Changli; Guo, Minxue; Wu, Minghui; Li, Jianjun; Jia, Lishan

2017-10-01

In this study, an efficient electro-flocculation process for Dunaliella salina with energy production by aluminum-air battery has been successfully applied. The formed aluminum hydroxide hydrates during discharging of battery were positively charged, which have a great potential for microalgae flocculation. The precipitation of aluminum hydroxide hydrates by algae also could improve the performance of aluminum-air battery. The harvesting efficiency could reach 97% in 20mins with energy production of 0.11kWh/kg. This discharging electro-flocculation (DEF) technology provides a new energy producing process to effectively harvest microalgae. Copyright © 2017 Elsevier Ltd. All rights reserved.

High surface adsorption properties of carbon-based nanomaterials are responsible for mortality, swimming inhibition, and biochemical responses in Artemia salina larvae

International Nuclear Information System (INIS)

Mesarič, Tina; Gambardella, Chiara; Milivojević, Tamara; Faimali, Marco; Drobne, Damjana; Falugi, Carla; Makovec, Darko; Jemec, Anita; Sepčić, Kristina

2015-01-01

Highlights: • Carbon-based nanomaterials adsorb onto the body surface of A. salina larvae. • Surface adsorption results in concentration–dependent inhibition of larval swimming. • Carbon-based nanomaterials induce no significant mortality of A. salina larvae. - Abstract: We investigated the effects of three different carbon-based nanomaterials on brine shrimp (Artemia salina) larvae. The larvae were exposed to different concentrations of carbon black, graphene oxide, and multiwall carbon nanotubes for 48 h, and observed using phase contrast and scanning electron microscopy. Acute (mortality) and behavioural (swimming speed alteration) responses and cholinesterase, glutathione-S-transferase and catalase enzyme activities were evaluated. These nanomaterials were ingested and concentrated in the gut, and attached onto the body surface of the A. salina larvae. This attachment was responsible for concentration–dependent inhibition of larval swimming, and partly for alterations in the enzyme activities, that differed according to the type of tested nanomaterials. No lethal effects were observed up to 0.5 mg/mL carbon black and 0.1 mg/mL multiwall carbon nanotubes, while graphene oxide showed a threshold whereby it had no effects at 0.6 mg/mL, and more than 90% mortality at 0.7 mg/mL. Risk quotients calculated on the basis of predicted environmental concentrations indicate that carbon black and multiwall carbon nanotubes currently do not pose a serious risk to the marine environment, however if uncontrolled release of nanomaterials continues, this scenario can rapidly change

High surface adsorption properties of carbon-based nanomaterials are responsible for mortality, swimming inhibition, and biochemical responses in Artemia salina larvae

Energy Technology Data Exchange (ETDEWEB)

Mesarič, Tina, E-mail: tina.mesaric84@gmail.com [Department of Biology, Biotechnical Faculty, University of Ljubljana (Slovenia); Gambardella, Chiara, E-mail: chiara.gambardella@ge.ismar.cnr.it [Institute of Marine Sciences, National Research Council, Genova (Italy); Milivojević, Tamara, E-mail: milivojevictamara@gmail.com [Department of Biology, Biotechnical Faculty, University of Ljubljana (Slovenia); Faimali, Marco, E-mail: marco.faimali@ismar.cnr.it [Institute of Marine Sciences, National Research Council, Genova (Italy); Drobne, Damjana, E-mail: damjana.drobne@bf.uni-lj.si [Department of Biology, Biotechnical Faculty, University of Ljubljana (Slovenia); Centre of Excellence in Nanoscience and Nanotechnology (CO Nanocentre), Ljubljana (Slovenia); Centre of Excellence in Advanced Materials and Technologies for the Future (CO NAMASTE), Ljubljana (Slovenia); Falugi, Carla, E-mail: carlafalugi@hotmail.it [Department of Earth, Environment and Life Sciences, University of Genova, Genova (Italy); Makovec, Darko, E-mail: darko.makovec@ijs.si [Jožef Stefan Institute, Jamova 39, 1000 Ljubljana (Slovenia); Jemec, Anita, E-mail: anita.jemec@bf.uni-lj.si [Department of Biology, Biotechnical Faculty, University of Ljubljana (Slovenia); Sepčić, Kristina, E-mail: kristina.sepcic@bf.uni-lj.si [Department of Biology, Biotechnical Faculty, University of Ljubljana (Slovenia)

2015-06-15

Highlights: • Carbon-based nanomaterials adsorb onto the body surface of A. salina larvae. • Surface adsorption results in concentration–dependent inhibition of larval swimming. • Carbon-based nanomaterials induce no significant mortality of A. salina larvae. - Abstract: We investigated the effects of three different carbon-based nanomaterials on brine shrimp (Artemia salina) larvae. The larvae were exposed to different concentrations of carbon black, graphene oxide, and multiwall carbon nanotubes for 48 h, and observed using phase contrast and scanning electron microscopy. Acute (mortality) and behavioural (swimming speed alteration) responses and cholinesterase, glutathione-S-transferase and catalase enzyme activities were evaluated. These nanomaterials were ingested and concentrated in the gut, and attached onto the body surface of the A. salina larvae. This attachment was responsible for concentration–dependent inhibition of larval swimming, and partly for alterations in the enzyme activities, that differed according to the type of tested nanomaterials. No lethal effects were observed up to 0.5 mg/mL carbon black and 0.1 mg/mL multiwall carbon nanotubes, while graphene oxide showed a threshold whereby it had no effects at 0.6 mg/mL, and more than 90% mortality at 0.7 mg/mL. Risk quotients calculated on the basis of predicted environmental concentrations indicate that carbon black and multiwall carbon nanotubes currently do not pose a serious risk to the marine environment, however if uncontrolled release of nanomaterials continues, this scenario can rapidly change.

Acclimation and tolerance of Artemia salina to copper salts

Energy Technology Data Exchange (ETDEWEB)

Saliba, L.J.; Krzyz, R.M.

1976-01-01

The brine shrimp Artemia salina L. was acclimated in sea water with cupric chloride, acetate, carbonate, and sulfate, each at concentrations of 0.1, 0.05 and 0.025 ppM Cu/sup + +/, together with sea water controls. Growth inhibition was observed in all four compounds, generally in direct relationship to the concentration. It was least in sulfate, and increased progressively in chloride, acetate and carbonate in that order. In toxicity tests, 2-week old larvae from each solution were exposed to concentrations of 10, 7.5, 5, 2.5 and 1 ppM Cu/sup + +/ of the same compounds, together with unacclimated larvae of the same age. Similar tests were held with 6-week old adults. Toxicity to unacclimated larvae and adults differed with the compounds, carbonate being the least toxic, followed by sulfate, chloride and acetate in increasing order. Larvae acclimated in chloride and sulfate showed an increased tolerance to 1 and 2.5 ppM Cu/sup + +/ compared to untreated controls. Tolerance was not enhanced from 5 ppM Cu/sup + +/ upwards. In both compounds, adults acclimated in 0.1 ppM Cu/sup + +/ showed an increased tolerance to concentrations between 1 and 7.5 ppM Cu/sup + +/ compared to controls. Considerable precipitation occurred with the high levels of this compound, thus effecting the ''final'' concentrations. No acclimation effect was observed in acetate for either larvae or adults. It is suggested that in A. salina, copper toxicity depends on the particular form of the metal, and that this difference is also evident in growth inhibition and in the potential acquisition of increased tolerance through exposure to low concentrations.

Pesticide Risk Communication, Risk Perception, and Self-Protective Behaviors among Farmworkers in California's Salinas Valley

Science.gov (United States)

Cabrera, Nolan L.; Leckie, James O.

2009-01-01

Agricultural pesticide use is the highest of any industry, yet there is little research evaluating farmworkers' understandings of the health risks chemical exposure poses. This study examines pesticide education, risk perception, and self-protective behaviors among farmworkers in California's Salinas Valley. Fifty current and former farmworkers…

Metabolic studies with NMR spectroscopy of the alga Dunaliella salina trapped within agarose beads.

Science.gov (United States)

Bental, M; Pick, U; Avron, M; Degani, H

1990-02-22

A technique for the entrapment of the unicellular algae Dunaliella salina in agarose beads and their perfusion during NMR measurements is presented. The trapped cells maintained their ability to proliferate under normal growth conditions, and remained viable and stable under steady-state conditions for long periods during NMR measurements. Following osmotic shock in the dark, prominent changes were observed in the intracellular level of ATP and polyphosphates, but little to no changes in the intracellular pH or orthoposphate content. When cells were subjected to hyperosmotic shock, the ATP level decreased. The content of NMR-visible polyphosphates decreased as well, presumably due to the production of longer, NMR-invisible structures. Following hypoosmotic shock, the ATP content increased and longer polyphosphates were broken down to shorter, more mobile polymers.

Cytotoxicity evaluation of gold nanoparticles on microalga Dunaliella salina in microplate test system

Science.gov (United States)

Chumakov, Daniil; Prilepskii, Artur; Dykman, Lev; Khlebtsov, Boris; Khlebtsov, Nikolai; Bogatyrev, Vladimir

2018-04-01

Gold nanoparticles are intensively studied in biomedicine. Assessment of their biocompatibility is highly important. Currently there is lack of evidence, concerning nanotoxicity of ultrasmall gold nanoparticles < 5 nm. Existing data are rather contradictory. The aim of that study was to evaluate the toxicity of 2 nm colloidal gold, using microalga Dunaliella salina. Cellular barriers of that microalga are very similar to animal cells so it might be considered as a valuable model for nanotoxicity testing. Chlorophyll content as a test-function was used. Spectrophotometric method for chlorophyll determination in vivo in suspensions of D.salina cultures was applied. Calculated EC50 48h value of ionic gold was 25.8 +/- 0.3 mg Au/L. EC50 value of phosphine-stabilized gold nanoclusters was 32.2 +/-1.1 mg Au/L. It was not possible to calculate EC50 for 15 nm citrate gold nanoparticles, as they were non-toxic at all concentrations tested. These results are confirmed by fluorescent -microscopic monitoring of the same probes. It was shown that 10-fold growth of phosphine-stabilized gold nanoparticles (from 2.3 +/- 0.9 nm to 21.1 +/- 7.5 nm) led to 7-fold decrease of their toxicity.

Comparative study on toxicity of ZnO and TiO2 nanoparticles on Artemia salina: effect of pre-UV-A and visible light irradiation.

Science.gov (United States)

Bhuvaneshwari, M; Sagar, Bhawana; Doshi, Siddharth; Chandrasekaran, N; Mukherjee, Amitava

2017-02-01

This study evaluated the toxicity potential of ZnO and TiO 2 nanoparticles under pre-UV-A irradiation and visible light condition on Artemia salina. The nanoparticle suspension was prepared in seawater medium and exposed under pre-UV-A (0.23 mW/cm 2 ) and visible light (0.18 mW/cm 2 ) conditions. The aggregation profiles of both nanoparticles (NPs) and dissolution of ZnO NPs under both irradiation conditions at various kinetic intervals (1, 24, 48 h) were studied. The 48-h LC 50 values were found to be 27.62 and 71.63 mg/L for ZnO NPs and 117 and 120.9 mg/L for TiO 2 NPs under pre-UV-A and visible light conditions. ZnO NPs were found to be more toxic to A. salina as compared to TiO 2 NPs. The enhanced toxicity was observed under pre-UV-A-irradiated ZnO NPs, signifying its phototoxicity. Accumulation of ZnO and TiO 2 NPs into A. salina depends on the concentration of particles and type irradiations. Elimination of accumulated nanoparticles was also evident under both irradiation conditions. Other than ZnO NPs, the dissolved Zn 2+ also had a significant effect on toxicity and accumulation in A. salina. Increased catalase (CAT) activity in A. salina indicates the generation of oxidative stress due to NP interaction. Thus, this study provides an understanding of the toxicity of photoreactive ZnO and TiO 2 NPs as related to the effects of pre-UV-A and visible light irradiation.

Estudo fitoquímico preliminar e bioensaio toxicológico frente a larvas de Artemia salina Leach. de extrato obtido de frutos de Solanum lycocarpum A. St.-Hill (Solanaceae

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Marcelo Gonzaga de Freitas Araújo

2010-11-01

Full Text Available

Neste trabalho avaliou-se o perfil fitoquímico e a toxicidade preliminar frente a larvas de Artemia salina do extrato etanólico de frutos de Solanum lycocarpum. O extrato foi submetido à analise fitoquímica preliminar para identificação das principais classes de metabolitos secundários presentes e testado frente a larvas de A. salina para obtenção das concentrações letais médias (CL50%. Os testes fitoquímicos demonstraram a presença de fenóis, taninos, saponinas, alcalóides e esteróides e triterpenos livres. O extrato foi fracionado em diferentes solventes para a avaliação da toxicidade frente à A. salina, apresentando considerável citotoxicidade encontrada na fração hidroalcoólica (CL50% = 285,546 µg/mL. Palavras-chave: Solanum lycocarpum, Artemia salina, triagem fitoquímica preliminar. ABSTRACT The phytochemical profile of ethanolic extract of Solanum lycocarpum fruits was analyzed and preliminary toxicity tests were performed against brine shrimp larvae. The extract was subjected to preliminary phytochemical analysis to identify the main classes of secondary metabolites and tested against the larvae of A. salina to obtain the median lethal concentrations (LC50%. The phytochemical tests showed the presence of phenols, tannins, saponins, alkaloids and free steroids. The extract was fractionated with various solvents for toxicity testing against the larvae and the hydroalcoholic fraction showed considerable cytotoxicity (CL50% = 285.546 g/mL. Keywords: Solanum lycocarpum, Artemia salina, phytochemical screening

Homogenous 96-plex PEA immunoassay exhibiting high sensitivity, specificity, and excellent scalability

DEFF Research Database (Denmark)

Assarsson, Erika; Lundberg, Martin; Holmquist, Göran

2014-01-01

reporters, shown potential to relieve the shortcomings of antibodies and their inherent cross-reactivity in multiplex protein quantification applications. The aim of the present study was to develop a robust 96-plex immunoassay based on the proximity extension assay (PEA) for improved high throughput...... detection of protein biomarkers. This was enabled by: (1) a modified design leading to a reduced number of pipetting steps compared to the existing PEA protocol, as well as improved intra-assay precision; (2) a new enzymatic system that uses a hyper-thermostabile enzyme, Pwo, for uniting the two probes......, such as serum and plasma, and also in xenografted mice and resuspended dried blood spots, consuming only 1 µL sample per test. All-in-all, the development of the current multiplex technique is a step toward robust high throughput protein marker discovery and research....

Parallel susceptibility testing of bacteria through culture-quantitative PCR in 96-well plates

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Jun Luo

2018-05-01

Full Text Available Objective: The methods combining culture and quantitative PCR(qPCR offer new solutions for rapid antibiotic susceptibility testing(AST. However, the multiple steps of DNA extraction and cold storage of PCR reagents needed make them unsuitable for rapid high throughput AST. In this study, a parallel culture-qPCR method was developed to overcome above problems. Method: In this method, bacteria culture and DNA extraction automatically and simultaneously completed through using a common PCR instrument as a controllable heating device. A lyophilized 16S rDNA targeted qPCR reagent was also developed, which was stable and could be kept at 4 °C for long time and at 37 °C for about two months. Result: Testing of 36 P. aeruginosa isolates and 28 S. aureus isolates showed that the method had good agreements with the standard broth microdilution method, with an overall agreement of 97.22% (95% CI, 85.83–99.51 for P. aeruginosa and 96.43% (95% CI, 79.76–99.81 for S. aureus. This method could test 12 samples against a panel of up to 7 antibiotics simultaneously in two 96-well PCR plates within 4 h, which greatly improves the testing efficiency of the culture-qPCR method. Conclusion: With rapidness to obtain results and the capabilities for automation and multiple-sample testing, the parallel culture-qPCR method would have great potentials in clinical labs. Keywords: Antibiotic susceptibility testing, Thermo-cold lysis, Lyophilized qPCR reagent, Quantitative PCR, Bacteria

Study of photosensitization reaction progress in a 96 well plate with photosensitizer rich condition using Talaporfin sodium

Science.gov (United States)

Ogawa, Emiyu; Takahashi, Mei; Arai, Tsunenori

2013-02-01

To quantitatively investigate photosensitization reaction in vitro against myocardial cells with photosensitizer rich condition in solution using Talaporfin sodium in the well of a 96 well plate, we studied photosensitization reaction progress in this well. We have proposed non-thermal conduction block of myocardium tissue using the photosensitization reaction with laser irradiation shortly after Talaporfin sodium injection. In above situation, the photosensitizer is located outside the myocardial cells in high concentration. To understand interaction of the photosensitization reaction in which the photosensitizer distributes outside cells, the photosensitization reaction progress in the well was studied. Talaporfin sodium (799.69 MW) solution and a 663 nm diode laser were used. The photosensitizer solution concentrations of 12.5-37.5 μM were employed. The photosensitizer fluorescence with 0.29 W/cm2 in irradiance, which was optimized in previous cell death study, was measured during the laser irradiation until 40 J/cm2. The photosensitizer solution absorbance and dissolved oxygen pressure after the laser irradiation were also measured. We found that the photosensitization reaction progress had 2 distinctive phases of different reaction rate: rapid photosensitization reaction consuming dissolved oxygen and gentle photosensitization reaction with oxygen diffusion from the solution-air boundary. The dissolved oxygen pressure and photosensitizer solution absorbance were 30% and 80% of the initial values after the laser irradiation, respectively. Therefore, oxygen was rate-controlling factor of the photosensitization reaction in the well with the photosensitizer rich condition. In the oxygen diffusion phase, the oxygen pressure was maintained around 40 mmHg until the laser irradiation of 40 J/cm2 and it is similar to that of myocardium tissue in vivo. We think that our 96 well plate in vitro system may simulate PDT in myocardial tissue with photosensitization reaction

Construcción macrotextual y cancionero amoroso (Un acercamiento analítico a "La voz a ti debida" de Pedro Salinas)

OpenAIRE

Corencia Cruz, Joaquín

2008-01-01

La tesis Construcción macrotextual y cancionero amoroso. Unacercamiento analítico a La voz a ti debida de Pedro Salinas. Fue leídapor Joaquín Corencia Cruz en la Sala de Juntas de la Facultad deFilología de la Universidad de Valencia (España) el 7 de noviembre de2008.El trabajo investigador se desarrolla a lo largo de nuevecapítulos. En el primero se produce una introducción a la tesisrevisando desde la trayectoria vital y profesional de Pedro Salinas acómo el macrotexto poético de La voz se ...

Development of a dual luciferase activity and fluorescamine protein assay adapted to a 384 micro-well plate format: Reducing variability in human luciferase transactivation cell lines aimed at endocrine active substances

Science.gov (United States)

Brennan, Jennifer; Tillitt, Donald E.

2018-01-01

There is a need to adapt cell bioassays to 384-well and 1536-well formats instead of the traditional 96-well format as high-throughput screening (HTS) demands increase. However, the sensitivity and performance of the bioassay must be re-verified in these higher micro-well plates, and verification of cell health must also be HT (high-throughput). We have adapted two commonly used human breast luciferase transactivation cell bioassays, the recently re-named estrogen agonist/antagonist screening VM7Luc4E2 cell bioassay (previously designated BG1Luc4E2) and the androgen/glucocorticoid screening MDA-kb2 cell bioassay, to 384-well formats for HTS of endocrine-active substances (EASs). This cost-saving adaptation includes a fast, accurate, and easy measurement of protein amount in each well via the fluorescamine assay with which to normalize luciferase activity of cell lysates without requiring any transfer of the cell lysates. Here we demonstrate that by accounting for protein amount in the cell lysates, antagonistic agents can easily be distinguished from cytotoxic agents in the MDA-kb2 and VM7Luc4E2 cell bioassays. Additionally, we demonstrate via the fluorescamine assay improved interpretation of luciferase activity in wells along the edge of the plate (the so-called “edge effect”), thereby increasing usable wells to the entire plate, not just interior wells.

Comparison of Five Assays for Detection of Clostridium difficile Toxin

Science.gov (United States)

Chapin, Kimberle C.; Dickenson, Roberta A.; Wu, Fongman; Andrea, Sarah B.

2011-01-01

Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. PMID:21704273

Evaluation of Alpha and Gamma Aluminum Oxide Nanoparticle Accumulation, Toxicity and Depuration in Artemia Salina Larvae

Science.gov (United States)

Ates, Mehmet; Demir, Veysel; Arslan, Zikri; Daniels, James; Farah, Ibrahim O.; Bogatu, Corneliu

2014-01-01

In this study, Artemia salina (crustacean filter feeders) larvae were used as a test model to investigate the toxicity of aluminum oxide nanoparticles (Al2O3 NPs) on marine microorganisms. The uptake, toxicity and elimination of α-Al2O3 (50 nm and 3.5 μm) and γ-Al2O3 (5 nm and 0.4 μm) NPs were studied. Twenty-four and ninety-six hour exposures of different concentrations of Al2O3 NPs to Artemia larvae were conducted in a seawater medium. When suspended in water, Al2O3 NPs aggregated substantially with the sizes ranging from 6.3 nm to > 0.3 μm for spherical NPs, and from 250 to 756 nm for rod-shaped NPs. The phase contrast microscope images revealed that NPs deposited inside the guts as aggregates. ICP-MS analysis showed that large particles (3.5 μm α-Al2O3) were not taken up by Artemia, while fine NPs (0.4 μm γ-Al2O3) and ultra-fine NPs (5 nm γ-Al2O3 and 50 nm α-Al2O3) accumulated substantially. Differences in toxicity were detected as changing with NP size and morphology. The malondialdehyde (MDA) levels indicated that smaller γ-Al2O3 (5 nm) NPs were more toxic than larger γ-Al2O3 (0.4 μm) particulates in 96 h. The highest mortality was measured as 34% in 96 h for γ-Al2O3 NPs (5 nm) at 100 mg/L (LC50 > 100 mg/L). γ-Al2O3 NPs were more toxic than α-Al2O3 NPs at in all conditions. PMID:24753078

The Cytotoxicity Study of Carboxymethyl Starch (CMS) of Sago Starch (Metro xylon sago) by Brine Shrimp Lethality Test (Artemia salina nauplii)

International Nuclear Information System (INIS)

Ibrahim Ijang; Fazliana Mohd Saaya; Zainon Othman

2014-01-01

CMS can be produced by substitution of the hydroxyl groups with sodium monochloroacetate in the presence of strong alkali. Carboxy methylation can be performed in water as a solvent or in a water-miscible organic solvent containing a small amount of water such as ethanol, isopropanol, methanol or toluene. The use of organic solvent will preserve the final product in the granular form and the side product can be washed out easily but some of them may be having potential toxicity and carcinogenic effect. In this study, CMS was investigated the level of toxicity by using brine shrimp lethality (BSLT). Brine shrimp test method was used to screen CMS for their biological activity. The screening results showed that the LC50, of CMS is more than 100 mg/ ml dose concentration. In conclusion, CMS is not cytotoxicity to Artemia salina nauplii and BSLT method is simple, inexpensive and convenient assay for the detection of cytotoxic compound. (author)

Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System.

Science.gov (United States)

Conway, Michael K; Gerger, Michael J; Balay, Erin E; O'Connell, Rachel; Hanson, Seth; Daily, Neil J; Wakatsuki, Tetsuro

2015-05-14

Continued advancement in pluripotent stem cell culture is closing the gap between bench and bedside for using these cells in regenerative medicine, drug discovery and safety testing. In order to produce stem cell derived biopharmaceutics and cells for tissue engineering and transplantation, a cost-effective cell-manufacturing technology is essential. Maintenance of pluripotency and stable performance of cells in downstream applications (e.g., cell differentiation) over time is paramount to large scale cell production. Yet that can be difficult to achieve especially if cells are cultured manually where the operator can introduce significant variability as well as be prohibitively expensive to scale-up. To enable high-throughput, large-scale stem cell production and remove operator influence novel stem cell culture protocols using a bench-top multi-channel liquid handling robot were developed that require minimal technician involvement or experience. With these protocols human induced pluripotent stem cells (iPSCs) were cultured in feeder-free conditions directly from a frozen stock and maintained in 96-well plates. Depending on cell line and desired scale-up rate, the operator can easily determine when to passage based on a series of images showing the optimal colony densities for splitting. Then the necessary reagents are prepared to perform a colony split to new plates without a centrifugation step. After 20 passages (~3 months), two iPSC lines maintained stable karyotypes, expressed stem cell markers, and differentiated into cardiomyocytes with high efficiency. The system can perform subsequent high-throughput screening of new differentiation protocols or genetic manipulation designed for 96-well plates. This technology will reduce the labor and technical burden to produce large numbers of identical stem cells for a myriad of applications.

Detection and removal of spatial bias in multiwell assays.

Science.gov (United States)

Lachmann, Alexander; Giorgi, Federico M; Alvarez, Mariano J; Califano, Andrea

2016-07-01

Multiplex readout assays are now increasingly being performed using microfluidic automation in multiwell format. For instance, the Library of Integrated Network-based Cellular Signatures (LINCS) has produced gene expression measurements for tens of thousands of distinct cell perturbations using a 384-well plate format. This dataset is by far the largest 384-well gene expression measurement assay ever performed. We investigated the gene expression profiles of a million samples from the LINCS dataset and found that the vast majority (96%) of the tested plates were affected by a significant 2D spatial bias. Using a novel algorithm combining spatial autocorrelation detection and principal component analysis, we could remove most of the spatial bias from the LINCS dataset and show in parallel a dramatic improvement of similarity between biological replicates assayed in different plates. The proposed methodology is fully general and can be applied to any highly multiplexed assay performed in multiwell format. ac2248@columbia.edu Supplementary data are available at Bioinformatics online. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

A Rapid Zika Diagnostic Assay to Measure Neutralizing Antibodies in Patients

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Chao Shan

2017-03-01

Full Text Available The potential association of microcephaly and other congenital abnormalities with Zika virus (ZIKV infection during pregnancy underlines the critical need for a rapid and accurate diagnosis. Due to the short duration of ZIKV viremia in infected patients, a serologic assay that detects antibody responses to viral infection plays an essential role in diagnosing patient specimens. The current serologic diagnosis of ZIKV infection relies heavily on the labor-intensive Plaque Reduction Neutralization Test (PRNT that requires more than one-week turnaround time and represents a major bottleneck for patient diagnosis. To overcome this limitation, we have developed a high-throughput assay for ZIKV and dengue virus (DENV diagnosis that can attain the “gold standard” of the current PRNT assay. The new assay is homogeneous and utilizes luciferase viruses to quantify the neutralizing antibody titers in a 96-well format. Using 91 human specimens, we showed that the reporter diagnostic assay has a higher dynamic range and maintains the relative specificity of the traditional PRNT assay. Besides the improvement of assay throughput, the reporter virus technology has also shortened the turnaround time to less than two days. Collectively, our results suggest that, along with the viral RT-PCR assay, the reporter virus-based serologic assay could be potentially used as the first-line test for clinical diagnosis of ZIKV infection as well as for vaccine clinical trials.

Influencia del medio ambiente en los materiales de construcción III. Dinámica salina

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Fernández Paris, José Manuel

1978-06-01

Full Text Available Las sales y la humedad son los dos factores que condicionan las alteraciones de los materiales de construcción. Los citados materiales, bien de por sí —piedras y ladrillos—, bien una vez endurecidos —morteros y hormigones—, están estructurados por partículas sólidas granulares, separadas por un sistema poroso, que constituye el medio -dinámico para el transporte de las disoluciones salinas procedentes del medio ambiente. La motilidad de las sales a través de la red de poros es función de tres variables físicoclimáticas: temperatura, humedad y condiciones de aireación. La dinámica salina se efectúa mediante procesos de difusión, unas veces de naturaleza química: difusión iónica, otras de naturaleza física: difusión capilar.

Pseudomonas salina sp. nov., isolated from a salt lake.

Science.gov (United States)

Zhong, Zhi-Ping; Liu, Ying; Hou, Ting-Ting; Liu, Hong-Can; Zhou, Yu-Guang; Wang, Fang; Liu, Zhi-Pei

2015-09-01

A Gram-staining-negative, facultatively aerobic bacterium, strain XCD-X85(T), was isolated from Xiaochaidan Lake, a salt lake (salinity 9.9%, w/v) in Qaidam basin, Qinghai province, China. Its taxonomic position was determined by using a polyphasic approach. Cells of strain XCD-X85(T) were non-endospore-forming rods, 0.4-0.6 μm wide and 1.0-1.6 μm long, and motile by means of a single polar flagellum. Strain XCD-X85(T) was catalase- and oxidase-positive. Growth was observed in the presence of 0-12.0% (w/v) NaCl (optimum, 1.0-2.0%) and at 4-35 °C (optimum, 25-30 °C) and pH 6.5-10.5 (optimum, pH 8.0-8.5). Strain XCD-X85(T) contained (>10%) summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), C12 : 0, C16 : 0 and summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) as the predominant fatty acids. The major respiratory quinone was ubiquinone 9 (Q-9). The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The DNA G+C content was 57.4 mol%. Phylogenetic trees based on 16S rRNA gene sequences showed that strain XCD-X85(T) was associated with the genus Pseudomonas, and showed highest 16S rRNA gene sequence similarities to Pseudomonas pelagia CL-AP6(T) (99.0%) and Pseudomonas bauzanensis BZ93(T) (96.8%). DNA-DNA relatedness of strain XCD-X85T to P. pelagia JCM 15562(T) was 19 ± 1%. On the basis of the data presented above, it is concluded that strain XCD-X85(T) represents a novel species of the genus Pseudomonas, for which the name Pseudomonas salina sp. nov. is proposed. The type strain is XCD-X85(T) ( = CGMCC 1.12482(T) = JCM 19469(T)).

Protocol: high throughput silica-based purification of RNA from Arabidopsis seedlings in a 96-well format

Directory of Open Access Journals (Sweden)

Salvo-Chirnside Eliane

2011-12-01

Full Text Available Abstract The increasing popularity of systems-based approaches to plant research has resulted in a demand for high throughput (HTP methods to be developed. RNA extraction from multiple samples in an experiment is a significant bottleneck in performing systems-level genomic studies. Therefore we have established a high throughput method of RNA extraction from Arabidopsis thaliana to facilitate gene expression studies in this widely used plant model. We present optimised manual and automated protocols for the extraction of total RNA from 9-day-old Arabidopsis seedlings in a 96 well plate format using silica membrane-based methodology. Consistent and reproducible yields of high quality RNA are isolated averaging 8.9 μg total RNA per sample (~20 mg plant tissue. The purified RNA is suitable for subsequent qPCR analysis of the expression of over 500 genes in triplicate from each sample. Using the automated procedure, 192 samples (2 × 96 well plates can easily be fully processed (samples homogenised, RNA purified and quantified in less than half a day. Additionally we demonstrate that plant samples can be stored in RNAlater at -20°C (but not 4°C for 10 months prior to extraction with no significant effect on RNA yield or quality. Additionally, disrupted samples can be stored in the lysis buffer at -20°C for at least 6 months prior to completion of the extraction procedure providing a flexible sampling and storage scheme to facilitate complex time series experiments.

Protocol: high throughput silica-based purification of RNA from Arabidopsis seedlings in a 96-well format.

Science.gov (United States)

Salvo-Chirnside, Eliane; Kane, Steven; Kerr, Lorraine E

2011-12-02

The increasing popularity of systems-based approaches to plant research has resulted in a demand for high throughput (HTP) methods to be developed. RNA extraction from multiple samples in an experiment is a significant bottleneck in performing systems-level genomic studies. Therefore we have established a high throughput method of RNA extraction from Arabidopsis thaliana to facilitate gene expression studies in this widely used plant model. We present optimised manual and automated protocols for the extraction of total RNA from 9-day-old Arabidopsis seedlings in a 96 well plate format using silica membrane-based methodology. Consistent and reproducible yields of high quality RNA are isolated averaging 8.9 μg total RNA per sample (~20 mg plant tissue). The purified RNA is suitable for subsequent qPCR analysis of the expression of over 500 genes in triplicate from each sample. Using the automated procedure, 192 samples (2 × 96 well plates) can easily be fully processed (samples homogenised, RNA purified and quantified) in less than half a day. Additionally we demonstrate that plant samples can be stored in RNAlater at -20°C (but not 4°C) for 10 months prior to extraction with no significant effect on RNA yield or quality. Additionally, disrupted samples can be stored in the lysis buffer at -20°C for at least 6 months prior to completion of the extraction procedure providing a flexible sampling and storage scheme to facilitate complex time series experiments.

Developmental toxicity of oxidized multi-walled carbon nanotubes on Artemia salina cysts and larvae: Uptake, accumulation, excretion and toxic responses.

Science.gov (United States)

Zhu, Song; Luo, Fei; Tu, Xiao; Chen, Wei-Chao; Zhu, Bin; Wang, Gao-Xue

2017-10-01

Using Artemia salina (A. salina) cysts (capsulated and decapsulated) and larvae [instar I (0-24 h), II (24-48 h) and III (48-72 h)] as experimental models, developmental toxicity of oxidized multi-walled carbon nanotubes (O-MWCNTs) was evaluated. Results revealed that hatchability of capsulated and decapsulated cysts was significantly decreased (p larvae in 600 mg/L. The EC 50 values for swimming inhibition of instar I, II and III were 535, 385 and 472 mg/L, respectively. Instar II showed the greatest sensitivity to O-MWCNTs, and followed by instar III, instar I, decapsulated cysts and capsulated cysts. Effects on hatchability, mortality and swimming were accounted for O-MWCNTs rather than metal catalyst impurities. Body length was decreased with the concentrations increased from 0 to 600 mg/L. O-MWCNTs attached onto the cysts, gill and body surface, resulting in irreversible damages. Reactive oxygen species, malondialdehyde content, total antioxidant capacity and antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) activities were increased following exposure, indicating that the effects were related to oxidative stress. O-MWCNTs were ingested and distributed in phagocyte, lipid vesicle and intestine. Most of the accumulated O-MWCNTs were excreted by A. salina at 72 h, but some still remained in the organism. Data of uptake kinetics showed that O-MWCNTs contents in A. salina were gradually increased from 1 to 48 h and followed by rapidly decreased from 48 to 72 h with a range from 5.5 to 28.1 mg/g. These results so far indicate that O-MWCNTs have the potential to affect aquatic organisms when released into the marine ecosystems. Copyright © 2017 Elsevier Ltd. All rights reserved.

Time-resolved immunofluorometric assay of serum ferritin

Energy Technology Data Exchange (ETDEWEB)

Yan, Yao [China Inst. of Atomic Energy, Beijing (China)

2007-06-15

This assay is a solid phase, two-site fluoroimmunometric assay based on the direct sandwish technique. Standards or samples containing ferritin are first reacted with immobilized anti-ferritin antibodies. Then the europium-lablled antibodies are reacted with the bound antigen. The range of this assay is 2-1000 ng/mL. The analytical sentivity is better than 0.05 ng/mL. The intra-assay variation and inter-assay variation are both below 5%; This kit was compared with Wallac DELFIA kit. The correlation is r=0.96. (authors)

Comparison of five assays for detection of Clostridium difficile toxin.

Science.gov (United States)

Chapin, Kimberle C; Dickenson, Roberta A; Wu, Fongman; Andrea, Sarah B

2011-07-01

Performance characteristics of five assays for detection of Clostridium difficile toxin were compared using fresh stool samples from patients with C. difficile infection (CDI). Assays were performed simultaneously and according to the manufacturers' instructions. Patients were included in the study if they exhibited clinical symptoms consistent with CDI. Nonmolecular assays included glutamate dehydrogenase antigen tests, with positive findings followed by the Premier Toxin A and B Enzyme Immunoassay (GDH/EIA), and the C. Diff Quik Chek Complete test. Molecular assays (PCR) included the BD GeneOhm Cdiff Assay, the Xpert C. difficile test, and the ProGastro Cd assay. Specimens were considered true positive if results were positive in two or more assays. For each method, the Youden index was calculated and cost-effectiveness was analyzed. Of 81 patients evaluated, 26 (32.1%) were positive for CDI. Sensitivity of the BD GeneOhm Cdiff assay, the Xpert C. difficile test, the ProGastro Cd assay, C. Diff Quik Chek Complete test, and two-step GDH/EIA was 96.2%, 96.2%, 88.5%, 61.5%, and 42.3%, respectively. Specificity of the Xpert C. difficile test was 96.4%, and for the other four assays was 100%. Compared with nonmolecular methods, molecular methods detected 34.7% more positive specimens. Assessment of performance characteristics and cost-effectiveness demonstrated that the BD GeneOhm Cdiff assay yielded the best results. While costly, the Xpert C. difficile test required limited processing and yielded rapid results. Because of discordant results, specimen processing, and extraction equipment requirements, the ProGastro Cd assay was the least favored molecular assay. The GDH/EIA method lacked sufficient sensitivity to be recommended. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

Uso da solução salina para manutenção de acessos venosos em adultos: uma revisão

OpenAIRE

Oliveira,Francimar Tinoco de; Silva,Lolita Dopico da

2006-01-01

Trata-se de uma revisão bibliográfica sobre a prática da salinização em cateteres venosos periféricos. O estudo objetivou analisar os artigos científicos sobre o uso de solução salina na manutenção da permeabilidade de cateteres venosos periféricos em adultos, indexados no MedLine e LILACS no período de 1995 a 2005. A amostra consistiu de 5 artigos os quais foram analisados quanto à procedência e periódico de publicação, ao delineamento do estudo, a amostragem, e ao efeito da solução salina. ...

SALT EFFECTS ON EGGS AND NAUPLII OF ARTEMIA SALINA L

Science.gov (United States)

Boone, Eleanor; Baas-Becking, L. G. M.

1931-01-01

Eggs of Artemia salina L., the brine shrimp, are easily obtainable in large quantities. Ecdysis takes place in two stages: (a) extrusion of the inner membrane, and (b) ecdysis of the nauplius from that membrane. The conditions which allow for the former are much more varied than those for the latter. Nauplii form in only solutions of a few sodium salts; and, in Mg, Ca, and Sr salts, potassium is very toxic. The possible environment for the nauplii (1 M total molarity) has been ascertained for chlorides of Na, K, Mg, and Ca. The facts observed account for the peculiar distribution of the organism. PMID:19872620

The comet assay: assessment of in vitro and in vivo DNA damage.

Science.gov (United States)

Bajpayee, Mahima; Kumar, Ashutosh; Dhawan, Alok

2013-01-01

Rapid industrialization and pursuance of a better life have led to an increase in the amount of chemicals in the environment, which are deleterious to human health. Pesticides, automobile exhausts, and new chemical entities all add to air pollution and have an adverse effect on all living organisms including humans. Sensitive test systems are thus required for accurate hazard identification and risk assessment. The Comet assay has been used widely as a simple, rapid, and sensitive tool for assessment of DNA damage in single cells from both in vitro and in vivo sources as well as in humans. Already, the in vivo comet assay has gained importance as the preferred test for assessing DNA damage in animals for some international regulatory guidelines. The advantages of the in vivo comet assay are its ability to detect DNA damage in any tissue, despite having non-proliferating cells, and its sensitivity to detect genotoxicity. The recommendations from the international workshops held for the comet assay have resulted in establishment of guidelines. The in vitro comet assay conducted in cultured cells and cell lines can be used for screening large number of compounds and at very low concentrations. The in vitro assay has also been automated to provide a high-throughput screening method for new chemical entities, as well as environmental samples. This chapter details the in vitro comet assay using the 96-well plate and in vivo comet assay in multiple organs of the mouse.

A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents

International Nuclear Information System (INIS)

Zhu, Xiaoming; Fu, Afu; Luo, Kathy Qian

2012-01-01

Highlights: ► An endothelial cell apoptosis assay using FRET-based biosensor was developed. ► The fluorescence of the cells changed from green to blue during apoptosis. ► This method was developed into a high-throughput assay in 96-well plates. ► This assay was applied to screen vascular disrupting agents. -- Abstract: In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening.

Effect of mixing rate on Beta-carotene production and extraction by dunaliella salina in two-phase bioreactors

NARCIS (Netherlands)

Hejazi, M.; Andrysiewicz, E.; Tramper, J.; Wijffels, R.H.

2003-01-01

beta-Carotene has many applications in the food, cosmetic, and pharmaceutical industries; Dunaliella salina is currently the main source for natural beta-carotene. We have investigated the effect of mixing rate and whether it leads to the facilitated release of beta-carotene from the cells of

MS transport assays for γ-aminobutyric acid transporters--an efficient alternative for radiometric assays.

Science.gov (United States)

Schmitt, Sebastian; Höfner, Georg; Wanner, Klaus T

2014-08-05

Transport assays for neurotransmitters based on radiolabeled substrates are widely spread and often indispensable in basic research and the drug development process, although the use of radioisotopes is inherently coupled to issues concerning radioactive waste and safety precautions. To overcome these disadvantages, we developed mass spectrometry (MS)-based transport assays for γ-aminobutyric acid (GABA), which is the major inhibitory neurotransmitter in the central nervous system (CNS). These "MS Transport Assays" provide all capabilities of [(3)H]GABA transport assays and therefore represent the first substitute for the latter. The performance of our approach is demonstrated for GAT1, the most important GABA transporter (GAT) subtype. As GABA is endogenously present in COS-7 cells employed as hGAT1 expression system, ((2)H6)GABA was used as a substrate to differentiate transported from endogenous GABA. To record transported ((2)H6)GABA, a highly sensitive, short, robust, and reliable HILIC-ESI-MS/MS quantification method using ((2)H2)GABA as an internal standard was developed and validated according to the Center for Drug Evaluation and Research (CDER) guidelines. Based on this LC-MS quantification, a setup to characterize hGAT1 mediated ((2)H6)GABA transport in a 96-well format was established, that enables automated processing and avoids any sample preparation. The K(m) value for ((2)H6)GABA determined for hGAT1 is in excellent agreement with results obtained from [(3)H]GABA uptake assays. In addition, the established assay format enables efficient determination of the inhibitory potency of GAT1 inhibitors, is capable of identifying those inhibitors transported as substrates, and furthermore allows characterization of efflux. The approach described here combines the strengths of LC-MS/MS with the high efficiency of transport assays based on radiolabeled substrates and is applicable to all GABA transporter subtypes.

A High Sensitivity Micro Format Chemiluminescence Enzyme Inhibition Assay for Determination of Hg(II

Directory of Open Access Journals (Sweden)

Kanchanmala Deshpande

2010-06-01

Full Text Available A highly sensitive and specific enzyme inhibition assay based on alcohol oxidase (AlOx and horseradish peroxidase (HRP for determination of mercury Hg(II in water samples has been presented. This article describes the optimization and miniaturization of an enzymatic assay using a chemiluminescence reaction. The analytical performance and detection limit for determination of Hg(II was optimized in 96 well plates and further extended to 384 well plates with a 10-fold reduction in assay volume. Inhibition of the enzyme activity by dissolved Hg(II was found to be linear in the range 5–500 pg.mL−1 with 3% CVin inter-batch assay. Due to miniaturization of assay in 384 well plates, Hg(II was measurable as low as 1 pg.mL−1 within15 min. About 10-fold more specificity of the developed assay for Hg(II analysis was confirmed by challenging with interfering divalent metal ions such as cadmium Cd(II and lead Pb(II. Using the proposed assay we could successfully demonstrate that in a composite mixture of Hg(II, Cd(II and Pb(II, inhibition by each metal ion is significantly enhanced in the presence of the others. Applicability of the proposed assay for the determination of the Hg(II in spiked drinking and sea water resulted in recoveries ranging from 100–110.52%.

Fine structure and function of the alimentary epithelium in Artemia salina nauplii

Energy Technology Data Exchange (ETDEWEB)

Hootman, S R; Conte, F P

1974-01-01

The fine structure of the alimentary tract in the second instar nauplius of the brine shrimp, Artemia salina, has been described. The foregut and hindgut of the larva are composed of cuboidal epithelium which is cuticularized. The epithelium of the midgut and gastric caeca is columnar and is characterized by apical microvilli, basal membrane infolds, and abundant mitochondria. The structural characteristics of the midgut cells correlate with previous physiological and biochemical evidence on both adult and larval brine shrimp which indicates that the midgut plays an important role in absorption and osmoregulation in these animals.

Interference from ordinarily used solvents in the outcomes of Artemia salina lethality test

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Sahgal Geethaa

2013-01-01

Full Text Available Methanol, ethanol, Tween 20 and dimethyl sulfoxide (DMSO are widely used as dissolving agents in Artemia salina lethality test (aka brine shrimp lethality test [BSLT] to screen the pharmaceutical properties of natural products. Nevertheless, there is lack of toxicity level of these solvents against brine shrimp. High concentration of these organic solvent might be toxic for this zoology invertebrate and interfere in the experimental outcomes. To avoid this, permissible concentration of the solvents used in BSLT was identified. BSLT was performed to evaluate the toxicity effect of Tween 20, methanol, ethanol and DMSO at 24 h post-treatment time point against A. salina. The suggested maximum working concentration (v/v for DMSO, methanol, ethanol was found to be 1.25% and that for Tween 20 was 0.16%. LC 50 for the solvents were 8.5% (DMSO, 6.4% (methanol, 3.4% (ethanol and 2.5% (Tween 20. The findings have shown a toxicity level among the solvents in descending order as Tween 20 > ethanol > methanol > DMSO. DMSO is a safer solvent to be used in BSLT compared with other tested solvents, whereas Tween 20 has been shown to be the most stringent solvent among the tested solvents. The findings are resourcefully useful to avoid interference of solvents in the assessment of natural products using BSLT.

A rapid enzymatic assay for high-throughput screening of adenosine-producing strains

Science.gov (United States)

Dong, Huina; Zu, Xin; Zheng, Ping; Zhang, Dawei

2015-01-01

Adenosine is a major local regulator of tissue function and industrially useful as precursor for the production of medicinal nucleoside substances. High-throughput screening of adenosine overproducers is important for industrial microorganism breeding. An enzymatic assay of adenosine was developed by combined adenosine deaminase (ADA) with indophenol method. The ADA catalyzes the cleavage of adenosine to inosine and NH3, the latter can be accurately determined by indophenol method. The assay system was optimized to deliver a good performance and could tolerate the addition of inorganic salts and many nutrition components to the assay mixtures. Adenosine could be accurately determined by this assay using 96-well microplates. Spike and recovery tests showed that this assay can accurately and reproducibly determine increases in adenosine in fermentation broth without any pretreatment to remove proteins and potentially interfering low-molecular-weight molecules. This assay was also applied to high-throughput screening for high adenosine-producing strains. The high selectivity and accuracy of the ADA assay provides rapid and high-throughput analysis of adenosine in large numbers of samples. PMID:25580842

The monoclonal S9.6 antibody exhibits highly variable binding affinities towards different R-loop sequences.

Directory of Open Access Journals (Sweden)

Fabian König

Full Text Available The monoclonal antibody S9.6 is a widely-used tool to purify, analyse and quantify R-loop structures in cells. A previous study using the surface plasmon resonance technology and a single-chain variable fragment (scFv of S9.6 showed high affinity (0.6 nM for DNA-RNA and also a high affinity (2.7 nM for RNA-RNA hybrids. We used the microscale thermophoresis method allowing surface independent interaction studies and electromobility shift assays to evaluate additional RNA-DNA hybrid sequences and to quantify the binding affinities of the S9.6 antibody with respect to distinct sequences and their GC-content. Our results confirm high affinity binding to previously analysed sequences, but reveals that binding affinities are highly sequence specific. Our study presents R-loop sequences that independent of GC-content and in different sequence variations exhibit either no binding, binding affinities in the micromolar range and as well high affinity binding in the nanomolar range. Our study questions the usefulness of the S9.6 antibody in the quantitative analysis of R-loop sequences in vivo.

Groundwater-quality data in the Monterey–Salinas shallow aquifer study unit, 2013: Results from the California GAMA Program

Science.gov (United States)

Goldrath, Dara A.; Kulongoski, Justin T.; Davis, Tracy A.

2016-09-01

Groundwater quality in the 3,016-square-mile Monterey–Salinas Shallow Aquifer study unit was investigated by the U.S. Geological Survey (USGS) from October 2012 to May 2013 as part of the California State Water Resources Control Board Groundwater Ambient Monitoring and Assessment (GAMA) Program’s Priority Basin Project. The GAMA Monterey–Salinas Shallow Aquifer study was designed to provide a spatially unbiased assessment of untreated-groundwater quality in the shallow-aquifer systems in parts of Monterey and San Luis Obispo Counties and to facilitate statistically consistent comparisons of untreated-groundwater quality throughout California. The shallow-aquifer system in the Monterey–Salinas Shallow Aquifer study unit was defined as those parts of the aquifer system shallower than the perforated depth intervals of public-supply wells, which generally corresponds to the part of the aquifer system used by domestic wells. Groundwater quality in the shallow aquifers can differ from the quality in the deeper water-bearing zones; shallow groundwater can be more vulnerable to surficial contamination.Samples were collected from 170 sites that were selected by using a spatially distributed, randomized grid-based method. The study unit was divided into 4 study areas, each study area was divided into grid cells, and 1 well was sampled in each of the 100 grid cells (grid wells). The grid wells were domestic wells or wells with screen depths similar to those in nearby domestic wells. A greater spatial density of data was achieved in 2 of the study areas by dividing grid cells in those study areas into subcells, and in 70 subcells, samples were collected from exterior faucets at sites where there were domestic wells or wells with screen depths similar to those in nearby domestic wells (shallow-well tap sites).Field water-quality indicators (dissolved oxygen, water temperature, pH, and specific conductance) were measured, and samples for analysis of inorganic

97 Etude éco-biologique d'Artémia salina des zones humides de l ...

African Journals Online (AJOL)

Grenelle

l'aviculture de poissons marins [1]. En Algérie, les travaux effectués sur ce crustacé sont relativement rares. Dans le présent ..... dépassent pour certains les normes établies par l'OMS, ce qui peut provoquer le comportement et la dynamique de la reproduction de l'Artémia salina, sans oublier aussi l'action anthropique qui ...

Pastas de Rhodomonas salina (Cryptophyta como alimento para Brachionus plicatilis (Rotifera

Directory of Open Access Journals (Sweden)

Miguel Guevara

2011-12-01

Full Text Available Pastas de Rhodomonas salina, obtenidas mediante centrifugación y floculación con quitosano y preservadas con o sin vitamina C, a -20°C fueron evaluadas bioquímicamente y proporcionadas como alimento al rotífero Brachionus plicatilis. Las pastas microalgales: (1 centrifugada y con vitamina C (CV, (2 centrifugada y sin vitamina C (C, (3 floculada y con vitamina C (FV y (4 floculada y sin adición de vitamina C (F; mantuvieron sus contenidos de proteínas y lípidos totales similares al cultivo control, con valores de 40.0±2.32% y 12.0±1.45%, respectivamente. La relación feofitina a/clorofila a fue similar (0.09-0.11 entre las pastas centrifugadas y el cultivo control, pero mayor en las pastas floculadas (1.28-1.48. Las pastas centrifugadas presentaron porcentajes de PUFAs totales, EPA y DHA similares al cultivo control (PUFAs: 47%, EPA: 4% y DHA: 4.7% y superiores al de las pastas floculadas. Las pastas obtenidas por centrifugación indujeron un crecimiento del rotífero igual al obtenido con el alimento control (densidad máxima: 320rotíferos/mL; tasa instantánea de crecimiento: 0.23rotíferos/día, fecundidad: 1.49huevos/ hembra y productividad: 43x103rotíferos/L/día. Se concluye que la pasta de R. salina centrifugada y congelada a -20°C, durante cuatro semanas, sin adición de vitamina C, mantiene su calidad nutricional similar a la del alga fresca y puede ser usada como alimento de Brachionus plicatilis.

Development of a novel 96-well format for liquid-liquid microextraction and its application in the HPLC analysis of biological samples.

Science.gov (United States)

Borijihan, Guirong; Li, Youxin; Gao, Jianguo; Bao, James J

2014-05-01

A novel 96-well liquid-liquid microextraction system combined with modern HPLC was developed and used for the simultaneous analysis of 96 biological samples. The system made use of hollow fibers, a 96-well plate, and a plastic base with a center hole and a side hole. One end of the hollow fiber was sealed, while the other end was attached to one of the holes positioned at the center for the plastic base. The needle was inserted into the liquid from inside or outside of the hollow fiber through the center or the side holes, respectively. The system was tested with plasma samples containing three compounds, acidic indomethacin, neutral dexamethasone, and basic propafenone. Some parameters, such as the kind and dimension of hollow fiber, pH and salt concentration of the donor phase, the selection of organic solvent for the acceptor phase, and the extraction time were investigated. Under the optimization conditions, the Log D and drug concentration of indomethacin, dexamethasone, and propafenone in plasma and urine samples were analyzed. Then, the methodology was validated. The results demonstrated that ng/mL levels could be exactly and rapidly analyzed by our system, which was equipped with an auto-injection sampler, making sample analysis more convenient. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Biotechnological potential of Synechocystis salina co-cultures with selected microalgae and cyanobacteria: Nutrients removal, biomass and lipid production.

Science.gov (United States)

Gonçalves, Ana L; Pires, José C M; Simões, Manuel

2016-01-01

Cultivation of microalgae and cyanobacteria has been the focus of several research studies worldwide, due to the huge biotechnological potential of these photosynthetic microorganisms. However, production of these microorganisms is still not economically viable. One possible alternative to improve the economic feasibility of the process is the use of consortia between microalgae and/or cyanobacteria. In this study, Chlorella vulgaris, Pseudokirchneriella subcapitata and Microcystis aeruginosa were co-cultivated with Synechocystis salina to evaluate how dual-species cultures can influence biomass and lipid production and nutrients removal. Results have shown that the three studied consortia achieved higher biomass productivities than the individual cultures. Additionally, nitrogen and phosphorus consumption rates by the consortia provided final concentrations below the values established by European Union legislation for these nutrients. In the case of lipid productivities, higher values were determined when S. salina was co-cultivated with P. subcapitata and M. aeruginosa. Copyright © 2015 Elsevier Ltd. All rights reserved.

Selection of well labelled insulin fractions for radioimmunoassay use

Energy Technology Data Exchange (ETDEWEB)

Awh, O D; Kim, J R [Korea Atomic Energy Research Inst., Seoul (Republic of Korea)

1980-06-01

Selection methods of well labelled insulin fractions based on two different criteria were compared to establish an efficient low level RIA of insulin and to elucidate the correlation between the immunoreactivity and the charcoal-adsorptivity of the radioiodine labelled insulin. The result indicated that the selection of well labelled insulin fractions by means of a charcoal-adsorption test is inappropriate. Generally, the distribution of radioactivity, antibody-bindability, and charcoal-adsorptivity of the labelled insulin was not consistent with each other. Thus, the selection should be carried out for every labelling batch to get the utmost assay reliability by antibody-bindability but not by charcoal-adsorptivity. By using the well selected labelled insulin fractions based on antibody-binding, a correct assay for a reference serum was possible, and by extending the incubation time up to 96 hrs, a sharp dose response curve could be obtained even in the range of below 5 ..mu..U/ml standard insulin doses.

Time-lapse 3-D measurements of a glucose biosensor in multicellular spheroids by light sheet fluorescence microscopy in commercial 96-well plates.

Science.gov (United States)

Maioli, Vincent; Chennell, George; Sparks, Hugh; Lana, Tobia; Kumar, Sunil; Carling, David; Sardini, Alessandro; Dunsby, Chris

2016-11-25

Light sheet fluorescence microscopy has previously been demonstrated on a commercially available inverted fluorescence microscope frame using the method of oblique plane microscopy (OPM). In this paper, OPM is adapted to allow time-lapse 3-D imaging of 3-D biological cultures in commercially available glass-bottomed 96-well plates using a stage-scanning OPM approach (ssOPM). Time-lapse 3-D imaging of multicellular spheroids expressing a glucose Förster resonance energy transfer (FRET) biosensor is demonstrated in 16 fields of view with image acquisition at 10 minute intervals. As a proof-of-principle, the ssOPM system is also used to acquire a dose response curve with the concentration of glucose in the culture medium being varied across 42 wells of a 96-well plate with the whole acquisition taking 9 min. The 3-D image data enable the FRET ratio to be measured as a function of distance from the surface of the spheroid. Overall, the results demonstrate the capability of the OPM system to measure spatio-temporal changes in FRET ratio in 3-D in multicellular spheroids over time in a multi-well plate format.

Comparative evaluation of impact of Zn and ZnO nanoparticles on brine shrimp (Artemia salina) larvae: effects of particle size and solubility on toxicity.

Science.gov (United States)

Ates, Mehmet; Daniels, James; Arslan, Zikri; Farah, Ibrahim O; Rivera, Hilsamar Félix

2013-01-01

Brine shrimp (Artemia salina) larvae were exposed to different sizes of zinc (Zn) and zinc oxide (ZnO) nanoparticles (NPs) to evaluate their toxicity in marine aquatic ecosystems. Acute exposure was conducted in seawater with 10, 50 and 100 mg L(-1) concentrations of the NPs for 24 h and 96 h. Phase contrast microscope images confirmed the accumulation of the NPs inside the guts. Artemia were unable to eliminate the ingested particles, which was thought to be due to the formation of massive particles in the guts. Although the suspensions of the NPs did not exhibit any significant acute toxicity within 24 h, mortalities increased remarkably in 96 h and escalated with increasing concentration of NP suspension to 42% for Zn NPs (40-60 nm) (LC50∼ 100 mg L(-1)) and to about 34% for ZnO NPs (10-30 nm) (LC50 > 100 mg L(-1)). The suspensions of Zn NPs were more toxic to Artemia than those of ZnO NPs under comparable regimes. This effect was attributed to higher Zn(2+) levels (ca. up to 8.9 mg L(-1)) released to the medium from Zn NPs in comparison to that measured in the suspensions of ZnO NPs (ca. 5.5 mg L(-1)). In addition, the size of the nanopowders appeared to contribute to the observed toxicities. Although the suspensions possessed aggregates of comparable sizes, smaller Zn NPs (40-60 nm) were relatively more toxic than larger Zn NPs (80-100 nm). Likewise, the suspensions of 10-30 nm ZnO NPs caused higher toxicity than those of 200 nm ZnO NPs. Lipid peroxidation levels were substantially higher in 96 h (p < 0.05), indicating that the toxic effects were due to the oxidative stress.

A novel, colorimetric neutralization assay for measuring antibodies to influenza viruses.

Science.gov (United States)

Lehtoranta, Liisa; Villberg, Anja; Santanen, Riitta; Ziegler, Thedi

2009-08-01

A colorimetric cell proliferation assay for measuring neutralizing antibodies to influenza viruses in human sera is described. Following a 90-min incubation, the serum-virus mixture was transferred to Madin-Darby canine kidney cells cultured in 96-well plates. After further incubation for three days, a tetrazolium salt was added to the wells. Cellular mitochondrial dehydrogenases cleave the tetrazolium salt to formazan, and the resulting color change is read by a spectrophotometer. The absorbance values correlate directly to the number of viable cells in the assay well and thus also to the neutralizing activity of influenza-specific antibodies present in the serum. With the few hands-on manipulations required, this assay allows simultaneous testing of a considerable number of sera, offers opportunities for automation, and is suitable for use under biosafety level-3 conditions. The test was used to study the antibody response after the administration of seasonal, inactivated, trivalent influenza vaccine. Antibody titers determined by the neutralization test in pre- and post-vaccination serum pairs were compared with those obtained by the hemagglutination inhibition assay. The neutralization test yielded higher pre- and post-vaccination titers and a larger number of significant increases in post-vaccination antibody titer than the hemagglutination inhibition test. This new test format could serve as a valuable laboratory tool for influenza vaccine studies.

Tolerance induction to stress caused by lavender (Lavandula angustifolia extracts in the microalga Dunaliella salina through metabolic modifications and β-carotene production

Directory of Open Access Journals (Sweden)

Sahar Mazang-Ghasemi

2016-03-01

Full Text Available The inhibitory or stimulatory effect of one plant on other plant species through the released chemical compounds into the environment, known as allelochemicals, is called allelopathy. In the present study, effect of aqueous extracts of lavender (Lavandula angustifolia leaves on growth pattern and ability of β-carotene production in unicellular green alga, Dunaliella salina was investigated. Based on these results, phenolics content and antioxidant capacity of the extracts was relatively low. Treated cultures with extract concentrations of 1, 2, 3 and 4 % showed that the process of cell division was ceased because of extracts phytotoxicity. However, the chlorophyll and β-carotene concentration pronouncedly increased in phytotoxin-treated cells, so that the highest of these values were detected in 3 % extracts-exposed cells. Percentage change of all three fresh weight, total sugar and protein between 0 and 48 h of phytotoxins treatment was significantly increased concurrently with increasing dose of extracts. These results suggest that D. salina tolerate allelochemicals-induced stress by metabolic modifications. Therefore, phytotoxins-tolerance mechanisms in D. salina are associated with physiological and metabolic adjustments by allocating the carbon flux to the synthesis of the sugars, chlorophyll and β-carotene, which induce phytotoxins stress tolerance in this alga.

Analysis of impact of temperature and saltwater on Nannochloropsis salina bio-oil production by ultra high resolution APCI FT-ICR MS

KAUST Repository

Sanguineti, Michael Mario; Hourani, Nadim; Witt, Matthí as; Sarathy, Mani; Thomsen, Laurenz A.; Kuhnert, Nikolai

2015-01-01

Concentrated Nannochloropsis salina paste was reconstituted in distilled water and synthetic saltwater and processed at 250°C and 300°C via hydrothermal liquefaction. The resulting bio-oils yielded a diverse distribution of product classes

Different strategies of osmoadaptation in the closely related marine myxobacteria Enhygromyxa salina SWB007 and Plesiocystis pacifica SIR-1.

Science.gov (United States)

Amiri Moghaddam, Jamshid; Boehringer, Nils; Burdziak, Amal; Kunte, Hans-Jörg; Galinski, Erwin A; Schäberle, Till F

2016-02-03

Only a few myxobacteria are known to date which are classified as marine due to their salt-dependency. In this study, the salt tolerance mechanism of these bacteria was investigated. Therefore, a growth medium was designed, in which the mutated Escherichia coli strain BKA13 served as sole food source for the predatory, heterotrophic myxobacteria. This enabled measurement of the osmolytes without any background and revealed that the closely related strains Enhygromyxa salina SWB007 and Plesiocystis pacifica SIR-1 developed different strategies to handle salt stress. P. pacifica SIR-1, which was grown between 1-4 % NaCl, relies solely on the accumulation of amino acids, while E. salina SWB007, which was grown between 0.5-3 % NaCl, employs, beside betaine, hydroxyectoine as the major compatible solute. In accordance with this analysis, only in the latter strain a gene locus was identified which codes for genes corresponding to the biosynthesis of betaine, ectoine, and hydroxyectoine.

Effect of salinity on metal mobility in Sečovlje salina sediment (northern Adriatic, Slovenia)

Science.gov (United States)

Kovač, N.; Ramšak, T.; Glavaš, N.; Dolenec, M.; Rogan Šmuc, N.

2016-12-01

Saline sediment (saline healing mud or "fango") from the Sečovlje Salina (northern Adriatic, Slovenia) is traditionally used in the coastal health resorts as a virgin material for medical treatment, wellness and relax purposes. Therapeutic qualities of the healing mud depend on its mineralogical composition and physical, mineralogical, geochemical and biological properties. Their microbial and potentially toxic elements contamination are the most important features affecting user safety. However, the degree of metal toxicity (and its regulation) for natural healing mud is still under discussion. Therefore, the influence of the overlying water salinity on the mobility of heavy metals (and some other geochemical characteristic) was studied for saline sediments of the Sečovlje Salina. Experiments takes place in tanks under defined conditions i.e. at day (21 °C): night (16 °C) cycle for three months. Sediment was covered with water of different salinities (36, 155, 323 g NaCl L-1 and distillate water) and mixed/stirred every week during the experimental period. At the same time, the evaporated water was replaced with distilled water. The mud samples were analyzed, at the beginning and at the end of experiment, for mineral (XRD), elemental composition (ICP-MS) and organic content (% TOC, % TN). Geochemical analysis of the aqueous phase (content of cations and anions) have also been carried out in an accredited Canadian laboratory Actlabs (Activation Laboratories, Canada). Salinity and maturation of sediment does not significantly affect its mineral composition. The samples taken at the end of the experiment have higher percent of water but lower organic carbon concentration. Concentrations of investigated elements are comparable to that in surface sediments from Central Adriatic Sea. In the water phase, concentrations of most elements (As, Ba, Cu, Mo, Mn, Ni, Sr, Sb) rise from the beginning to the end of the experiment, whereas the metal (potentially toxic elements

The small ethylene response factor ERF96 is involved in the regulation of the abscisic acid response in Arabidopsis

Directory of Open Access Journals (Sweden)

Xiaoping eWang

2015-11-01

Full Text Available Ethylene regulates many aspects of plant growth and development including seed germination, leaf senescence, and fruit ripening, and of plant responses to environmental stimuli including both biotic and abiotic stresses. Ethylene Response Factors (ERFs are plant-specific transcription factors and are a subfamily of the AP2 (APETALA2/ERF transcription factor family. The function of many members in this large gene family remains largely unknown. ERF96, a member of the Group IX ERF family transcription factors, has recently been shown to be a transcriptional activator that is involved in plant defense response in Arabidopsis. Here we provide evidence that ERF96 is a positive regulator of abscisic acid (ABA responses. Bioinformatics analysis indicated that there are a total four small ERFs in Arabidopsis including ERF95, ERF96, ERF97 and ERF98, and that ERF96 forms a cluster with ERF95 and ERF97. By using quantitative RT-PCR, we found that ERF96 is expressed in all tissues and organs examined except roots, with relatively high expression in flowers and seeds. Results from the protoplast transfection assay results indicated that the EDLL motif-containing C-terminal domain is responsible for ERF96’s transcriptional activity. Although loss-of-function mutant of ERF96 was morphologically similar to wild type plants, transgenic plants overexpressing ERF96 had smaller rosette size and were delayed in flowering time. In ABA sensitivity assays, we found that ERF96 overexpression plants were hypersensitive to ABA in terms of ABA inhibition of seed germination, early seedling development and root elongation. Consistent with these observations, elevated transcript levels of some ABA-responsive genes including RD29A, ABI5, ABF3, ABF4, P5CS and COR15A were observed in the transgenic plants in the presence of ABA. However, in the absence of ABA treatment, the transcript levels of these ABA-responsive genes remained largely unchanged. Our experiments also showed

Gp96 Peptide Antagonist gp96-II Confers Therapeutic Effects in Murine Intestinal Inflammation

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Claudia A. Nold-Petry

2017-12-01

Full Text Available BackgroundThe expression of heat shock protein gp96 is strongly correlated with the degree of tissue inflammation in ulcerative colitis and Crohn’s disease, thereby leading us to the hypothesis that inhibition of expression via gp96-II peptide prevents intestinal inflammation.MethodsWe employed daily injections of gp96-II peptide in two murine models of intestinal inflammation, the first resulting from five daily injections of IL-12/IL-18, the second via a single intrarectal application of TNBS (2,4,6-trinitrobenzenesulfonic acid. We also assessed the effectiveness of gp96-II peptide in murine and human primary cell culture.ResultsIn the IL-12/IL-18 model, all gp96-II peptide-treated animals survived until day 5, whereas 80% of placebo-injected animals died. gp96-II peptide reduced IL-12/IL-18-induced plasma IFNγ by 89%, IL-1β by 63%, IL-6 by 43% and tumor necrosis factor (TNF by 70% compared to controls. The clinical assessment Disease Activity Index of intestinal inflammation severity was found to be significantly lower in the gp96-II-treated animals when compared to vehicle-injected mice. gp96-II peptide treatment in the TNBS model limited weight loss to 5% on day 7 compared with prednisolone treatment, whereas placebo-treated animals suffered a 20% weight loss. Histological disease severity was reduced equally by prednisolone (by 40% and gp96-II peptide (35%. Mice treated with either gp96-II peptide or prednisolone exhibited improved endoscopic scores compared with vehicle-treated control mice: vascularity, fibrin, granularity, and translucency scores were reduced by up to 49% by prednisolone and by up to 30% by gp96-II peptide. In vitro, gp96-II peptide reduced TLR2-, TLR4- and IL-12/IL-18-induced cytokine expression in murine splenocytes, with declines in constitutive IL-6 (54%, lipopolysaccharide-induced TNF (48%, IL-6 (81% and in Staphylococcus epidermidis-induced TNF (67% and IL-6 (81%, as well as IL-12/IL-18-induced IFNγ (75%. gp

Application of 3D Printing Technology in Increasing the Diagnostic Performance of Enzyme-Linked Immunosorbent Assay (ELISA) for Infectious Diseases.

Science.gov (United States)

Singh, Harpal; Shimojima, Masayuki; Shiratori, Tomomi; An, Le Van; Sugamata, Masami; Yang, Ming

2015-07-08

Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients. The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems. Utilizing available 3D printing platforms, a '3D well' was designed and developed to have an increased surface area compared to those of 96-well plates. The ease and rapidity of the development of the 3D well prototype provided an opportunity for its rapid validation through the diagnostic performance of ELISA in infectious disease without modifying current laboratory practices for ELISA. The improved sensitivity of the 3D well of up to 2.25-fold higher compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization and Lab-On-a-Chip platforms to reduce time, volume of reagents and samples needed for such assays in the laboratory diagnosis of infectious and other diseases including applications in other disciplines.

A simple micro-extraction plate assay for automated LC-MS/MS analysis of human serum 25-hydroxyvitamin D levels.

Science.gov (United States)

Geib, Timon; Meier, Florian; Schorr, Pascal; Lammert, Frank; Stokes, Caroline S; Volmer, Dietrich A

2015-01-01

This short application note describes a simple and automated assay for determination of 25-hydroxyvitamin D (25(OH)D) levels in very small volumes of human serum. It utilizes commercial 96-well micro-extraction plates with commercial 25(OH)D isotope calibration and quality control kits. Separation was achieved using a pentafluorophenyl liquid chromatography column followed by multiple reaction monitoring-based quantification on an electrospray triple quadrupole mass spectrometer. Emphasis was placed on providing a simple assay that can be rapidly established in non-specialized laboratories within days, without the need for laborious and time consuming sample preparation steps, advanced calibration or data acquisition routines. The analytical figures of merit obtained from this assay compared well to established assays. To demonstrate the applicability, the assay was applied to analysis of serum samples from patients with chronic liver diseases and compared to results from a routine clinical immunoassay. Copyright © 2015 John Wiley & Sons, Ltd.

PH-sensing 96-well microtitre plates for the characterization of acid production by dairy starter cultures.

Science.gov (United States)

John, Gernot T; Goelling, Detlef; Klimant, Ingo; Schneider, Holger; Heinzle, Elmar

2003-08-01

A new method for characterization of acid production by dairy starter cultures is presented. Microplates with integrated optical pH sensors are developed. Two fluorophores, a pH-sensitive and a pH-insensitive one are immobilised at the bottom of a polystyrene 96-well microtitre plate. The pH-insensitive fluorophore serves as an internal reference and makes calibration unnecessary. The sensor measures pH accurately in optically well-defined media. Particles and fluorophores contained in the bulk medium disturbed the measurements. Despite these disturbances it was possible to clearly sense differences in inoculum type and in inoculum sizes of cultures of Lactococcus lactis and of Streptococcus thermophilus at 30 and 37 degrees C. Besides a pH-related signal there is information about other changes during milk fermentation. The cultivation results were compared with those from the established CINAC-method. From this comparison it can be concluded that the new method can be used reliably to characterize particularly a large number of strains for screening purposes but also for quality control.

Performance of seven serological assays for diagnosing tularemia

Science.gov (United States)

2014-01-01

Background Tularemia is a rare zoonotic disease caused by the Gram-negative bacterium Francisella tularensis. Serology is frequently the preferred diagnostic approach, because the pathogen is highly infectious and difficult to cultivate. The aim of this retrospective study was to determine the diagnostic accuracy of tularemia specific tests. Methods The Serazym®Anti-Francisella tularensis ELISA, Serion ELISA classic Francisella tularensis IgG/IgM, an in-house ELISA, the VIRapid® Tularemia immunochromatographic test, an in-house antigen microarray, and a Western Blot (WB) assay were evaluated. The diagnosis tularemia was established using a standard micro-agglutination assay. In total, 135 sera from a series of 110 consecutive tularemia patients were tested. Results The diagnostic sensitivity and diagnostic specificity of the tests were VIRapid (97.0% and 84.0%), Serion IgG (96.3% and 96.8%), Serion IgM (94.8% and 96.8%), Serazym (97.0% and 91.5%), in-house ELISA (95.6% and 76.6%), WB (93.3% and 83.0%), microarray (91.1% and 97.9%). Conclusions The diagnostic value of the commercial assays was proven, because the diagnostic accuracy was >90%. The diagnostic sensitivity of the in-house ELISA and the WB were acceptable, but the diagnostic accuracy was <90%. Interestingly, the antigen microarray test was very specific and had a very good positive predictive value. PMID:24885274

Desenvolvimento de mudas de pinhão-manso irrigadas com água salina

Directory of Open Access Journals (Sweden)

Fábio Santos Matos

2013-08-01

Full Text Available O uso de água salina na irrigação torna-se importante alternativa diante daescassez de água de boa qualidade em todo o mundo. O pinhão-manso (Jatropha curcas L. possui baixa exigência hídrica, sobrevive e apresenta produção satisfatória em solos de baixa fertilidade. No entanto, a sua produção é maior em cultivos irrigados, o que reforça a necessidade de desenvolvimento de pesquisas para uso de água salina. Este trabalho objetivou avaliar o efeito de diferentes níveis de salinidade da água de irrigação nas características morfofisiológicas de mudas de pinhão-manso. Para isso, foi conduzido experimento em casa de vegetação com interceptação de 50 % da radiação solar, localizada na Universidade Estadual de Goiás, Ipameri, Goiás. O experimento foi conduzido em vasos com capacidade de 4 L de solo, utilizando-se o delineamento inteiramente casualizado, com quatro tratamentos e cinco repetições. As plantas foram irrigadas diariamente com 150 mL de água não salina, durante os 30 primeiros dias após a germinação das sementes. Do 31º ao 50º dia, as plantas foram submetidas a quatro tratamentos: plantas diariamente irrigadas com água de condutividade elétrica igual a 0,5; 8; 16 e 24 dS m-1. Aos 50 dias após a germinação, analisaram-se as seguintes características nas mudas de pinhão-manso: número de folhas; altura de planta; diâmetro de ramo; teor relativo de água; área foliar; clorofila total; razões de massa radicular, massa caulinar, massa foliar e parte aérea/sistema radicular; e biomassa total. Os resultados evidenciaram que as mudas de pinhão-manso irrigadas com água de condutividade elétrica 8 dS m-1 não apresentaram redução do crescimento vegetativo. Todavia, a água de irrigação com condutividade elétrica 16 dS m-1 causou redução no crescimento vegetativo e elevou a senescência e abscisão foliar. Água com condutividade elétrica elétrica de pinhão-manso na fase de mudas.

Salinity-Induced Palmella Formation Mechanism in Halotolerant Algae Dunaliella salina Revealed by Quantitative Proteomics and Phosphoproteomics

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Sijia Wei

2017-05-01

Full Text Available Palmella stage is critical for some unicellular algae to survive in extreme environments. The halotolerant algae Dunaliella salina is a good single-cell model for studying plant adaptation to high salinity. To investigate the molecular adaptation mechanism in salinity shock-induced palmella formation, we performed a comprehensive physiological, proteomics and phosphoproteomics study upon palmella formation of D. salina using dimethyl labeling and Ti4+-immobilized metal ion affinity chromatography (IMAC proteomic approaches. We found that 151 salinity-responsive proteins and 35 salinity-responsive phosphoproteins were involved in multiple signaling and metabolic pathways upon palmella formation. Taken together with photosynthetic parameters and enzyme activity analyses, the patterns of protein accumulation and phosphorylation level exhibited the mechanisms upon palmella formation, including dynamics of cytoskeleton and cell membrane curvature, accumulation and transport of exopolysaccharides, photosynthesis and energy supplying (i.e., photosystem II stability and activity, cyclic electron transport, and C4 pathway, nuclear/chloroplastic gene expression regulation and protein processing, reactive oxygen species homeostasis, and salt signaling transduction. The salinity-responsive protein–protein interaction (PPI networks implied that signaling and protein synthesis and fate are crucial for modulation of these processes. Importantly, the 3D structure of phosphoprotein clearly indicated that the phosphorylation sites of eight proteins were localized in the region of function domain.

Improved assay for measuring heparin binding to bull sperm

International Nuclear Information System (INIS)

Miller, D.J.; Ax, R.L.

1988-01-01

The binding of heparin to sperm has been used to study capacitation and to rank relative fertility of bulls. Previous binding assays were laborious, used 10 7 sperm per assay point, and required large amounts of radiolabeled heparin. A modified heparin-binding assay is described that used only 5 x 10 4 cells per incubation well and required reduced amounts of [ 3 H] heparin. The assay was performed in 96-well Millititer plates, enabling easy incubation and filtering. Dissociation constants and concentrations of binding sites did not differ if analyzed by Scatchard plots, Woolf plots, or by log-logit transformed weighted nonlinear least squares regression, except in the case of outliers. In such cases, Scatchard analysis was more sensitive to outliers. Nonspecific binding was insignificant using nonlinear logistic fit regression and a proportion graph. The effects were tested of multiple free-thawing of sperm in either a commercial egg yolk extender, 40 mM Tris buffer with 8% glycerol, or 40 mM Tris buffer without glycerol. Freeze-thawing in extender did not affect the dissociation constant or the concentration of binding sites. However, freeze-thawing three times in 40 mM Tris reduced the concentration of binding sites and lowered the dissociation constant (raised the affinity). The inclusion of glycerol in the 40 mM Tris did not significantly affect the estimated dissociation constant or the concentration of binding sites as compared to 40 mM Tris without glycerol

Prospecção fitoquímica de Sonchus oleraceus e sua toxicidade sobre o microcrustáceo Artemia salina Phytochemical prospecting of Sonchus oleraceus and its toxicity to Artemia salina

OpenAIRE

J.M. Lima; C.A. Silva; M.B. Rosa; J.B. Santos; T.G. Oliveira; M.B. Silva

2009-01-01

A espécie vegetal Sonchus oleraceus é uma planta daninha presente em diversas culturas no Brasil e de utilização na medicina popular. Neste trabalho, realizou-se a prospecção fitoquímica dessa espécie com extratos em etanol, água e diclorometano, bem como testes de toxicidade sobre o microcrustáceo Artemia salina. O extrato aquoso apresentou em sua composição açúcares redutores, compostos fenólicos, taninos, flavonóides e cumarinas. No extrato etanólico, observaram-se os mesmos compostos qual...

Synergistic Use of Gold Nanoparticles (AuNPs) and “Capillary Enzyme-Linked Immunosorbent Assay (ELISA)” for High Sensitivity and Fast Assays

Science.gov (United States)

Kim, Wan-Joong; Cho, Hyo Young; Jeong, Bongjin; Byun, Sangwon; Huh, JaeDoo; Kim, Young Jun

2017-01-01

Using gold nanoparticles (AuNPs) on “capillary enzyme-linked immunosorbent assay (ELISA)”, we produced highly sensitive and rapid assays, which are the major attributes for point-of-care applications. First, in order to understand the size effect of AuNPs, AuNPs of varying diameters (5 nm, 10 nm, 15 nm, 20 nm, 30 nm, and 50 nm) conjugated with Horseradish Peroxidase (HRP)-labeled anti-C reactive protein (antiCRP) (AuNP•antiCRP-HRP) were used for well-plate ELISA. AuNP of 10 nm produced the largest optical density, enabling detection of 0.1 ng/mL of CRP with only 30 s of incubation, in contrast to 10 ng/mL for the ELISA run in the absence of AuNP. Then, AuNP of 10 nm conjugated with antiCRP-HRP (AuNP•antiCRP-HRP) was used for “capillary ELISA” to detect as low as 0.1 ng/mL of CRP. Also, kinetic study on both 96-well plates and in a capillary tube using antiCRP-HRP or AuNP•antiCRP-HRP showed a synergistic effect between AuNP and the capillary system, in which the fastest assay was observed from the “AuNP capillary ELISA”, with its maximum absorbance reaching 2.5 min, while the slowest was the typical well-plate ELISA with its maximum absorbance reaching in 13.5 min. PMID:29278402

Application of 3D Printing Technology in Increasing the Diagnostic Performance of Enzyme-Linked Immunosorbent Assay (ELISA) for Infectious Diseases

Science.gov (United States)

Singh, Harpal; Shimojima, Masayuki; Shiratori, Tomomi; An, Le Van; Sugamata, Masami; Yang, Ming

2015-01-01

Enzyme-linked Immunosorbent Assay (ELISA)-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients. The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems. Utilizing available 3D printing platforms, a ‘3D well’ was designed and developed to have an increased surface area compared to those of 96-well plates. The ease and rapidity of the development of the 3D well prototype provided an opportunity for its rapid validation through the diagnostic performance of ELISA in infectious disease without modifying current laboratory practices for ELISA. The improved sensitivity of the 3D well of up to 2.25-fold higher compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization and Lab-On-a-Chip platforms to reduce time, volume of reagents and samples needed for such assays in the laboratory diagnosis of infectious and other diseases including applications in other disciplines. PMID:26184194

Application of 3D Printing Technology in Increasing the Diagnostic Performance of Enzyme-Linked Immunosorbent Assay (ELISA for Infectious Diseases

Directory of Open Access Journals (Sweden)

Harpal Singh

2015-07-01

Full Text Available Enzyme-linked Immunosorbent Assay (ELISA-based diagnosis is the mainstay for measuring antibody response in infectious diseases and to support pathogen identification of potential use in infectious disease outbreaks and clinical care of individual patients. The development of laboratory diagnostics using readily available 3D printing technologies provides a timely opportunity for further expansion of this technology into immunodetection systems. Utilizing available 3D printing platforms, a ‘3D well’ was designed and developed to have an increased surface area compared to those of 96-well plates. The ease and rapidity of the development of the 3D well prototype provided an opportunity for its rapid validation through the diagnostic performance of ELISA in infectious disease without modifying current laboratory practices for ELISA. The improved sensitivity of the 3D well of up to 2.25-fold higher compared to the 96-well ELISA provides a potential for the expansion of this technology towards miniaturization and Lab-On-a-Chip platforms to reduce time, volume of reagents and samples needed for such assays in the laboratory diagnosis of infectious and other diseases including applications in other disciplines.

Assembly of the Lactuca sativa, L. cv. Tizian draft genome sequence reveals differences within major resistance complex 1 as compared to the cv. Salinas reference genome.

Science.gov (United States)

Verwaaijen, Bart; Wibberg, Daniel; Nelkner, Johanna; Gordin, Miriam; Rupp, Oliver; Winkler, Anika; Bremges, Andreas; Blom, Jochen; Grosch, Rita; Pühler, Alfred; Schlüter, Andreas

2018-02-10

Lettuce (Lactuca sativa, L.) is an important annual plant of the family Asteraceae (Compositae). The commercial lettuce cultivar Tizian has been used in various scientific studies investigating the interaction of the plant with phytopathogens or biological control agents. Here, we present the de novo draft genome sequencing and gene prediction for this specific cultivar derived from transcriptome sequence data. The assembled scaffolds amount to a size of 2.22 Gb. Based on RNAseq data, 31,112 transcript isoforms were identified. Functional predictions for these transcripts were determined within the GenDBE annotation platform. Comparison with the cv. Salinas reference genome revealed a high degree of sequence similarity on genome and transcriptome levels, with an average amino acid identity of 99%. Furthermore, it was observed that two large regions are either missing or are highly divergent within the cv. Tizian genome compared to cv. Salinas. One of these regions covers the major resistance complex 1 region of cv. Salinas. The cv. Tizian draft genome sequence provides a valuable resource for future functional and transcriptome analyses focused on this lettuce cultivar. Copyright © 2017 Elsevier B.V. All rights reserved.

Executive summary of a draft report on the geology and salt deposits of the Salina Salt Basin

International Nuclear Information System (INIS)

1978-01-01

The study discussed is the first phase of a program for the geologic evaluation of the Silurian-age bedded salt of the Salina Group. The Salina Salt Basin, as used in this study, includes those portions of the Appalachian and Michigan basins that are underlain by the Salina Group. The full draft report consists of a regional reconnaissance, identification of study areas in New York and Ohio which are deserving of a more thorough evaluation, and a program plan to accomplish that evaluation. The entire draft report is in two volumes, contains 1068 pages and 204 figures, and has a bibliography that consists of over 1100 separate entries. This summary has been prepared for the benefit of those who wish to review the results of this phase of the evaluation but who do not want to go through the exhaustive detail that is present in the full report. The regional reconnaissance was accomplished by a very thorough and extensive literature review, addressing the following topics: depth of salt, thickness, stratigraphy, tectonics, structure, seismicity, hydrology, erosion and denudation, and mineral resources. Before further technical evaluation proceeds, the draft report and the proposed program are being subjected to a thorough evaluation by a number of groups, including appropriate state agencies. This rather extensive review process is being conducted to ensure that the program is performed entirely in the open and subject to continuous public surveillance. This report does not represent the first work that has been done in this region with regard to evaluating the salt deposits for waste disposal. Previous efforts have been limited, however, and have been done by individual consultants. At the present time, the U.S. Geological Survey is also participating in the technical evaluation; their results will be issued separately. In addition to the technical evaluations, environmental surveys will also be conducted as an integral part of this thorough evaluation program

Salt-stimulation of caesium accumulation in the euryhaline green microalga Chlorella salina: potential relevance to the development of a biological Cs-removal process

Energy Technology Data Exchange (ETDEWEB)

Avery, S. V.; Codd, G. A.; Gadd, G. M. [Department of Biological Sciences, University of Dundee, Dundee DD1 4HN (United Kingdom)

1993-07-01

Accumulation of Cs{sup +} by Chlorella salina was 28-fold greater in cells incubated in the presence than in the absence of 0.5 M-NaCl. An approximate 70% removal of external Cs{sup +} resulted after 15 h incubation of cells with 50 μ;M-CsCl and 0.5 M-NaCl. LiCl also had a stimulatory effect on Cs{sup +} uptake, although mannitol did not. Cs{sup +} influx increased with increasing external NaCl concentration and was maximal between 25-500 mM-NaCl at approximately 4 nmol Cs{sup +} h−1 (10{sup 6} cells){sup −1}. Little effect on Cs{sup +} uptake resulted from the presence of Mg{sup 2+} or Ca{sup 2+} or from varying the external pH, and Cs{sup +} was relatively non-toxic towards C. salina. At increasing cell densities (from 4 × 10{sup 5} to 1 × 10{sup 7} cells ml{sup +1}), decreasing amounts of Cs{sup +} were accumulated per cell although the rate of Cs{sup +} removal from the external medium was still greatest at the higher cell densities examined. Freely suspended C. salina and cell-loaded alginate microbeads accumulated similar levels of Cs{sup +}, however, 46% of total Cs{sup +} uptake was attributable to the calcium-alginate matrix in the latter case. When Cs{sup +}-loaded cells were subjected to hypoosmotic shock, loss of cellular Cs{sup +} occurred allowing easy Cs{sup +} recovery. This loss exceeded 90% of cellular Cs{sup +} when cells were washed with solutions containing ≤ 50 mM-NaCl between consecutive Cs{sup +} uptake periods; these cells subsequently lost their ability to accumulate large amounts of Cs{sup +}. Maximal Cs{sup +} uptake (approximately 85.1% removal after three 15 h incubations) occurred when cells were washed with a solution containing 500 mM-NaCl and 200 mM-KCl between incubations. The relevance of these results to the possible use of C. salina in a salt-dependent biological Cs-removal process is discussed. (author)

Effects of salinity, commercial salts, and water type on cultivation of the cryptophyte microalgae Rhodomonas salina and the calanoid Copepod Acartia tonsa

DEFF Research Database (Denmark)

Jepsen, Per Meyer; Thoisen, Christina V.; Carron-Cabaret, Thibaut

2018-01-01

Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research was to investi......Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research...... was to investigate the applicability of commercial salts and clarify the potential effects on the cultivation of the microalga Rhodomonas salina and the copepod Acartia tonsa. Three commercial salts were tested, Red Sea Salt (RS), Red Sea – Coral Pro Salt (CP), and Blue Treasure Salt. R. salina was cultured...

National Uranium Resource Evaluation: Salina Quadrangle, Utah

International Nuclear Information System (INIS)

Lupe, R.D.; Campbell, J.A.; Franczyk, K.J.; Luft, S.J.; Peterson, F.; Robinson, K.

1982-09-01

Two stratigraphic units, the Late Jurassic Salt Wash Member of the Morrison Formation and the Triassic Chinle Formation, were determined to be favorable for the occurrence of uranium deposits that meet the minimum size and grade requirements of the US Department of Energy in the Salina 1 x 2 0 Quadrangle, Utah. Three areas judged favorable for the Salt Wash Member are the Tidwell and Notom districts, and the Henry Mountains mineral belt. The criteria used to establish favorability were the presence of: (1) fluvial sandstone beds deposited by low-energy streams; (2) actively moving major and minor structures such as the Paradox basin and the many folds within it; (3) paleostream transport directions approximately perpendicular to the trend of many of the paleofolds; (4) presence of favorable gray lacustrine mudstone beds; and (5) known uranium occurrences associated with the favorable gray mudstones. Four favorable areas have been outlined for the Chinle Formation. These are the San Rafael Swell, Inter River, and the Orange Cliffs subareas and the Capitol Reef area. The criteria used to establish these areas are: the sandstone-to-mudstone ratios and the geographic distribution of the Petrified Forest Member of the Chinle Formation which is considered as the probable source for the uranium

78 FR 37790 - In the Matter of: Mario Salinas-Lucio, Inmate Number #61687-279, FCI La Tuna, Federal Corrections...

Science.gov (United States)

2013-06-24

... DEPARTMENT OF COMMERCE Bureau of Industry and Security In the Matter of: Mario Salinas-Lucio... Order with the Under Secretary of Commerce for Industry and Security. The appeal must be filed within 45....S.C. 793, 794 or 798; section 4(b) of the Internal Security Act of 1950 (50 U.S.C. 783(b)), or...

Gyrodactylus salinae n. sp. (Platyhelminthes: Monogenea infecting the south European toothcarp Aphanius fasciatus (Valenciennes (Teleostei, Cyprinodontidae from a hypersaline environment in Italy

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Huyse Tine

2011-06-01

Full Text Available Abstract Background Historically, non-native species of Gambusia (Poeciliidae have been used to control larval stages of the Asian tiger mosquito, Stegomyia albopicta Reinert, Harbach et Kitching, 2004 throughout Italy. The potential utility of indigenous populations of Aphanius fasciatus (Valenciennes (Teleostei: Cyprinodontidae as an appropriate alternative biological control is currently being explored. A sub-sample of ten fish collected from Cervia Saline, Italy (salinity 65 ppt; 30°C to assess their reproductive capability in captivity, harboured a moderate infection of Gyrodactylus von Nordmann, 1832 (Platyhelminthes, Monogenea. A subsequent morphological and molecular study identified this as being a new species. Results Gyrodactylus salinae n. sp. is described from the skin, fins and gills of A. fasciatus. Light and scanning electron microscopical (SEM examination of the opisthaptoral armature and their comparison with all other recorded species suggested morphological similarities to Gyrodactylus rugiensoides Huyse et Volckaert, 2002 from Pomatoschistus minutus (Pallas. Features of the ventral bar, however, permit its discrimination from G. rugiensoides. Sequencing of the nuclear ribosomal DNA internal transcribed spacers 1 and 2 and the 5.8S rRNA gene and a comparison with all species listed in GenBank confirmed they are unique and represent a new species (most similar to Gyrodactylus anguillae Ergens, 1960, 8.3% pair-wise distance based on 5.8S+ITS2. This represents the first species of Gyrodactylus to be described from Aphanius and, to date, has the longest ITS1 (774 bp sequenced from any Gyrodactylus. Additional sampling of Cervia Saline throughout the year, found G. salinae n. sp. to persist in conditions ranging from 35 ppt and 5°C in December to 65 ppt and 30°C in July, while in captivity a low level of infection was present, even in freshwater conditions (0 ppt. Conclusions The ability of G. salinae n. sp. to tolerate a wide

Premarket evaluations of the IMDx C. difficile for Abbott m2000 Assay and the BD Max Cdiff Assay.

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Stellrecht, K A; Espino, A A; Maceira, V P; Nattanmai, S M; Butt, S A; Wroblewski, D; Hannett, G E; Musser, K A

2014-05-01

Clostridium difficile-associated diarrhea is a well-recognized complication of antibiotic use. Historically, diagnosing C. difficile has been difficult, as antigen assays are insensitive and culture-based methods require several days to yield results. Nucleic acid amplification tests (NAATs) are quickly becoming the standard of care. We compared the performance of two automated investigational/research use only (IUO/RUO) NAATs for the detection of C. difficile toxin genes, the IMDx C. difficile for Abbott m2000 Assay (IMDx) and the BD Max Cdiff Assay (Max). A prospective analysis of 111 stool specimens received in the laboratory for C. difficile testing by the laboratory's test of record (TOR), the BD GeneOhm Cdiff Assay, and a retrospective analysis of 88 specimens previously determined to be positive for C. difficile were included in the study. One prospective specimen was excluded due to loss to follow-up discrepancy analysis. Of the remaining 198 specimens, 90 were positive by all three methods, 9 were positive by TOR and Max, and 3 were positive by TOR only. One negative specimen was initially inhibitory by Max. The remaining 95 specimens were negative by all methods. Toxigenic C. difficile culture was performed on the 12 discrepant samples. True C. difficile-positive status was defined as either positive by all three amplification assays or positive by toxigenic culture. Based on this definition, the sensitivity and specificity were 96.9% and 95% for Max and 92.8% and 100% for IMDx. In summary, both highly automated systems demonstrated excellent performance, and each has individual benefits, which will ensure that they will both have a niche in clinical laboratories.

Approaches about prescriptions and corporal practices in schools in Pirapora, Januária and Salinas (1906-1927

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ELISÂNGELA CHAVES

2012-01-01

Full Text Available The text analyses the proposal of the corporal education in schools in the north of Minas Gerais during 1906 and 1927, especially in Pirapora, januária and Salinas. As main source, the research made use of oral reports and other documents directly related to school memory to understand how physical activities were realized as habit and attitude-forming, in a historic moment in which Brazil used to cherish the modernizing speech.

Pro-neurogenic effects of andrographolide on RSC96 Schwann cells in vitro

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Xu, Fuben; Wu, Huayu; Zhang, Kun; Lv, Peizhen; Zheng, Li; Zhao, Jinmin

2016-01-01

Nerve regeneration remains a challenge to the treatment of peripheral nerve injury. Andrographolide (Andro) is the main active constituent of Andrographis paniculata, which has been applied in the treatment of several diseases, including inflammation, in ancient China. Andro has been reported to facilitate the reduction of edema and to exert analgesic effects in the treatment of various diseases. These findings suggest that Andro may be considered a promising anti-inflammatory agent that may suppress destruction and accelerate proliferation of Schwann cells following peripheral nerve injury. In the present study, the effects of Andro on RSC96 cells were investigated in vitro. The RSC96 cell line is a spontaneously immortalized rat Schwann cell line, which was originally derived from a long-term culture of rat primary Schwann cells. RSC96 cells were treated with a range of 0 to 50 µM Andro prior to the MTT assay. Cell proliferation, morphology, synthesis and nerve-specific gene expression were performed to detect the effect of Andro on RSC96 cells. The results of the present study demonstrated that the recommended doses of Andro ranged between 0.78 and 12.5 µM, among which the most obvious response was observed when used at 3.125 µM (P<0.05). DNA content was improved in Andro groups compared with the control group (P<0.05). In addition, Andro was able to promote the gene expression of glial cell line-derived neurotrophic factor, brain-derived neurotrophic factor, ciliary neurotrophic factor, and the specific Schwann cell marker S100β (P<0.05). The results of a viability assay, hematoxylin-eosin staining, and immunohistochemistry were also improved in Andro groups. These results indicated that Andro may accelerate proliferation of RSC96 cells in vitro, whilst maintaining the Schwann cell phenotype; therefore, the present study may provide valuable evidence for the further exploration of the effects of Andro on peripheral nerves. PMID:27599453

Laboratory scale photobioreactor for high production of microalgae Rhodomonas salina used as food for intensive copepod cultures

DEFF Research Database (Denmark)

Thuy, Minh Vu Thi; Jepsen, Per Meyer; Hansen, Benni Winding

Introduction Microalgae are essential feeds for many cultured molluscs, larvae of marine fishes, crustaceans as well as other important live feeds including rotifers, Artemia and copepods (Muller-Feuga, 2000). Microalgae are grown either in open culture systems (ponds) or closed systems (photobio......Introduction Microalgae are essential feeds for many cultured molluscs, larvae of marine fishes, crustaceans as well as other important live feeds including rotifers, Artemia and copepods (Muller-Feuga, 2000). Microalgae are grown either in open culture systems (ponds) or closed systems...... for copepods (Støttrup and Jensen, 1990; Zhang et al., 2013). Despite the benefit of using R. salina in cultivation of copepods, to our knowledge, there is no report on the production of this microalga at industrial scale to supply sufficient food for mass production of copepods. We intend to conduct the basic...... was cultivated continuously at temperature of 20ºC and salinity of 30ppt in two tubular PBRs with addition of CO2. The experiment was run two times and each PBR in 18 - 30 days. Periodically, the algae were sampled for analyzing the growth, biochemical composition and production. An exponential light model...

Analysis of protein stability and ligand interactions by thermal shift assay.

Science.gov (United States)

Huynh, Kathy; Partch, Carrie L

2015-02-02

Purification of recombinant proteins for biochemical assays and structural studies is time-consuming and presents inherent difficulties that depend on the optimization of protein stability. The use of dyes to monitor thermal denaturation of proteins with sensitive fluorescence detection enables rapid and inexpensive determination of protein stability using real-time PCR instruments. By screening a wide range of solution conditions and additives in a 96-well format, the thermal shift assay easily identifies conditions that significantly enhance the stability of recombinant proteins. The same approach can be used as an initial low-cost screen to discover new protein-ligand interactions by capitalizing on increases in protein stability that typically occur upon ligand binding. This unit presents a methodological workflow for small-scale, high-throughput thermal denaturation of recombinant proteins in the presence of SYPRO Orange dye. Copyright © 2015 John Wiley & Sons, Inc.

Thermodynamic equilibrium solubility measurements in simulated fluids by 96-well plate method in early drug discovery.

Science.gov (United States)

Bharate, Sonali S; Vishwakarma, Ram A

2015-04-01

An early prediction of solubility in physiological media (PBS, SGF and SIF) is useful to predict qualitatively bioavailability and absorption of lead candidates. Despite of the availability of multiple solubility estimation methods, none of the reported method involves simplified fixed protocol for diverse set of compounds. Therefore, a simple and medium-throughput solubility estimation protocol is highly desirable during lead optimization stage. The present work introduces a rapid method for assessment of thermodynamic equilibrium solubility of compounds in aqueous media using 96-well microplate. The developed protocol is straightforward to set up and takes advantage of the sensitivity of UV spectroscopy. The compound, in stock solution in methanol, is introduced in microgram quantities into microplate wells followed by drying at an ambient temperature. Microplates were shaken upon addition of test media and the supernatant was analyzed by UV method. A plot of absorbance versus concentration of a sample provides saturation point, which is thermodynamic equilibrium solubility of a sample. The established protocol was validated using a large panel of commercially available drugs and with conventional miniaturized shake flask method (r(2)>0.84). Additionally, the statistically significant QSPR models were established using experimental solubility values of 52 compounds. Copyright © 2015 Elsevier Ltd. All rights reserved.

Composição química e toxicidade frente Aedes aegypti L. e Artemia salina Leach do óleo essencial das folhas de Myrcia sylvatica (G. Mey. DC.

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C.S. ROSA

2016-03-01

Full Text Available RESUMO A dengue está entre as doenças virais de propagação vetorial mais importante no mundo, causando sérios impactos de morbidade e mortalidade. Desta forma, o presente trabalho teve como objetivo analisar a composição química e a toxicidade do óleo essencial de Myrcia sylvatica (G. Mey D.C. frente Aedes aegypti e Artemia salina. Folhas de M. sylvatica foram coletadas no Parque Nacional da Chapada das Mesas, no município de Carolina (MA no mês de fevereiro de 2012. O óleo foi obtido por hidrodestilação e sua composição química foi determinada por cromatografia gasosa acoplada à espectrometria de massa (CG/EM. O bioensaio frente Artemia salina e às larvas de 3° estádio de Aedes aegypti foram realizados em diferentes concentrações. Os dados de mortalidade foram avaliados por regressão linear para determinar os valores de CL50. Obteve-se 0,5% de rendimento, sendo o (E-cariofileno o constituinte majoritário. O óleo essencial apresentou uma CL50 = 79,44 µg/mL frente A. salina, sendo considerado altamente tóxico. No entanto, este óleo não demonstrou efeito sobre as larvas de A. aegypti. Considerando que o teste de Artemia salina tem correlação com atividades biológicas de grande interesse terapêutico como antitumoral, o óleo essencial das folhas de M. sylvatica demonstrou potencial para desenvolvimento de produtos farmacêuticos.

Depositional environment, sand provenance, and diagenesis of the Basal Salina Formation (lower Eocene), northwestern Peru

Science.gov (United States)

Marsaglia, K. M.; Carozzi, A. V.

The Basal Salina Formation is a lower Eocene transgressive sequence consisting of interbedded shales, siltstones, and conglomeratic sandstones. This formation occurs in the Talara basin of northwestern Peru and is one of a series of complexly faulted hydrocarbon-producing formations within this extensional forearc basin. These sediments were probably deposited in a fan-delta complex that developed along the ancestral Amotape Mountains during the early Eocene. Most of the sediment was derived from the low-grade metamorphic and plutonic rocks that comprise the Amotape Mountains, and their sedimentary cover. Detrital modes of these sandstones reflect the complex tectonic history of the area, rather than the overall forearc setting. Unlike most forearc sediments, these are highly quartzose, with only minor percentages of volcanic detritus. This sand is variably indurated and cemented by chlorite, quartz, calcite, and kaolinite. Clay-mineral matrix assemblages show gradational changes with depth, from primarily detrital kaolinite to diagenetic chlorite and mixed-layered illite/smectite. Basal Salina sandstones exhibit a paragenetic sequence that may be tied to early meteoric influx or late-stage influx of thermally driven brines associated with hydrocarbon migration. Much of the porosity is secondary, resulting from a first-stage dissolution of silicic constituents (volcanic lithic fragments, feldspar, and fibrous quartz) and a later dissolution of surrounding carbonate cement. Types of pores include skeletal grains, grain molds, elongate pores, and fracture porosity. Measured porosity values range up to 24% and coarser samples tend to be more porous. Permeability is enhanced by fractures and deterred by clay-mineral cements and alteration residues.

Bioatividade de três espécies vegetais nativas da Floresta Atlântica brasileira frente ao microcrustáceo Artemia salina

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W.M Pompilho

2014-09-01

Full Text Available Este trabalho teve por objetivo a investigação fitoquímica e propriedades antioxidantes de extratos das folhas de Trigynaea oblongifolia Schltdl (Annonaceae, Ottonia frutescens Trel (Piperaceae, e Bathysa australis (St Hill Hooz (Rubiaceae, bem como avaliar, in vitro, a letalidade frente ao microcrustáceo Artemia salina Leach. Os extratos foram preparados por maceração em metanol 10% (p/v por sete dias, à temperatura ambiente. A atividade antioxidante dos extratos foi determinada pela metodologia que utiliza o radical estável DPPH. A toxicidade dos extratos foi avaliada frente ao microcrustáceo A. salina. Os extratos de O. frutescens e B. australis apresentaram as seguintes classes de metabólitos secundários: Alcalóides, Antraquinonas, Cumarinas, Polifenóis (Taninos, Saponinas. Nos extratos de T. oblongifolia, além dos metabólitos citados anteriormente, foi detectada a presença de Flavonóides. A atividade antioxidante, observada em 30 minutos na concentração de 24 µg/mL de extrato, foi de: O. frutescens - 38,3%, T. oblongifolia - 32,3%, e B. australis - 32,1%. A Concentração Letal, CL50, dos extratos em A. salina foi de: O. frutescens - 149,75 ± 1,02 µg/mL, T. oblongifolia - 148,8 ± 1,74 µg/mL, e B. australis - 684 ± 9,04 µg/mL. Neste contexto, destacamos as espécies, nativas da Floresta Atlântica, O. frutescens e T. oblongifolia de grande potencial na bioprospecção de moléculas biologicamente ativas.

Comparing ImmunoCard with two EIA assays for Clostridium difficile toxins.

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Chan, Edward L; Seales, Diane; Drum, Hong

2009-01-01

To compare three Clostridium difficile EIA kits for the detection of C. difficile toxins from clinical specimens. A total of 287 fresh and stored stool specimens were tested using all three assays. Stools with discrepant results were sent to a reference laboratory for tissue cytotoxin assay. Trinity Medical Center, a community hospital with network hospitals. Patients with diarrhea submitted stools for detection of C. difficile toxins. Of the 287 stool specimens, 116 were positive and 171 negative for C. difficile toxins. The sensitivity, specificity, and positive and negative predictive values of Meridian EIA assay were 99.1, 97.7, 96.6, and 99.4%; ImmunoCard were 100, 98.2, 97.5, and 100%; BioStar OIA assay were 94, 98.8, 98.2, and 96% respectively. ImmunoCardprovides the best sensitivity (100%) for C. difficile toxins A and B detection. The BioStar OIA rapid test missed seven positive stool specimens possibly due to failure to detect toxin B. ImmunoCard has slightly higher predictive values, shorter turnaround time and greater convenience compared to the Meridian EIA Assay. ImmunoCard may be cost effective not only in smaller laboratories, but also in high volume laboratories, when used on a STAT basis or single request.

Avaliação da bioatividade dos extratos de cúrcuma (Curcuma longa L., Zingiberaceae em Artemia salina e Biomphalaria glabrata

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Carlos R. M. da Silva Filho

Full Text Available A cúrcuma é o rizoma limpo, em boas condições, seco e moído da Curcuma longa L., uma planta herbácea da família Zingiberaceae. Visando novas alternativas para o controle da esquistossomose, os extratos de Curcuma longa L. foram testados para a avaliação da atividade moluscicida contra caramujos adultos da espécie Biomphalaria glabrata, e toxicidade (ensaio de letalidade com Artemia salina. A oleoresina e o óleo essencial de cúrcuma foram ativos contra Artemia salina (CL50 = 80,43 e CL50 = 319,82 μg/mL, respectivamente e também ativos contra os indivíduos adultos de Biomphalaria glabrata (CL50 = 58,3 e CL50 = 46,73 μg/mL, respectivamente. A partir dos resultados obtidos pôde ser concluído que ambos os extratos podem constituir uma alternativa no controle da população desses caramujos e na redução da esquistossomose.

Abelhas (Hymenoptera: apoidea visitantes das flores de goiaba em pomar comercial in Salinas, MG Bee diversity in a commercial guava orchard in Salinas, Minas Gerais State, Brazil

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Rosemeire Alves Guimarães

2009-01-01

Full Text Available As abelhas são responsáveis por cerca de 80% a 100% da polinização de culturas agrícolas, especialmente aquelas relacionadas com a produção de sementes e frutos. A investigação da diversidade de abelhas em pomares de goiaba pode ser subsídio para estratégias de incremento da produtividade. Nesta perspectiva, o objetivo deste estudo foi identificar a diversidade de abelhas visitantes das flores de goiaba (Psidium guajava, em pomar comercial em Salinas (MG. O trabalho foi desenvolvido em maio de 2005 e foram coletadas as abelhas visitantes das flores nos horários entre 6h e 18h, totalizando-se 44 horas de coleta. Coletaram-se 705 abelhas de 17 espécies, sendo Trigona spinipes a mais freqüente e dominante na cultura da goiaba. Apis mellifera, Melipona quadrifasciata e Tetragonisca angustula foram consideradas acessórias. Aproximadamente 84% dos indivíduos foram coletados da manhã, de 6h às 10h.Pollination is an important factor in agricultural systems, especially in growing fruits and seed production, which depend greatly on bee visiting during blossom season; highly successful gains within these activities varies between 80 and nearly 100 per cent, owing to the bees. The assessment of bee diversity in commercial orchards of guava may contribute to a more desirable strategic design and consequent improvement of production. The aim of the study was identify the diversity of visiting bees to guava flowers (Psidium guajava in a commercial orchard in Salinas, State of Minas Gerais, Brazil. The work was carried during blossom season of May - 2005. Field works occurred between 6:00 am to 6:00 pm, counting with 44 hours of collection, when 705 bees were collected. The richness observed was of 17 species, the most frequent and dominant being Trigona spinipes. Among the collection there were some considered accessory species: Apis mellifera, Melipona quadrifasciata and Tetragonisca angustula. Most of individual bees have been captured

The efficiency of a new hydrodynamic cavitation pilot system on Artemia salina cysts and natural population of copepods and bacteria under controlled mesocosm conditions.

Science.gov (United States)

Cvetković, Martina; Grego, Mateja; Turk, Valentina

2016-04-15

A study of the efficiency of hydrodynamic cavitation and separation was carried out to evaluate an innovative, environmentally safe and acceptable system for ballast water treatment for reducing the risk of introducing non-native species worldwide. Mesocosm experiments were performed to assess the morphological changes and viability of zooplankton (copepods), Artemia salina cysts, and the growth potential of marine bacteria after the hydrodynamic cavitation treatment with a different number of cycles. Our preliminary results confirmed the significant efficiency of the treatment since more than 98% of the copepods and A. salina cysts were damaged, in comparison with the initial population. The efficiency increased with the number of the hydrodynamic cavitation cycles, or in combination with a separation technique for cysts. There was also a significant decrease in bacterial abundance and growth rate, compared to the initial number and growth potential. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

Analytical characteristics and comparative evaluation of Aptima HCV quant Dx assay with the Abbott RealTime HCV assay and Roche COBAS AmpliPrep/COBAS TaqMan HCV quantitative test v2.0.

Science.gov (United States)

Worlock, A; Blair, D; Hunsicker, M; Le-Nguyen, T; Motta, C; Nguyen, C; Papachristou, E; Pham, J; Williams, A; Vi, M; Vinluan, B; Hatzakis, A

2017-04-04

The Aptima HCV Quant Dx assay (Aptima assay) is a fully automated quantitative assay on the Panther® system. This assay is intended for confirmation of diagnosis and monitoring of HCV RNA in plasma and serum specimens. The purpose of the testing described in this paper was to evaluate the performance of the Aptima assay. The analytical sensitivity, analytical specificity, precision, and linearity of the Aptima assay were assessed. The performance of the Aptima assay was compared to two commercially available HCV assays; the Abbott RealTime HCV assay (Abbott assay, Abbott Labs Illinois, USA) and the Roche COBAS Ampliprep/COBAS Taqman HCV Quantitative Test v2.0 (Roche Assay, Roche Molecular Systems, Pleasanton CA, USA). The 95% Lower Limit of Detection (LoD) of the assay was determined from dilutions of the 2nd HCV WHO International Standard (NIBSC 96/798 genotype 1) and HCV positive clinical specimens in HCV negative human plasma and serum. Probit analysis was performed to generate the 95% predicted detection limits. The Lower Limit of Quantitation (LLoQ) was established for each genotype by diluting clinical specimens and the 2nd HCV WHO International Standard (NIBSC 96/798 genotype 1) in HCV negative human plasma and serum. Specificity was determined using 200 fresh and 536 frozen HCV RNA negative clinical specimens including 370 plasma specimens and 366 serum specimens. Linearity for genotypes 1 to 6 was established by diluting armored RNA or HCV positive clinical specimens in HCV negative serum or plasma from 8.08 log IU/mL to below 1 log IU/mL. Precision was tested using a 10 member panel made by diluting HCV positive clinical specimens or spiking armored RNA into HCV negative plasma and serum. A method comparison was conducted against the Abbott assay using 1058 clinical specimens and against the Roche assay using 608 clinical specimens from HCV infected patients. In addition, agreement between the Roche assay and the Aptima assay using specimens with low

Application of a novel combination of near-infrared spectroscopy and a humidity-controlled 96-well plate to the characterization of the polymorphism of imidafenacin.

Science.gov (United States)

Uchida, Hiroshi; Yoshinaga, Tokuji; Mori, Hirotoshi; Otsuka, Makoto

2010-11-01

This study aimed to apply a currently available chemometric near-infrared spectroscopy technique to the characterization of the polymorphic properties of drug candidates. The technique requires only small quantities of samples and is therefore applicable to drugs in the early stages of development. The combination of near-infrared spectroscopy and a patented 96-well plate divided into 32 individual, humidity-controlled, three-well compartments was used in the characterization of a hygroscopic drug, imidafenacin, which has two polymorphs and one pseudo-polymorph. Characterization was also conducted with powder X-ray diffraction and thermal analysis. The results were compared with those from routinely used conventional analyses. Both the microanalysis and conventional analysis successfully characterised the substance (transformation and relative stability among the two polymorphs and a pseudo-polymorph) depending on the storage conditions. Near-infrared spectroscopic analyses utilizing a humidity-controlled 96-well plate required only small amounts of the sample for characterization under the various conditions of relative humidity. Near-infrared microanalysis can be applied to polymorphic studies of small quantities of a drug candidate. The results also suggest that the method will predict the behaviors of a hygroscopic candidate in solid pharmaceutical preparations at the early stages of drug development. © 2010 The Authors. JPP © 2010 Royal Pharmaceutical Society of Great Britain.

Aplicación del bioensayo de Artemia salina en el análisis general de plantas medicinales brasileñas

OpenAIRE

Montanher, Ana Beatriz Pimentel; Pizzolatti, Moacir Geraldo; Brighente, Inês Maria Costa

2002-01-01

Extractos de once especies de la flora brasileña fueron estudiados para evaluar la actividad citotóxica por el test de Artemia salina. Las plantas seleccionadas fueron: Baccharis pseudotenuifolia, Baccharis ligustrina, Baccharis platypoda, Baccharis coridifolia, Polygala paniculata, Polygala sabulosa, Croton celtidifolius, Cyathea phalerata, Trichilia catigua, Eugenia uniflora y Schinus molle. Los resultados obtenidos para los extractos crudos de B. pseudotenuifolia, B. ligustrina, B. c...

Solid-phase peptide quantitation assay using labeled monoclonal antibody and glutaraldehyde fixation

International Nuclear Information System (INIS)

Kasprzyk, P.G.; Cuttitta, F.; Avis, I.; Nakanishi, Y.; Treston, A.; Wong, H.; Walsh, J.H.; Mulshine, J.L.

1988-01-01

A solid-phase radioimmunoassay utilizing iodinated peptide-specific monoclonal antibody as a detection system instead of labeled peptide has been developed. Regional specific monoclonal antibodies to either gastrin-releasing peptide or gastrin were used as models to validate the general application of our modified assay. Conditions for radioactive labeling of the monoclonal antibody were determined to minimize oxidant damage, which compromises the sensitivity of other reported peptide quantitation assays. Pretreatment of 96-well polyvinyl chloride test plates with a 5% glutaraldehyde solution resulted in consistent retention of sufficient target peptide on the solid-phase matrix to allow precise quantitation. This quantitative method is completed within 1 h of peptide solid phasing. Pretreatment of assay plates with glutaraldehyde increased binding of target peptide and maximized antibody binding by optimizing antigen presentation. The hypothesis that glutaraldehyde affects both peptide binding to the plate and orientation of the peptide was confirmed by analysis of several peptide analogs. These studies indicate that peptide binding was mediated through a free amino group leaving the carboxy-terminal portion of the target peptide accessible for antibody binding. It was observed that the length of the peptide also affects the amount of monoclonal antibody that will bind. Under the optimal conditions, results from quantitation of gastrin-releasing peptide in relevant samples agree well with those from previously reported techniques. Thus, we report here a modified microplate assay which may be generally applied for the rapid and sensitive quantitation of peptide hormones

Development and evaluation of a blocking enzyme-linked immunosorbent assay and virus neutralization assay to detect antibodies to viral hemorrhagic septicemia virus

Science.gov (United States)

Wilson, Anna; Goldberg, Tony; Marcquenski, Susan; Olson, Wendy; Goetz, Frederick; Hershberger, Paul; Hart, Lucas M.; Toohey-Kurth, Kathy

2014-01-01

Viral hemorrhagic septicemia virus (VHSV) is a target of surveillance by many state and federal agencies in the United States. Currently, the detection of VHSV relies on virus isolation, which is lethal to fish and indicates only the current infection status. A serological method is required to ascertain prior exposure. Here, we report two serologic tests for VHSV that are nonlethal, rapid, and species independent, a virus neutralization (VN) assay and a blocking enzyme-linked immunosorbent assay (ELISA). The results show that the VN assay had a specificity of 100% and sensitivity of 42.9%; the anti-nucleocapsid-blocking ELISA detected nonneutralizing VHSV antibodies at a specificity of 88.2% and a sensitivity of 96.4%. The VN assay and ELISA are valuable tools for assessing exposure to VHSV.

Preliminary results of the Artemia salina experiments in biostack on LDEF

International Nuclear Information System (INIS)

Graul, E.H.; Ruether, W.; Hiendl, C.O.

1992-01-01

The mosaic egg of the brine shrimp, Artemia salina, resting in blastula or gastrula state represents a system that during further development, proceeds without any further development to the larval stage, the free swimming nauplius. Therefore, injury to a single cell of the egg will be manifest in the larvae. In several experiments, it was shown that the passage of a single heavy ion through the shrimp egg damaged a cellular area large enough to disturb either embryogenesis or further development of the larvae, or the integrity of the adult individual. Emergence from the egg shell was heavily disturbed by the heavy ions as was hatching. Additional late effects, due to a hit by a heavy ion, are delayed of growth and of sexual maturity, and reduced fertility. Anomalies in the body and the extremities could be observed more frequently for the nauplii which had developed from eggs hit by heavy ions

Effects of UV/Ag-TiO2/O3 advanced oxidation on unicellular green alga Dunaliella salina: implications for removal of invasive species from ballast water.

Science.gov (United States)

Wu, Donghai; You, Hong; Du, Jiaxuan; Chen, Chuan; Jin, Darui

2011-01-01

The UV/Ag-TiO2/O3 process was investigated for ballast water treatment using Dunaliella salina as an indicator. Inactivation curves were obtained, and the toxicity of effluent was determined. Compared with individual unit processes using ozone or UV/Ag-TiO2, the inactivation efficiency of D. salina by the combined UV/Ag-TiO2/O3 process was enhanced. The presence of ozone caused an immediate decrease in chlorophyll a (chl-a) concentration. Inactivation efficiency and ch1-a removal efficiency were positively correlated with ozone dose and ultraviolet intensity. The initial total residual oxidant (TRO) concentration of effluent increased with increasing ozone dose, and persistence of TRO resulted in an extended period of toxicity. The results suggest that UV/Ag-TiO2/O3 has potential for ballast water treatment.

Composição química, atividade antibacteriana in vitro e toxicidade em Artemia salina do óleo essencial das inflorescências de Ocimum gratissimum L., Lamiaceae Chemical composition, antibacterial activity in vitro and brine-shrimp toxicity of the essential oil from inflorescences of Ocimum gratissimum L., Lamiaceae.

Directory of Open Access Journals (Sweden)

Lenise L. Silva

2010-11-01

Full Text Available O óleo essencial das inflorescências de Ocimum gratissimum L., Lamiaceae foi obtido por hidrodestilação e analisado por CG/EM. Os constituintes majoritários identificados foram eugenol (81,94% e γ-muuroleno (12,58%. O óleo essencial das inflorescências demonstrou atividade antibacteriana frente a todas as cepas bacterianas testadas pelo método de microdiluição em caldo. Merece destaque a atividade verificada frente às cepas resistentes de Enterococcus faecalis, Staphylococcus aureus e Escherichia coli. Os valores obtidos de concentração inibitória mínima (CIM e a concentração bactericida mínima (CBM variaram, respectivamente, entre 0,5-2 mg/mL e 1-4 mg/mL. Valores de CL50 de 233,8 (200,7-272,0 µg/mL para o óleo essencial e 186,1 (144,1-228,5 µg/mL para o eugenol, utilizado como controle positivo, foram observados frente à Artemia salina L.The essential oil obtained by hydrodistillation of the inflorescences of Ocimum gratissimum L. was analyzed by GC/MS. The main constituents were eugenol (81.94% and γ-muurolene (12.58%. Antibacterial activity was shown against all assayed strains by the broth microdilution method. It's worth noting the activity against resistant strains of Enterococcus faecalis, Staphylococcus aureus and Escherichia coli. Minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC values ranged between 0.5-2 mg/mL and 1-4 mg/mL, respectively. Preliminary toxicity assayed by the brine-shrimp (Artemia salina L. test showed LC50 values of 233.8 (200.7 - 272.0 µg/mL and 186.1 (144.1 - 228.5 µg/mL, respectively for the essential oil and eugenol (positive control.

High sensitivity, high surface area Enzyme-linked Immunosorbent Assay (ELISA).

Science.gov (United States)

Singh, Harpal; Morita, Takahiro; Suzuki, Yuma; Shimojima, Masayuki; Le Van, An; Sugamata, Masami; Yang, Ming

2015-01-01

Enzyme-linked immunosorbent assays (ELISA) are considered the gold standard in the demonstration of various immunological reactions with an application in the detection of infectious diseases such as during outbreaks or in patient care. This study aimed to produce an ELISA-based diagnostic with an increased sensitivity of detection compared to the standard 96-well method in the immunologic diagnosis of infectious diseases. A '3DStack' was developed using readily available, low cost fabrication technologies namely nanoimprinting and press stamping with an increased surface area of 4 to 6 times more compared to 96-well plates. This was achieved by stacking multiple nanoimprinted polymer sheets. The flow of analytes between the sheets was enhanced by rotating the 3DStack and confirmed by Finite-Element (FE) simulation. An Immunoglobulin G (IgG) ELISA for the detection of antibodies in human serum raised against Rubella virus was performed for validation. An improved sensitivity of up to 1.9 folds higher was observed using the 3DStack compared to the standard method. The increased surface area of the 3DStack developed using nanoimprinting and press stamping technologies, and the flow pattern between sheets generated by rotating the 3DStack were potential contributors to a more sensitive ELISA-based diagnostic device.

Assessment of a recombinant androgen receptor binding assay: initial steps towards validation.

Science.gov (United States)

Freyberger, Alexius; Weimer, Marc; Tran, Hoai-Son; Ahr, Hans-Jürgen

2010-08-01

Despite more than a decade of research in the field of endocrine active compounds with affinity for the androgen receptor (AR), still no validated recombinant AR binding assay is available, although recombinant AR can be obtained from several sources. With funding from the European Union (EU)-sponsored 6th framework project, ReProTect, we developed a model protocol for such an assay based on a simple AR binding assay recently developed at our institution. Important features of the protocol were the use of a rat recombinant fusion protein to thioredoxin containing both the hinge region and ligand binding domain (LBD) of the rat AR (which is identical to the human AR-LBD) and performance in a 96-well plate format. Besides two reference compounds [dihydrotestosterone (DHT), androstenedione] ten test compounds with different affinities for the AR [levonorgestrel, progesterone, prochloraz, 17alpha-methyltestosterone, flutamide, norethynodrel, o,p'-DDT, dibutylphthalate, vinclozolin, linuron] were used to explore the performance of the assay. At least three independent experiments per compound were performed. The AR binding properties of reference and test compounds were well detected, in terms of the relative ranking of binding affinities, there was good agreement with published data obtained from experiments using recombinant AR preparations. Irrespective of the chemical nature of the compound, individual IC(50)-values for a given compound varied by not more than a factor of 2.6. Our data demonstrate that the assay reliably ranked compounds with strong, weak, and no/marginal affinity for the AR with high accuracy. It avoids the manipulation and use of animals, as a recombinant protein is used and thus contributes to the 3R concept. On the whole, this assay is a promising candidate for further validation. Copyright 2009 Elsevier Inc. All rights reserved.

La sedimentación salina actual en las lagunas de La Mancha: una síntesis.

OpenAIRE

Peña Zarzuelo, Antonio de la; Marfil, R.

1986-01-01

[ES] La Mancha es una región natural de más de 30.000 Km2, caracterizada por una topografía extraordinariamente plana y un clima de tipo semiárido (Cuadro 1, Fig. 2), en la que existen numerosas lagunas salinas (Fig. 1), la mayoría de las cuales, por su régimen anual, pueden ser consideradas como «playa-lakes» (Fig. 3). Desde el punto de vista hidroquimico sus salmueras están integradas por: a) aniones: SO4 y Cl, con CO3 y CO 3H subordinados, y b) cationes: Mg 2+ y ...

Evaluation of the bioactivities of some Myanmar medicinal plants using brine shrimp (Artemia salina) toxicity test

International Nuclear Information System (INIS)

Sabai; Khin Khin Win Aung; Nwe Ni Thin; Kyi Shwe; Tin Myint Htwe

2001-01-01

For a variety of toxic substances, brine shrimp larvae (Artemia salina) are usually used as a simple bioassay method and it is also applied for natural product research. The brine shrimp larvae (nauplii) are obtained by natural hatching method from Artemia cysts. By using the larvae, the results from these experiments lead to the lethal dose, LD 50 values of extracts of selected medicinal plants. Activities of a broad range of plant extracts are manifested as toxicity to the brine shrimp. Screening results with six plant extracts are compared with pure caffeine. This method is rapid, reliable, inexpensive and convenient. (author)

A High-Content Assay for Biosensor Validation and for Examining Stimuli that Affect Biosensor Activity.

Science.gov (United States)

Slattery, Scott D; Hahn, Klaus M

2014-12-01

Biosensors are valuable tools used to monitor many different protein behaviors in vivo. Demand for new biosensors is high, but their development and characterization can be difficult. During biosensor design, it is necessary to evaluate the effects of different biosensor structures on specificity, brightness, and fluorescence responses. By co-expressing the biosensor with upstream proteins that either stimulate or inhibit the activity reported by the biosensor, one can determine the difference between the biosensor's maximally activated and inactivated state, and examine response to specific proteins. We describe here a method for biosensor validation in a 96-well plate format using an automated microscope. This protocol produces dose-response curves, enables efficient examination of many parameters, and unlike cell suspension assays, allows visual inspection (e.g., for cell health and biosensor or regulator localization). Optimization of single-chain and dual-chain Rho GTPase biosensors is addressed, but the assay is applicable to any biosensor that can be expressed or otherwise loaded in adherent cells. The assay can also be used for purposes other than biosensor validation, using a well-characterized biosensor as a readout for effects of upstream molecules. Copyright © 2014 John Wiley & Sons, Inc.

Storage conditions affect oxidative stability and nutritional composition of freeze-dried Nannochloropsis salina

DEFF Research Database (Denmark)

Safafar, Hamed; Langvad, Sten; Møller, Peter

2017-01-01

composition of microalgae biomass. In order to investigate the worsening of the nutritional quality of freeze dried biomass, a multifactorial storage experiment was conducted on a high EPA (eicosapentaenoic acid) Nannochloropsis salina biomass. The storage time (0–56 days), storage temperature (5, 20,and 40...... °C and packaging conditions (under vacuum and ambient pressure)used as main factors. During the 56 days of storage, both time and temperature strongly influenced the oxidation reactions which result in deterioration of bioactive compounds such as carotenoids, tocopherols, and EPA. Lipid deterioration......, or cosmetics requires the knowledge of the optimum storage conditions to prevent the value-added compounds from deterioration. Results of this study improve our understanding of the chemical deterioration under different storage conditions and can help the producers/customers to extend the shelf life...

Spatial scale and the diversity of benthic cyanobacteria and diatoms in a salina

DEFF Research Database (Denmark)

Nübel, U.; Garcia-Pichel, F.; Kühl, Michael

1999-01-01

We characterized the richness of benthic cyanobacteria and diatoms in a salina system using traditional and molecular biological methods. After determining the different morphotypes and 16S rRNA genes present in various localities within this hypersaline system, an analysis of the increase......, and for the estimation of the average degree of dissemination of community members within the system. We found interesting differences between analyses based on morphotypes or 16S rRNA genes. The cumulative number of rRNA gene sequences exceeded that of morphotypes by more than two-fold. This indicates that many...... organisms possessing distinct 16S rRNA gene sequences could not be distinguished on the basis of morphology. Thus, some of the apparently widely distributed morphotypes may in fact conceal several ecologically independent genotypes....

Influence of PbS nanoparticle polymer coating on their aggregation behavior and toxicity to the green algae Dunaliella salina

Energy Technology Data Exchange (ETDEWEB)

Zamani, Hajar [Department of Biology, Faculty of Sciences, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of); Moradshahi, Ali, E-mail: moradshahi@susc.ac.ir [Department of Biology, Faculty of Sciences, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of); Jahromi, Hamed Dehdashti; Sheikhi, Mohammad Hosein [Nanotechnology Research Institute, Shiraz University, Shiraz 71454 (Iran, Islamic Republic of)

2014-09-15

Highlights: • Lead sulfide nanoparticles (PbS NPs) are toxic to D. salina. • Gum-Arabic coating alters the toxicity of PbS NPs. • Cell-NPs agglomerates and lipid peroxidation could explain the toxicity of PbS NPs. • Shading effect and dissolution do not seem to contribute to the toxicity of PbS NPs. • Particle–particle interaction was reduced by coating; therefore, PbS NPs were stabilized in the culture media. - Abstract: The potential hazards of nanoparticles (NPs) to the environment and to living organisms need to be considered for a safe development of nanotechnology. In the present study, the potential toxic effects of uncoated and gum Arabic-coated lead sulfide nanoparticles (GA-coated PbS NPs) on the growth, lipid peroxidation, reducing capacity and total carotenoid content of the hypersaline unicellular green algae Dunaliella salina were investigated. Coatings of PbS NPs with GA, as confirmed by Fourier transform infrared spectroscopy, reduced the toxicity of PbS NPs. Uncoated PbS NP toxicity to D. salina was attributed to higher algal cell-NP agglomerate formation, higher lipid peroxidation, lower content of total reducing substances and lower total carotenoid content. Low levels of Pb{sup 2+} in the growth culture media indicate that PbS NP dissolution does not occur in the culture. Also, the addition of 100 μM Pb{sup 2+} to the culture media had no significant (P > 0.05) effect on algal growth. The shading of light (shading effect) by PbS NPs, when simulated using activated charcoal, did not contribute to the overall toxic effect of PbS NPs which was evident by insignificant (P > 0.05) reduction in the growth and antioxidant capacity of the algae. When PbS NP aggregation in culture media (without algal cells) was followed for 60 min, uncoated form aggregated rapidly reaching aggregate sizes with hydrodynamic diameter of over 2500 nm within 60 min. Effective particle–particle interaction was reduced in the GA-coated NPs. Aggregates of about

Influence of PbS nanoparticle polymer coating on their aggregation behavior and toxicity to the green algae Dunaliella salina

International Nuclear Information System (INIS)

Zamani, Hajar; Moradshahi, Ali; Jahromi, Hamed Dehdashti; Sheikhi, Mohammad Hosein

2014-01-01

Highlights: • Lead sulfide nanoparticles (PbS NPs) are toxic to D. salina. • Gum-Arabic coating alters the toxicity of PbS NPs. • Cell-NPs agglomerates and lipid peroxidation could explain the toxicity of PbS NPs. • Shading effect and dissolution do not seem to contribute to the toxicity of PbS NPs. • Particle–particle interaction was reduced by coating; therefore, PbS NPs were stabilized in the culture media. - Abstract: The potential hazards of nanoparticles (NPs) to the environment and to living organisms need to be considered for a safe development of nanotechnology. In the present study, the potential toxic effects of uncoated and gum Arabic-coated lead sulfide nanoparticles (GA-coated PbS NPs) on the growth, lipid peroxidation, reducing capacity and total carotenoid content of the hypersaline unicellular green algae Dunaliella salina were investigated. Coatings of PbS NPs with GA, as confirmed by Fourier transform infrared spectroscopy, reduced the toxicity of PbS NPs. Uncoated PbS NP toxicity to D. salina was attributed to higher algal cell-NP agglomerate formation, higher lipid peroxidation, lower content of total reducing substances and lower total carotenoid content. Low levels of Pb 2+ in the growth culture media indicate that PbS NP dissolution does not occur in the culture. Also, the addition of 100 μM Pb 2+ to the culture media had no significant (P > 0.05) effect on algal growth. The shading of light (shading effect) by PbS NPs, when simulated using activated charcoal, did not contribute to the overall toxic effect of PbS NPs which was evident by insignificant (P > 0.05) reduction in the growth and antioxidant capacity of the algae. When PbS NP aggregation in culture media (without algal cells) was followed for 60 min, uncoated form aggregated rapidly reaching aggregate sizes with hydrodynamic diameter of over 2500 nm within 60 min. Effective particle–particle interaction was reduced in the GA-coated NPs. Aggregates of about 440 nm

Cultural heritage and food identity: The pre-Hispanic salt of Zapotitlán Salinas, Mexico

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Renard, Marie-Christine

2016-06-01

Full Text Available Salt production in Zapotitlán de las Salinas (Puebla, Mexico dates back to pre-Hispanic times when the Popolocas inhabiting the Tehuacán Valley paid it as tribute to the Aztecs. The technique to obtain salt has changed little over the past 500 years and know-how continues to be transmitted from generation to generation of salters (salineros. It is a resource that is deeply anchored in the identity of the inhabitants of Zapotitlán and regional cuisine. Salt has endured over the centuries as a perennial resource and constitutes a source of income for its owners. However, despite these historical and cultural factors of territorial anchorage, salters have not attained the level of organization necessary to obtain a fair value in the market. Failure to appreciate this product has led to the abandonment of a large percentage of the saltworks that once existed. This essay will analyze the socio-economic and cultural constraints that have prevented this community from attaining the level of territorial governance necessary to enhance the market value of Zapotitlán salt on the market but how, with the depletion of other economic options, its people are returning to the salt, with new strategies. Lastly, the paper will conclude with a consideration of its future potential.La producción de sal de Zapotitlán de las Salinas (Puebla, México se remonta a épocas prehispánicas cuando los popolocas, moradores del valle de Tehuacán, lo tributaban a los aztecas. La técnica para la obtención de la sal ha cambiado poco desde hace 500 años y el saber-hacer se sigue transmitiendo entre las generaciones de salineros. Es un recurso profundamente anclado en la identidad de los habitantes de Zapotitlán y en la gastronomía regional. Su existencia ha perdurado en el transcurso de los siglos por ser un recurso perenne y constituir una renta para sus dueños. Sin embargo, a pesar de los factores históricos y culturales de anclaje territorial, los salineros no

Gamma-H2AX biodosimetry for use in large scale radiation incidents: comparison of a rapid ‘96 well lyse/fix’ protocol with a routine method

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Jayne Moquet

2014-03-01

Full Text Available Following a radiation incident, preliminary dose estimates made by γ-H2AX foci analysis can supplement the early triage of casualties based on clinical symptoms. Sample processing time is important when many individuals need to be rapidly assessed. A protocol was therefore developed for high sample throughput that requires less than 0.1 ml blood, thus potentially enabling finger prick sampling. The technique combines red blood cell lysis and leukocyte fixation in one step on a 96 well plate, in contrast to the routine protocol, where lymphocytes in larger blood volumes are typically separated by Ficoll density gradient centrifugation with subsequent washing and fixation steps. The rapid ‘96 well lyse/fix’ method reduced the estimated sample processing time for 96 samples to about 4 h compared to 15 h using the routine protocol. However, scoring 20 cells in 96 samples prepared by the rapid protocol took longer than for the routine method (3.1 versus 1.5 h at zero dose; 7.0 versus 6.1 h for irradiated samples. Similar foci yields were scored for both protocols and consistent dose estimates were obtained for samples exposed to 0, 0.2, 0.6, 1.1, 1.2, 2.1 and 4.3 Gy of 250 kVp X-rays at 0.5 Gy/min and incubated for 2 h. Linear regression coefficients were 0.87 ± 0.06 (R2 = 97.6% and 0.85 ± 0.05 (R2 = 98.3% for estimated versus actual doses for the routine and lyse/fix method, respectively. The lyse/fix protocol can therefore facilitate high throughput processing for γ-H2AX biodosimetry for use in large scale radiation incidents, at the cost of somewhat longer foci scoring times.

A protein chip membrane-capture assay for botulinum neurotoxin activity

International Nuclear Information System (INIS)

Marconi, Severine; Ferracci, Geraldine; Berthomieu, Maelys; Kozaki, Shunji; Miquelis, Raymond; Boucraut, Jose; Seagar, Michael

2008-01-01

Botulinum neurotoxins A and B (BoNT/A and B) are neuromuscular blocking agents which inhibit neurotransmission by cleaving the intra-cellular presynaptic SNARE proteins SNAP-25 and VAMP2, localized respectively in plasma membrane and synaptic vesicles. These neurotoxins are both dangerous pathogens and powerful therapeutic agents with numerous clinical and cosmetic applications. Consequently there is a need for in vitro assays of their biological activity to screen for potential inhibitors and to replace the widely used in vivo mouse assay. Surface plasmon resonance (SPR) was used to measure membrane vesicle capture by antibodies against SNAP-25 and VAMP2. Substrate cleavage by BoNTs modified capture providing a method to assay toxin activity. Firstly using synaptic vesicles as a substrate, a comparison of the EC 50 s for BoNT/B obtained by SPR, ELISA or flow cytometry indicated similar sensitivity although SPR assays were more rapid. Sonication of brain or neuronal cultures generated plasma membrane fragments with accessible intra-cellular epitopes adapted to measurement of BoNT/A activity. SPR responses were proportional to antigen concentration permitting detection of as little as 4 pM SNAP-25 in crude lysates. BoNT/A activity was assayed using monoclonal antibodies that specifically recognize a SNAP-25 epitope generated by the proteolytic action of the toxin. Incubation of intact primary cultured neurons with BoNT/A yielded an EC 50 of 0.5 pM. The SPR biosensor method was sensitive enough to monitor BoNT/A and B activity in cells cultured in a 96-well format providing an alternative to experimental animals for toxicological assays

Identifikasi Metabolit Sekunder Ekstrak Etil Asetat Biji Alpukat (Persea americana Mill. dan Uji Toksisitas Terhadap Larva Udang Artemia salina Leach.

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Nurdia Asdar

2014-07-01

Full Text Available Identification of metabolite compound in acetone Extract from Fruits Avocado seed (Percea americana Mill and toxicity test against Artemia salina Leach. Aim of this research is to identify the secondary metabolite compound in aceton extract from fruits Avocado seed (Percea americana Mill and to determine the potential of secondary metabolites from an avocado extract as anticancer. The compound was obtained from Identification process in several stages, namely extraction, fractination, purification and identification. The identification process was color test, TLC, spectroscopy IR. The toxicity test by Brine Shrimp Lethality Test (BSLT to Artemia salina Leach. The compound obtained from this research was yellow-white crystalline needle-shaped, the purity test with TLC analysis showed a stain in three eluen system on the stain test on three eluent systems with Rf proportions as follows; 0,15 from chloroform:ethyl acetat (6:4, 0,33 from methanol:chloroform (1:9, 0,75 for acetone:ethyl acetat, and positively to the reagent Wagner and gave brown precipitate for indicate as alcaloid group. This result is supported by spectroscopy from compound.While toxicity tests showed condensed acetone extracts  and pure compounds are toxic with LC50 value of each is 20.61 mg/mL and 39,81 mg/mL.

Validation of 2 commercial Neospora caninum antibody enzyme linked immunosorbent assays

Science.gov (United States)

Wu, John T.Y.; Dreger, Sally; Chow, Eva Y.W.; Bowlby, Evelyn E.

2002-01-01

Abstract This is a validation study of 2 commercially available enzyme linked immunosorbent assays (ELISA) for the detection of antibodies against Neospora caninum in bovine serum. The results of the reference sera (n = 30) and field sera from an infected beef herd (n = 150) were tested by both ELISAs and the results were compared statistically. When the immunoblotting results of the reference bovine sera were compared to the ELISA results, the same identity score (96.67%) and kappa values (K) (0.93) were obtained for both ELISAs. The sensitivity and specificity values for the IDEXX test were 100% and 93.33% respectively. For the Biovet test 93.33% and 100% were obtained. The corresponding positive (PV+) and negative predictive (PV−) values for the 2 assays were 93.75% and 100% (IDEXX), and 100% and 93.75% (Biovet). In the 2nd study, competitive inhibition ELISA (c-ELISA) results on bovine sera from an infected herd were compared to the 2 sets of ELISA results. The identity scores of the 2 ELISAs were 98% (IDEXX) and 97.33% (Biovet). The K values calculated were 0.96 (IDEXX) and 0.95 (Biovet). For the IDEXX test the sensitivity and specificity were 97.56% and 98.53%, whereas for the Biovet assay 95.12% and 100% were recorded, respectively. The corresponding PV+ and PV− values were 98.77% and 97.1% (IDEXX), and 100% and 94.44% (Biovet). Our validation results showed that the 2 ELISAs worked equally well and there was no statistically significant difference between the performance of the 2 tests. Both tests showed high reproducibility, repeatability and substantial agreement with results from 2 other laboratories. A quality assurance based on the requirement of the ISO/IEC 17025 standards has been adopted throughout this project for test validation procedures. PMID:12418782

Nivel de conocimiento sobre tuberculosis pulmonar y actitud de los pacientes, centro salud San Juan de Salinas, 2017

OpenAIRE

Atuncar Mendoza, Pamela

2017-01-01

El objetivo del estudios de investigación fue determinar la relación que existe entre el nivel de conocimiento sobretuberculosis pulmonar y la actitud hacia el tratamiento detuberculosis pulmonar de los pacientes dela Estrategia Sanitaria Nacional de Prevención y Control de la Tuberculosis, Centro de Salud San Juan de Salinas, San Martinde Porres, año 2017. En cuanto a lametodología fue de tipo de investigación aplicada, nivel descriptivo correccional,la población estuvo conformada por 50 p...

Development of a heavy metals enzymatic-based assay using papain

International Nuclear Information System (INIS)

Shukor, Yunus; Baharom, Nor Azlan; Rahman, Fadhil Abd.; Abdullah, Mohd. Puad; Shamaan, Nor Aripin; Syed, Mohd. Arif

2006-01-01

A heavy metals enzymatic-based assay using papain was developed. Papain was assayed using the Casein-coomassie-dye-binding assay. The assay is sensitive to several heavy metals. The IC 50 (concentration of toxicant giving 50% inhibition) of Hg 2+ , Ag 2+ , Pb 2 , Zn 2+ is 0.39, 0.40, 2.16, 2.11 mg l -1 , respectively. For Cu 2+ and Cd 2+ the LOQ (limits of quantitation) is 0.004 and 0.1 mg l -1 , respectively. The IC 50 and LOQ values were found to be generally comparable to several other enzymatic and bioassays tests such as: immobilized urease, 15-min Microtox TM , 48 h Daphnia magna, and 96 h Rainbow trout. The papain assay is xenobiotics tolerant, has a wide pH for optimum activity, is temperature stable, and has a relatively quick assay time. The papain assay was used to identify polluted water samples from industrial sources in Penang, Malaysia. We found one site where the assay gave a positive toxic response. The toxicity of the site was confirmed using Atomic Emission Spectrometry analysis

Broad-range (pan) Salmonella and Salmonella serotype typhi-specific real-time PCR assays: potential tools for the clinical microbiologist.

Science.gov (United States)

Farrell, John J; Doyle, Laura J; Addison, Rachel M; Reller, L Barth; Hall, Geraldine S; Procop, Gary W

2005-03-01

We describe broad-range salmonellae (ie, Salmonella) and Salmonella serotype Typhi-specific LightCycler (Roche Diagnostics, Indianapolis, IN) real-time polymerase chain reaction assays. We validated these with a battery of 280 bacteria, 108 of which were salmonellae representing 20 serotypes. In addition, 298 isolates from 170 clinical specimens that were suspected to possibly represent Salmonella were tested with the pan- Salmonella assay. Finally, the pan-Salmonella assay also was used to test DNA extracts from 101 archived, frozen stool specimens, 55 of which were culture-positive for salmonellae. Both assays were 100% sensitive and specific when cultured isolates of the battery were tested. The pan- Salmonella assay also characterized correctly all salmonellae on the primary isolation agar and was 96% sensitive (53/55) and 96% specific (49/51) when nucleic acid extracts from direct stool specimens were tested. These assays represent potential tools the clinical microbiologist could use to screen suspect isolates or stool specimens for Salmonella.

Evaluation of Culture Conditions to Obtain Fatty Acids from Saline Microalgae Species: Dunaliella salina, Sinecosyfis sp., and Chroomonas sp.

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D. A. Castilla Casadiego

2016-01-01

Full Text Available The use of the saline microalgae, Dunaliella salina, Sinecosyfis sp., and Chroomonas sp., was explored as an alternative source for the production of fatty acids using fertilizer and glycerol as culture media. The nutrient medium used contained “Nutrifoliar,” a commercial fertilizer, and/or glycerol, in natural sea water. The microalgae were placed in cultures with different conditions. The parameters that favored the largest production of fatty acids were 24 hours of agitation and illumination, 1620 L/day of air supply, 2.25 L of air/min, and a temperature of 32°C using “Nutrifoliar” as the culture media. Results indicated that, from 3 g of microalgae in wet base of Chroomonas sp., 54.43 mg of oil was produced. The chromatographic characterization of oil obtained revealed the presence of essential fatty acids such as 9,12,15-octadecatrienoic acid (omega-3 and 4,7,10-hexadecatrienoic acid (omega-6 from the species Dunaliella salina. On the other hand, 9,12-octadecadienoic acid (omega-6 and cis-11-eicosenoic acid (omega-9 were identified from the species Chroomonas sp. The temperature variations played an important role in the velocity of growth or the production of the algae biomass, the amount of oil, and the ability to produce fatty acids.

[Immunocytochemical studies on the phase of differentiation of hatching gland cells in brine shrimp, Artemia salina].

Science.gov (United States)

Li, Ling; Fan, Ting Jun; Wang, Xiao Feng; Cong, Ri Shan; Yu, Qiu Tao; Zhong, Qi Wang

2004-04-01

Hatching enzyme (HE), synthesized in hatching gland cells (HGCs), plays vital roles in animal hatching. Immunocytochemical techniques employing anti-GST-UVS.2 antiserum, prepared from Xenopus HE and with specificity to brine shrimp HE, were first used to investigate the differentiation and variability of hatching gland cells (HGCs) in the hatching process of embryos of brine shrimp, Artemia salina, in this study. HGCs with immunoreactivity to anti-GST-UVS.2 antiserum were identified, for the first time, in brine shrimp embryos during hatching process. Immunocytochemical staining results showed that, (1) HE-positive immunoreactivity is really specific to Artemia HE, and its appearance and disappearance are closely correlated with the hatching process of Artemia salina. (2) Artemia HGCs, first appeared in embryos 5 hours before hatching and disappeared 4 hours after hatching, were also a transient type of cells, with an existence period of 9 hours. (3) The head portion of Artemia embryo is probably the initial position of HE secretion, and likely to be the main position of HE secretion as well. The detailed process and mechanism need to be studied. (4) The appearance of HGCs is in a synchronous mode from places all over the embryos, and their disappearance is also in a synchronous mode. (5) The number of HGCs increased gradually along with embryo development process and reached a maximum number at hatching. Contrarily, the number of HGCs decreased gradually after hatching, and HGCs disappeared 5 hours after hatching. However, the intensity of HE-positive reaction was almost at the same level at the period of HGCs'presence. (6) Artemia HGCs were distributed throughout the body of embryos at all time during their presence. Therefore, it can concluded that Artemia HGCs, as a transient type of cells, first appeared in embryos 4 hours before hatching and disappeared in embryos 5 hours after hatching, and with distinguished patterns of appearance, disappearance and

Rainfall leaching is critical for long-term use of recycled water in the Salinas Valley

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Belinda E. Platts

2014-07-01

Full Text Available In 1998, Monterey County Water Recycling Projects began delivering water to 12,000 acres in the northern Salinas Valley. Two years later, an ongoing study began assessing the effects of the recycled water on soil salinity. Eight sites are receiving recycled water and a control site is receiving only well water. In data collected from 2000 to 2012, soil salinity of the 36-inch-deep profile was on average approximately double that of the applied water, suggesting significant leaching from applied water (irrigation or rainfall. In this study, we investigated some of the soil water hydrology factors possibly controlling the soil salinity results. Using soil water balance modeling, we found that rainfall had more effect on soil salinity than did leaching from irrigation. Increasing applied water usually only correlated significantly with soil salinity parameters in the shallow soil profile (1 to 12 inches depth and at 24 to 36 inches at sites receiving fairly undiluted recycled water. Winter rains, though, had a critical effect. Increasing rainfall depths were significantly correlated with decreasing soil salinity of the shallow soil at all test sites, though this effect also diminished with increased soil depth. When applied water had high salinity levels, winter rainfall in this area was inadequate to prevent soil salinity from increasing.

Processing recommendations for using low-solids digestate as nutrient solution for poly-ß-hydroxybutyrate production with Synechocystis salina.

Science.gov (United States)

Meixner, K; Fritz, I; Daffert, C; Markl, K; Fuchs, W; Drosg, B

2016-12-20

Within the last decades, environmental pollution with persistent plastics steadily increased; therefore the production of biodegradable materials like poly-ß-hydroxybutyrate (PHB) is essential. Currently, PHB is produced with heterotrophic bacteria from crops. This leads to competition with food and feed production, which can be avoided by using photoautotrophic cyanobacteria, as Synechocystis salina, synthesizing PHB from CO 2 at nutrient limitation. This study aims to increase the economic efficiency of PHB production with cyanobacteria by using nutrients from anaerobic digestate. First, growth and PHB production of S. salina in digestate fractions (supernatant and permeate, with/without precipitating agents) and dilutions thereof and then the scale-up (photobioreactor, 200 L working volume) were evaluated. With precipitated and centrifuged digestate diluted 1/3 the highest biomass (1.55gL -1 ) and PHB concentrations (95.4mgL -1 ), being 78% of those in mineral media, were achieved. In the photobioreactor-experiments biomass (1.63gL -1 ) and PHB concentrations (88.7mgL -1 ), being 79% and 72% of those in mineral medium, were reached, but in a cultivation time 10days longer than in mineral medium. The possibility to use digestate as sustainable and low cost nutrient solution for microalgae cultivation and photoautotrophic PHB production, instead of applying it on fields or processing it to achieve discharge limits, makes this application a highly valid option. Copyright © 2016 Elsevier B.V. All rights reserved.

PHYTOCHEMICAL STUDY, MICROBIOLOGICAL AND CYTOTOXICITYTY ACTIVITY IN Artemia salina LEACH, AERIAL PARTS OF Petiveria alliacea L. PHYTOLACCACEAE.

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Darllen Suzanny Barbosa de Oliveira

2013-12-01

Full Text Available This study aimed to phytochemical study of the aerial parts of Petiveria alliacea L. (young branches and leaves, cytotoxicity evaluation (LC50 of leaves in relation to Artemia salina L. and antimicrobial activity, which were used strains of gram-positive bacterium: Staphylococcus aureus and gram-negative: Klebsiella pneumoniae and Escherichia coli, these submitted to the methanol crude extract (MCE of leaves, at the following concentrations: 25.50 and 100mg/mL. It was possible to identify in the young branches of P. alliacea L. the presence of organic acids, phenols and tannins in the alkaloids leaves, steroids and triterpenoids, saponins, phenols and tannins, where the activities of these metabolites match with some information alleged by the population. The leaves’ Methanol Crude Extract showed LC50=1709.77μg/mL, being nontoxic at the tested concentrations, whereas for plant extract in relation to A.salina are considered nontoxic when LC50>1000μg/mL. The antimicrobial activity of the Methanol Crude Extract of the leaves showed inhibition only for the bacteria Escherichia coli at 100mg/mL concentration, and this activity may be related to the presence of phenols and tannins in the extract. The obtained results turn the species promising in search of secondary metabolites, but there is the need of further studies to identify its main active ingredients. Palavras-chave: Mucuracaa, Phytochemical Screening, Class of Compounds. DOI: http://dx.doi.org/10.18561/2179-5746/biotaamazonia.v3n3p76-82

A priori estimation of accuracy and of the number of wells to be employed in limiting dilution assays

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J.G. Chaui-Berlinck

2000-08-01

Full Text Available The use of limiting dilution assay (LDA for assessing the frequency of responders in a cell population is a method extensively used by immunologists. A series of studies addressing the statistical method of choice in an LDA have been published. However, none of these studies has addressed the point of how many wells should be employed in a given assay. The objective of this study was to demonstrate how a researcher can predict the number of wells that should be employed in order to obtain results with a given accuracy, and, therefore, to help in choosing a better experimental design to fulfill one's expectations. We present the rationale underlying the expected relative error computation based on simple binomial distributions. A series of simulated in machina experiments were performed to test the validity of the a priori computation of expected errors, thus confirming the predictions. The step-by-step procedure of the relative error estimation is given. We also discuss the constraints under which an LDA must be performed.

Pollution by polycyclic aromatic hydrocarbons (PAH's) in sediments and organisms from Salina Cruz Port, Oaxaca, Mexico

International Nuclear Information System (INIS)

Botello, A.V.; Villanueva, S.; Diaz, G.; Pica, Y.

1995-01-01

The presence and levels of polycyclic aromatic hydrocarbons (PAH's) in [sediments and biota from the Port of Salina Cruz, Oaxaca; were evaluated by means of gas capillary chromatography using columns of high resolution. The results show a seasonal variability of the PAH's concentrations in sediments being higher in the port area and lower in oceanic sediments. The increase of the PAH's levels in Crassostrea iridiscens and Penaeus stylirostris is important and related to the bioaccumulation process. The presence of PAH's conformed by 4 y 5 benzene rings in these species must be noted specially because they have carcinogenic properties and their effects on the local fisheries should be considered. (Author)

Antifungal chemical compounds identified using a C. elegans pathogenicity assay.

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Julia Breger

2007-02-01

Full Text Available There is an urgent need for the development of new antifungal agents. A facile in vivo model that evaluates libraries of chemical compounds could solve some of the main obstacles in current antifungal discovery. We show that Candida albicans, as well as other Candida species, are ingested by Caenorhabditis elegans and establish a persistent lethal infection in the C. elegans intestinal track. Importantly, key components of Candida pathogenesis in mammals, such as filament formation, are also involved in nematode killing. We devised a Candida-mediated C. elegans assay that allows high-throughput in vivo screening of chemical libraries for antifungal activities, while synchronously screening against toxic compounds. The assay is performed in liquid media using standard 96-well plate technology and allows the study of C. albicans in non-planktonic form. A screen of 1,266 compounds with known pharmaceutical activities identified 15 (approximately 1.2% that prolonged survival of C. albicans-infected nematodes and inhibited in vivo filamentation of C. albicans. Two compounds identified in the screen, caffeic acid phenethyl ester, a major active component of honeybee propolis, and the fluoroquinolone agent enoxacin exhibited antifungal activity in a murine model of candidiasis. The whole-animal C. elegans assay may help to study the molecular basis of C. albicans pathogenesis and identify antifungal compounds that most likely would not be identified by in vitro screens that target fungal growth. Compounds identified in the screen that affect the virulence of Candida in vivo can potentially be used as "probe compounds" and may have antifungal activity against other fungi.

Performance of the Xpert HIV-1 Viral Load Assay: a Systematic Review and Meta-analysis.

Science.gov (United States)

Nash, Madlen; Huddart, Sophie; Badar, Sayema; Baliga, Shrikala; Saravu, Kavitha; Pai, Madhukar

2018-04-01

Viral load (VL) is the preferred treatment-monitoring approach for HIV-positive patients. However, more rapid, near-patient, and low-complexity assays are needed to scale up VL testing. The Xpert HIV-1 VL assay (Cepheid, Sunnyvale, CA) is a new, automated molecular test, and it can leverage the GeneXpert systems that are being used widely for tuberculosis diagnosis. We systematically reviewed the evidence on the performance of this new tool in comparison to established reference standards. A total of 12 articles (13 studies) in which HIV patient VLs were compared between Xpert HIV VL assay and a reference standard VL assay were identified. Study quality was generally high, but substantial variability was observed in the number and type of agreement measures reported. Correlation coefficients between Xpert and reference assays were high, with a pooled Pearson correlation ( n = 8) of 0.94 (95% confidence interval [CI], 0.89, 0.97) and Spearman correlation ( n = 3) of 0.96 (95% CI, 0.86, 0.99). Bland-Altman metrics ( n = 11) all were within 0.35 log copies/ml of perfect agreement. Overall, Xpert HIV-1 VL performed well compared to current reference tests. The minimal training and infrastructure requirements for the Xpert HIV-1 VL assay make it attractive for use in resource-constrained settings, where point-of-care VL testing is most needed. Copyright © 2018 Nash et al.

High surface adsorption properties of carbon-based nanomaterials are responsible for mortality, swimming inhibition, and biochemical responses in Artemia salina larvae.

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Mesarič, Tina; Gambardella, Chiara; Milivojević, Tamara; Faimali, Marco; Drobne, Damjana; Falugi, Carla; Makovec, Darko; Jemec, Anita; Sepčić, Kristina

2015-06-01

We investigated the effects of three different carbon-based nanomaterials on brine shrimp (Artemia salina) larvae. The larvae were exposed to different concentrations of carbon black, graphene oxide, and multiwall carbon nanotubes for 48 h, and observed using phase contrast and scanning electron microscopy. Acute (mortality) and behavioural (swimming speed alteration) responses and cholinesterase, glutathione-S-transferase and catalase enzyme activities were evaluated. These nanomaterials were ingested and concentrated in the gut, and attached onto the body surface of the A. salina larvae. This attachment was responsible for concentration-dependent inhibition of larval swimming, and partly for alterations in the enzyme activities, that differed according to the type of tested nanomaterials. No lethal effects were observed up to 0.5mg/mL carbon black and 0.1mg/mL multiwall carbon nanotubes, while graphene oxide showed a threshold whereby it had no effects at 0.6 mg/mL, and more than 90% mortality at 0.7 mg/mL. Risk quotients calculated on the basis of predicted environmental concentrations indicate that carbon black and multiwall carbon nanotubes currently do not pose a serious risk to the marine environment, however if uncontrolled release of nanomaterials continues, this scenario can rapidly change. Copyright © 2015 Elsevier B.V. All rights reserved.

Cytotoxicity Test Based on Human Cells Labeled with Fluorescent Proteins: Fluorimetry, Photography, and Scanning for High-Throughput Assay.

Science.gov (United States)

Kalinina, Marina A; Skvortsov, Dmitry A; Rubtsova, Maria P; Komarova, Ekaterina S; Dontsova, Olga A

2018-06-01

High- and medium-throughput assays are now routine methods for drug screening and toxicology investigations on mammalian cells. However, a simple and cost-effective analysis of cytotoxicity that can be carried out with commonly used laboratory equipment is still required. The developed cytotoxicity assays are based on human cell lines stably expressing eGFP, tdTomato, mCherry, or Katushka2S fluorescent proteins. Red fluorescent proteins exhibit a higher signal-to-noise ratio, due to less interference by medium autofluorescence, in comparison to green fluorescent protein. Measurements have been performed on a fluorescence scanner, a plate fluorimeter, and a camera photodocumentation system. For a 96-well plate assay, the sensitivity per well and the measurement duration were 250 cells and 15 min for the scanner, 500 cells and 2 min for the plate fluorimeter, and 1000 cells and less than 1 min for the camera detection. These sensitivities are similar to commonly used MTT (tetrazolium dye) assays. The used scanner and the camera had not been previously applied for cytotoxicity evaluation. An image processing scheme for the high-resolution scanner is proposed that significantly diminishes the number of control wells, even for a library containing fluorescent substances. The suggested cytotoxicity assay has been verified by measurements of the cytotoxicity of several well-known cytotoxic drugs and further applied to test a set of novel bacteriotoxic compounds in a medium-throughput format. The fluorescent signal of living cells is detected without disturbing them and adding any reagents, thus allowing to investigate time-dependent cytotoxicity effects on the same sample of cells. A fast, simple and cost-effective assay is suggested for cytotoxicity evaluation based on mammalian cells expressing fluorescent proteins and commonly used laboratory equipment.

Crescimento, consumo hídrico e composição mineral de alface cultivada em hidroponia com águas salinas

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Dalva Paulus

2012-02-01

Full Text Available Objetivou-se, com este trabalho, avaliar o crescimento, o consumo hídrico e a extração de nutrientes, por alface (Lactuca sativa L. em sistema hidropônico NFT (fluxo laminar de nutrientes, com a utilização de águas salinas no preparo da solução nutritiva e reposição da lâmina diária evapotranspirada. Foram conduzidos dois experimentos, em ambiente protegido. O delineamento experimental foi em blocos ao acaso, sendo estudados os efeitos de cinco níveis de salinidade da água, utilizando-se NaCl. O aumento da salinidade da água reduziu, de forma linear, o crescimento e o consumo hídrico da alface. A salinidade reduziu a concentração de macronutrientes na parte aérea, mas não se observaram sintomas de deficiência nutricional. O rendimento (massa de matéria fresca da parte aérea foi reduzido entre 6 e 6,5% por dS m-1, no primeiro e segundo cultivo, respectivamente. Os resultados obtidos em sistema de cultivo NFT podem indicar a possibilidade do uso da água salina como alternativa para produção de hortaliças, para produtores que têm disponibilidade restrita de água doce, porém, com redução de produtividade.

Ethanol fermentation with Kluyveromyces marxianus from Jerusalem artichoke grown in salina and irrigated with a mixture of seawater and freshwater.

Science.gov (United States)

Yuan, W J; Zhao, X Q; Ge, X M; Bai, F W

2008-12-01

To study fuel ethanol fermentation with Kluyveromyces marxianus ATCC8554 from Jerusalem artichoke (Helianthus tuberosus) grown in salina and irrigated with a mixture of seawater and freshwater. The growth and ethanol fermentation of K. marxianus ATCC8554 were studied using inulin as substrate. The activity of inulinase, which attributes to the hydrolysis of inulin, the main carbohydrate in Jerusalem artichoke, was monitored. The optimum temperatures were 38 degrees C for growth and inulinase production, and 35 degrees C for ethanol fermentation. Aeration was not necessary for ethanol fermentation with the K. marxianus from inulin. Then, the fresh Jerusalem artichoke tubers grown in salina and irrigated with 25% and 50% seawater were further examined for ethanol fermentation with the K. marxianus, and a higher ethanol yield was achieved for the Jerusalem artichoke tuber irrigated with 25% seawater. Furthermore, the dry meal of the Jerusalem artichoke tubers irrigated with 25% seawater was examined for ethanol fermentation at three solid concentrations of 200, 225 and 250 g l(-1), and the highest ethanol yield of 0.467, or 91.5% of the theoretical value of 0.511, was achieved for the slurry with a solid concentration of 200 g l(-1). Halophilic Jerusalem artichoke can be used for fuel ethanol production. Halophilic Jerusalem artichoke, not competing with grain crops for arable land, is a sustainable feedstock for fuel ethanol production.

Estudo de processos de produção de etanol anidro através da destilação extrativa salina com recuperação do agente de separação

OpenAIRE

Eliana Luci Ligero

1999-01-01

Resumo: : Uma técnica amplamente utilizada na produção de etanol é a destilação azeotrópica benzeno, cuja propriedade cancerígena é bastante conhecida. Um processo alternativo, ecologicamente viável, é a destilação extrativa salina. O emprego de um sal solúvel na desidratação do etanol possibilita uma redução no consumo energético, além de produzir um etanol livre do agente de separação. Uma etapa importante na produção de etanol anidro através da destilação extrativa salina é a recuperação d...

Validation and application of a high-performance liquid chromatography-tandem mass spectrometric method for simultaneous quantification of lopinavir and ritonavir in human plasma using semi-automated 96-well liquid-liquid extraction.

Science.gov (United States)

Wang, Perry G; Wei, Jack S; Kim, Grace; Chang, Min; El-Shourbagy, Tawakol

2006-10-20

Kaletra is an important antiretroviral drug, which has been developed by Abbott Laboratories. It is composed of lopinavir (low-pin-a-veer) and ritonavir (ri-toe-na-veer). Both have been proved to be human immunodeficiency virus (HIV) protease inhibitors and have substantially reduced the morbidity and mortality associated with HIV-1 infection. We have developed and validated an assay, using liquid chromatography coupled with atmospheric pressure chemical ionization tandem mass spectrometry (LC/MS/MS), for the routine quantification of lopinavir and ritonavir in human plasma, in which lopinavir and ritonavir can be simultaneously analyzed with high throughput. The sample preparation consisted of liquid-liquid extraction with a mixture of hexane: ethyl acetate (1:1, v/v), using 100 microL of plasma. Chromatographic separation was performed on a Waters Symmetry C(18) column (150 mm x 3.9 mm, particle size 5 microm) with reverse-phase isocratic using mobile phase of 70:30 (v/v) acetonitrile: 2 mM ammonium acetate aqueous solution containing 0.01% formic acid (v/v) at a flow rate of 1.0 mL/min. A Waters symmetry C(18) guard column (20 mm x 3.9 mm, particle size 5 microm) was connected prior to the analytical column, and a guard column back wash was performed to reduce the analytical column contamination using a mixture of tetrahydrofuran (THF), methanol and water (45:45:10, v/v/v). The analytical run was 4 min. The use of a 96-well plate autosampler allowed a batch size up to 73 study samples. A triple-quadrupole mass spectrometer was operated in a positive ion mode and multiple reaction monitoring (MRM) was used for drug quantification. The method was validated over the concentration ranges of 19-5,300 ng/mL for lopinavir and 11-3,100 ng/mL for ritonavir. A-86093 was used as an internal standard (I.S.). The relative standard deviation (RSD) were <6% for both lopinavir and ritonavir. Mean accuracies were between the designed limits (+/-15%). The robust and rapid LC

Micropatterned comet assay enables high throughput and sensitive DNA damage quantification.

Science.gov (United States)

Ge, Jing; Chow, Danielle N; Fessler, Jessica L; Weingeist, David M; Wood, David K; Engelward, Bevin P

2015-01-01

The single cell gel electrophoresis assay, also known as the comet assay, is a versatile method for measuring many classes of DNA damage, including base damage, abasic sites, single strand breaks and double strand breaks. However, limited throughput and difficulties with reproducibility have limited its utility, particularly for clinical and epidemiological studies. To address these limitations, we created a microarray comet assay. The use of a micrometer scale array of cells increases the number of analysable comets per square centimetre and enables automated imaging and analysis. In addition, the platform is compatible with standard 24- and 96-well plate formats. Here, we have assessed the consistency and sensitivity of the microarray comet assay. We showed that the linear detection range for H2O2-induced DNA damage in human lymphoblastoid cells is between 30 and 100 μM, and that within this range, inter-sample coefficient of variance was between 5 and 10%. Importantly, only 20 comets were required to detect a statistically significant induction of DNA damage for doses within the linear range. We also evaluated sample-to-sample and experiment-to-experiment variation and found that for both conditions, the coefficient of variation was lower than what has been reported for the traditional comet assay. Finally, we also show that the assay can be performed using a 4× objective (rather than the standard 10× objective for the traditional assay). This adjustment combined with the microarray format makes it possible to capture more than 50 analysable comets in a single image, which can then be automatically analysed using in-house software. Overall, throughput is increased more than 100-fold compared to the traditional assay. Together, the results presented here demonstrate key advances in comet assay technology that improve the throughput, sensitivity, and robustness, thus enabling larger scale clinical and epidemiological studies. © The Author 2014. Published by

Methodological considerations for using umu assay to assess photo-genotoxicity of engineered nanoparticles

DEFF Research Database (Denmark)

Cupi, Denisa; Baun, Anders

2016-01-01

In this study we investigated the feasibility of high-throughput (96-well plate) umu assay to test the genotoxic effect of TiO2 engineered nanoparticles (ENPs) under UV light (full spectrum) and visible light (455nm). Exposure of TiO2 ENPs to up to 60min of UV light induced a photocatalytic...... production of ROS. However, UV light itself caused cytotoxic damage to Salmonella typhimurium at exposures >15min and a genotoxic effect at exposures >0.5min; and use of UV filters did not lower this effect. No genotoxicity of TiO2 ENPs was observed under visible light conditions at concentrations up to 100...

GPC1 Regulated by miR-96-5p, Rather than miR-182-5p, in Inhibition of Pancreatic Carcinoma Cell Proliferation

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Chunlong Li

2014-04-01

Full Text Available To determine the relationships between miR-96-5p/-182-5p and GPC1 in pancreatic cancer (PC, we conducted the population and in vitro studies. We followed 38 pancreatic cancer patients, measured and compared the expression of miR-96-5p/-182-5p, GPC1, characteristics and patients’ survival time of different miR-96-5p/-182-5p expression levels in PC tissues. In an in vitro study, we investigated the proliferation, cycle and apotosis in cells transfected with mimics/inhibitors of the two miRNAs, and determine their effects on GPC1 by dual-luciferase assay. In the follow-up study, we found that the expressions of miR-96-5p/-182-5p were lower/higher in PC tissues; patients with lower/higher levels of miR-96-5p/-182-5p suffered poorer characteristics and decreased survival time. In the in vitro study, the expressions of miR-96-5p/-182-5p were different in cells. Proliferation of cells transfected with miR-96-5p mimics/inhibitors was lower/higher in Panc-1/BxPC-3; when transfected with miR-182-5p mimics/inhibitors, proliferation of cells were higher/lower in AsPC-1/Panc-1. In a cell cycle study, panc-1 cells transfected with miR-96-5p mimics was arrested at G0/G1; BxPC-3 cells transfected with miR-96-5p inhibitors showed a significantly decrease at G0/G1; AsPC-1 cells transfected with miR-182-5p mimics was arrested at S; Panc-1 cells transfected with miR-182-5p inhibitors showed a decrease at S. MiR-96-5p mimics increased the apoptosis rate in Panc-1 cells, and its inhibitors decreased the apoptosis rate in BxPC-3. Dual luciferase assay revealed that GPC1 was regulated by miR-96-5p, not -182-5p. We found that miR-96-5p/-182-5p as good markers for PC; miR-96-5p, rather than -182-5p, inhibits GPC1 to suppress proliferation of PC cells.

Avaliação da atividade tóxica em Artemia salina e Biomphalaria glabrata de extratos de quatro espécies do gênero Eleocharis (Cyperaceae

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A.L.T.G. Ruiz

Full Text Available O gênero Eleocharis R. Br. compreende cerca de 200 espécies, ocorrendo em ambientes úmidos tais como brejos e margens de rios e lagos. Procurando novos agentes moluscicidas, os extratos de Eleocharis acutangula (Roxb. Schult., Eleocharis interstincta (Vahl Roem. & Schult., Eleocharis maculosa (Vahl Roem. & Schult. e Eleocharis sellowiana Kunth foram testados para atividade moluscicida, contra caramujos adultos e desovas, e toxicidade (ensaio de letalidade com Artemia salina. O extrato hexânico de Eleocharis acutangula (parte subterrânea fresca foi ativo contra Artemia salina (CL50 = 476,00 mg/mL, enquanto os demais extratos apresentaram CL50 >> 10³ mg/mL, sugerindo baixa toxicidade. O extrato hidro-etanólico de Eleocharis sellowiana (parte subterrânea fresca foi ativo contra desovas de Biomphalaria glabrata (CL50 = 24,27 mg/mL mas inativo contra indivíduos adultos. Os demais extratos testados não apresentaram atividade moluscicida.

Temperature-dose relationships with aflatoxin M1 in milk on the brine shrimp (Artemia salina larvae

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Lejla Duraković

2011-06-01

Full Text Available Temperature-dose relationships with aflatoxin M1 (AFM1 were studied using the brine shrimp Artemia salina larvae as an biological indicator in the temperature range from 20 °C to 40 °C. Increase in the incubation temperature resulted in sensitivity increase by the brine shrimp to AFM1. Optimum sensitivity occured at 30 °C. Positive results were obtained at 0.18 μg AFM1 x L-1 of whole pasteurized milk with a mortality of over 15%. Greater than 90 % mortality occurred at dose levels of 0.9 μg AFM1 x L-1 and above. The test can be conducted during 30-60 hours.

Ground-Water Quality Data in the Monterey Bay and Salinas Valley Basins, California, 2005 - Results from the California GAMA Program

Science.gov (United States)

Kulongoski, Justin T.; Belitz, Kenneth

2007-01-01

Ground-water quality in the approximately 1,000-square-mile Monterey Bay and Salinas Valley study unit was investigated from July through October 2005 as part of the California Ground-Water Ambient Monitoring and Assessment (GAMA) program. The study was designed to provide a spatially unbiased assessment of raw ground-water quality, as well as a statistically consistent basis for comparing water quality throughout California. Samples were collected from 94 public-supply wells and 3 monitoring wells in Monterey, Santa Cruz, and San Luis Obispo Counties. Ninety-one of the public-supply wells sampled were selected to provide a spatially distributed, randomized monitoring network for statistical representation of the study area. Six wells were sampled to evaluate changes in water chemistry: three wells along a ground-water flow path were sampled to evaluate lateral changes, and three wells at discrete depths from land surface were sampled to evaluate changes in water chemistry with depth from land surface. The ground-water samples were analyzed for volatile organic compounds (VOCs), pesticides, pesticide degradates, nutrients, major and minor ions, trace elements, radioactivity, microbial indicators, and dissolved noble gases (the last in collaboration with Lawrence Livermore National Laboratory). Naturally occurring isotopes (tritium, carbon-14, helium-4, and the isotopic composition of oxygen and hydrogen) also were measured to help identify the source and age of the sampled ground water. In total, 270 constituents and water-quality indicators were investigated for this study. This study did not attempt to evaluate the quality of water delivered to consumers; after withdrawal from the ground, water typically is treated, disinfected, and (or) blended with other waters to maintain water quality. In addition, regulatory thresholds apply to treated water that is served to the consumer, not to raw ground water. In this study, only six constituents, alpha radioactivity, N

Assessment of a robust model protocol with accelerated throughput for a human recombinant full length estrogen receptor-alpha binding assay: protocol optimization and intralaboratory assay performance as initial steps towards validation.

Science.gov (United States)

Freyberger, Alexius; Wilson, Vickie; Weimer, Marc; Tan, Shirlee; Tran, Hoai-Son; Ahr, Hans-Jürgen

2010-08-01

Despite about two decades of research in the field of endocrine active compounds, still no validated human recombinant (hr) estrogen receptor-alpha (ERalpha) binding assay is available, although hr-ERalpha is available from several sources. In a joint effort, US EPA and Bayer Schering Pharma with funding from the EU-sponsored 6th framework project, ReProTect, developed a model protocol for such a binding assay. Important features of this assay are the use of a full length hr-ERalpha and performance in a 96-well plate format. A full length hr-ERalpha was chosen, as it was considered to provide the most accurate and human-relevant results, whereas truncated receptors could perform differently. Besides three reference compounds [17beta-estradiol, norethynodrel, dibutylphthalate] nine test compounds with different affinities for the ERalpha [diethylstilbestrol (DES), ethynylestradiol, meso-hexestrol, equol, genistein, o,p'-DDT, nonylphenol, n-butylparaben, and corticosterone] were used to explore the performance of the assay. Three independent experiments per compound were performed on different days, and dilutions of test compounds from deep-frozen stocks, solutions of radiolabeled ligand and receptor preparation were freshly prepared for each experiment. The ERalpha binding properties of reference and test compounds were well detected. As expected dibutylphthalate and corticosterone were non-binders in this assay. In terms of the relative ranking of binding affinities, there was good agreement with published data obtained from experiments using a human recombinant ERalpha ligand binding domain. Irrespective of the chemical nature of the compound, individual IC(50)-values for a given compound varied by not more than a factor of 2.5. Our data demonstrate that the assay was robust and reliably ranked compounds with strong, weak, and no affinity for the ERalpha with high accuracy. It avoids the manipulation and use of animals, i.e., the preparation of uterine cytosol as

Status and understanding of groundwater quality in the Monterey-Salinas Shallow Aquifer Study Unit, 2012–13: California GAMA Priority Basin Project

Science.gov (United States)

Burton, Carmen; Wright, Michael

2018-05-30

Groundwater quality in the approximately 7,820-square-kilometer (km2) Monterey-Salinas Shallow Aquifer (MS-SA) study unit was investigated from October 2012 to May 2013 as part of the second phase of the Priority Basin Project of the Groundwater Ambient Monitoring and Assessment (GAMA) Program. The study unit is in the central coast region of California in the counties of Santa Cruz, Monterey, and San Luis Obispo. The GAMA Priority Basin Project is being conducted by the California State Water Resources Control Board in cooperation with the U.S. Geological Survey and the Lawrence Livermore National Laboratory.The MS-SA study was designed to provide a statistically robust assessment of untreated-groundwater quality in the shallow aquifer systems. The assessment was based on water-quality samples collected by the U.S. Geological Survey from 100 groundwater sites and 70 household tap sites, along with ancillary data such as land use and well-construction information. The shallow aquifer systems were defined by the depth interval of wells associated with domestic supply. The MS-SA study unit consisted of four study areas—Santa Cruz (210 km2), Pajaro Valley (360 km2), Salinas Valley (2,000 km2), and Highlands (5,250 km2).This study had two primary components: the status assessment and the understanding assessment. The first primary component of this study—the status assessment—assessed the quality of the groundwater resource indicated by data from samples analyzed for volatile organic compounds (VOCs), pesticides, and naturally present inorganic constituents, such as major ions and trace elements. The status assessment is intended to characterize the quality of groundwater resources in the shallow aquifer system of the MS-SA study unit, not the treated drinking water delivered to consumers by water purveyors. As opposed to the public wells, however, water from private wells, which often tap the shallow aquifer, is usually consumed without any treatment. The second

Pomada orgânica natural ou solução salina isotônica no tratamento de feridas limpas induzidas em ratos

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Rahal Sheila Canevese

2001-01-01

Full Text Available Foram utilizados 40 ratos Wistar, machos, com peso inicial médio de 225 gramas, separados em dois grupos. Após serem anestesiados com éter, foi induzida uma ferida limpa na região torácica dorsal cranial, mediante a utilização de molde metálico de 2,0 x 1,5cm. No grupo I, foi utilizada pomada contendo óleo de fígado de bacalhau, extrato de confrei, extrato cítrico, óxido de zinco e veículo (Creamex®; no grupo II, a ferida foi apenas limpa com solução salina isotônica. Para a avaliação histológica, foram submetidos à eutanásia cinco animais de cada grupo no 3º, 7º, 14º e 21º dia de pós-operatório. O grupo tratado com a pomada orgânica apresentou padrão cicatricial de qualidade superior em relação ao grupo tratado com salina isotônica, representado por formação de fibras colágenas, neoformação de vasos e reepitelização completa da epiderme.

A Multiplexed Assay That Monitors Effects of Multiple Compound Treatment Times Reveals Candidate Immune-Enhancing Compounds.

Science.gov (United States)

Zhao, Ziyan; Henowitz, Liza; Zweifach, Adam

2018-05-01

We previously developed a flow cytometry assay that monitored lytic granule exocytosis in cytotoxic T lymphocytes stimulated by contacting beads coated with activating anti-CD3 antibodies. That assay was multiplexed in that responses of cells that did or did not receive the activating stimulus were distinguished via changes in light scatter accompanying binding of cells to beads, allowing us to discriminate compounds that activate responses on their own from compounds that enhance responses in cells that received the activating stimulus, all within a single sample. Here we add a second dimension of multiplexing by developing means to assess in a single sample the effects of treating cells with test compounds for different times. Bar-coding cells before adding them to test wells lets us determine compound treatment time while also monitoring activation status and response amplitude at the point of interrogation. This multiplexed assay is suitable for screening 96-well plates. We used it to screen compounds from the National Cancer Institute, identifying several compounds that enhance anti-LAMP1 responses. Multiple-treatment-time (MTT) screening enabled by bar-coding and read via high-throughput flow cytometry may be a generally useful method for facilitating the discovery of compounds of interest.

IDENTIFIKASI DAN UJI AKTIVITAS ANTIKANKER EKSTRAK SPONS Ianthella basta TERHADAP LARVA Artemia salina L.

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Ni Wayan Sri Sukmarianti

2013-05-01

Full Text Available The main purpose of this research is to determine the toxicity of extracts sponge Ianthella basta against Artemia salina larvae and to identify the chemical compounds contained in those toxic isolates. The preliminary test of the anticancer activity has conducted by Brine Shrimp Letalithy (BST test. The results showed that the chloroform extract was the most toxic with LC50 value of 22,39 ppm. Futher, the cloroform extract was separated and purified by coloumn chromatography using eluent of solvent mixture of chloroform : ethyl acetate : n-hexane by 7 : 2 : 1 and 4 fractions were obtained. The most toxic fraction was the fraction C with LC50 value of 35,36 ppm. Based on the GC-MS results, the toxic isolate is allegedly containing chemicals compound of hexadecanoic methyl ester and hexadecanoic acid.

Influence of surface geometry on the culture of human cell lines: A comparative study using flat, round-bottom and v-shaped 96 well plates.

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Sara Shafaie

Full Text Available In vitro cell based models have been invaluable tools for studying cell behaviour and for investigating drug disposition, toxicity and potential adverse effects of administered drugs. Within this drug discovery pipeline, the ability to assess and prioritise candidate compounds as soon as possible offers a distinct advantage. However, the ability to apply this approach to a cell culture study is limited by the need to provide an accurate, in vitro-like, microenvironment in conjunction with a low cost and high-throughput screening (HTS methodology. Although the geometry and/or alignment of cells has been reported to have a profound influence on cell growth and differentiation, only a handful of studies have directly compared the growth of a single cell line on different shaped multiwell plates the most commonly used substrate for HTS, in vitro, studies. Herein, the impact of various surface geometries (flat, round and v-shaped 96 well plates, as well as fixed volume growth media and fixed growth surface area have been investigated on the characteristics of three commonly used human cell lines in biopharmaceutical research and development, namely ARPE-19 (retinal epithelial, A549 (alveolar epithelial and Malme-3M (dermal fibroblastic cells. The effect of the surface curvature on cells was characterised using a combination of a metabolic activity assay (CellTiter AQ/MTS, LDH release profiles (CytoTox ONE and absolute cell counts (Guava ViaCount, respectively. In addition, cell differentiation and expression of specific marker proteins were determined using flow cytometry. These in vitro results confirmed that surface topography had a significant effect (p < 0.05 on cell activity and morphology. However, although specific marker proteins were expressed on day 1 and 5 of the experiment, no significant differences were seen between the different plate geometries (p < 0.05 at the later time point. Accordingly, these results highlight the impact of

40 CFR 96.73 - Notifications.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Notifications. 96.73 Section 96.73 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET... Reporting § 96.73 Notifications. The NOX authorized account representative for a NOX Budget unit shall...

A role for programmed cell death in the microbial loop.

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Mónica V Orellana

Full Text Available The microbial loop is the conventional model by which nutrients and minerals are recycled in aquatic eco-systems. Biochemical pathways in different organisms become metabolically inter-connected such that nutrients are utilized, processed, released and re-utilized by others. The result is that unrelated individuals end up impacting each others' fitness directly through their metabolic activities. This study focused on the impact of programmed cell death (PCD on a population's growth as well as its role in the exchange of carbon between two naturally co-occurring halophilic organisms. Flow cytometric, biochemical, ¹⁴C radioisotope tracing assays, and global transcriptomic analyses show that organic algal photosynthate released by Dunalliela salina cells undergoing PCD complements the nutritional needs of other non-PCD D. salina cells. This occurs in vitro in a carbon limited environment and enhances the growth of the population. In addition, a co-occurring heterotroph Halobacterium salinarum re-mineralizes the carbon providing elemental nutrients for the mixoheterotrophic chlorophyte. The significance of this is uncertain and the archaeon can also subsist entirely on the lysate of apoptotic algae. PCD is now well established in unicellular organisms; however its ecological relevance has been difficult to decipher. In this study we found that PCD in D. salina causes the release of organic nutrients such as glycerol, which can be used by others in the population as well as a co-occurring halophilic archaeon. H. salinarum also re-mineralizes the dissolved material promoting algal growth. PCD in D. salina was the mechanism for the flow of dissolved photosynthate between unrelated organisms. Ironically, programmed death plays a central role in an organism's own population growth and in the exchange of nutrients in the microbial loop.

45 CFR 96.65 - Discovery.

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2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Discovery. 96.65 Section 96.65 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION BLOCK GRANTS Hearing Procedure § 96.65 Discovery. The use of interrogatories, depositions, and other forms of discovery shall not be allowed. ...

50 CFR 27.96 - Advertising.

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2010-10-01

... 50 Wildlife and Fisheries 6 2010-10-01 2010-10-01 false Advertising. 27.96 Section 27.96 Wildlife and Fisheries UNITED STATES FISH AND WILDLIFE SERVICE, DEPARTMENT OF THE INTERIOR (CONTINUED) THE NATIONAL WILDLIFE REFUGE SYSTEM PROHIBITED ACTS Other Disturbing Violations § 27.96 Advertising. Except as...

40 CFR 96.355 - Banking.

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2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Banking. 96.355 Section 96.355 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET... Allowance Tracking System § 96.355 Banking. (a) CAIR NOX Ozone Season allowances may be banked for future...

40 CFR 96.55 - Banking.

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2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Banking. 96.55 Section 96.55 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET... Tracking System § 96.55 Banking. (a) NOX allowances may be banked for future use or transfer in a...

40 CFR 96.255 - Banking.

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2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Banking. 96.255 Section 96.255 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET... Tracking System § 96.255 Banking. (a) CAIR SO2 allowances may be banked for future use or transfer in a...

40 CFR 96.155 - Banking.

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2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Banking. 96.155 Section 96.155 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS (CONTINUED) NOX BUDGET... Tracking System § 96.155 Banking. (a) CAIR NOX allowances may be banked for future use or transfer in a...

Obtención de glicerol a partir de la Microalga Dunaliella Salina

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Lisethy Hernández Nazario

2000-08-01

Full Text Available En el campo de la tecnología farmacéutica, el glicerol es un disolvente ampliamente utilizado en virtud de sus propiedades físico-químicas en la formulación de diferentes formas farmacéuticas. Se investigaron las posibilidades de obtención de glicerol como un subproducto del proceso de extracción de ß-carotenos a partir de cultivos de Dunaliella salina, desarrollados bajo régimen autotrófico en el Centro de Investigaciones de Energía Solar. El flujo tecnológico propuesto comprende el tratamiento de la biomasa con hidróxido de calcio, la filtración del producto resultante, la extracción del ß-carotenos con un solvente insoluble en agua y, por último, la separación del glicerol neutralizando convenientemente del filtrado con ácido. El rendimiento de glicerol fue del 4-5 %, valor susceptible de ser incrementado mediante la inducción metabólica de los cultivos.

32 CFR 96.4 - Policy.

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2010-07-01

... OF CRIMINAL HISTORY RECORD INFORMATION BY THE MILITARY SERVICES § 96.4 Policy. Section 503 of title... 32 National Defense 1 2010-07-01 2010-07-01 false Policy. 96.4 Section 96.4 National Defense... campaigns to obtain enlistments. It is the policy of the Department of Defense that the Military Services...

Reposição de volume na sepse com solução salina hipertônica Sepsis volume reposition with hypertonic saline solution

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Gilberto Friedman

2008-09-01

Full Text Available Esta revisão discute os efeitos hemodinâmicos e imunomoduladores da solução hipertônica em choque experimental e em pacientes com sepse. Comentamos sobre os mecanismos de ação da solução hipertônica, recorrendo a dados sobre choque hemorrágico e séptico. Atuações específicas da solução salina hipertônica aplicáveis a sepse grave e choque séptico são enfatizadas. Os dados disponíveis corroboram os benefícios em potencial da infusão de solução salina hipertônica em vários aspetos da fisiopatologia da sepse, inclusive hipoperfusão dos tecidos, consumo reduzido de oxigênio, disfunção endotelial, depressão miocárdica e presença de um amplo elenco de citocinas próinflamatórias e várias espécies de oxidantes. Uma terapia que, ao mesmo tempo, bloqueie os componentes prejudiciais da sepse terá um impacto no seu tratamento. Estudos prospectivos adequadamente desenhados poderão no futuro comprovar o papel benéfico da solução salina hipertônica.The present review discusses the hemodynamic and immune-modulatory effects of hypertonic saline in experimental shock and in patients with sepsis. We comment on the mechanisms of action of hypertonic saline, calling upon data in hemorrhagic and septic shock. Specific actions of hypertonic saline applicable to severe sepsis and septic shock are highlighted. Data available support potential benefits of hypertonic saline infusion in various aspects of the pathophysiology of sepsis, including tissue hypoperfusion, decreased oxygen consumption, endothelial dysfunction, cardiac depression, and the presence of a broad array of pro-inflammatory cytokines and various oxidant species. A therapy that simultaneously blocks the damaging components of sepsis will have an impact on the management of sepsis. Proper designed prospective studies may prove a beneficial role for hypertonic saline solution in the future.

IL MUSEO SALINAS, PARADOSSO DEL WEB MARKETING CULTURALE La comunicazione archeologica ai tempi dei social media, a museo chiuso

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Elisa Bonacini

2016-11-01

Full Text Available The aim of this paper is to offer the case study of the archaeological museum Antonino Salinas in Palermo, that - closed for many years for the restoration of the architectural complex that houses it, the seventeenth-century Olivella’ s House of the Philippine Fathers - has invented new strategies of the archaeological communication on the social media. After two years, waiting for its reopening, the museum has been able to create a real brand and to renovate the museum’s reputation. The solutions adopted by the Museum could “inspire” other institutions to adopt those web marketing cultural strategies.

Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

International Nuclear Information System (INIS)

Pastrana, Diana V.; Buck, Christopher B.; Pang, Y.-Y. S.; Thompson, Cynthia D.; Castle, Philip E.; FitzGerald, Peter C.; Krueger Kjaer, Susanne; Lowy, Douglas R.; Schiller, John T.

2004-01-01

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies

A colorimetric assay of 1-aminocyclopropane-1-carboxylate (ACC) based on ninhydrin reaction for rapid screening of bacteria containing ACC deaminase.

Science.gov (United States)

Li, Z; Chang, S; Lin, L; Li, Y; An, Q

2011-08-01

1-Aminocyclopropane-1-carboxylate (ACC) deaminase activity is an efficient marker for bacteria to promote plant growth by lowering ethylene levels in plants. We aim to develop a method for rapidly screening bacteria containing ACC deaminase, based on a colorimetric ninhydrin assay of ACC. A reliable colorimetric ninhydrin assay was developed to quantify ACC using heat-resistant polypropylene chimney-top 96-well PCR plates, having the wells evenly heated in boiling water, preventing accidental contamination from boiling water and limiting evaporation. With this method to measure bacterial consumption of ACC, 44 ACC-utilizing bacterial isolates were rapidly screened out from 311 bacterial isolates that were able to grow on minimal media containing ACC as the sole nitrogen source. The 44 ACC-utilizing bacterial isolates showed ACC deaminase activities and belonged to the genus Burkholderia, Pseudomonas or Herbaspirillum. Determination of bacterial ACC consumption by the PCR-plate ninhydrin-ACC assay is a rapid and efficient method for screening bacteria containing ACC deaminase from a large number of bacterial isolates. The PCR-plate ninhydrin-ACC assay extends the utility of the ninhydrin reaction and enables a rapid screening of bacteria containing ACC deaminase from large numbers of bacterial isolates. © 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology.

Validation and modification of dried blood spot-based glycosylated hemoglobin assay for the longitudinal aging study in India.

Science.gov (United States)

Hu, Peifeng; Edenfield, Michael; Potter, Alan; Kale, Varsha; Risbud, Arun; Williams, Sharon; Lee, Jinkook; Bloom, David E; Crimmins, Eileen; Seeman, Teresa

2015-01-01

This study aims to validate a modified dried blood spot (DBS)-based glycosylated hemoglobin (HbA1c) assay protocol, after a pretest in India showed poor correlation between the original DBS-based protocol and venous results. The original protocol was tested on different chemistry analyzers and then simplified at the University of Washington (UW). A second pretest was conducted in India to validate the modified assay protocol, using 44 quality control specimens. Data from UW indicated that, using the original protocol, the correlation coefficients between DBS and venous results were above 0.98 on both Bio-Rad and Olympus chemistry analyzers. The protocol worked equally well on filter paper, with or without pre-treatment, and when the recommended amount of blood spot material, or less, was used. A second pretest of the modified protocol confirmed that DBS-based levels from both Olympus and Roche chemistry analyzers were well correlated with DBS results from UW (correlation coefficients were above 0.96), as well as with venous values (correlation coefficients were above 0.94). The DBS-based HbA1c values are highly correlated with venous results. The pre-treatment of filter paper does not appear to be necessary. The poor results from the first pretest are probably due to factors unrelated to the protocol, such as problems with the chemistry analyzer or assay reagents. © 2015 Wiley Periodicals, Inc.

Performance of the new automated Abbott RealTime MTB assay for rapid detection of Mycobacterium tuberculosis complex in respiratory specimens.

Science.gov (United States)

Chen, J H K; She, K K K; Kwong, T-C; Wong, O-Y; Siu, G K H; Leung, C-C; Chang, K-C; Tam, C-M; Ho, P-L; Cheng, V C C; Yuen, K-Y; Yam, W-C

2015-09-01

The automated high-throughput Abbott RealTime MTB real-time PCR assay has been recently launched for Mycobacterium tuberculosis complex (MTBC) clinical diagnosis. This study would like to evaluate its performance. We first compared its diagnostic performance with the Roche Cobas TaqMan MTB assay on 214 clinical respiratory specimens. Prospective analysis of a total 520 specimens was then performed to further evaluate the Abbott assay. The Abbott assay showed a lower limit of detection at 22.5 AFB/ml, which was more sensitive than the Cobas assay (167.5 AFB/ml). The two assays demonstrated a significant difference in diagnostic performance (McNemar's test; P = 0.0034), in which the Abbott assay presented significantly higher area under curve (AUC) than the Cobas assay (1.000 vs 0.880; P = 0.0002). The Abbott assay demonstrated extremely low PCR inhibition on clinical respiratory specimens. The automated Abbott assay required only very short manual handling time (0.5 h), which could help to improve the laboratory management. In the prospective analysis, the overall estimates for sensitivity and specificity of the Abbott assay were both 100 % among smear-positive specimens, whereas the smear-negative specimens were 96.7 and 96.1 %, respectively. No cross-reactivity with non-tuberculosis mycobacterial species was observed. The superiority in sensitivity of the Abbott assay for detecting MTBC in smear-negative specimens could further minimize the risk in MTBC false-negative detection. The new Abbott RealTime MTB assay has good diagnostic performance which can be a useful diagnostic tool for rapid MTBC detection in clinical laboratories.

Variación espacial y temporal de la diversidad de hormigas en el Jardín Botánico del valle de Zapotitlán de las Salinas, Puebla Spatial and temporal variation of the diversity ants in the Botanic Garden from Zapotitlán de las Salinas Valley, Puebla

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Rafael Guzmán-Mendoza

2010-08-01

Full Text Available Debido al escaso conocimiento que se tiene de las hormigas de las zonas áridas de México, se evaluó la biodiversidad de este taxón en el Jardín Botánico de Zapotitlán de las Salinas, Puebla. Se realizaron 2 colectas, una en época de lluvias (agosto de 2004 y otra en la temporada de secas (febrero de 2005. En ambas ocasiones se seleccionaron 2 sitios contrastantes en estructura vegetal y se colocaron 10 trampas de caída para cada sitio. La captura fue más abundante en temporada de lluvias. El sitio con menor diversidad y cobertura vegetal mostró una mayor diversidad de hormigas durante el estudio. Comparado con otras zonas áridas de México, el valle de Zapotitlán de las Salinas resultó ser una de las zonas semiáridas con mayor riqueza en especies de hormigas; con 12 nuevos registros se incrementó a 27 el número de especies, sólo una por debajo de otras localidades cercanas al valle. Los datos sugieren que con un mayor esfuerzo de muestreo y la aplicación combinada de otras técnicas de recolección, la riqueza de especies en el área podría elevarse considerablemente.The knowledge about ants of arid zones of Mexico is very poor. For this reason we assessed the biodiversity of this taxon in 2 sites with different vegetation structure. Ten pitfall traps were placed in each site, and 2 surveys were conducted, 1 in August (rainy season, 2004 and another in February (dry season, 2005. The capture was more abundant in the rainy season; however, during the study, a higher ant diversity was observed in the site with low vegetation cover and few plant diversity. Compared with other arid zones of Mexico, the Zapotitlán de las Salinas Valley is the site with the highest ant species richness. The new records have increased richness to 27 ant species, which is very close to the number of species registered in other localities near Zapotitlan. Our data suggest that increasing sampling effort and application of different sampling

Relationship between the radioisotopic footpad assay and other immunological assays in tumor bearing rats

International Nuclear Information System (INIS)

Mizushima, Yutaka; Takeichi, Noritoshi; Minami, Akio; Kasai, Masaharu; Itaya, Toshiyuki

1981-01-01

KMT-17, a fibrosarcoma induced by 3-methylcholanthrene in a WKA rat, is a sensitive tumor to various kinds of immunological assays and is a suitable model tumor for the study of the immune status in tumor bearing hosts. The antitumor immune response of KMT-17 bearing rats was studied by a radioisotopic footpad assay (FPA) in comparison with other in vivo and in vitro assays. Delayed hypersensitivity to tumor antigens measured by the FPA was observed from the 8th day after transplantation of KMT-17 cells, reached a peak on the 12 - 15th day, and then declined in the late stage on the 17th day. The kinetics of the FPA correlated well with those of an in vivo Winn assay and of an in vitro lymphocyte cytotoxicity assay ( 51 Cr-release assay). The appearance of an antitumor antibody detected by a complement dependent cytotoxicity test also correlated well with the kinetics of the FPA. A growth inhibition assay (GIA) for non-specific cell-mediated immunity also showed similar kinetics to that of the FPA. The delayed hypersensitivity footpad reaction to tumor cell extracts measured by this FPA was tumor-specific. These results suggest that the FPA is a simple and reliable in vivo assay for evaluating antitumor immunity in tumor bearing hosts. (author)

Evaluation of the Alexon-Trend ProSpecT Campylobacter Microplate Assay

Science.gov (United States)

Tolcin, Rita; LaSalvia, Margaret M.; Kirkley, Barbara A.; Vetter, Emily A.; Cockerill, Franklin R.; Procop, Gary W.

2000-01-01

We evaluated stool specimens known to contain or be free of Campylobacter by traditional culture, using the ProSpecT Campylobacter microplate assay (Alexon-Trend, Ramsey, Minn.). This rapid enzyme immunoassay for the detection of Campylobacter-specific antigens demonstrated 96% sensitivity and 99% specificity and is an acceptable alternative method of Campylobacter detection. PMID:11015419

Novel microwell-based spectrophotometric assay for determination of atorvastatin calcium in its pharmaceutical formulations

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Abdel-Rahman Hamdy M

2011-10-01

Full Text Available Abstract The formation of a colored charge-transfer (CT complex between atorvastatin calcium (ATR-Ca as a n-electron donor and 2, 3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ as a π-electron acceptor was investigated, for the first time. The spectral characteristics of the CT complex have been described, and the reaction mechanism has been proved by computational molecular modeling. The reaction was employed in the development of a novel microwell-based spectrophotometric assay for determination of ATR-Ca in its pharmaceutical formulations. The proposed assay was carried out in 96-microwell plates. The absorbance of the colored-CT complex was measured at 460 nm by microwell-plate absorbance reader. The optimum conditions of the reaction and the analytical procedures of the assay were established. Under the optimum conditions, linear relationship with good correlation coefficient (0.9995 was found between the absorbance and the concentration of ATR-Ca in the range of 10-150 μg/well. The limits of detection and quantitation were 5.3 and 15.8 μg/well, respectively. No interference was observed from the additives that are present in the pharmaceutical formulation or from the drugs that are co-formulated with ATR-Ca in its combined formulations. The assay was successfully applied to the analysis of ATR-Ca in its pharmaceutical dosage forms with good accuracy and precision. The assay described herein has great practical value in the routine analysis of ATR-Ca in quality control laboratories, as it has high throughput property, consumes minimum volume of organic solvent thus it offers the reduction in the exposures of the analysts to the toxic effects of organic solvents, and reduction in the analysis cost by 50-fold. Although the proposed assay was validated for ATR-Ca, however, the same methodology could be used for any electron-donating analyte for which a CT reaction can be performed.

Highly parallel and short-acting amplification with locus-specific primers to detect single nucleotide polymorphisms by the DigiTag2 assay.

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Nao Nishida

Full Text Available The DigiTag2 assay enables analysis of a set of 96 SNPs using Kapa 2GFast HotStart DNA polymerase with a new protocol that has a total running time of about 7 hours, which is 6 hours shorter than the previous protocol. Quality parameters (conversion rate, call rate, reproducibility and concordance were at the same levels as when genotype calls were acquired using the previous protocol. Multiplex PCR with 192 pairs of locus-specific primers was available for target preparation in the DigiTag2 assay without the optimization of reaction conditions, and quality parameters had the same levels as those acquired with 96-plex PCR. The locus-specific primers were able to achieve sufficient (concentration of target amplicon ≥5 nM and specific (concentration of unexpected amplicons <2 nM amplification within 2 hours, were also able to achieve detectable amplifications even when working in a 96-plex or 192-plex form. The improved DigiTag2 assay will be an efficient platform for screening an intermediate number of SNPs (tens to hundreds of sites in the replication analysis after genome-wide association study. Moreover, highly parallel and short-acting amplification with locus-specific primers may thus facilitate widespread application to other PCR-based assays.

Brine shrimp bioassay: importance of correct taxonomic identification of Artemia (Anostraca) species.

Science.gov (United States)

Ruebhart, David R; Cock, Ian E; Shaw, Glen R

2008-08-01

Despite the common use of the brine shrimp bioassay in toxicology, there is confusion in the literature regarding citation of the correct taxonomic identity of the Artemia species used. The genus Artemia, once thought to be represented by a single species Artemia salina, is now known to be composed of several bisexual species as well as parthenogenetic populations. Artemia franciscana is the best studied of the Artemia species and is considered to represent the vast majority of studies in which Artemia is used as an experimental test organism. We found that in studies referring to the use of A. salina, the zoogeography of the cyst harvest site indicated that the species used was actually A. franciscana. Those performing bioassays with Artemia need to exercise diligence in assigning correct species identification, as the identity of the test organism is an important parameter in assuring the validity of the results of the assay.

First secondary metabolites from Herissantia crispa L (Brizicky) and the toxicity activity against Artemia salina Leach

International Nuclear Information System (INIS)

Costa, Danielly Albuquerque da; Matias, Wemerson Neves; Lima, Igara Oliveira; Xavier, Aline Lira; Costa, Vivian Bruna Machado; Diniz, Margareth de Fatima Formiga Melo; Agra, Maria de Fatima; Batista, Leonia Maria; Souza, Maria de Fatima Vanderlei de; Silva, Davi Antas e

2009-01-01

The phytochemical investigation of Herissantia crispa led to the isolation of seven compounds, identified as: sitosterol 3-O-β-D-glucopyranoside, stigmasterol 3-O-β-D-glucopyranoside, 3,5,7,4'-tetrahydroxyflavone (kaempferol), 3,5,7,3',4'-pentahydroxyflavone (quercetin), unpublished in the genus Herissantia, besides β-sitosterol, kaempferol 3-O-β-D-(6''-E-p-coumaroil) (tiliroside) glucopyranoside and kaempferol 3,7-di-O-α-L-ramnopyranoside (lespedin), described for the first time in the species. The structural determination of the compounds was made by means of spectroscopy methods such as Infrared Spectroscopy, 1 H and 13 C Nuclear Magnetic Resonance, with the aid of two dimensional techniques, and by comparison with literature data. The toxicity activity of the MeOH extract and lespedin on Artemia salina Leach. was also carried out. (author)

The laboratory environmental algae pond simulator (LEAPS) photobioreactor: Validation using outdoor pond cultures of Chlorella sorokiniana and Nannochloropsis salina

Energy Technology Data Exchange (ETDEWEB)

Huesemann, M.; Williams, P.; Edmundson, S.; Chen, P.; Kruk, R.; Cullinan, V.; Crowe, B.; Lundquist, T.

2017-09-01

A bench-scale photobioreactor system, termed Laboratory Environmental Algae Pond Simulator (LEAPS), was designed and constructed to simulate outdoor pond cultivation for a wide range of geographical locations and seasons. The LEAPS consists of six well-mixed glass column photobioreactors sparged with CO2-enriched air to maintain a set-point pH, illuminated from above by a programmable multicolor LED lighting (0 to 2,500 µmol/m2-sec), and submerged in a temperature controlled water-bath (-2 °C to >60 °C). Measured incident light intensities and water temperatures deviated from the respective light and temperature set-points on average only 2.3% and 0.9%, demonstrating accurate simulation of light and temperature conditions measured in outdoor ponds. In order to determine whether microalgae strains cultured in the LEAPS exhibit the same linear phase biomass productivity as in outdoor ponds, Chlorella sorokiniana and Nannochloropsis salina were cultured in the LEAPS bioreactors using light and temperature scripts measured previously in the respective outdoor pond studies. For Chlorella sorokiniana, the summer season biomass productivity in the LEAPS was 6.6% and 11.3% lower than in the respective outdoor ponds in Rimrock, Arizona, and Delhi, California; however, these differences were not statistically significant. For Nannochloropsis salina, the winter season biomass productivity in the LEAPS was statistically significantly higher (15.2%) during the 27 day experimental period than in the respective outdoor ponds in Tucson, Arizona. However, when considering only the first 14 days, the LEAPS biomass productivity was only 9.2% higher than in the outdoor ponds, a difference shown to be not statistically significant. Potential reasons for the positive or negative divergence in LEAPS performance, relative to outdoor ponds, are discussed. To demonstrate the utility of the LEAPS in predicting productivity, two other strains – Scenedesmus obliquus and Stichococcus minor

Diversidade de abelhas visitantes das flores de Citrus em pomares de laranjeira e tangerineira Diversity of flowers visiting bees of Citrus in salinas, state of minas gerais

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Eduardo Tigre do Nascimento

2011-03-01

Full Text Available A presença de abelhas em culturas de valor comercial, no período de florescimento, é importante não apenas para garantir a polinização cruzada, quando indispensável, mas também para incrementar a produção de frutos e sementes. Nessa perspectiva, o presente trabalho teve a finalidade de identificar as abelhas visitantes das flores em pomar de laranjeira (Citrus sinensis e tangerineira (Citrus reticulata em Salinas - MG. O trabalho foi conduzido na fazenda experimental Santa Isabel, da Escola Agrotécnica Federal de Salinas - MG, em pomares de laranjeira com as variedades, pera, baía e baianinha, e de tangerineira, variedade poncã. As coletas das abelhas visitantes das culturas da laranjeira e tangerineira foram feitas nas floradas principais, entre os meses de agosto e outubro. Foram coletadas as abelhas que visitaram as flores nos horários entre 06 h e 18 h, totalizando 52 horas na laranjeira e 50 horas na tangerineira. No pomar de laranjeira, foram coletados 5.045 espécimes com riqueza de 12 espécies, e na tangerineira, 1.428 espécimes com 20 espécies. As espécies de abelhas de maior predominância e dominância em todos os horários amostrados foram Apis mellifera e Trigona spinipes na cultura da laranjeira e A. mellifera, T. spinipes e Tetragonisca angustula na cultura da tangerineira.The presence of bees on cultures with commercial value during flowering is important to ensure the cross-pollination, when necessary, but also to increase the production of fruits and seeds. This work aimed to identify flowers' visiting bees on orange and tangerine orchards in Salinas - MG. The work was leaded on Santa Isabel experimental farm of the Federal Agro-technical School of Salinas - MG, on orange orchards (Citrus sinensis with pear, bahia and baianinha varieties in the spacing of 5,0 x 6,0, and on tangerine orchards (C. reticulate using poncã variety in the spacing of 5,0 x 5,0. The sampling of visiting bees from orange and tangerine

21 CFR 1250.96 - Rodent control.

Science.gov (United States)

2010-04-01

... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Rodent control. 1250.96 Section 1250.96 Food and... SANITATION Sanitation Facilities and Conditions on Vessels § 1250.96 Rodent control. Vessels shall be... of rodent control. ...

Influência da irrigação intraperitoneal de solução salina em diferentes temperaturas na formação de aderências e prevenção de hipotermia após pneumoperitônio em ratos

Directory of Open Access Journals (Sweden)

Oswaldo Lourenço de Molla Neto

Full Text Available Este estudo experimental em ratos avalia a influência da irrigação da cavidade peritoneal com solução salina isotônica (0,9 %, em diferentes temperaturas, na formação de aderências peritoneais e prevenção de hipotermia após pneumoperitônio. Foram utilizados 80 ratos divididos em quatro grupos de 20 animais: grupo controle (G1 sem irrigação, grupos com irrigação a temperatura ambiente 22,0°C (G2, a 35,0°C (G3 e a 45,0°C (G4. A análise da hipotermia foi realizada através da monitorização da temperatura retal em três diferentes momentos: após a anestesia (T1, cinco minutos depois da insuflação de dióxido de carbono (T2 e cinco minutos após a irrigação com solução salina (T3. Os animais foram sacrificados no 28º dia de pós-operatório. Observaram-se aderências nos grupos com irrigação, sendo que, com salina à temperatura de 45,0ºC houve maior formação de aderências (30,0% , porém, esta diferença não foi significante. No G2 ocorreu uma queda significante na temperatura média retal quando comparada aos demais grupos, demonstrando que a hipotermia na cirurgia laparoscópica pode ser reduzida com o uso de solução salina aquecida.

Crescimento, consumo hídrico e composição mineral de alface cultivada em hidroponia com águas salinas

OpenAIRE

Paulus,Dalva; Paulus,Eloi; Nava,Gilmar Antônio; Moura,Cláudia Andrade

2012-01-01

Objetivou-se, com este trabalho, avaliar o crescimento, o consumo hídrico e a extração de nutrientes, por alface (Lactuca sativa L.) em sistema hidropônico NFT (fluxo laminar de nutrientes), com a utilização de águas salinas no preparo da solução nutritiva e reposição da lâmina diária evapotranspirada. Foram conduzidos dois experimentos, em ambiente protegido. O delineamento experimental foi em blocos ao acaso, sendo estudados os efeitos de cinco níveis de salinidade da água, utilizando-se Na...

22 CFR 9.6 - Derivative classification.

Science.gov (United States)

2010-04-01

... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Derivative classification. 9.6 Section 9.6 Foreign Relations DEPARTMENT OF STATE GENERAL SECURITY INFORMATION REGULATIONS § 9.6 Derivative classification. (a) Definition. Derivative classification is the incorporating, paraphrasing, restating or...

A quantitative comet infection assay for influenza virus

Science.gov (United States)

Lindsay, Stephen M.; Timm, Andrea; Yin, John

2011-01-01

Summary The virus comet assay is a cell-based virulence assay used to evaluate an antiviral drug or antibody against a target virus. The comet assay differs from the plaque assay in allowing spontaneous flows in 6-well plates to spread virus. When implemented quantitatively the comet assay has been shown to have an order-of-magnitude greater sensitivity to antivirals than the plaque assay. In this study, a quantitative comet assay for influenza virus is demonstrated, and is shown to have a 13-fold increase in sensitivity to ribavirin. AX4 cells (MDCK cells with increased surface concentration of α2–6 sialic acid, the influenza virus receptor) have reduced the comet size variability relative to MDCK cells, making them a better host cell for use in this assay. Because of enhanced antiviral sensitivity in flow-based assays, less drug is required, which could lead to lower reagent costs, reduced cytotoxicity, and fewer false-negative drug screen results. The comet assay also serves as a readout of flow conditions in the well. Observations from comets formed at varying humidity levels indicate a role for evaporation in the mechanism of spontaneous fluid flow in wells. PMID:22155578

Radiometric assays for glycerol, glucose, and glycogen

International Nuclear Information System (INIS)

Bradley, D.C.; Kaslow, H.R.

1989-01-01

We have developed radiometric assays for small quantities of glycerol, glucose and glycogen, based on a technique described by Thorner and Paulus for the measurement of glycerokinase activity. In the glycerol assay, glycerol is phosphorylated with [32P]ATP and glycerokinase, residual [32P]ATP is hydrolyzed by heating in acid, and free [32P]phosphate is removed by precipitation with ammonium molybdate and triethylamine. Standard dose-response curves were linear from 50 to 3000 pmol glycerol with less than 3% SD in triplicate measurements. Of the substances tested for interference, only dihydroxyacetone gave a slight false positive signal at high concentration. When used to measure glycerol concentrations in serum and in media from incubated adipose tissue, the radiometric glycerol assay correlated well with a commonly used spectrophotometric assay. The radiometric glucose assay is similar to the glycerol assay, except that glucokinase is used instead of glycerokinase. Dose response was linear from 5 to 3000 pmol glucose with less than 3% SD in triplicate measurements. Glucosamine and N-acetylglucosamine gave false positive signals when equimolar to glucose. When glucose concentrations in serum were measured, the radiometric glucose assay agreed well with hexokinase/glucose-6-phosphate dehydrogenase (H/GDH)-based and glucose oxidase/H2O2-based glucose assays. The radiometric method for glycogen measurement incorporates previously described isolation and digestion techniques, followed by the radiometric assay of free glucose. When used to measure glycogen in mouse epididymal fat pads, the radiometric glycogen assay correlated well with the H/GDH-based glycogen assay. All three radiometric assays offer several practical advantages over spectral assays

Microbead agglutination based assays

KAUST Repository

Kodzius, Rimantas

2013-01-21

We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

A simple fluorescence based assay for quantification of human immunodeficiency virus particle release

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Heuser Anke-Mareil

2010-04-01

Full Text Available Abstract Background The assembly and release of human immunodeficiency virus (HIV particles from infected cells represent attractive, but not yet exploited targets for antiretroviral therapy. The availability of simple methods to measure the efficiency of these replication steps in tissue culture would facilitate the identification of host factors essential for these processes as well as the screening for lead compounds acting as specific inhibitors of particle formation. We describe here the development of a rapid cell based assay for quantification of human immunodeficiency virus type 1 (HIV-1 particle assembly and/or release. Results Using a fluorescently labelled HIV-derivative, which carries an eYFP domain within the main viral structural protein Gag in the complete viral protein context, the release of virus like particles could be monitored by directly measuring the fluorescence intensity of the tissue culture supernatant. Intracellular Gag was quantitated in parallel by direct fluorescence analysis of cell lysates, allowing us to normalize for Gag expression efficiency. The assay was validated by comparison with p24 capsid ELISA measurements, a standard method for quantifying HIV-1 particles. Optimization of conditions allowed the robust detection of particle amounts corresponding to 50 ng p24/ml in medium by fluorescence spectroscopy. Further adaptation to a multi-well format rendered the assay suitable for medium or high throughput screening of siRNA libraries to identify host cell factors involved in late stages of HIV replication, as well as for random screening approaches to search for potential inhibitors of HIV-1 assembly or release. Conclusions The fast and simple fluorescence based quantification of HIV particle release yielded reproducible results which were comparable to the well established ELISA measurements, while in addition allowing the parallel determination of intracellular Gag expression. The protocols described here

45 CFR 96.41 - General determination.

Science.gov (United States)

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false General determination. 96.41 Section 96.41 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION BLOCK GRANTS Direct Funding of Indian Tribes and Tribal Organizations § 96.41 General determination. (a) The Department has determined...

22 CFR 96.31 - Corporate structure.

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2010-04-01

... 22 Foreign Relations 1 2010-04-01 2010-04-01 false Corporate structure. 96.31 Section 96.31 Foreign Relations DEPARTMENT OF STATE LEGAL AND RELATED SERVICES ACCREDITATION OF AGENCIES AND APPROVAL OF... Approval Licensing and Corporate Governance § 96.31 Corporate structure. (a) The agency qualifies for...

12 CFR 545.96 - Agency office.

Science.gov (United States)

2010-01-01

... 12 Banks and Banking 5 2010-01-01 2010-01-01 false Agency office. 545.96 Section 545.96 Banks and Banking OFFICE OF THRIFT SUPERVISION, DEPARTMENT OF THE TREASURY FEDERAL SAVINGS ASSOCIATIONS-OPERATIONS § 545.96 Agency office. (a) General. A Federal savings association may establish or maintain an agency...

7 CFR 1948.96 - Audit requirements.

Science.gov (United States)

2010-01-01

... Program § 1948.96 Audit requirements. (a) Audit requirements for Site Development and Acquisition Grants will be made in accordance with FmHA Instruction 1942-G. (b) Audits for planning grants made in... 7 Agriculture 13 2010-01-01 2009-01-01 true Audit requirements. 1948.96 Section 1948.96...

First secondary metabolites from Herissantia crispa L (Brizicky) and the toxicity activity against Artemia salina Leach

Energy Technology Data Exchange (ETDEWEB)

Costa, Danielly Albuquerque da; Matias, Wemerson Neves; Lima, Igara Oliveira; Xavier, Aline Lira; Costa, Vivian Bruna Machado; Diniz, Margareth de Fatima Formiga Melo; Agra, Maria de Fatima; Batista, Leonia Maria; Souza, Maria de Fatima Vanderlei de [Universidade Federal da Paraiba (UFPB), Joao Pessoa, PB (Brazil). Lab. de Tecnologia Farmaceutica Prof. Delby Fernandes de Medeiros; Silva, Davi Antas e [Universidade Federal Rural de Pernambuco, Serra Talhada, PE (Brazil). Dept. de Quimica

2009-07-01

The phytochemical investigation of Herissantia crispa led to the isolation of seven compounds, identified as: sitosterol 3-O-{beta}-D-glucopyranoside, stigmasterol 3-O-{beta}-D-glucopyranoside, 3,5,7,4'-tetrahydroxyflavone (kaempferol), 3,5,7,3',4'-pentahydroxyflavone (quercetin), unpublished in the genus Herissantia, besides {beta}-sitosterol, kaempferol 3-O-{beta}-D-(6''-E-p-coumaroil) (tiliroside) glucopyranoside and kaempferol 3,7-di-O-{alpha}-L-ramnopyranoside (lespedin), described for the first time in the species. The structural determination of the compounds was made by means of spectroscopy methods such as Infrared Spectroscopy, {sup 1}H and {sup 13}C Nuclear Magnetic Resonance, with the aid of two dimensional techniques, and by comparison with literature data. The toxicity activity of the MeOH extract and lespedin on Artemia salina Leach. was also carried out. (author)

El turismo de naturaleza en espacios naturales. El caso del parque regional de las Salinas y Arenales de San Pedro del Pinatar

OpenAIRE

Ballesteros Pelegrín, Gustavo Alfonso

2014-01-01

Los espacios naturales atraen a turistas que buscan el contacto con la naturaleza, surgiendo nuevos productos turísticos. Surgen, de este modo, nuevos productos turísticos. En la oferta de turismo de naturaleza del Parque Regional de las Salinas y Arenales de San Pedro del Pinatar destaca: deportes en la naturaleza, aventura y ecoturismo. Este último se divide en: turismo ornitológico y fotográfico, que junto al tradicional de sol y playa y de salud, se ve favorecido por su situación geográfi...

45 CFR 96.47 - Primary care.

Science.gov (United States)

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Primary care. 96.47 Section 96.47 Public Welfare... and Tribal Organizations § 96.47 Primary care. Applications for direct funding of Indian tribes and tribal organizations under the primary care block grant must comply with 42 CFR Part 51c (Grants for...

Estimating the wound healing ability of bioactive milk proteins using an optimized cell based assay

DEFF Research Database (Denmark)

Nyegaard, Steffen; Andreasen, Trine; Rasmussen, Jan Trige

Milk contains many different proteins of which the larger constituents like the caseins and major whey constituents are well characterized. We have for some time been studying the structure and function of proteins associated with the milk fat globule membrane like lactadherin, MUC1/15, xanthine...... oxidoreductase along with minor whey constituents like osteopontin, EPV20 etc. The enterocyte migration rate is a key parameter in maintaining intestinal homeostasis and intestinal repair when recovering from infection or intestinal diseases like Crohns and ulcerative colitis. We developed a novel in vitro wound...... healing assay to determine the bioactive effects of various milk proteins using human small intestine cells grown on extracellular matrix. Silicone inserts are placed in a 96-well plate and enterocytes seeded around it, creating a monolayer with a cell free area. In current ongoing experiments, various...

Label-Free, LC-MS-Based Assays to Quantitate Small-Molecule Antagonist Binding to the Mammalian BLT1 Receptor.

Science.gov (United States)

Chen, Xun; Stout, Steven; Mueller, Uwe; Boykow, George; Visconti, Richard; Siliphaivanh, Phieng; Spencer, Kerrie; Presland, Jeremy; Kavana, Michael; Basso, Andrea D; McLaren, David G; Myers, Robert W

2017-08-01

We have developed and validated label-free, liquid chromatography-mass spectrometry (LC-MS)-based equilibrium direct and competition binding assays to quantitate small-molecule antagonist binding to recombinant human and mouse BLT1 receptors expressed in HEK 293 cell membranes. Procedurally, these binding assays involve (1) equilibration of the BLT1 receptor and probe ligand, with or without a competitor; (2) vacuum filtration through cationic glass fiber filters to separate receptor-bound from free probe ligand; and (3) LC-MS analysis in selected reaction monitoring mode for bound probe ligand quantitation. Two novel, optimized probe ligands, compounds 1 and 2, were identified by screening 20 unlabeled BLT1 antagonists for direct binding. Saturation direct binding studies confirmed the high affinity, and dissociation studies established the rapid binding kinetics of probe ligands 1 and 2. Competition binding assays were established using both probe ligands, and the affinities of structurally diverse BLT1 antagonists were measured. Both binding assay formats can be executed with high specificity and sensitivity and moderate throughput (96-well plate format) using these approaches. This highly versatile, label-free method for studying ligand binding to membrane-associated receptors should find broad application as an alternative to traditional methods using labeled ligands.

Crescimento e fitomassa da beterraba sob irrigação suplementar com água de diferentes concentrações salinas

Directory of Open Access Journals (Sweden)

Daniella Pereira dos Santos

Full Text Available RESUMO O uso da água salina na agricultura é uma alternativa viável, tendo-se em vista o aumento da demanda de água doce. Objetivou-se, com este trabalho, avaliar o crescimento e a produção de fitomassa da beterraba, sob irrigação com água de diferentes concentrações salinas, em experimento em condição de campo, no Campus da Universidade Federal de Alagoas, em Arapiraca. Os tratamentos foram cinco níveis de condutividade elétrica (1,0; 2,0; 3,0; 4,0 e 5,0 dS m-1. O delineamento foi em blocos casualizados, com quatro repetições. O máximo rendimento da beterraba aos 27 dias após aplicação dos tratamentos salinos foi obtido com uma salinidade de 3,0 dS m-1, para as variáveis altura de planta (AP, diâmetro do caule (DC, comprimento da raiz (CR, fitomassa seca da parte aérea (FSPA e fitomassa seca total (FST. Aos 42 dias após aplicação dos tratamentos salinos, as variáveis fitomassa fresca da parte aérea (FFPA, fitomassa fresca da raiz (FFR, fitomassa fresca total (FFT, fitomassa seca da parte aérea (FSPA e fitomassa seca total (FST aumentaram com o aumento da salinidade da água. A chuva pode ter influenciado os resultados obtidos para as avaliações, realizadas aos 42 dias após aplicação dos tratamentos salinos.

Enterocin 96, a Novel Class II Bacteriocin Produced by Enterococcus faecalis WHE 96, Isolated from Munster Cheese▿

Science.gov (United States)

Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd

2009-01-01

Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains. PMID:19411428

Enterocin 96, a novel class II bacteriocin produced by Enterococcus faecalis WHE 96, isolated from Munster cheese.

Science.gov (United States)

Izquierdo, Esther; Wagner, Camille; Marchioni, Eric; Aoude-Werner, Dalal; Ennahar, Saïd

2009-07-01

Enterococcus faecalis WHE 96, a strain isolated from soft cheese based on its anti-Listeria activity, produced a 5,494-Da bacteriocin that was purified to homogeneity by ultrafiltration and cation-exchange and reversed-phase chromatographies. The amino acid sequence of this bacteriocin, named enterocin 96, was determined by Edman degradation, and its structural gene was sequenced, revealing a double-glycine leader peptide. After a comparison with other bacteriocins, it was shown that enterocin 96 was a new class II bacteriocin that showed very little similarity with known structures. Enterocin 96 was indeed a new bacteriocin belonging to class II bacteriocins. The activity spectrum of enterocin 96 covered a wide range of bacteria, with strong activity against most gram-positive strains but very little or no activity against gram-negative strains.

Changes in the Structure of the Microbial Community Associated with Nannochloropsis salina following Treatments with Antibiotics and Bioactive Compounds

Science.gov (United States)

Geng, Haifeng; Tran-Gyamfi, Mary B.; Lane, Todd W.; Sale, Kenneth L.; Yu, Eizadora T.

2016-01-01

Open microalgae cultures host a myriad of bacteria, creating a complex system of interacting species that influence algal growth and health. Many algal microbiota studies have been conducted to determine the relative importance of bacterial taxa to algal culture health and physiological states, but these studies have not characterized the interspecies relationships in the microbial communities. We subjected Nanochroloropsis salina cultures to multiple chemical treatments (antibiotics and quorum sensing compounds) and obtained dense time-series data on changes to the microbial community using 16S gene amplicon metagenomic sequencing (21,029,577 reads for 23 samples) to measure microbial taxa-taxa abundance correlations. Short-term treatment with antibiotics resulted in substantially larger shifts in the microbiota structure compared to changes observed following treatment with signaling compounds and glucose. We also calculated operational taxonomic unit (OTU) associations and generated OTU correlation networks to provide an overview of possible bacterial OTU interactions. This analysis identified five major cohesive modules of microbiota with similar co-abundance profiles across different chemical treatments. The Eigengenes of OTU modules were examined for correlation with different external treatment factors. This correlation-based analysis revealed that culture age (time) and treatment types have primary effects on forming network modules and shaping the community structure. Additional network analysis detected Alteromonadeles and Alphaproteobacteria as having the highest centrality, suggesting these species are “keystone” OTUs in the microbial community. Furthermore, we illustrated that the chemical tropodithietic acid, which is secreted by several species in the Alphaproteobacteria taxon, is able to drastically change the structure of the microbiota within 3 h. Taken together, these results provide valuable insights into the structure of the microbiota

Caracterización de bacterias halófilas productoras de amilasas aisladas de las Salinas de San Blas en Junín

Directory of Open Access Journals (Sweden)

Pamela Elizabeth Canales Mormontoy

2014-07-01

Full Text Available Título en español: Caracterización de bacterias halófilas productoras de amilasas aisladas de las Salinas de San Blas en Junín Título en ingles: Characterization of halophilic bacteria producing amylase isolated from San Blas Salterns in Junin Título corto: Bacterias halófilas amilolíticas de las Salinas de San Blas Resumen:  El objetivo de este estudio fue caracterizar bacterias halófilas con actividad amilolítica provenientes de las Salinas de San Blas-Junín, ubicadas en los Andes peruanos aproximadamente a 4100 m de altitud. Este estudio se realizó con 34 bacterias aisladas de muestras de suelos las cuales se cultivaron en agar agua de sales (SW 5 % conteniendo extracto de levadura 0,5 % y almidón 1 %. El 41 % de bacterias mostró la capacidad de hidrolizar almidón, éstas fueron caracterizadas mediante pruebas fisiológicas y bioquímicas convencionales. Tres bacterias fueron Gram-negativas y once Gram-positivas. El 21 % (3/14 creció en un amplio rango de concentración de sales, entre 5 y 20 %. El 14 % (2/14 de las bacterias presentó actividad lipolítica, proteolítica y nucleolítica, y el 29 % (4/14, presentó actividad proteolítica y nucleolítica. Las bacterias se identificaron mediante los perfiles de restricción de los genes ribosómicos 16S amplificados, las enzimas usadas fueron Hae III, BstU I, Hinf I y Cfo I. Los genes ribosómicos 16S de siete bacterias que presentaron perfiles de ADN diferentes se amplificaron, secuenciaron y analizaron mediante programas bioinformáticos. Del análisis fenotípico y molecular de las 14 bacterias amilolíticas se obtuvieron dos grupos, uno perteneciente al género Halomonas (3 y el otro, al género Bacillus (11. Las bacterias amilolíticas caracterizadas podrían ser de potencial uso a nivel industrial.  Palabras clave: Salinas de San Blas, amilasas, genes ribosómicos 16S, ARDRA, Bacillus, Halomonas. Abstract: The aim of this study was to characterize halophilic

Encapsulación de moléculas pequeñas mediante la precipitación salina de poliuretanos catioméricos

OpenAIRE

Fernández d’Arlas, Borja; Corcuera, María Ángeles; Eceiza, Arantxa

2015-01-01

En este trabajo se estudia un copolímero de poliuretano catiomérico (PU) con alta proporción de uretano como agente encapsulante de fármacos modelos (FM) mediante la encapsulación inducida por precipitación salina del PU y FM a pH < pI del PU. Mediante espectroscopia UV-Vis se ha estimado el porcentaje de encapsulación de varios FMs proponiéndose un modelo semi-empírico para determinar la distribución observada en las eficiencias de encapsulación, E, en función de su volumen molar...

Atributos biológicos de dos suelos de Quibor con aplicación de abono orgánico y soluciones salinas

OpenAIRE

Mendoza, Betty; Florentino, Adriana; Hernández-Hernández, Rosa Mary; Aciego, Juan; Torres, Duilio; Vera, Elena

2013-01-01

La evaluación del efecto de la aplicación de abono orgánico y soluciones salinas sobre los atributos biológicos de suelos de Quibor estado Lara, Venezuela, uno bajo manejo convencional (CV) y otro bajo manejo conservacionista (CS), se realizó mediante un ensayo de invernadero. Se utilizaron tres dosis de materia orgánica (MO): 0, 15 y 30 Mg ha-1 y cuatro soluciones de riego (SR): testigo con agua (T), sulfato de calcio (SC), cloruro de sodio (CN) y la mezcla de sulfato de calcio con cloruro d...

Altered lipid accumulation in Nannochloropsis salina CCAP849/3 following EMS and UV induced mutagenesis

Directory of Open Access Journals (Sweden)

T.A. Beacham

2015-09-01

Full Text Available Microalgae have potential as a chemical feed stock in a range of industrial applications. Nannochloropsis salina was subject to EMS mutagenesis and the highest lipid containing cells selected using fluorescence-activated cell sorting. Assessment of growth, lipid content and fatty acid composition identified mutant strains displaying a range of altered traits including changes in the PUFA content and a total FAME increase of up to 156% that of the wild type strain. Combined with a reduction in growth this demonstrated a productivity increase of up to 76%. Following UV mutagenesis, lipid accumulation of the mutant cultures was elevated to more than 3 fold that of the wild type strain, however reduced growth rates resulted in a reduction in overall productivity. Changes observed are indicative of alterations to the regulation of the omega 6 Kennedy pathway. The importance of these variations in physiology for industrial applications such as biofuel production is discussed.

The 96th Amino Acid of the Coat Protein of Cucumber Green Mottle Mosaic Virus Affects Virus Infectivity

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Zhenwei Zhang

2017-12-01

Full Text Available Cucumber green mottle mosaic virus (CGMMV is one of the most devastating viruses infecting members of the family Cucurbitaceae. The assembly initiation site of CGMMV is located in the coding region of the coat protein, which is not only involved in virion assembly but is also a key factor determining the long-distance movement of the virus. To understand the effect of assembly initiation site and the adjacent region on CGMMV infectivity, we created a GTT deletion mutation in the GAGGTTG assembly initiation site of the infectious clone of CGMMV, which we termed V97 (deletion mutation at residue 97 of coat protein, followed by the construction of the V94A and T104A mutants. We observed that these three mutations caused mosaic after Agrobacterium-mediated transformation in Nicotiana benthamiana, albeit with a significant delay compared to the wild type clone. The mutants also had a common spontaneous E96K mutation in the coat protein. These results indicated that the initial assembly site and the sequence of the adjacent region affected the infectivity of the virus and that E96 might play an essential role in this process. We constructed two single point mutants—E96A and E96K—and three double mutants—V94A-E96K, V97-E96K and T104A-E96K—to further understand the role of E96 in CGMMV pathogenesis. After inoculation in N. benthamiana, E96A showed delayed systemic symptoms, but the E96K and three double mutants exhibited typical symptoms of mosaic at seven days post-infection. Then, sap from CGMMV-infected N. benthamiana leaves was mechanically inoculated on watermelon plants. We confirmed that E96 affected CGMMV infection using double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA, reverse transcription-polymerase chain reaction (RT-PCR, and sequencing, which further confirmed the successful infection of the related mutants, and that E96K can compensate the effect of the V94, V97, and T104 mutations on virus infectivity. In

First results with a radioreceptor-assay (TRAK-Assay) for TSH-receptor-autoantibodies

International Nuclear Information System (INIS)

Becker, W.; Reiners, C.; Boerner, W.

1983-01-01

A new radioreceptor-assay (TRAK-assay) for autoantibodies against TSH-receptors was tested in 48 untreated thyrotoxic patients (26 regional autonomies, 22 toxic diffuse goiters). None of the 26 patients with regional autonomy showed positive autoantibody-titers. 4 patients with toxic diffuse goiter and thyrotoxic exophthalmos were TRAK-positive. Positive titers of microsomal and thyreoglobulin autoantibodies could be seen in 8 of 9 patients with positive TRAK-titers. In accordance with the conventional methods for detecting thyroid-stimulating immunoglobulins the new TRAK-assay seems to be suited for differentiating between immunogenic toxic diffuse goiter (Graves' disease) and goiter with disseminated autonomy as well as for prediction of relapse. (orig.) [de

A new assay for cytotoxic lymphocytes, based on a radioautographic readout of 111In release, suitable for rapid, semi-automated assessment of limit-dilution cultures

International Nuclear Information System (INIS)

Shortman, K.; Wilson, A.

1981-01-01

A new assay for cytotoxic T lymphocytes is described, of general application, but particularly suitable for rapid, semi-automated assessment of multiple microculture tests. Target cells are labelled with high efficiency and to high specific activity with the oxine chelate of 111 indium. After a 3-4 h incubation of test cells with 5 X 10 3 labelled target cells in V wells of microtitre trays, samples of the supernatant are spotted on paper (5 μl) or transferred to soft-plastic U wells (25-50 μl) and the 111 In release assessed by radioautography. Overnight exposure of X-ray film with intensifying screens at -70 0 C gives an image which is an intense dark spot for maximum release, a barely visible darkening with the low spontaneous release, and a definite positive with 10% specific lysis. The degree of film darkening, which can be quantitated by microdensitometry, shows a linear relationship with cytotoxic T lymphocyte dose up to the 40% lysis level. The labelling intensity and sensitivity can be adjusted over a wide range, allowing a single batch of the short half-life isotope to serve for 2 weeks. The 96 assays from a single tray are developed simultaneously on a single small sheet of film. Many trays can be processed together, and handling is rapid if 96-channel automatic pipettors are used. The method allows rapid visual scanning for positive and negative limit dilution cultures in cytotoxic T cell precursor frequency and specificity studies. In addition, in conjunction with an automated densitometer designed to scan microtitre trays, the method provides an efficient alternative to isotope counting in routine cytotoxic assays. (Auth.)

Characterization of the commercially-available fluorescent chloroquine-BODIPY conjugate, LynxTag-CQGREEN, as a marker for chloroquine resistance and uptake in a 96-well plate assay.

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Cheryl C Y Loh

Full Text Available Chloroquine was a cheap, extremely effective drug against Plasmodium falciparum until resistance arose. One approach to reversing resistance is the inhibition of chloroquine efflux from its site of action, the parasite digestive vacuole. Chloroquine accumulation studies have traditionally relied on radiolabelled chloroquine, which poses several challenges. There is a need for development of a safe and biologically relevant substitute. We report here a commercially-available green fluorescent chloroquine-BODIPY conjugate, LynxTag-CQGREEN, as a proxy for chloroquine accumulation. This compound localized to the digestive vacuole of the parasite as observed under confocal microscopy, and inhibited growth of chloroquine-sensitive strain 3D7 more extensively than in the resistant strains 7G8 and K1. Microplate reader measurements indicated suppression of LynxTag-CQGREEN efflux after pretreatment of parasites with known reversal agents. Microsomes carrying either sensitive- or resistant-type PfCRT were assayed for uptake; resistant-type PfCRT exhibited increased accumulation of LynxTag-CQGREEN, which was suppressed by pretreatment with known chemosensitizers. Eight laboratory strains and twelve clinical isolates were sequenced for PfCRT and Pgh1 haplotypes previously reported to contribute to drug resistance, and pfmdr1 copy number and chloroquine IC50s were determined. These data were compared with LynxTag-CQGREEN uptake/fluorescence by multiple linear regression to identify genetic correlates of uptake. Uptake of the compound correlated with the logIC50 of chloroquine and, more weakly, a mutation in Pgh1, F1226Y.

Development of novel, 384-well high-throughput assay panels for human drug transporters: drug interaction and safety assessment in support of discovery research.

Science.gov (United States)

Tang, Huaping; Shen, Ding Ren; Han, Yong-Hae; Kong, Yan; Balimane, Praveen; Marino, Anthony; Gao, Mian; Wu, Sophie; Xie, Dianlin; Soars, Matthew G; O'Connell, Jonathan C; Rodrigues, A David; Zhang, Litao; Cvijic, Mary Ellen

2013-10-01

Transporter proteins are known to play a critical role in affecting the overall absorption, distribution, metabolism, and excretion characteristics of drug candidates. In addition to efflux transporters (P-gp, BCRP, MRP2, etc.) that limit absorption, there has been a renewed interest in influx transporters at the renal (OATs, OCTs) and hepatic (OATPs, BSEP, NTCP, etc.) organ level that can cause significant clinical drug-drug interactions (DDIs). Several of these transporters are also critical for hepatobiliary disposition of bilirubin and bile acid/salts, and their inhibition is directly implicated in hepatic toxicities. Regulatory agencies took action to address transporter-mediated DDI with the goal of ensuring drug safety in the clinic and on the market. To meet regulatory requirements, advanced bioassay technology and automation solutions were implemented for high-throughput transporter screening to provide structure-activity relationship within lead optimization. To enhance capacity, several functional assay formats were miniaturized to 384-well throughput including novel fluorescence-based uptake and efflux inhibition assays using high-content image analysis as well as cell-based radioactive uptake and vesicle-based efflux inhibition assays. This high-throughput capability enabled a paradigm shift from studying transporter-related issues in the development space to identifying and dialing out these concerns early on in discovery for enhanced mechanism-based efficacy while circumventing DDIs and transporter toxicities.

Evaluation of toxic effects with transition metal ions, EDTA, SBTI and acrylic polymers on Aedes aegypti (L., 1762 (Culicidae and Artemia salina (Artemidae

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Eduardo José de Arruda

2011-06-01

Full Text Available This work aimed to evaluate the toxicity of some insecticides compounds on Aedes aegypti and Artemia salina larvae. Bioassays were carried out to evaluate the toxic effect after of 24 and 72 h using the compounds or associations. The LC10, LC50 and LC90 values were obtained and utilized for toxicity comparations. For Ae. aegypti, LC50 were 32.65 mg L-1 in 24 h for Na2[EDTA-Cu(II] and total mortality in 72 h for SAP-Na2[EDTA-Cu(II].

45 CFR 96.123 - Assurances.

Science.gov (United States)

2010-10-01

... submit an annual report as required under § 96.122(d) and § 96.130(e); (6) Pregnant women are provided... Federal laws and regulations, including those relating to lobbying (45 CFR Part 93), drug-free workplace (45 CFR 76.600), discrimination (PHS Act Sec. 1947), false statements or failure to disclose certain...

Comportamiento geoquímico de las formaciones salinas bajo el efecto de la temperatura y la irradiación

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Pueyo, J. J.

1993-08-01

Full Text Available The production of radiation induced defects, as well as fluid migration (brine and gases, are the most relevant phenomena due to the combined effects of heat and irradiation in repositories emplaced in rock salt. In order to gain knowledge of these phenomena, laboratory experiments on samples coming from the Sallent Mine (Barcelona have been performed. The water content ranges between 0.04 and 1.00 %. Nevertheless, smaller releases of brine are expected, since part of it is present in fluid inclusions of a size below 100 µm. The chemical composition of the brine, which is a key issue in the study of container corrosion, is in the case of the fluid inc1usions enriched in magnesium (1.15 mol/l and potassium (0.31 mol/l. The study of the gas content reveals the absence of toxic gases and the scarcity of hydrogen and methane. Finally, the amount of radiation induced defects, which are controlled by the mineralogical composition of the rock salt, is similar to those obtained with the Jain-Lidiard model.La generación de defectos cristalinos y la migración de las fases fluidas (salmueras y gases, son los efectos más relevantes de la irradiación y la temperatura sobre una formación salina, susceptible de albergar un almacenamiento de residuos radiactivos. Para determinar dichos efectos se ha puesto a punto la metodología necesaria y se ha verificado sobre muestras salinas de la mina de Sallent (Barcelona. El contenido en agua oscila entre 0,04 y 1 %. Sin embargo, la cantidad de salmuera susceptible de migrar será inferior a la total, debido a que parte de esta se encuentra en inclusiones fluidas inferiores a 100 µm. La composición química de la salmuera, factor clave para el estudio de corrosión de contenedores, es de carácter magnésico (1,15 moles/litro y potásico (0,31 moles/litro en el caso de las inclusiones. El análisis de los gases presentes indica la ausencia de gases tóxicos y concentraciones muy bajas de hidrógeno y metano

Endogenous Locus Reporter Assays.

Science.gov (United States)

Liu, Yaping; Hermes, Jeffrey; Li, Jing; Tudor, Matthew

2018-01-01

Reporter gene assays are widely used in high-throughput screening (HTS) to identify compounds that modulate gene expression. Traditionally a reporter gene assay is built by cloning an endogenous promoter sequence or synthetic response elements in the regulatory region of a reporter gene to monitor transcriptional activity of a specific biological process (exogenous reporter assay). In contrast, an endogenous locus reporter has a reporter gene inserted in the endogenous gene locus that allows the reporter gene to be expressed under the control of the same regulatory elements as the endogenous gene, thus more accurately reflecting the changes seen in the regulation of the actual gene. In this chapter, we introduce some of the considerations behind building a reporter gene assay for high-throughput compound screening and describe the methods we have utilized to establish 1536-well format endogenous locus reporter and exogenous reporter assays for the screening of compounds that modulate Myc pathway activity.

The value of microscopic-observation drug susceptibility assay in the diagnosis of tuberculosis and detection of multidrug resistance.

Science.gov (United States)

Sertel Şelale, Denİz; Uzun, Meltem

2018-01-01

Inexpensive, rapid, and reliable tests for detecting the presence and drug susceptibility of Mycobacterium tuberculosis complex (MTBC) are urgently needed to control the transmission of tuberculosis. In this study, we aimed to assess the accuracy and speed of the microscopic-observation drug susceptibility (MODS) assay in the identification of MTBC and detection of multidrug resistance. Sputum samples from patients suspected to have tuberculosis were simultaneously tested with MODS and conventional culture [Löwenstein-Jensen (LJ) culture, BACTEC MGIT™ 960 (MGIT) system], and drug susceptibility testing (MGIT system) methods. A total of 331 sputum samples were analyzed. Sensitivity and specificity of MODS assay for detection of MTBC strains were 96% and 98.8%, respectively. MODS assay detected multidrug resistant MTBC isolates with 92.3% sensitivity and 96.6% specificity. Median time to culture positivity was similar for MGIT (8 days) and MODS culture (8 days), but was significantly longer with LJ culture (20 days) (p tuberculosis and detection of multidrug resistance. © 2017 APMIS. Published by John Wiley & Sons Ltd.

Envelhecimento acelerado em sementes de azevém com e sem solução salina e saturada Accelerated aging of ryegrass seeds submitted to saturated salt solution

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Lilian Madruga de Tunes

2011-01-01

Full Text Available O presente trabalho teve por objetivo analisar a metodologia do teste de envelhecimento acelerado para avaliação do potencial fisiológico de sementes de azevém, bem como verificar a possibilidade do uso de solução não saturada e saturada de NaCl como opção para controle da absorção de água pelas sementes durante a realização do teste, sem reduzir sua sensibilidade. Foram utilizados quatro lotes de sementes, submetidos aos testes de germinação, emergência de plântulas, índice de velocidade de emergência e envelhecimento acelerado, empregando-se os períodos de exposição de 24, 48, 72 e 96h, com e sem solução salina (NaCl. A utilização de solução não saturada e saturada de NaCl diminui a absorção de água pelas sementes de azevém durante o teste de envelhecimento acelerado, acarretando uma taxa de deterioração menos acentuada e resultados menos drásticos e mais uniformes. A opção 24h com solução não saturada (SNS só conseguiu estratificar os lotes em dois grupos, não devendo ser indicada como promissora. Assim, deve-se usar a opção 48h com solução saturada de NaCl (SSS, que estratificou em três lotes.The objective of the present study was to evaluate the methodology of the accelerated aging test to evaluate the physiological potential of s ryegrass seeds, as well as verify the possibility of the use of unsaturated and saturated solution of NaCl as an option for control of water uptake by seeds during the test without reducing its sensitivity. Four lots of ryegrass seeds were tested for germination, seedling emergence, speed emergence and accelerated aging (periods of 24, 48, 72 and 96h, with or without the use of saturated and saline solution of NaCl. The use of unsaturated and saturated solution of NaCl reduces water absorption by ryegrass seeds during the accelerated aging test, resulting in a rate of deterioration is less pronounced ,less drastic and more uniform results. Option 24h with

Efficacy and safety of maraviroc versus efavirenz, both with zidovudine/lamivudine: 96-week results from the MERIT study.

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Sierra-Madero, Juan; Di Perri, Giovanni; Wood, Robin; Saag, Michael; Frank, Ian; Craig, Charles; Burnside, Robert; McCracken, Jennifer; Pontani, Dennis; Goodrich, James; Heera, Jayvant; Mayer, Howard

2010-01-01

The MERIT study evaluated maraviroc versus efavirenz, both with zidovudine/lamivudine, in treatment-naïve patients with CCR5-tropic (R5) HIV-1. Post hoc analyses previously assessed week 48 outcomes in patients rescreened with R5 virus by a more sensitive tropism assay. Week 96 efficacy (post hoc, n = 614) and safety (n = 721) were assessed. Proportions of subjects <50 copies/mL (58.8% maraviroc, 62.7% efavirenz) and time to loss of virologic response (TLOVR) responders (<50 copies/mL: 60.5% vs 60.7%) were similar. Maraviroc recipients had greater CD4 increases (+ 212 vs + 171 cells/mm(3)) and fewer adverse event discontinuations (6.1% vs 15.5%), malignancies, and category C events. Week 96 data confirm week 48 observations in MERIT.

Tratamiento del prolapso rectal en la infancia con infiltración de solución salina al 16,5 %

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Carlos Ramírez Pérez

Full Text Available Introducción: cuando falla el tratamiento médico conservador en los pacientes pediátricos con prolapso rectal se impone la infiltración perirrectal con sustancias irritantes. En la década de los 90 esas infiltraciones en nuestro centro se hacían con glicerina, pero escaseó en el mercado, y ante tal problema, se buscó otro agente infiltrante alternativo. En el presente trabajo se describe la experiencia con la utilización de solución salina al 16,5 %. Métodos: se realizó un estudio de intervención, longitudinal, prospectivo, experimental, del tipo ensayo clínico no controlado. Consta de una segunda parte en la que se utilizó la aleatorización con un grupo control para la validación. El universo estuvo constituido por 27 pacientes y la muestra, por 16 pacientes con prolapso rectal, que fueron atendidos en el servicio de gastroenterología del Hospital Pediátrico Provincial de Holguín en el quinquenio 2003-2007. Las variables desarrolladas fueron: concentración idónea para el tratamiento, cantidad de sustancia a infiltrar, complicaciones, recurrencia, número de infiltraciones y curación al año o más. Los resultados se exponen en forma de tablas porcentuales. Resultados: la eficacia con la utilización de solución salina al 16,5 % fue del 100 %, todos los pacientes curaron, y ninguno presentó recurrencia, por lo que no fueron necesarias 2 o más sesiones de tratamiento. Se comentó de un niño que, luego de fallar la infiltración con leche materna en primera opción y fallido también el cerclaje, finalmente resolvió con este método. Las complicaciones fueron relativamente pocas (18,9 %: un absceso, una celulitis y una retención urinaria con necesidad de sondaje durante 2 semanas, sin secuelas posteriores. Se realizó aleatorización con igual número de pacientes en quienes se usó la glicerina, los resultados del uso de ambas sustancias fueron muy parecidos, y el número de complicaciones fue ligeramente mayor en el

49 CFR 176.96 - Materials of construction.

Science.gov (United States)

2010-10-01

... 49 Transportation 2 2010-10-01 2010-10-01 false Materials of construction. 176.96 Section 176.96 Transportation Other Regulations Relating to Transportation PIPELINE AND HAZARDOUS MATERIALS SAFETY... Requirements for Barges § 176.96 Materials of construction. Barges used to transport hazardous materials must...

A Novel Assay for Easy and Rapid Quantification of Helicobacter pylori Adhesion.

Science.gov (United States)

Skindersoe, Mette E; Rasmussen, Lone; Andersen, Leif P; Krogfelt, Karen A

2015-06-01

Reducing adhesion of Helicobacter pylori to gastric epithelial cells could be a new way to counteract infections with this organism. We here present a novel method for quantification of Helicobacter pylori adhesion to cells. Helicobacter pylori is allowed to adhere to AGS or MKN45g cells in a 96-well microtiter plate. Then wells are added saponin, which lyses the cells without affecting the bacteria. After addition of alamarBlue(®) (resazurin) and 1- to 2-hour incubation, fluorescence measurements can be used to quantify the number of adherent bacteria. By use of the method, we demonstrate that adhesion of both a sabA and babA deletion mutant of H. pylori is significantly reduced compared to the wild type. The method offers a number of applications and may be used to compare the adherence potential of different strains of H. pylori to either cells or different materials or to screen for potential anti-adhesive compounds. The results presented here suggest that this easy and reproducible assay is well suited for quantitative investigation of H. pylori adhesion. © 2015 John Wiley & Sons Ltd.

Multiplexing a high-throughput liability assay to leverage efficiencies.

Science.gov (United States)

Herbst, John; Anthony, Monique; Stewart, Jeremy; Connors, David; Chen, Taosheng; Banks, Martyn; Petrillo, Edward W; Agler, Michele

2009-06-01

In order to identify potential cytochrome P-450 3A4 (drug-metabolizing enzyme) inducers at an early stage of the drug discovery process, a cell-based transactivation high-throughput luciferase reporter assay for the human pregnane X receptor (PXR) in HepG2 cells has been implemented and multiplexed with a viability end point for data interpretation, as part of a Lead Profiling portfolio of assays. As a routine part of Lead Profiling operations, assays are periodically evaluated for utility as well as for potential improvements in technology or process. We used a recent evaluation of our PXR-transactivation assay as a model for the application of Lean Thinking-based process analysis to lab-bench assay optimization and automation. This resulted in the development of a 384-well multiplexed homogeneous assay simultaneously detecting PXR transactivation and HepG2 cell cytotoxicity. In order to multiplex fluorescent and luminescent read-outs, modifications to each assay were necessary, which included optimization of multiple assay parameters such as cell density, plate type, and reagent concentrations. Subsequently, a set of compounds including known cytotoxic compounds and PXR inducers were used to validate the multiplexed assay. Results from the multiplexed assay correlate well with those from the singleplexed assay formats measuring PXR transactivation and viability separately. Implementation of the multiplexed assay for routine compound profiling provides improved data quality, sample conservation, cost savings, and resource efficiencies.

A histidine-rich protein 2-based malaria drug sensitivity assay for field use.

Science.gov (United States)

Noedl, Harald; Attlmayr, Bernhard; Wernsdorfer, Walther H; Kollaritsch, Herwig; Miller, Robert S

2004-12-01

With the spread of antimalarial drug resistance, simple and reliable tools for the assessment of antimalarial drug resistance, particularly in endemic regions and under field conditions, have become more important than ever before. We therefore developed a histidine-rich protein 2 (HRP2)-based drug sensitivity assay for testing of fresh isolates of Plasmodium falciparum in the field. In contrast to the HRP2 laboratory assay, the field assay uses a procedure that further simplifies the handling and culturing of malaria parasites by omitting centrifugation, washing, the use of serum, and dilution with uninfected red blood cells. A total of 40 fresh Plasmodium falciparum isolates were successfully tested for their susceptibility to dihydroartemisinin, mefloquine, quinine, and chloroquine (50% inhibitory concentration [IC50] = 3.43, 61.89, 326.75, and 185.31 nM, respectively). Results very closely matched those obtained with a modified World Health Organization schizont maturation assay (R2 = 0.96, P < 0.001; mean log difference at IC50 = 0.054).

Estudio de caracterización de emprendedores de la Parroquia Salinas de la Provincia de Bolívar - Ecuador

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Jorge Goyes Noboa

2016-09-01

Salinas de Guaranda. Se han identificado variables y factores sociales, económicos, de financiamiento, organizacionales, estructuras administrativas, económicas, cultura organizacional, formas asociativas, motivación para crear negocios, estructuras de negocios predominantes que impulsan la economía social y solidaria presente en la zona. El mayor número de emprendimientos está ligado directa e indirectamente (66% de negocios a las organizaciones sociales y fundaciones salineras, son negocios relativamente jóvenes (menos de 10 años de existencia que han surgido por la necesidad de generar actividades económicas que apoyen al ingreso familiar y comunitario, con lo que afianza el modelo de desarrollo social vigente.

Regulatory Project Manager for Salina and Permian Basins for the NWTS [National Waste Terminal Storage] Program: Final techical report

International Nuclear Information System (INIS)

1986-12-01

The identification of candidate sites for nuclear waste repositories involves geological and environmental studies to characterize potential sites. These investigations include the collection and analysis of detailed geological and environmental data and comparison of the data against predetermined site performance criteria, i.e., geologic characteristics, environmental protection, and socioeconomic impacts. The work summarized in this final technical report encompasses mainly ''environmental characterization'' studies in the Permian Basin in the Texas Panhandle during the period of 1977-86; in the earlier phase of the contract, regional environmental work was also done in the Salina Basin (1977-79) and certain licensing support activities and safety analyses were conducted (1977-82). Considerable regulatory support work was also performed during 1986. 9 figs., 2 tabs

Performance evaluation of four type-specific commercial assays for detection of herpes simplex virus type 1 antibodies in a Middle East and North Africa population.

Science.gov (United States)

Aldisi, Rana S; Elsidiq, Malaz S; Dargham, Soha R; Sahara, Afifah S; Al-Absi, Enas S; Nofal, Mariam Y; Mohammed, Layla I; Abu-Raddad, Laith J; Nasrallah, Gheyath K

2018-03-22

The number of diagnostic assays for the detection of herpes simplex virus type 1 (HSV-1) antibodies has increased over the years. However, their performance characteristics could vary among global populations. To investigate performance of two commercial ELISA kits, HerpeSelect ® 1 ELISA and Euroimmun Anti-HSV-1 (gC1) ELISA (IgG); and two commercial immunoblot (IB)/Western blot (WB) assays, HerpeSelect ® 1 and 2 Immunoblot IgG, and Euroimmun Anti-HSV-1/HSV-2 gG2 Euroline-WB (IgG/IgM); in detecting HSV-1 antibodies in a Middle East and North Africa (MENA) population. Blood specimens were collected from blood donors in Doha, Qatar, June 2013-2016. Twenty specimens were randomly selected from 10 MENA nationalities (Egypt, Iran, Jordan, Lebanon, Pakistan, Palestine, Qatar, Sudan, Syria, and Yemen; total = 200), and tested for HSV-1 antibodies. Across all six comparisons between assays, positive percent agreement ranged between 95.7% (95% CI: 91.4-98.3%) and 100.0% (95% CI: 97.8-100.0%). Negative percent agreement ranged between 86.2% (95% CI: 68.3-96.1%) and 96.2% (95% CI: 80.4-99.9%). Overall percent agreement ranged between 95.7% (95% CI: 91.7-97.8%) and 99.4% (95% CI: 96.7-99.9%). Cohen's kappa statistic ranged between 0.84 (95% CI: 0.73-0.95) and 0.98 (95% CI: 0.93-1.00). Compared against IB/WB, HerpeSelect ® and Euroimmun had sensitivities and specificities >96% and >86%, respectively. Positive and negative predictive values were >97% and >83%, respectively. The assays showed excellent concordance with one another, and with a high kappa statistic. The ELISA kits demonstrated robust diagnostic performance compared to the IB/WB assays. These findings support the assays' utility in clinical diagnosis and research in MENA populations. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

Performance characteristics of bioassay, radioenzymatic assay, homogeneous enzyme immunoassay, and high-performance liquid chromatographic determination of serum gentamicin

International Nuclear Information System (INIS)

Delaney, C.J.; Opheim, K.E.; Smith, A.L.; Plorde, J.J.

1982-01-01

We compared the accuracy, precision, and between-method error of the microbiological assay, the radioenzymatic assay, the homogeneous enzyme immunoassay, and the high-performance liquid chromatographic assay for the quantitation of gentamicin in serum. Precision and accuracy were evaluated by reference samples prepared to contain 0.0 to 32.7 micrograms of gentamicin per ml. Correlations between the methods utilized patient sera with gentamicin concentrations ranging from 0.6 to 13.3 micrograms/ml. All methods were reliable within acceptable limits for routine clinical use; intermethod correlation coefficients exceeded 0.96. Relative to the microbiological assay, the alternative methods offer the advantage of rapid analysis. The elapsed times for acquiring data on a set of 10 specimens under routine operating conditions were 0.5 h by the enzyme immunoassay, 4 h by the radioenzymatic assay, 5 h by the high-performance liquid chromatographic assay, and 10 h by the microbiological assay

33 CFR 96.130 - Incorporation by reference.

Science.gov (United States)

2010-07-01

... (ISM) Code by Administrations, November 23, 1995—96.320, 96.440 Resolution A.739(18), Guidelines for.../code_of_federal_regulations/ibr_locations.html. (b) The material approved for incorporation by...), International Management Code for the Safe Operation of Ships and for Pollution Prevention, November 4, 1993—96...

Assay-specific decision limits for two new automated parathyroid hormone and 25-hydroxyvitamin D assays.

Science.gov (United States)

Souberbielle, Jean-Claude; Fayol, Véronique; Sault, Corinne; Lawson-Body, Ethel; Kahan, André; Cormier, Catherine

2005-02-01

The recent development of nonradioactive automated assays for serum parathyroid hormone (PTH) and 25-hydroxyvitamin D (25OHD) has made measurement of these two hormones possible in many laboratories. In this study, we compared two new assays for PTH and 25OHD adapted on an automated analyzer, the LIAISON, with two manual immunoassays used worldwide. We studied 228 osteoporotic patients, 927 healthy individuals, 38 patients with primary hyperparathyroidism, and 167 hemodialyzed patients. Serum PTH was measured with the Allegro and the LIAISON assays, and 25OHD was measured with DiaSorin RIA and the LIAISON assay. Regression analysis was used to calculate decision thresholds for the LIAISON assays that were equivalent to those of the Allegro PTH and DiaSorin 25OHD assays. The 25OHD concentrations obtained with the LIAISON assay and the RIA in osteoporotic patients were well correlated (r = 0.83; P 50 nmol/L as eligible for the reference population for the LIAISON PTH assay. In this group, the 3rd-97th percentile interval for LIAISON PTH was 3-51 ng/L. Considering upper reference limits of 46 and 51 ng/L for the Allegro and LIAISON assays, respectively, the frequency of above-normal PTH concentrations in patients with primary hyperparathyroidism was similar in both assays. Regression analysis between serum PTH measured by the Allegro and LIAISON assays in 167 hemodialyzed patients and the corresponding Bland-Altman analysis of these data suggest that the LIAISON PTH assay tends to read higher than the Allegro assay at low concentrations but lower at high concentrations (>300 ng/L). Because clinical decision limits for both PTH and 25OHD should be assay specific, we propose equivalences between these assays and two manual assays used worldwide. These assay-specific decision limits should help potential users of the LIAISON PTH and 25OHD assays.

The Indigenous World, 1995-96 = El Mundo Indigena, 1995-96.

Science.gov (United States)

Jensen, Marianne, Comp.

This annual publication examines political, legal, social, and educational issues concerning indigenous peoples around the world during 1995-96. Part I highlights news events and ongoing situations in specific countries, including threats to indigenous territories, human rights violations, political victories, developments at the United Nations,…

Development of direct competitive biomimetic immunosorbent assay based on quantum dot label for determination of trichlorfon residues in vegetables.

Science.gov (United States)

Liu, Qiurui; Jiang, Mingdi; Ju, Zeliang; Qiao, Xuguang; Xu, Zhixiang

2018-06-01

A direct competitive biomimetic immunosorbent assay method based on molecularly imprinted polymer was developed for the determination of trichlorfon. A CdSe/ZnS quantum dot label was used as the marker. The hydrophilic imprinted film was synthesized directly on the surface of a 96-well plate, and characterized by Fourier-transform infrared spectroscopy and thermo-gravimetric analyses. The method exhibited high stability, selectivity, and sensitivity. Under optimal conditions, the limits of detection and sensitivity of the biomimetic immunosorbent assay method were 9.0 μg L -1 and 5.0 mg L -1 (0.1 mg kg -1 and 62.5 mg kg -1 for vegetable sample), respectively. Low cross-reactivity values of 19.2% and 15.6% were obtained for the structural analogues. Spinach and rape samples spiked with trichlorfon were extracted and determined by this method with recoveries ranging from 83.6% to 91.1%. The method was applied for the detection of trichlorfon residues in leek and cucumber samples, and results correlated well with those obtained using GC. Copyright © 2018 Elsevier Ltd. All rights reserved.

Probe colorimeter for quantitating enzyme-linked immunosorbent assays and other colorimetric assays performed with microplates.

Science.gov (United States)

Ackerman, S B; Kelley, E A

1983-03-01

The performance of a fiberoptic probe colorimeter (model PC800; Brinkmann Instruments, Inc., Westbury, N.Y.) for quantitating enzymatic or colorimetric assays in 96-well microtiter plates was compared with the performances of a spectrophotometer (model 240; Gilford Instrument Laboratories, Inc., Oberlin, Ohio) and a commercially available enzyme immunoassay reader (model MR590; Dynatech Laboratories, Inc., Alexandria, Va.). Alkaline phosphatase-p-nitrophenyl phosphate in 3 M NaOH was used as the chromophore source. Six types of plates were evaluated for use with the probe colorimeter; they generated reproducibility values (100% coefficient of variation) ranging from 91 to 98% when one individual made 24 independent measurements on the same dilution of chromophore on each plate. Eleven individuals each performed 24 measurements with the colorimeter on either a visually light (absorbance of 0.10 at 420 nm) or a dark (absorbance of 0.80 at 420 nm) dilution of chromophore; reproducibilities averaged 87% for the light dilution and 97% for the dark dilution. When one individual measured the same chromophore sample at least 20 times in the colorimeter, in the spectrophotometer or in the enzyme immunoassay reader, reproducibility for each instrument was greater than 99%. Measurements of a dilution series of chromophore in a fixed volume indicated that the optical responses of each instrument were linear in a range of 0.05 to 1.10 absorbance units.

45 CFR 96.136 - Independent peer review.

Science.gov (United States)

2010-10-01

... influence the quality of the services provided. (d) As part of the independent peer review, the reviewers... 45 Public Welfare 1 2010-10-01 2010-10-01 false Independent peer review. 96.136 Section 96.136... Abuse Prevention and Treatment Block Grant § 96.136 Independent peer review. (a) The State shall for the...

Emergência e crescimento inicial de plântulas de albízia submetidas à irrigação com água salina

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Mayky F. P. de Lima

2015-02-01

Full Text Available Objetivou-se, com este trabalho, avaliar o efeito de diferentes concentrações de sais na água de irrigação sob a emergência e o crescimento inicial de plântulas de albízia (Albizia lebbeck (L. Benth.. O experimento foi desenvolvido em casa de vegetação do departamento de Ciências Vegetais da Universidade Federal Rural do Semiárido, Mossoró, RN, usando-se o delineamento inteiramente casualizado, com seis tratamentos e quatro repetições, com 24 sementes em cada parcela. Os tratamentos foram constituídos das concentrações salinas 0,68 (água de abastecimento; 1,5; 3,0; 4,5; 6,0 e 7,5 dS m-1, obtidas através da adição de NaCl em água. As variáveis avaliadas foram: porcentagem de emergência de plântulas, índice de velocidade de emergência, comprimento da parte aérea, comprimento de raiz, número de folíolos, condutividade elétrica acumulada do substrato, índice de conteúdo de clorofila, massa seca da raiz, do caule, das folhas e total. O aumento da salinidade na água de irrigação interfere negativamente na emergência e no crescimento inicial de plântulas de albízia, sendo recomendado água com concentração salina até 1,5 dS m-1, caracterizando esta espécie como glicófita, sensível à salinidade.

Development of an automatic high-throughput assay for tetracycline determination by using Eu2O3 nanoparticles and dry-reagent technology.

Science.gov (United States)

Aguilar-Vázquez, L; Aguilar-Caballos, M P; Gómez-Hens, A

2014-02-01

The usefulness of europium oxide nanoparticles (Eu2O3 NPs) as analytical reagent for the direct determination of organic compounds is described for the first time. Tetracycline, which forms a luminescent chelate with europium, has been chosen as a model analyte. Dry reagent chemistry is used in a 96-well format, which considerably speeds up the determination and contributes to its automation. The NPs are immobilized onto polystyrene wells by adding a volume of a Eu2O3 NP dispersion in 2-propanol to each well and drying in an oven until they dry completely. At the moment of analysis, a standard or sample volume (200 μL) in the appropriate medium is added, and the mixture shaken for 15 min at 37°C. The method allows the determination of tetracycline in the range 20-1000 ng mL(-1), with a detection limit of 8 ng mL(-1). The inter-assay and intra-assay precision, which were assayed at two different tetracycline concentrations and expressed as relative standard deviation, were in the ranges of 6.5-8.2% and 9.2-12.7%, respectively. The study of the selectivity of the system showed that the method is adequate for tetracycline determination in agri-food samples, since most of antibiotics assayed did not interfere the determination. Only other tetracycline antibiotics provided luminescent signal when reacting to Eu2O3 NPs. The method has been applied to the determination of tetracycline in calf urine and in honey samples obtaining recovery values in the ranges of 85.0-110.0% and 99.7-116.7%, respectively. © 2013 Published by Elsevier B.V.

Aproximações sobre prescrições e práticas corporais nos Grupos escolares de Pirapora, Januária e Salinas: a educação dos corpos sertanejos - (1906-1927 * Approaches about prescriptions and corporal practices in schools in Pirapora, Januária and Salinas

Directory of Open Access Journals (Sweden)

ELISÂNGELA CHAVES

2012-05-01

Full Text Available Normal 0 21 false false false PT-BR X-NONE X-NONE MicrosoftInternetExplorer4 /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabela normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:14.0pt; font-family:"Times New Roman","serif"; mso-fareast-font-family:Calibri; mso-fareast-theme-font:minor-latin; mso-fareast-language:EN-US;} O texto analisa a proposta da educação corporal de grupos escolares localizados no norte de Minas Gerais no período de 1906 a 1927, notadamente nos municípios de Pirapora, Januária e Salinas. A pesquisa tomou como fonte principal relatos orais e demais documentos diretamente relacionados com a memória escolar para compreender como as atividades físicas foram percebidas como modeladoras de hábitos e atitudes, em um momento histórico em que Brasil prezava pelo discurso modernizador. Palavras-chave: Educação corporal – Memória – Desenvolvimento.   Abstract: The text analyses the proposal of the corporal education in schools in the north of Minas Gerais during 1906 and 1927, especially in Pirapora, januária and Salinas. As main source, the research made use of oral reports and other documents directly related to school memory to understand how physical activities were realized as habit and attitude-forming, in a historic moment in which Brazil used to cherish the modernizing speech. Keywords: Corporal education – Memory – Depevolpment.

[Teacher enhancement at Supercomputing `96

Energy Technology Data Exchange (ETDEWEB)

NONE

1998-02-13

The SC`96 Education Program provided a three-day professional development experience for middle and high school science, mathematics, and computer technology teachers. The program theme was Computers at Work in the Classroom, and a majority of the sessions were presented by classroom teachers who have had several years experience in using these technologies with their students. The teachers who attended the program were introduced to classroom applications of computing and networking technologies and were provided to the greatest extent possible with lesson plans, sample problems, and other resources that could immediately be used in their own classrooms. The attached At a Glance Schedule and Session Abstracts describes in detail the three-day SC`96 Education Program. Also included is the SC`96 Education Program evaluation report and the financial report.

Upregulation of miR-96 enhances cellular proliferation of prostate cancer cells through FOXO1.

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Benedikta S Haflidadóttir

Full Text Available Aberrant expression of miR-96 in prostate cancer has previously been reported. However, the role and mechanism of action of miR-96 in prostate cancer has not been determined. In this study, the diagnostic and prognostic properties of miR-96 expression levels were investigated by qRT-PCR in two well documented prostate cancer cohorts. The miR-96 expression was found to be significantly higher in prostate cancer patients and correlate with WHO grade, and decreased overall survival time; patients with low levels of miR-96 lived 1.5 years longer than patients with high miR-96 levels. The therapeutic potential was further investigated in vitro, showing that ectopic levels of miR-96 enhances growth and cellular proliferation in prostate cancer cells, implying that miR-96 has oncogenic properties in this setting. We demonstrate that miR-96 expression decreases the transcript and protein levels of FOXO1 by binding to one of two predicted binding sites in the FOXO1 3'UTR sequence. Blocking this binding site completely inhibited the growth enhancement conveyed by miR-96. This finding was corroborated in a large external prostate cancer patient cohort where miR-96 expression inversely correlated to FOXO1 expression. Taken together these findings indicate that miR-96 plays a key role in prostate cancer cellular proliferation and can enhance prostate cancer progression. This knowledge might be utilized for the development of novel therapeutic tools for prostate cancer.

High-throughput screening of cellulase F mutants from multiplexed plasmid sets using an automated plate assay on a functional proteomic robotic workcell

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Qureshi Nasib

2006-05-01

Full Text Available Abstract Background The field of plasmid-based functional proteomics requires the rapid assay of proteins expressed from plasmid libraries. Automation is essential since large sets of mutant open reading frames are being cloned for evaluation. To date no integrated automated platform is available to carry out the entire process including production of plasmid libraries, expression of cloned genes, and functional testing of expressed proteins. Results We used a functional proteomic assay in a multiplexed setting on an integrated plasmid-based robotic workcell for high-throughput screening of mutants of cellulase F, an endoglucanase from the anaerobic fungus Orpinomyces PC-2. This allowed us to identify plasmids containing optimized clones expressing mutants with improved activity at lower pH. A plasmid library of mutagenized clones of the celF gene with targeted variations in the last four codons was constructed by site-directed PCR mutagenesis and transformed into Escherichia coli. A robotic picker integrated into the workcell was used to inoculate medium in a 96-well deep well plate, combining the transformants into a multiplexed set in each well, and the plate was incubated on the workcell. Plasmids were prepared from the multiplexed culture on the liquid handler component of the workcell and used for in vitro transcription/translation. The multiplexed expressed recombinant proteins were screened for improved activity and stability in an azo-carboxymethylcellulose plate assay. The multiplexed wells containing mutants with improved activity were identified and linked back to the corresponding multiplexed cultures stored in glycerol. Spread plates were prepared from the glycerol stocks and the workcell was used to pick single colonies from the spread plates, prepare plasmid, produce recombinant protein, and assay for activity. The screening assay and subsequent deconvolution of the multiplexed wells resulted in identification of improved Cel

Efeito alelopático e toxicidade frente à Artemia salina Leach dos extatos do fruto de Euterpe edulis Martius Allelopathic effects and toxicity against Artemia salina Leach of extracts of the fruit of Euterpe edulis Martius

Directory of Open Access Journals (Sweden)

Cristina Peitz de Lima

2011-06-01

Full Text Available Alelopatia é um processo envolvendo metabólitos secundários produzidos por plantas que infl uenciam o crescimento e desenvolvimento de sistemas agrícolas. Devido à toxicidade dos herbicidas sintéticos para o meio ambiente e para a saúde humana tem-se aumentado o interesse na exploração da alelopatia como uma alternativa para o controle de plantas daninhas. O presente trabalho avaliou efeito dos extratos dos frutos de Euterpe edulis Martius sobre o desenvolvimento de cipselas e plântulas de Lactuca sativa Linné, foram determinados o índice de velocidade de germinação, o crescimento da radícula e do hipocótilo. Para a avaliação da toxicidade dos extratos foi realizado o ensaio de toxicidade frente ao microcrustáceo Artemia salina Leach determinando-se a CL50 e percentual de mortalidade. A fração remanescente demonstrou efeito alelopático, pois todas as concentrações alteraram os valores do índice de velocidade de germinação e as concentrações de 0,2 e 0,4 mg inibiram tanto o crescimento da radícula quanto o crescimento do hipocótilo. No ensaio de toxicidade todos os extratos apresentaram CL50 superior a 1000 ppm e 0% de mortalidade das artemias, indicando a não toxicidade dos extratos.Allelopathy is a process involving secondary metabolites produced by plants that influence growth and development of agricultural systems. Because of the toxicity of synthetic herbicides to the environment and human health, there has been increased interest in exploiting allelopathy as an alternative for weed control. This study evaluated the eff ect of extracts of Euterpe edulis Martius fruits on the development of cypselae and seedlings of Lactuca sativa Linné; the germination speed index, radicle and hypocotyl growth were determined. To evaluate the toxicity of the extracts the toxicity test against Artemia salina Leach was used, where the LC50 and mortality rate were determined. Th e remaining fraction showed allelopathic effect

Towards non-invasive 3D hepatotoxicity assays with optical coherence phase microscopy

Science.gov (United States)

Nelson, Leonard J.; Koulovasilopoulos, Andreas; Treskes, Philipp; Hayes, Peter C.; Plevris, John N.; Bagnaninchi, Pierre O.

2015-03-01

Three-dimensional tissue-engineered models are increasingly recognised as more physiologically-relevant than standard 2D cell culture for pre-clinical drug toxicity testing. However, many types of conventional toxicity assays are incompatible with dense 3D tissues. This study investigated the use of optical coherence phase microscopy (OCPM) as a novel approach to assess cell death in 3D tissue culture. For 3D micro-spheroid formation Human hepatic C3A cells were encapsulated in hyaluronic acid gels and cultured in 100μl MEME/10%FBS in 96-well plates. After spheroid formation the 3D liver constructs were exposed to acetaminophen on culture day 8. Acetaminophen hepatotoxicity in 3D cultures was evaluated using standard biochemical assays. An inverted OCPM in common path configuration was developed with a Callisto OCT engine (Thorlabs), centred at 930nm and a custom scanning head. Intensity data were used to perform in-depth microstructural imaging. In addition, phase fluctuations were measured by collecting several successive B scans at the same location, and statistics on the first time derivative of the phase, i.e. time fluctuations, were analysed over the acquisition time interval to retrieve overall cell viability. OCPM intensity (cell cluster size) and phase fluctuation statistics were directly compared with biochemical assays. In this study, we investigated optical coherence phase tomography to assess cell death in a 3d liver model after exposure to a prototypical hepatotoxin, acetaminophen. We showed that OCPM has the potential to assess noninvasively and label-free drug toxicity in 3D tissue models.

31 CFR 9.6 - Confidential information.

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2010-07-01

... 9.6 Money and Finance: Treasury Office of the Secretary of the Treasury EFFECTS OF IMPORTED ARTICLES ON THE NATIONAL SECURITY § 9.6 Confidential information. Information submitted in confidence which... marked “Business Confidential.” [40 FR 50717, Oct. 31, 1975] ...

Real-time pcr (qpcr) assay for rhizoctonia solani anastomoses group ag2-2 iiib

International Nuclear Information System (INIS)

Abbas, S.J.; Ahmad, B.

2014-01-01

Rhizoctonia solani anastomosis group AG2-2 IIIB is a severe sugar beet and maize pathogen. It causes crown and root rot disease which leads to yield losses world-wide. The soil-borne pathogen is difficult to detect and quantify by conventional methods. We developed a real-time PCR (qPCR) assay for the quantification of genomic DNA of Rhizoctonia solani AG2-2 IIIB based on the ITS region of rDNA genes. The limit of quantification of the assay is 1.8 pg genomic DNA. The amplification efficiency was 96.4. The assay will be helpful in the diagnoses of Rhizoctonia solani infection of sugar beet and maize roots and in the quantification of R. solani AG2-2 IIIB inoculum in plant debris and soil. (author)

Operating characteristics of a qualitative troponin assay for the diagnosis of acute coronary syndrome.

Science.gov (United States)

Farsi, Davood; Pishbin, Elham; Abbasi, Saeed; Hafezimoghadam, Peyman; Fathi, Marzieh; Zare, Mohammad Amin

2013-04-01

The troponin I serum level is widely used in acute coronary syndrome patients for their classification. The qualitative assay is faster and more available than the quantitative assay. The objective was to determine the operating characteristics of a qualitative troponin I assay compared with a quantitative method. This is a prospective observational study and patients suspected to have acute coronary syndrome were enrolled. A rapid troponin I test and a quantitative assay were carried out for each patient on arrival and 6 h after admission. A total of 262 patients were enrolled. The degree of agreement between the second rapid qualitative and quantitative troponin I was excellent (κ=0.946; 95% confidence interval, 0.903-0.989). The sensitivity, specificity, negative predictive value, and positive predictive value of the rapid qualitative troponin I test were 92.6, 100, 96.8, and 100%, respectively. In conclusion, this study reveals an excellent agreement between quantitative and qualitative bedside assays 6 h after admission in a sample of Iranian patients in the emergency department.

Un carnaval para el yo lésbico: Los cuentos de Gilda Salinas

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Elena Madrigal

2011-01-01

Full Text Available Gilda Salinas rompe las unidades cuentísticas tradicionales al hacer de cada texto de Del destete al desempance. Cuentos lésbicos y un colado un episodio en la vida de una lesbiana que, con “voz propia”, narra sus peripecias nocturnas por la ciudad de México, de los años setenta a la actualidad. Al ubicar las acciones en sitios de diversión lésbica que no existen más, la voz, cual cronista, rescata un ámbito de la vida homosexual a la vez que fija bromas y códigos lingüísticos que por su origen oral hubieran corrido el riesgo de perderse con las generaciones que los animaron. Sus estrategias carnavalizan el tema lésbico a la vez que legitiman la validez de las búsquedas expresivas y de comportamiento del ser lesbiana en un tiempo y en un lugar. Su aportación en los planos del género literario y de la formación de constantes en la narrativa homosexual mexicana indudablemente enriquece los medios expresivos, la visibilidad, la diversidad, las vidas y las historias de la y las lesbianas.

Comparison of Six Automated Treponema-Specific Antibody Assays.

Science.gov (United States)

Park, Borae G; Yoon, Jihoon G; Rim, John Hoon; Lee, Anna; Kim, Hyon-Suk

2016-01-01

Six different Treponema (TP)-specific immunoassays were compared to the fluorescent treponemal antibody absorption (FTA-ABS) test. A total of 615 samples were tested. The overall percent agreement, analytical sensitivity, and analytical specificity of each assay compared to the FTA-ABS test were as follows: Architect Syphilis TP, 99.2%, 96.8%, and 100%; Cobas Syphilis, 99.8%, 99.4%, and 100%; ADVIA Centaur Syphilis, 99.8%, 99.4%, and 100%; HISCL Anti-TP assay kit, 99.7%, 98.7%, and 100%; Immunoticles Auto3 TP, 99.0%, 97.5%, and 99.6%; Mediace TPLA, 98.0%, 98.1%, and 98.0%. All results that were discrepant between the TP-specific assays were associated with samples from noninfectious cases (11 immunoassay false positives and 7 from previous syphilis cases). Our study demonstrated that TP-specific immunoassays generally showed high sensitivities, specificities, and percentages of agreement compared to FTA-ABS, with rare cases of false-positive or false-negative results. Therefore, most TP-specific immunoassays are acceptable for use in screening for syphilis. However, it is important to perform a thorough review of a patient's clinical and treatment history for interpreting the results of syphilis serology. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

27 CFR 21.96 - Ammonia, aqueous.

Science.gov (United States)

2010-04-01

... 27 Alcohol, Tobacco Products and Firearms 1 2010-04-01 2010-04-01 false Ammonia, aqueous. 21.96 Section 21.96 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE BUREAU, DEPARTMENT... Ammonia, aqueous. (a) Alkalinity. Strongly alkaline to litmus. (b) Ammonia content. 27 to 30 percent by...

Avaliação da hiperresponsividade brônquica à solução salina hipertônica em crianças e adolescentes Bronchial hyperresponsiveness to hypertonic saline challenge in children and adolescents

Directory of Open Access Journals (Sweden)

Paulo Kussek

2006-06-01

Full Text Available OBJETIVO: Avaliar a hiperresponsividade brônquica à solução salina hipertônica a 4,5% como método alternativo a outros agentes broncoconstritores e sua relação com a sensibilização alérgica do paciente. MÉTODOS: Estudo transversal, experimental, com 85 indivíduos assim distribuídos: 45 no grupo de asmáticos e 17 no grupo controle não asmáticos e não alérgicos, que completaram o teste. Para nebulizar a solução salina hipertônica foi utilizado um nebulizador ultra-sônico de grande volume, sucessivamente durante 0,5, 1, 2, 4 e 8 minutos até haver queda > 15% em relação ao volume expiratório forçado no primeiro segundo basal. A dosagem de imunoglobulina E específica ao Dermatophagoides pteronyssinus por ImmunoCap foi considerada positiva quando > 0,35 kU/L. RESULTADOS: No grupo de asmáticos, 36 apresentaram queda média do volume expiratório forçado no primeiro segundo de 27,4% após nebulização de solução salina hipertônica. Nenhum do grupo controle (imunoglobulina E OBJECTIVE: To assess airway hyperresponsiveness to 4.5% hypertonic saline solution in comparison to that obtained through challenge with other bronchoconstriction agents and in relation to patient allergic sensitization. METHODS: A cross-sectional, experimental study was conducted, initially involving 85 subjects. After exclusions, the final sample consisted of 62 patients, divided into two groups: a study group of those with asthma (n = 45 and a control group of those with no asthma or allergies (n = 17. Hypertonic saline was nebulized using an ultrasonic nebulizer and administered successively for 0.5, 1, 2, 4 and 8 minutes until a drop in forced expiratory volume in one second of = 15% was achieved in relation to the baseline value. The level of specific immunoglobulin E to Dermatophagoides pteronyssinus level was determined by ImmunoCAP assay and was considered positive when > 0.35 kU/L. RESULTS: In the 36 asthma group subjects presenting a

Loop-mediated isothermal amplification assay for rapid and sensitive detection of sheep pox and goat pox viruses in clinical samples.

Science.gov (United States)

Venkatesan, G; Balamurugan, V; Bhanuprakash, V; Singh, R K; Pandey, A B

2016-06-01

A Loop-mediated isothermal amplification (LAMP) assay targeting the highly conserved DNA polymerase gene of capripox virus genome was developed and evaluated for rapid detection of sheep pox and goat pox viruses. The optimized LAMP assay is found specific and sensitive for amplification of target DNA with a diagnostic sensitivity and specificity of 96.6% and 100% respectively compared to quantitative PCR. The detection rate of LAMP, PCR and Q-PCR assays is found to be 81.5%, 67% and 83% respectively. This LAMP assay has the potential for rapid clinical diagnosis and surveillance of sheep pox and goat pox in field diagnostic laboratories. Copyright © 2016 Elsevier Ltd. All rights reserved.

[Comparison of the performance of the ECLusys anti-HCV reagent with the Lumipulse f and HISCL 2000-i HCVAb assays].

Science.gov (United States)

Sugiura, Aya; Iwahara, Kunihiro; Suga, Yasuyuki; Uchiyama, Sachinori; Maekawa, Masato

2012-09-01

We compared the ECLusys Anti-HCV (ECL) reagent to the Lumipulse f (LPf) and HISCL (HIS) HCV assays. In a correlation test using 210 routine clinical specimens measured using the Lumipulse method (96 positive and 114 negative), most of the results were consistent for all specimens. In a dilution sensitivity test using three different routine positive specimens, the ECL assay enabled detection at higher levels of sensitivity than either the LPf or the HIS assay. Moreover, when the distribution of the cut-off index (C.O.I.) values of the routine LPf negative specimens were compared to those on the ECL and HIS assays, it was found that on the ECL assay, most of the specimens had cut-off index values < 0.1, indicating a more clear-cut distribution. In a specificity test using high RF positive specimens(n = 33), pregnancy specimens (n = 35), cytomegalovirus (CMV) antibody positive specimens (n = 36), and high M protein positive specimens (n = 21), the ECL assay yielded positive results for a CMV antibody positive specimen and three high M protein positive specimens. Further testing using samples from the same patients collected on different days than these four samples resulted in a second positive result for the CMV positive specimen, and single antigen measurement yielded a Core/NS3 positive result, as well, suggesting past infection. However, since negative results were obtained for the three M protein positive specimens, the possibility of this being a ECLusys non-specific reaction could not be ruled out. The above results confirmed that the ECL assay provides superior fundamental performance, and possesses test performance nearly identical to that of the existing measurement methods that are widely used at a large number of facilities, and would therefore be a suitable assay for use in routine HCV antibody screening.

Semi-automated limit-dilution assay and clonal expansion of all T-cell precursors of cytotoxic lymphocytes

Energy Technology Data Exchange (ETDEWEB)

Wilson, A.; Chen, W.F.; Scollay, R.; Shortman, K. (Walter and Eliza Hall Inst. of Medical Research, Parkville (Australia))

1982-08-13

A limit-dilution microculture system is described, where almost all precursor T cells of the cytotoxic lineage (CTL-p) develop into extended clones of cytotoxic T cells (CTL), which are then detected with a new radio-autographic /sup 111/In-release assay. The principle is to polyclonally activate all T cells with concanavalin A, to expand the resultant clones over an 8-9 day period in cultures saturated with growth factors, then to detect all clones with cytotoxic function by phytohaemagglutinin mediated lysis of P815 tumour cells. The key variables for obtaining high cloning efficiency are the use of flat-bottomed 96-well culture trays, the use of appropriately irradiated spleen filler cells, and the inclusion of a T-cell growth factor supplement. Cultures are set up at input levels of around one T cell per well. Forty percent of T cells then form CTL clones readily detected by the cytotoxic assay. The lytic activity of the average clone is equivalent to 3000 CTL, but clone size appears to be much larger. The precursor cells are predominantly if not entirely from the Lyt 2/sup +/ T-cell subclass and almost all cells of this subclass form cytolytic clones. Analysis of the frequency of positive cultures shows a good fit to the expected Poisson distribution, with no evidence of the CTL-p frequency estimates being distorted by helper or suppressor effects.

Semi-automated limit-dilution assay and clonal expansion of all T-cell precursors of cytotoxic lymphocytes

International Nuclear Information System (INIS)

Wilson, A.; Chen, W.-F.; Scollay, R.; Shortman, K.

1982-01-01

A limit-dilution microculture system is described, where almost all precursor T cells of the cytotoxic lineage (CTL-p) develop into extended clones of cytotoxic T cells (CTL), which are then detected with a new radio-autographic 111 In-release assay. The principle is to polyclonally activate all T cells with concanavalin A, to expand the resultant clones over an 8-9 day period in cultures saturated with growth factors, then to detect all clones with cytotoxic function by phytohaemagglutinin mediated lysis of P815 tumour cells. The key variables for obtaining high cloning efficiency are the use of flat-bottomed 96-well culture trays, the use of appropriately irradiated spleen filler cells, and the inclusion of a T-cell growth factor supplement. Cultures are set up at input levels of around one T cell per well. Forty percent of T cells then form CTL clones readily detected by the cytotoxic assay. The lytic activity of the average clone is equivalent to 3000 CTL, but clone size appears to be much larger. The precursor cells are predominantly if not entirely from the Lyt 2 + T-cell subclass and almost all cells of this subclass form cytolytic clones. Analysis of the frequency of positive cultures shows a good fit to the expected Poisson distribution, with no evidence of the CTL-p frequency estimates being distorted by helper or suppressor effects. (Auth.)

29 CFR 96.12 - Audit requirements.

Science.gov (United States)

2010-07-01

.... (b) The audit requirements contained in 29 CFR part 99 shall be followed for audits of all fiscal... 29 Labor 1 2010-07-01 2010-07-01 true Audit requirements. 96.12 Section 96.12 Labor Office of the Secretary of Labor AUDIT REQUIREMENTS FOR GRANTS, CONTRACTS, AND OTHER AGREEMENTS Audits of States, Local...

Effect of the addition of Lactobacillus delbrueckii subsp. delbrueckii on the gut microbiota composition and contribution to the well-being of European sea bass (Dicentrarchus labrax, L.)

OpenAIRE

Silvi, Stefania; Nardi, Miria; Sulpizio, Roberto; Orpianesi, Carla; Caggiano, Massimo; Carnevali, Oliana; Cresci, Alberto

2011-01-01

The present study aimed to test the effects of probiotic treatment on gut microbiota and the contribution to the well-being of European sea bass (Dicentrarchus labrax, L.). A bacterial strain of Lactobacillus delbrueckii subsp. delbrueckii (AS13B), isolated from adult European sea bass gut, was administered during sea bass development using Brachionus plicatilis and/or Artemia salina as carriers. The effective strain colonization and modulation of the gut microbiota, the mortality and the cor...

Chemosensitivity testing of primary human renal cell carcinoma by a tetrazolium based microculture assay (MTT).

Science.gov (United States)

Mickisch, G; Fajta, S; Keilhauer, G; Schlick, E; Tschada, R; Alken, P

1990-01-01

MTT staining procedures have been used in chemosensitivity testing of established cell lines of human and other sources as well as of human leukaemias, but only limited information on its application in primary solid human tumors is presently available. We have evaluated MTT staining in primary human Renal Cell Carcinomas (RCCs), studied various factors interfering with the optimal use, and finally applied it in subsequent chemosensitivity testing. The method depends on the conversion of a water-soluble tetrazolium salt (MTT) to a purple colored formazan precipitate, a reaction effected by enzymes active only in living cells. Single cell suspensions of RCCs were obtained either by enzymatic dispersion or by mechanical dissagregation, filtered through gauze, and purified by Ficoll density centrifugation. Tests were carried out in 96-well microculture plates. 10(4) viable tumor cells per well at 4 h incubation time with 20 micrograms MTT/100 microliters total medium volume yielded best results. Formazan crystals were dissolved with DMSO, and the plates were immediately measured on a microculture plate reader at 540 nm. Under these criteria, linearity of the system could be demonstrated. For chemosensitivity testing, cells were continuously exposed to a number of drugs prior to the MTT staining procedure. Reproducibility of results was assessed and confirmed by culturing RCCs in flasks additionally, resubmitting them after 1, 2, and 4 weeks to the MTT assay. We conclude that the semiautomated MTT assay offers a valid, rapid, reliable and simple method to determine the degree of chemoresistance in primary human RCCs.

Development of a rapid loop-mediated isothermal amplification assay for diagnosis and assessment of cure of Leishmania infection.

Science.gov (United States)

Verma, Sandeep; Singh, Ruchi; Sharma, Vanila; Bumb, Ram Avtar; Negi, Narendra Singh; Ramesh, V; Salotra, Poonam

2017-03-23

Leishmaniasis is a spectrum of diseases with great relevance to public health. Conventional diagnostic methods are time consuming, needing trained personnel. A robust, rapid and cost effective diagnostic test is warranted for on-time diagnosis and field application. We have developed a loop mediated isothermal amplification (LAMP) assay with primers (n = 6) based on Leishmania donovani kDNA for detection of Leishmania infection, using a closed tube to prevent cross-contamination. The assay was used to detect Leishmania infection in biological samples obtained from patients of visceral leishmaniasis (VL), post kala-azar dermal leishmaniasis (PKDL) and cutaneous leishmaniasis (CL). The assay was positive for L. donovani, L. tropica and L. major parasites, with the highest sensitivity towards L. donovani (1 fg DNA). The high sensitivity of the assay for detection of L. donovani was reflected in its ability to detect parasite DNA within 30 min of amplification time with a threshold detection limit of ≥25 copies per reaction. The assay detected parasite in 64 of 66 VL blood samples (sensitivity, 96.9%; 95% CI: 89.6-99.2%), 15 of 15 VL bone marrow aspirate samples (sensitivity, 100%; 95% CI:79.6-100%), 65 of 67 PKDL tissue biopsy samples (sensitivity, 97%; 95% CI:89.7-99.2%). The assay was evaluated in a few cases of CL wherein it was found positive in 8 of 10 tissue biopsies (sensitivity, 80%; 95% CI: 49-94.3%). The assay was negative in all control blood (n = 76) and tissue biopsy (n = 24) samples (specificity, 100%; 95% CI: 96.3-100%). Further, the assay was evaluated for its utility in assessment of cure in treated VL and PKDL patients. The assay detected parasite DNA in 2 of 20VL blood samples and 2 of 21 PKDL tissue samples. Out of 4 cases that were positive for parasite DNA at post treatment stage, 2 patients (1VL and 1 PKDL) returned with relapse. The study demonstrated a Leishmania genus specific closed tube LAMP assay for reliable and rapid

Assays for calcitonin receptors

International Nuclear Information System (INIS)

Teitelbaum, A.P.; Nissenson, R.A.; Arnaud, C.D.

1985-01-01

The assays for calcitonin receptors described focus on their use in the study of the well-established target organs for calcitonin, bone and kidney. The radioligand used in virtually all calcitonin binding studies is 125 I-labelled salmon calcitonin. The lack of methionine residues in this peptide permits the use of chloramine-T for the iodination reaction. Binding assays are described for intact bone, skeletal plasma membranes, renal plasma membranes, and primary kidney cell cultures of rats. Studies on calcitonin metabolism in laboratory animals and regulation of calcitonin receptors are reviewed

Development and Validation of a Novel Dual Luciferase Reporter Gene Assay to Quantify Ebola Virus VP24 Inhibition of IFN Signaling

Directory of Open Access Journals (Sweden)

Elisa Fanunza

2018-02-01

Full Text Available The interferon (IFN system is the first line of defense against viral infections. Evasion of IFN signaling by Ebola viral protein 24 (VP24 is a critical event in the pathogenesis of the infection and, hence, VP24 is a potential target for drug development. Since no drugs target VP24, the identification of molecules able to inhibit VP24, restoring and possibly enhancing the IFN response, is a goal of concern. Accordingly, we developed a dual signal firefly and Renilla luciferase cell-based drug screening assay able to quantify IFN-mediated induction of Interferon Stimulated Genes (ISGs and its inhibition by VP24. Human Embryonic Kidney 293T (HEK293T cells were transiently transfected with a luciferase reporter gene construct driven by the promoter of ISGs, Interferon-Stimulated Response Element (ISRE. Stimulation of cells with IFN-α activated the IFN cascade leading to the expression of ISRE. Cotransfection of cells with a plasmid expressing VP24 cloned from a virus isolated during the last 2014 outbreak led to the inhibition of ISRE transcription, quantified by a luminescent signal. To adapt this system to test a large number of compounds, we performed it in 96-well plates; optimized the assay analyzing different parameters; and validated the system by calculating the Z′- and Z-factor, which showed values of 0.62 and 0.53 for IFN-α stimulation assay and VP24 inhibition assay, respectively, indicative of robust assay performance.

Effects of Light and Temperature on Fatty Acid Production in Nannochloropsis Salina

Energy Technology Data Exchange (ETDEWEB)

Van Wagenen, Jonathan M.; Miller, Tyler W.; Hobbs, Samuel J.; Hook, Paul W.; Crowe, Braden J.; Huesemann, Michael H.

2012-03-12

Accurate prediction of algal biofuel yield will require empirical determination of physiological responses to the climate, particularly light and temperature. One strain of interest, Nannochloropsis salina, was subjected to ranges of light intensity (5-850 {mu}mol m{sup -2} s{sup -1}) and temperature (13-40 C); exponential growth rate, total fatty acids (TFA) and fatty acid composition were measured. The maximum acclimated growth rate was 1.3 day{sup -1} at 23 C and 250 {mu}mol m{sup -2} s{sup -1}. Fatty acids were detected by gas chromatography with flame ionization detection (GC-FID) after transesterification to corresponding fatty acid methyl esters (FAME). A sharp increase in TFA containing elevated palmitic acid (C16:0) and palmitoleic acid (C16:1) during exponential growth at high light was observed, indicating likely triacylglycerol accumulation due to photo-oxidative stress. Lower light resulted in increases in the relative abundance of unsaturated fatty acids; in thin cultures, increases were observed in palmitoleic and eicosapentaenoeic acids (C20:5{omega}3). As cultures aged and the effective light intensity per cell converged to very low levels, fatty acid profiles became more similar and there was a notable increase of oleic acid (C18:1{omega}9). The amount of unsaturated fatty acids was inversely proportional to temperature, demonstrating physiological adaptations to increase membrane fluidity. This data will improve prediction of fatty acid characteristics and yields relevant to biofuel production.

«Cada página lograda es una letra al más allá». Estudio de la correspondencia Américo Castro-Pedro Salinas

OpenAIRE

Zamarreño Méndez, Fabio

2016-01-01

Este trabajo explora la correspondencia entre Pedro Salinas y Américo Castro a partir de una edición, que, a causa de la legislación vigente respecto de los derechos de autor, no puede ser publicada. Por tanto, aquí encontramos el estudio introductorio, que analiza ese epistolario desde todas las perspectivas: comenzando con un repaso teórico del subgénero epistolar, pronto se explican las dificultades en torno a la reconstrucción cronológica del epistolario. Temáticamente, se analizan difere...

A high-throughput fluorescence-based assay system for appetite-regulating gene and drug screening.

Directory of Open Access Journals (Sweden)

Yasuhito Shimada

Full Text Available The increasing number of people suffering from metabolic syndrome and obesity is becoming a serious problem not only in developed countries, but also in developing countries. However, there are few agents currently approved for the treatment of obesity. Those that are available are mainly appetite suppressants and gastrointestinal fat blockers. We have developed a simple and rapid method for the measurement of the feeding volume of Danio rerio (zebrafish. This assay can be used to screen appetite suppressants and enhancers. In this study, zebrafish were fed viable paramecia that were fluorescently-labeled, and feeding volume was measured using a 96-well microplate reader. Gene expression analysis of brain-derived neurotrophic factor (bdnf, knockdown of appetite-regulating genes (neuropeptide Y, preproinsulin, melanocortin 4 receptor, agouti related protein, and cannabinoid receptor 1, and the administration of clinical appetite suppressants (fluoxetine, sibutramine, mazindol, phentermine, and rimonabant revealed the similarity among mechanisms regulating appetite in zebrafish and mammals. In combination with behavioral analysis, we were able to evaluate adverse effects on locomotor activities from gene knockdown and chemical treatments. In conclusion, we have developed an assay that uses zebrafish, which can be applied to high-throughput screening and target gene discovery for appetite suppressants and enhancers.

A micromethod for the assay of cellular secretory physiology: Application to rabbit parietal cells

International Nuclear Information System (INIS)

Adrian, T.E.; Goldenring, J.R.; Oddsdottir, M.; Zdon, M.J.; Zucker, K.A.; Lewis, J.J.; Modlin, I.M.

1989-01-01

A micromethod for investigating secretory physiology in isolated cells was evaluated. The method utilized a specially designed polycarbonate incubation chamber to provide constant oxygenation to cells incubating in a 96-well microtiter plate. Cells were rapidly separated from media by vacuum filtration. Isolated parietal cells were utilized to demonstrate the versatility of the method for assay of intracellular accumulation of [ 14 C]-aminopyrine, secretion of intrinsic factor into the medium, and assay of intracellular cAMP. Histamine stimulated the uptake of [ 14 C]aminopyrine and intrinsic factor secretion in a sustained and linear fashion. At the end of the 2-h period uptake of aminopyrine and secretion of intrinsic factor were increased 17- and 5-fold, respectively. This response to histamine was accompanied by a rapid and sustained 3-fold rise in intracellular cyclic AMP. In contrast, carbamylcholine caused a transient increase in [ 14 C]aminopyrine accumulation and intrinsic factor secretion which was most pronounced during the first 10 min and had almost ceased by 30 min. Carbamylcholine had no effect on intracellular cAMP levels. This new method, which can handle 400 replicates using parietal cells from the fundic mucosa of a single rabbit, is suitable for studying the time course of intracellular events which accompany general secretory processes

El turismo de naturaleza en espacios naturales. El caso del Parque Regional de las Salinas y Arenales de San Pedro del Pinatar

Directory of Open Access Journals (Sweden)

Gustavo A. Ballesteros Pelegrín

2014-01-01

Full Text Available Los espacios naturales atraen a turistas que buscan el contacto con la naturaleza, surgiendo nuevos productos turísticos. Surgen, de este modo, nuevos productos turísticos. En la oferta de turismo de naturaleza del Parque Regional de las Salinas y Arenales de San Pedro del Pinatar destaca: deportes en la naturaleza, aventura y ecoturismo. Este último se divide en: turismo ornitológico y fotográfico, que junto al tradicional de sol y playa y de salud, se ve favorecido por su situación geográfica, condiciones naturales y amplia oferta hotelera. Sin embargo, el turismo cultural basado en la tradición salinera y pesquera no ha sido desarrollado.

46 CFR 96.40-1 - Pilot boarding equipment.

Science.gov (United States)

2010-10-01

... 46 Shipping 4 2010-10-01 2010-10-01 false Pilot boarding equipment. 96.40-1 Section 96.40-1... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Pilot Boarding Equipment § 96.40-1 Pilot boarding... boat or other vessel. (b) Each vessel must have suitable pilot boarding equipment available for use on...

Potential use of an ultrasound antifouling technology as a ballast water treatment system

Science.gov (United States)

Estévez-Calvar, Noelia; Gambardella, Chiara; Miraglia, Francesco; Pavanello, Giovanni; Greco, Giuliano; Faimali, Marco; Garaventa, Francesca

2018-03-01

The aim of this study was to investigate, at a laboratory scale, the potentialities of an ultrasound-based treatment initially designed to eliminate fouling, as a ballast water treatment system. Therefore, early life stages of three different zooplanktonic species (Amphibalanus amphitrite, Brachionus plicatilis and Artemia salina) were exposed to ultrasound waves (20-22 kHz). The experimental set up included static assays with variations of time exposure (30 s, 60 s and 30 s on/60 s off/30 s on), material of tanks (stainless steel, galvanized steel and plastic) and position of the ultrasound source. Results showed that the treatment efficacy increased from 30 to 60 s and no differences were registered between 60 s-continuous exposure and pulse exposure. The highest efficacy was observed in Experiment I (metal-to-metal contact assay) with a mortality value of 93-95% for B. plicatilis and A. salina. It consisted of organisms located inside stainless steel tubes that were located in direct contact with the ultrasound source and treated for 60 s. Further, we found that, generally, A. amphitrite and B. plicatilis were the most resistant species to the ultrasound treatment whereas A. salina was the most sensitive. We further discuss that US may unlikely be used for commercial vessels, but may be used to treat ballast water in smaller ballast tanks as on board of mega yachts.

Expression of 9-cis-EPOXYCAROTENOID DIOXYGENASE4 Is Essential for Thermoinhibition of Lettuce Seed Germination but Not for Seed Development or Stress Tolerance[C][W

Science.gov (United States)

Huo, Heqiang; Dahal, Peetambar; Kunusoth, Keshavulu; McCallum, Claire M.; Bradford, Kent J.

2013-01-01

Thermoinhibition, or failure of seeds to germinate at warm temperatures, is common in lettuce (Lactuca sativa) cultivars. Using a recombinant inbred line population developed from a lettuce cultivar (Salinas) and thermotolerant Lactuca serriola accession UC96US23 (UC), we previously mapped a quantitative trait locus associated with thermoinhibition of germination to a genomic region containing a gene encoding a key regulated enzyme in abscisic acid (ABA) biosynthesis, 9-cis-EPOXYCAROTENOID DIOXYGENASE4 (NCED4). NCED4 from either Salinas or UC complements seeds of the Arabidopsis thaliana nced6-1 nced9-1 double mutant by restoring germination thermosensitivity, indicating that both NCED4 genes encode functional proteins. Transgenic expression of Salinas NCED4 in UC seeds resulted in thermoinhibition, whereas silencing of NCED4 in Salinas seeds led to loss of thermoinhibition. Mutations in NCED4 also alleviated thermoinhibition. NCED4 expression was elevated during late seed development but was not required for seed maturation. Heat but not water stress elevated NCED4 expression in leaves, while NCED2 and NCED3 exhibited the opposite responses. Silencing of NCED4 altered the expression of genes involved in ABA, gibberellin, and ethylene biosynthesis and signaling pathways. Together, these data demonstrate that NCED4 expression is required for thermoinhibition of lettuce seeds and that it may play additional roles in plant responses to elevated temperature. PMID:23503626

Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence.

Science.gov (United States)

Xie, Yicheng; Wahab, Laith; Gill, Jason J

2018-04-12

Bacteriophages, which are the natural predators of bacteria, have re-emerged as an attractive alternative to combat antibiotic resistant bacteria. Phages are highly specific at the species and strain level and measurement of the phage host range plays an important role in utilizing the phage as antimicrobials. The most common method for phage host range determination has been to spot phage lysates on soft agar overlays and observe plaque formation. In this study, a liquid culture-based assay was developed in a 96-well microtiter plate format to measure the phage host range and virulence for a collection of 15 Salmonella phages against a panel of 20 Salmonella strains representing 11 serovars. This method was compared to a traditional spot method. The majority of the host range results from two methods were in agreement including in cases where a bacterial strain was insensitive to the phage. Each method produced a false-negative result in 19/300 (6%) of the measured phage-host combinations when compared to the other method. The spot method tended to indicate greater phage sensitivity than the microtiter assay even though direct comparisons of the response magnitude between the two methods is difficult since they operate on different mechanisms. The microtiter plate assay was able to provide data on both the phage host range and virulence in greater resolution in a high-throughput format.

Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence

Directory of Open Access Journals (Sweden)

Yicheng Xie

2018-04-01

Full Text Available Bacteriophages, which are the natural predators of bacteria, have re-emerged as an attractive alternative to combat antibiotic resistant bacteria. Phages are highly specific at the species and strain level and measurement of the phage host range plays an important role in utilizing the phage as antimicrobials. The most common method for phage host range determination has been to spot phage lysates on soft agar overlays and observe plaque formation. In this study, a liquid culture-based assay was developed in a 96-well microtiter plate format to measure the phage host range and virulence for a collection of 15 Salmonella phages against a panel of 20 Salmonella strains representing 11 serovars. This method was compared to a traditional spot method. The majority of the host range results from two methods were in agreement including in cases where a bacterial strain was insensitive to the phage. Each method produced a false-negative result in 19/300 (6% of the measured phage-host combinations when compared to the other method. The spot method tended to indicate greater phage sensitivity than the microtiter assay even though direct comparisons of the response magnitude between the two methods is difficult since they operate on different mechanisms. The microtiter plate assay was able to provide data on both the phage host range and virulence in greater resolution in a high-throughput format.

The FOCON96 1.0 computer code

International Nuclear Information System (INIS)

Merle-Szeremeta, A.; Thomassin, A.

1999-01-01

The Institute of Protection and Nuclear Safety (I.P.S.N.) has developed a computer code, FOCON96 1.0 to calculate the dosimetric consequences of atmospheric radioactive releases from nuclear installations after several years of usual operation. This communication describes the principal characteristics of FOCON96 1.0 and its functionalities. The principal elements of a comparison between FOCON96 1.0 and PC-CREAM ( European computer code developed by the N.R.P.B. and answering the same criteria) are given here. (N.C.)

Draw your assay: Fabrication of low-cost paper-based diagnostic and multi-well test zones by drawing on a paper.

Science.gov (United States)

Oyola-Reynoso, Stephanie; Heim, Andrew P; Halbertsma-Black, Julian; Zhao, C; Tevis, Ian D; Çınar, Simge; Cademartiri, Rebecca; Liu, Xinyu; Bloch, Jean-Francis; Thuo, Martin M

2015-11-01

Interest in low-cost diagnostic devices has recently gained attention, in part due to the rising cost of healthcare and the need to serve populations in resource-limited settings. A major challenge in the development of such devices is the need for hydrophobic barriers to contain polar bio-fluid analytes. Key approaches in lowering the cost in diagnostics have centered on (i) development of low-cost fabrication techniques/processes, (ii) use of affordable materials, or, (iii) minimizing the need for high-tech tools. This communication describes a simple, low-cost, adaptable, and portable method for patterning paper and subsequent use of the patterned paper in diagnostic tests. Our approach generates hydrophobic regions using a ball-point pen filled with a hydrophobizing molecule suspended in a solvent carrier. An empty ball-point pen was filled with a solution of trichloro perfluoroalkyl silane in hexanes (or hexadecane), and the pen used to draw lines on Whatman® chromatography 1 paper. The drawn regions defined the test zones since the trichloro silane reacts with the paper to give a hydrophobic barrier. The formation of the hydrophobic barriers is reaction kinetic and diffusion-limited, ensuring well defined narrow barriers. We performed colorimetric glucose assays and enzyme-linked immuno-sorbent assay (ELISA) using the created test zones. To demonstrate the versatility of this approach, we fabricated multiple devices on a single piece of paper and demonstrated the reproducibility of assays on these devices. The overall cost of devices fabricated by drawing are relatively lower (

A Continuous, Fluorogenic Sirtuin 2 Deacylase Assay

DEFF Research Database (Denmark)

Galleano, Iacopo; Schiedel, Matthias; Jung, Manfred

2016-01-01

and kinetic insight regarding sirtuin inhibitors, it is important to have access to efficient assays. In this work, we report readily synthesized fluorogenic substrates enabling enzyme-economical evaluation of SIRT2 inhibitors in a continuous assay format as well as evaluation of the properties of SIRT2...

Pharmacological and toxicological evaluation of Urtica dioica.

Science.gov (United States)

Dar, Sabzar Ahmad; Ganai, Farooq Ahmad; Yousuf, Abdul Rehman; Balkhi, Masood-Ul-Hassan; Bhat, Towseef Mohsin; Sharma, Poonam

2013-02-01

Medicinal plants are a largely unexplored source of drug repository. Urtica dioica L. (Urticaceae) is used in traditional medicine to treat diverse conditions. The present study describes the antidiabetic, antiinflammatory, antibacterial activity, and toxicological studies of Urtica dioica. U. dioica leaves were subjected to solvent extraction with hexane, chloroform, ethyl acetate, methanol, and aqueous, respectively, and screened for antidiabetic (300 mg/kg bw by glucose tolerance test; GTT), antiinflammatory (200 mg/kg bw by rat paw edema assay) and antibacterial activities [by disc-diffusion and minimum inhibitory concentration (MIC) assays]. Toxicological studies were carried on Artemia salina and Wistar rats; phytochemical analyses were carried out, using chromatographic and spectroscopic techniques. The aqueous extract of U. dioica (AEUD) significantly (p 1000 μg/mL each on A. salina. Our results showed that the U. dioica leaves are an interesting source of bioactive compounds, justifying their use in folk medicine, to treat various diseases.

Genetic point-of-care diagnosis of Mycoplasma pneumoniae infection using LAMP assay.

Science.gov (United States)

Kakuya, Fujio; Kinebuchi, Takahiro; Fujiyasu, Hiroaki; Tanaka, Ryosuke; Kano, Hiroki

2014-08-01

Mycoplasma pneumoniae (MP) is a major pathogen of lower respiratory tract infection (LRTI) in children. A rapid diagnostic method during the acute phase is required for the prescription of effective antibiotics. A prospective, single-centered study was conducted on community-acquired LRTI in children. We regarded the day of fever onset as the first day of illness. In part 1, we studied 191 patients with signs of LRTI. We compared diagnostic reliability using loop-mediated isothermal amplification (LAMP) assay and serological testing at the first visit. In part 2, we evaluated the clinical characteristics of 117 patients with positive LAMP assay. In part 1, 31 patients met the definite MP infection criteria. LAMP assay had a sensitivity of 96.8% and specificity of 100%, whereas enzyme immunoassay had a sensitivity of 38.7% and specificity of 76.9%, and particle agglutination test had a sensitivity of 19.4% and specificity of 93.1%. In part 2, of 106 patients with fever, 100 patients were diagnosed by the day 7 of illness. The diagnosis was made a mean of 3.5 ± 2.1 days after the onset of fever. LAMP assay had excellent sensitivity and specificity for the detection of acute MP infection at the first visit. This assay can diagnose MP infection during the very acute phase. LAMP assay is appropriate for genetic point-of-care diagnosis of MP infection in hospital laboratories. © 2014 Japan Pediatric Society.

Geology of the Delta, Escalante, Price, Richfield, and Salina 10 x 20 quadrangles, Utah

International Nuclear Information System (INIS)

Thayer, P.A.

1981-11-01

The National Uranium Resource Evaluation (NURE) program was established to evaluate domestic uranium resources in the continental United States and to identify areas favorable for uranium exploration. The Grand Junction Office of the Department of Energy is responsible for administering the program. The Savannah River Laboratory (SRL) is responsible for hydrogeochemical and stream-sediment reconnaissance (HSSR) of 3.9 million km 2 (1,500,000 mi 2 ) in 37 eastern and western states. This document provides geologic and mineral resources reports for the Delta, Escalante, Price, Richfield, and Salina 1 0 x 2 0 National Topographic Map Series quadrangles, Utah. The purpose of these reports is to provide background geologic and mineral resources information to aid in the interpretation of NURE geochemical reconnaissance data. Except for the Escalante Quadrangle, each report is accompanied by a geologic map and a mineral locality map (Plates 1-8, in pocket). The US Geological Survey previously published a 1 0 x 2 0 geologic map of the Escalante Quadrangle and described the uranium deposits in the area (Hackman and Wyant, 1973). NURE hydrogeochemical and stream-sediment reconnaissance data for these quadrangles have been issued previously in some of the reports included in the references

Genotoxic effects of boric acid and borax in zebrafish, Danio rerio using alkaline comet assay.

Science.gov (United States)

Gülsoy, Nagihan; Yavas, Cüneyd; Mutlu, Özal

2015-01-01

The present study is conducted to determine the potential mechanisms of Boron compounds, boric acid (BA) and borax (BX), on genotoxicity of zebrafish Danio rerio for 24, 48, 72 and 96-hours acute exposure (level:1, 4, 16, 64 mg/l BA and BX) in semi-static bioassay experiment. For that purpose, peripheral erythrocytes were drawn from caudal vein and Comet assay was applied to assess genotoxicity. Acute (96 hours) exposure and high concentrations of boric acid and borax increases % tail DNA and Olive tail moment. Genotoxicity was found for BA as concentration-dependent and BX as concentration and time dependent manner. In general, significant effects (P borax-induced genotoxicity in fish.

A 96-well-plate-based optical method for the quantitative and qualitative evaluation of Pseudomonas aeruginosa biofilm formation and its application to susceptibility testing.

Science.gov (United States)

Müsken, Mathias; Di Fiore, Stefano; Römling, Ute; Häussler, Susanne

2010-08-01

A major reason for bacterial persistence during chronic infections is the survival of bacteria within biofilm structures, which protect cells from environmental stresses, host immune responses and antimicrobial therapy. Thus, there is concern that laboratory methods developed to measure the antibiotic susceptibility of planktonic bacteria may not be relevant to chronic biofilm infections, and it has been suggested that alternative methods should test antibiotic susceptibility within a biofilm. In this paper, we describe a fast and reliable protocol for using 96-well microtiter plates for the formation of Pseudomonas aeruginosa biofilms; the method is easily adaptable for antimicrobial susceptibility testing. This method is based on bacterial viability staining in combination with automated confocal laser scanning microscopy. The procedure simplifies qualitative and quantitative evaluation of biofilms and has proven to be effective for standardized determination of antibiotic efficiency on P. aeruginosa biofilms. The protocol can be performed within approximately 60 h.

Evaluation of the Chagas Stat-Paktm Assay for Detection of Trypanosoma cruzi Antibodies in Wildlife Reservoirs

Science.gov (United States)

Yabsley, Michael J.; Brown, Emily L.; Roellig, Dawn M.

2010-01-01

An immunochromatographic assay (Chagas Stat-Pak™) was evaluated for the detection of Trypanosoma cruzi antibodies in 4 species of wildlife reservoirs. Antibodies to T. cruzi were detected in raccoons (Procyon lotor) (naturally and experimentally infected) and degus (Octodon degu) (experimentally-infected) using the Chagas Stat-Pak. In naturally exposed wild raccoons, the Chagas Stat-Pak had a sensitivity and specificity of 66.7–80.0% and 96.3%, respectively. Compared with indirect immunofluorescent antibody assay results, serocon-version as determined by Chagas Stat-Pak was delayed for experimentally infected raccoons, but occurred sooner in experimentally infected degus. The Chagas Stat-Pak did not detect antibodies in naturally or experimentally infected Virginia opossums (Didelphis virginiana) or in experimentally infected short-tailed opossums (Monodelphis domestica). These data suggest that the Chagas Stat-Pak might be useful in field studies of raccoons and degus when samples would not be available for more-conventional serologic assays. Because this assay did not work on either species of marsupial, the applicability of the assay should be examined before it is used in other wild species. PMID:19016578

Genetic variation for lettuce seed thermoinhibition is associated with temperature-sensitive expression of abscisic Acid, gibberellin, and ethylene biosynthesis, metabolism, and response genes.

Science.gov (United States)

Argyris, Jason; Dahal, Peetambar; Hayashi, Eiji; Still, David W; Bradford, Kent J

2008-10-01

Lettuce (Lactuca sativa 'Salinas') seeds fail to germinate when imbibed at temperatures above 25 degrees C to 30 degrees C (termed thermoinhibition). However, seeds of an accession of Lactuca serriola (UC96US23) do not exhibit thermoinhibition up to 37 degrees C in the light. Comparative genetics, physiology, and gene expression were analyzed in these genotypes to determine the mechanisms governing the regulation of seed germination by temperature. Germination of the two genotypes was differentially sensitive to abscisic acid (ABA) and gibberellin (GA) at elevated temperatures. Quantitative trait loci associated with these phenotypes colocated with a major quantitative trait locus (Htg6.1) from UC96US23 conferring germination thermotolerance. ABA contents were elevated in Salinas seeds that exhibited thermoinhibition, consistent with the ability of fluridone (an ABA biosynthesis inhibitor) to improve germination at high temperatures. Expression of many genes involved in ABA, GA, and ethylene biosynthesis, metabolism, and response was differentially affected by high temperature and light in the two genotypes. In general, ABA-related genes were more highly expressed when germination was inhibited, and GA- and ethylene-related genes were more highly expressed when germination was permitted. In particular, LsNCED4, a gene encoding an enzyme in the ABA biosynthetic pathway, was up-regulated by high temperature only in Salinas seeds and also colocated with Htg6.1. The temperature sensitivity of expression of LsNCED4 may determine the upper temperature limit for lettuce seed germination and may indirectly influence other regulatory pathways via interconnected effects of increased ABA biosynthesis.

AN ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) METHOD FOR THE URINARY BIOMONITORING OF 2,4-DICHLOROPHRENOCYACETIC ACID (2,4-D)

Science.gov (United States)

An enzyme-linked immunosorbent assay (ELISA) method was developed to quantitatively measure 2,4-dichlorophenoyacetic acid (2,4-D) in human urine. Samples were diluted (1:5) with phosphate-buffered saline, 0.05% Tween 20, with 0.02% sodium azide, and analyzed by a 96-microwekk pl...

A high content, high throughput cellular thermal stability assay for measuring drug-target engagement in living cells.

Science.gov (United States)

Massey, Andrew J

2018-01-01

Determining and understanding drug target engagement is critical for drug discovery. This can be challenging within living cells as selective readouts are often unavailable. Here we describe a novel method for measuring target engagement in living cells based on the principle of altered protein thermal stabilization / destabilization in response to ligand binding. This assay (HCIF-CETSA) utilizes high content, high throughput single cell immunofluorescent detection to determine target protein levels following heating of adherent cells in a 96 well plate format. We have used target engagement of Chk1 by potent small molecule inhibitors to validate the assay. Target engagement measured by this method was subsequently compared to target engagement measured by two alternative methods (autophosphorylation and CETSA). The HCIF-CETSA method appeared robust and a good correlation in target engagement measured by this method and CETSA for the selective Chk1 inhibitor V158411 was observed. However, these EC50 values were 23- and 12-fold greater than the autophosphorylation IC50. The described method is therefore a valuable advance in the CETSA method allowing the high throughput determination of target engagement in adherent cells.

45 CFR 96.127 - Requirements regarding tuberculosis.

Science.gov (United States)

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Requirements regarding tuberculosis. 96.127... Substance Abuse Prevention and Treatment Block Grant § 96.127 Requirements regarding tuberculosis. (a... Department of Health/Tuberculosis Control Officer, which address how the program— (1) Will, directly or...

BmNHR96 participate BV entry of BmN-SWU1 cells via affecting the cellular cholesterol level.

Science.gov (United States)

Dong, Xiao-Long; Liu, Tai-Hang; Wang, Wei; Pan, Cai-Xia; Du, Guo-Yu; Wu, Yun-Fei; Pan, Min-Hui; Lu, Cheng

2017-01-22

B.mori nucleopolyhedrovirus (BmNPV), which produces BV and ODV two virion phenotypes in its life cycle, caused the amount of economic loss in sericulture. But the mechanism of its infection was still unclear. In this study we characterized B.mori nuclear hormone receptor 96 (BmNHR96) as a NHR96 family member, which was localized in the nucleus. We also found BmNHR96 over-expression could enhance the entry of BV as well as cellular cholesterol level. Furthermore, we validated that BmNHR96 increased membrane fusion mediated by GP64, which could probably promote BV-infection. In summary, our study suggested that BmNHR96 plays an important role in BV infection and this function probably actualized by affecting cellular cholesterol level, and our results provided insights to the mechanisms of BV-infection of B.mori. Copyright © 2016 Elsevier Inc. All rights reserved.

Detection of Francisella tularensis-Specific Antibodies in Patients with Tularemia by a Novel Competitive Enzyme-Linked Immunosorbent Assay

Science.gov (United States)

Sharma, Neekun; Hotta, Akitoyo; Yamamoto, Yoshie; Fujita, Osamu; Uda, Akihiko; Morikawa, Shigeru; Yamada, Akio

2013-01-01

A novel competitive enzyme-linked immunosorbent assay (cELISA) was developed and evaluated for detection of antibodies against Francisella tularensis in humans. The assay is based on the ability of serum antibodies to inhibit the binding of monoclonal antibodies (MAbs) directed against F. tularensis lipopolysaccharide antigens. The assay was evaluated using serum samples of tularemia patients, inactivated F. tularensis-immunized rabbits, and F. tularensis-infected mice. Antibodies against F. tularensis were successfully detected in serum samples of tularemia patients as well as the immunized and infected animals. The cELISA method was compared to indirect ELISA (iELISA) and the commonly used microagglutination test (MA) using serum samples of 19 tularemia patients and 50 healthy individuals. The sensitivity and specificity of cELISA were 93.9 and 96.1%, respectively, in comparison to the iELISA. MA was less sensitive than cELISA with a sensitivity and specificity of only 81.8 and 98.0%, respectively. A high degree of correlation (R2 = 0.8226) was observed between cELISA and iELISA results. The novel cELISA developed in this study appears to be highly sensitive and specific for serodiagnosis of human tularemia. The potential of the MAb-based cELISA to be used in both human and animal samples emphasizes its usefulness for serological survey of tularemia among multiple animal species. PMID:23114700

Avaliação do potencial fisiológico de sementes de melancia pelo teste de envelhecimento acelerado

Directory of Open Access Journals (Sweden)

Aline Klug Radke

2017-10-01

Full Text Available Para uma análise plena da qualidade fisiológica de sementes, há necessidade de complementar informação provida do teste de germinação com testes de vigor, a fim de selecionar os lotes vigorosos para comercialização. Objetivou-se estudar metodologias do teste de envelhecimento acelerado tradicional e modificado, com solução salina saturada e não saturada, avaliando o potencial fisiológico de sementes de melancia. Foram utilizadas sementes de melancia cultivar Congo e Crimson Sweet, representadas por quatro e cincos lotes de sementes, respectivamente. Para a avaliação da qualidade das sementes de melancia foram utilizados as seguintes determinações: testes de germinação, primeira contagem de germinação, teste de frio, índice de velocidade de emergência, emergência de plântulas e procedimentos do teste de envelhecimento acelerado nas metodologias: tradicional, solução salina saturada (40g de NaCl por 100 mLde água e solução salina não saturada (11g de NaCl por 100 mLde água, a 41°C, por períodos de 48; 72 e 96 horas. O teste de envelhecimento acelerado, utilizando solução salina não saturada ou solução salina saturada e combinação 41°C por 72 horas, mostra-se adequado para avaliação do potencial fisiológico de sementes de melancia, constituindo-se em um teste promissor para avaliação da expressão do vigor.

Novel approach to high-throughput determination of endocrine disruptors using recycled diatomaceous earth as a green sorbent phase for thin-film solid-phase microextraction combined with 96-well plate system.

Science.gov (United States)

Kirschner, Nicolas; Dias, Adriana Neves; Budziak, Dilma; da Silveira, Cristian Berto; Merib, Josias; Carasek, Eduardo

2017-12-15

A sustainable approach to TF-SPME is presented using recycled diatomaceous earth, obtained from a beer purification process, as a green sorbent phase for the determination of bisphenol A (BPA), benzophenone (BzP), triclocarban (TCC), 4-methylbenzylidene camphor (4-MBC) and 2-ethylhexyl-p-methoxycinnamate (EHMC) in environmental water samples. TF-SPME was combined with a 96-well plate system allowing for high-throughput analysis due to the simultaneous extraction/desorption up to 96 samples. The proposed sorbent phase exhibited good stability in organic solvents, as well as satisfactory analytical performance. The optimized method consisted of 240 min of extraction at pH 6 with the addition of NaCl (15% w/v). A mixture of MeOH:ACN (50:50 v/v) was used for the desorption the analytes, using a time of 30 min. Limits of detection varied from 1 μg L -1 for BzP and TCC to 8 μg L -1 for the other analytes, and R 2 ranged from 0.9926 for 4-MBC to 0.9988 for BPA. This novel and straightforward approach offers an environmentally-friendly and very promising alternative for routine analysis. . The total sample preparation time per sample was approximately 2.8 min, which is a significant advantage when a large number of analytical run is required. Copyright © 2017 Elsevier B.V. All rights reserved.

Estudio del comercio electrónico como estrategia de negocios dentro de las empresas que brindan servicios de hospedaje en el Cantón Salinas, provincia de Santa Elena.

OpenAIRE

Mateo López, Blanca Annabell

2015-01-01

El presente trabajo tiene como objetivo analizar los mecanismos de promoción y comercialización, que utilizan los establecimientos que brindan servicios de hospedaje en Salinas, identificando los medios de comunicación que manejan para potenciar sus negocios. Al conocer que a nivel mundial existen varias estrategias soportadas en medios digitales por parte de los hoteles, se ha determinado la necesidad de los empresarios hoteleros de tener en sus empresas una página web, o mejoras...

ROS and calcium signaling mediated pathways involved in stress responses of the marine microalgae Dunaliella salina to enhanced UV-B radiation.

Science.gov (United States)

Zhang, Xinxin; Tang, Xuexi; Wang, Ming; Zhang, Wei; Zhou, Bin; Wang, You

2017-08-01

UV-B ray has been addressed to trigger common metabolic responses on marine microalgae, however, the upstream events responsible for these changes in marine microalgae are poorly understood. In the present study, a species of marine green microalgae Dunaliella salina was exposed to a series of enhanced UV-B radiation ranging from 0.25 to 1.00 KJ·m -2 per day. The role of ROS and calcium signaling in the D. salina responses to UV-B was discussed. Results showed that enhanced UV-B radiation markedly decreased the cell density in a dose-dependent manner, but the contents of protein and glycerol that were essential for cell growth increased. It suggested that it was cell division instead of cell growth that UV-B exerted negative effects on. The subcellular damages on nuclei and plasmalemma further evidenced the hypothesis. The nutrient absorption was affected with UV-B exposure, and the inhibition on PO 4 3- uptake was more serious compared to NO 3 - uptake. UV-B radiation promoted reactive oxygen species (ROS) formation and thiobarbituric acid reactive substances (TBARS) contents, decreased the redox status and altered the antioxidant enzyme activities. The addition of the ROS scavenger and the glutathione biosynthesis precursor N-acetyl-l-cysteine (NAC) alleviated the stress degree, implying ROS-mediated pathway was involved in the stress response to UV-B radiation. Transient increase in Ca 2+ -ATPase was triggered simultaneously with UV-B exposure. Meanwhile, the addition of an intracellular free calcium chelator aggravated the damage of cell division, but exogenous calcium and ion channel blocker applications did not, inferring that endogenously initiated calcium signaling played roles in response to UV-B. Cross-talk analysis showed a relatively clear relationship between ROS inhibition and Ca 2+ -ATPase suppression, and a relation between Ca 2+ inhibition and GPx activity change was also observed. It was thus presumed that ROS-coupled calcium signaling via the

High throughput comet assay to study genotoxicity of nanomaterials

Directory of Open Access Journals (Sweden)

Naouale El Yamani

2015-06-01

Full Text Available The unique physicochemical properties of engineered nanomaterials (NMs have accelerated their use in diverse industrial and domestic products. Although their presence in consumer products represents a major concern for public health safety, their potential impact on human health is poorly understood. There is therefore an urgent need to clarify the toxic effects of NMs and to elucidate the mechanisms involved. In view of the large number of NMs currently being used, high throughput (HTP screening technologies are clearly needed for efficient assessment of toxicity. The comet assay is the most used method in nanogenotoxicity studies and has great potential for increasing throughput as it is fast, versatile and robust; simple technical modifications of the assay make it possible to test many compounds (NMs in a single experiment. The standard gel of 70-100 μL contains thousands of cells, of which only a tiny fraction are actually scored. Reducing the gel to a volume of 5 μL, with just a few hundred cells, allows twelve gels to be set on a standard slide, or 96 as a standard 8x12 array. For the 12 gel format, standard slides precoated with agarose are placed on a metal template and gels are set on the positions marked on the template. The HTP comet assay, incorporating digestion of DNA with formamidopyrimidine DNA glycosylase (FPG to detect oxidised purines, has recently been applied to study the potential induction of genotoxicity by NMs via reactive oxygen. In the NanoTEST project we investigated the genotoxic potential of several well-characterized metal and polymeric nanoparticles with the comet assay. All in vitro studies were harmonized; i.e. NMs were from the same batch, and identical dispersion protocols, exposure time, concentration range, culture conditions, and time-courses were used. As a kidney model, Cos-1 fibroblast-like kidney cells were treated with different concentrations of iron oxide NMs, and cells embedded in minigels (12

Growth of mussels Mytilus edulis at algal (Rhodomonas salina) concentrations below and above saturation level for reduced filtration rate

DEFF Research Database (Denmark)

Riisgård, Hans Ulrik; Pleissner, Daniel; Larsen, Poul Scheel

2013-01-01

Average filtration and growth rates of groups of juvenile Mytilus edulis (n =2545 of 22-35 mm shell length) were measured at different concentrations of an algal cell monoculture in 9 laboratory experiments of duration 14-30 days, 4 experiments below and 5 above the limit of incipient saturation...... concentration (Csat ≈ 6000-7000 Rhodomonas salina cells ml-1). From a nearly constant filtration rate (F ≈ 30 ml min-1 for a 30 mm shell length) at measured algal concentrations below Csat the steady-state filtration rate decreased approximately as 1/C for increasing algal concentrations (C) above Csat...... is exceeded and then as partial valve closure and reduced filtration and growth rates along with production of pseudofaeces. A survey of naturally occurring phytoplankton biomass in the sea shows that this is generally below Csat except for the short spring bloom periods; hence mussels generally feed...

Determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts.

Science.gov (United States)

Tsukatani, Tadayuki; Suenaga, Hikaru; Ishiyama, Munetaka; Ezoe, Takatoshi; Matsumoto, Kiyoshi

2011-07-15

A method for the determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8)} via 2-methyl-1,4-napthoquinone (NQ) was developed. Measurement conditions were optimized for the microbiological determination of water-soluble vitamins, such as vitamin B(6), biotin, folic acid, niacin, and pantothenic acid, using microorganisms that have a water-soluble vitamin requirement. A linear relationship between absorbance and water-soluble vitamin concentration was obtained. The proposed method was applied to determine the concentration of vitamin B(6) in various foodstuffs. There was good agreement between vitamin B(6) concentrations determined after 24h using the WST-8 colorimetric method and those obtained after 48h using a conventional method. The results suggest that the WST-8 colorimetric assay is a useful method for the rapid determination of water-soluble vitamins in a 96-well microtiter plate. Copyright © 2011 Elsevier Ltd. All rights reserved.

Konsekvensanalyse af Kulturby 96

DEFF Research Database (Denmark)

Fridberg, Torben; Koch-Nielsen, Inger

1997-01-01

Rapporten om Kulturby 96 indeholder en beskrivelse af kulturbyårets aktiviteter og responsen herpå fra publikum, turister, presse og samarbejdspartnere i øvrigt. Der redegøres for projektets organisatoriske struktur, herunder Kulturbysekretariatets organisation og rolle. Rapporten indeholder...

Detection of Mycobacterium tuberculosis Complex in Paraffin-Embedded Tissues by the New Automated Abbott RealTime MTB Assay.

Science.gov (United States)

Fu, Yung-Chieh; Liao, I-Chuang; Chen, Hung-Mo; Yan, Jing-Jou

2016-07-01

The Abbott RealTime MTB assay, launched in June 2014, has been shown to have a competitive performance in the detection of the Mycobacterium tuberculosis (MTB) complex in respiratory specimens. The present study was conducted to investigate the usefulness of the Abbott MTB Realtime assay in the detection of MTB in formalin-fixed paraffin-embedded (FFPE) tissues. A total of 96 FFPE specimens obtained from microbiologically proven MTB cases (N=60) and nontuberculous Mycobacterium cases (N=36) were analyzed. The performance of the Abbott MTB Realtime assay was compared with that of the Roche Cobas TaqMan MTB assay. The overall sensitivity and specificity of the Abbott assay were 63.3% and 97.2%, respectively, compared with 11.7% and 100% for the Cobas assay. The detection rate of the Abbott assay was much higher among 37 acid-fast-positive specimens than among 23 acid-fast-negative specimens (89.3% versus 21.7%, respectively). The detection rate of the assay was higher among 29 resection specimens than among 31 small biopsy specimens (86.2% versus 41.9%, respectively). Our results suggest that the Abbott RealTime MTB assay can be used to differentiate MTB from nontuberculous mycobacterial infections in acid-fast-positive FFPE tissues. © 2016 by the Association of Clinical Scientists, Inc.

16 CFR 1.96 - Compromise of penalty.

Science.gov (United States)

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Compromise of penalty. 1.96 Section 1.96 Commercial Practices FEDERAL TRADE COMMISSION ORGANIZATION, PROCEDURES AND RULES OF PRACTICE GENERAL... may compromise any penalty or proposed penalty at any time, with leave of court when necessary, taking...

Artisanal salt production in Aveiro/Portugal - an ecofriendly process.

Science.gov (United States)

Rodrigues, Carolina M; Bio, Ana; Amat, Francisco; Vieira, Natividade

2011-11-04

Solar salinas are man-made systems exploited for the extraction of salt, by solar and wind evaporation of seawater. Salt production achieved by traditional methods is associated with landscapes and environmental and patrimonial values generated throughout history. Since the mid-twentieth century, this activity has been facing a marked decline in Portugal, with most salinas either abandoned or subjected to destruction, making it necessary to find a strategy to reverse this trend.It is, however, possible to generate revenue from salinas at several levels, not merely in terms of good quality salt production, but also by obtaining other products that can be commercialized, or by exploring their potential for tourism, and as research facilities, among others. Furthermore, with an adequate management, biodiversity can be restored to abandoned salinas, which constitute important feeding and breeding grounds for resident and migratory aquatic birds, many of which are protected by European Community Directives.The aims of this manuscript are to present a brief overview on the current state of sea salt exploitation in Portugal and to stress the importance of recovering these salinas for the conservation of this particular environment, for the regional economy, the scientific community and the general public. The Aveiro salina complex is presented in detail, to exemplify salina structure and functioning, as well as current problems and potential solutions for artisanal salinas.

40 CFR 96.340 - State trading budgets.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false State trading budgets. 96.340 Section...) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS CAIR NOX Ozone Season Allowance Allocations § 96.340 State trading budgets. (a) Except as provided in...

40 CFR 96.140 - State trading budgets.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false State trading budgets. 96.140 Section...) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS CAIR NOX Allowance Allocations § 96.140 State trading budgets. The State trading budgets for annual...

44 CFR 9.6 - Decision-making process.

Science.gov (United States)

2010-10-01

... 44 Emergency Management and Assistance 1 2010-10-01 2010-10-01 false Decision-making process. 9.6... HOMELAND SECURITY GENERAL FLOODPLAIN MANAGEMENT AND PROTECTION OF WETLANDS § 9.6 Decision-making process... protection decision-making process to be followed by the Agency in applying the Orders to its actions. While...

Burnup calculation code system COMRAD96

International Nuclear Information System (INIS)

Suyama, Kenya; Masukawa, Fumihiro; Ido, Masaru; Enomoto, Masaki; Takyu, Shuiti; Hara, Toshiharu.

1997-06-01

COMRAD was one of the burnup code system developed by JAERI. COMRAD96 is a transfered version of COMRAD to Engineering Work Station. It is divided to several functional modules, 'Cross Section Treatment', 'Generation and Depletion Calculation', and 'Post Process'. It enables us to analyze a burnup problem considering a change of neutron spectrum using UNITBURN. Also it can display the γ Spectrum on a terminal. This report is the general description and user's manual of COMRAD96. (author)

Biobarcode assay for the oral anticoagulant acenocoumarol.

Science.gov (United States)

Broto, Marta; Salvador, J Pablo; Galve, Roger; Marco, M Pilar

2018-02-01

A novel approach for therapeutic drug monitoring of oral anticoagulants (OA) in clinical samples is reported, based on a NP-based biobarcode assay. The proposed strategy uses specific antibodies for acenocumarol (ACL) covalently bound to magnetic particles (pAb236-MP) and a bioconjugate competitor (hACL-BSA) linked to encoded polystyrene probes (hACL-BSA-ePSP) on a classical competitive immunochemical format. By using this scheme ACL can be detected in low nM range (LOD, 0.96 ± 0.26, N = 3, in buffer) even in complex samples such as serum or plasma (LOD 4 ± 1). The assay shows a high reproducibility (%CV 1.1 day-to-day) and is robust, as it is demonstrated by the fact that ACL can be quantified in complex biological samples with a very good accuracy (slope = 0.97 and R 2 = 0.91, of the linear regression obtained when analyzing spiked vs measured values). Moreover, we have demonstrated that the biobarcode approach has the potential to overcome one of the main challenges of the multiplexed diagnostic, which is the possibility to measure in a single run biomarker targets present at different concentration ranges. Thus, it has been proven that the signal and the detectability can be modulated by just modifying the oligonucleotide load of the encoded probes. This fact opens the door for combining in the same assay encoded probes with the necessary oligonucleotide load to achieve the detectability required for each biomarker target. Copyright © 2017 Elsevier B.V. All rights reserved.

Sida tuberculata (Malvaceae): a study based on development of extractive system and in silico and in vitro properties.

Science.gov (United States)

da Rosa, H S; Salgueiro, A C F; Colpo, A Z C; Paula, F R; Mendez, A S L; Folmer, V

2016-07-11

Sida tuberculata (Malvaceae) is a medicinal plant traditionally used in Brazil as an antimicrobial and anti-inflammatory agent. Here, we aimed to investigate the different extractive techniques on phytochemical parameters, as well as to evaluate the toxicity and antioxidant capacity of S. tuberculata extracts using in silico and in vitro models. Therefore, in order to determine the dry residue content and the main compound 20-hydroxyecdysone (20E) concentration, extracts from leaves and roots were prepared testing ethanol and water in different proportions. Extracts were then assessed by Artemia salina lethality test, and toxicity prediction of 20E was estimated. Antioxidant activity was performed by DPPH and ABTS radical scavenger assays, ferric reducing power assay, nitrogen derivative scavenger, deoxyribose degradation, and TBARS assays. HPLC evaluation detected 20E as main compound in leaves and roots. Percolation method showed the highest concentrations of 20E (0.134 and 0.096 mg/mL of extract for leaves and roots, respectively). All crude extracts presented low toxic potential on A. salina (LD50 >1000 µg/mL). The computational evaluation of 20E showed a low toxicity prediction. For in vitro antioxidant tests, hydroethanolic extracts of leaves were most effective compared to roots. In addition, hydroethanolic extracts presented a higher IC50 antioxidant than aqueous extracts. TBARS formation was prevented by leaves hydroethanolic extract from 0.015 and 0.03 mg/mL and for roots from 0.03 and 0.3 mg/mL on egg yolk and rat tissue, respectively (P<0.05). These findings suggest that S. tuberculata extracts are a considerable source of ecdysteroids and possesses a significant antioxidant property with low toxic potential.

EL4 cell-based colorimetric toxin neutralization activity assays for determination of neutralizing anti-ricin antibodies.

Science.gov (United States)

Lindsey, Changhong Y; Brown, J Edward; Torabazar, Nahid R; Smith, Leonard A

2013-01-01

A recombinant ricin toxin A-chain 1-33/44-198 vaccine (RVEc), developed at the United States Army Medical Research Institute of Infectious Diseases as a vaccine candidate, is under investigation in a phase 1 clinical study. To effectively evaluate the immunogenicity of this ricin vaccine and to eliminate the use of radioactive material, an EL4 cell-based colorimetric toxin neutralization activity (TNA) assay using a CellTiter 96 AQueous One Solution Cell Proliferation Assay Reagent has been developed, optimized, and applied in the vaccine efficacy studies. The TNA assay measures the protective neutralizing anti-ricin antibodies in animal sera by determining the cell viability after ricin exposure in the assay system and comparing it to a purified mouse polyclonal antiricin IgG standard curve. The standard curve of the anti-ricin TNA assay closely fits a four-parameter logistic regression model. The unknown test sample concentration was expressed as microg/mL, but not the 50% effective concentration (EC50), which was determined by most TNA assays. The neutralizing endpoint titers, not the 50% effective dilution (ED50), of human specimens were measured with the TNA assay in support of the clinical study of the RVEc vaccine. The optimal amount of ricin toxin, EL4 cells, and concentration of standards used in the assay system was established to minimize false-negative and false-positive results of serum specimens from the nonclinical and clinical studies of RVEc. The testing conditions were adjusted to optimize assay performance. The colorimetric TNA assay replaced a radioactive TNA assay previously used in the ricin vaccine studies.

45 CFR 96.18 - Participation by faith-based organizations.

Science.gov (United States)

2010-10-01

... 45 Public Welfare 1 2010-10-01 2010-10-01 false Participation by faith-based organizations. 96.18 Section 96.18 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION BLOCK GRANTS General Procedures § 96.18 Participation by faith-based organizations. The funds provided under this part...

High performance liquid chromatographic assay for the quantitation of total glutathione in plasma

Science.gov (United States)

Abukhalaf, Imad K.; Silvestrov, Natalia A.; Menter, Julian M.; von Deutsch, Daniel A.; Bayorh, Mohamed A.; Socci, Robin R.; Ganafa, Agaba A.

2002-01-01

A simple and widely used homocysteine HPLC procedure was applied for the HPLC identification and quantitation of glutathione in plasma. The method, which utilizes SBDF as a derivatizing agent utilizes only 50 microl of sample volume. Linear quantitative response curve was generated for glutathione over a concentration range of 0.3125-62.50 micromol/l. Linear regression analysis of the standard curve exhibited correlation coefficient of 0.999. Limit of detection (LOD) and limit of quantitation (LOQ) values were 5.0 and 15 pmol, respectively. Glutathione recovery using this method was nearly complete (above 96%). Intra-assay and inter-assay precision studies reflected a high level of reliability and reproducibility of the method. The applicability of the method for the quantitation of glutathione was demonstrated successfully using human and rat plasma samples.

Search for the β decay of 96Zr

Science.gov (United States)

Finch, S. W.; Tornow, W.

2016-01-01

96Zr and 48Ca are unique among double-β decay candidate nuclides in that they may also undergo single-β decay. In the case of 96Zr, the single-β decay mode is dominated by the fourth-forbidden β decay with a 119 keV Q value. A search was conducted for the β decay of 96Zr by observing the decay of the daughter 96Nb nucleus. Two coaxial high-purity germanium detectors were used in coincidence to detect the γ-ray cascade produced by the daughter nucleus as it de-excited to the ground state. The experiment was carried out at the Kimballton Underground Research Facility and produced 685.7 days of data with a 17.91 g enriched sample. No counts were seen above background, producing a limit of T1/2 > 2.4 ×1019 year. This is the first experimental search that is able to discern between the β decay and the double-β decay to an excited state of 96Zr.

Burnup calculation code system COMRAD96

Energy Technology Data Exchange (ETDEWEB)

Suyama, Kenya [Japan Atomic Energy Research Inst., Tokai, Ibaraki (Japan). Tokai Research Establishment; Masukawa, Fumihiro; Ido, Masaru; Enomoto, Masaki; Takyu, Shuiti; Hara, Toshiharu

1997-06-01

COMRAD was one of the burnup code system developed by JAERI. COMRAD96 is a transfered version of COMRAD to Engineering Work Station. It is divided to several functional modules, `Cross Section Treatment`, `Generation and Depletion Calculation`, and `Post Process`. It enables us to analyze a burnup problem considering a change of neutron spectrum using UNITBURN. Also it can display the {gamma} Spectrum on a terminal. This report is the general description and user`s manual of COMRAD96. (author)

Thermally activated demagnetization in (La0.97 Ca0.03)0.96Mn0.96 O3-δ

International Nuclear Information System (INIS)

Alonso, J.M.; Arroyo, A.; Cortes-Gil, R.; Garcia, M.A.; Gonzalez-Calbet, J.M.; Gonzalez, J.M.; Hernando, A.; Rojo, J.M.; Vallet-Regi, M.

2005-01-01

We report on the temperature dependence of the relaxation properties of a Mn perovskite with overall composition (La 0.97 Ca 0.03 ) 0.96 (Mn 4+ 0.07 Mn 3+ 0.93 ) 0.96 O 2.90 .Our data are correlated with the simultaneous occurrence in the sample of metallic double-exchange magnetically coupled, Mn 4+ clusters and of insulating superexchange coupled regions.

Trocas gasosas e conteúdo de carboidratos e compostos nitrogenados em pinhão-manso irrigado com águas residuária e salina

Directory of Open Access Journals (Sweden)

Antonio Evami Cavalcante Sousa

2012-10-01

Full Text Available O objetivo deste trabalho foi avaliar alterações nas trocas gasosas e nos conteúdos de carboidratos e compostos nitrogenados, em pinhão-manso (Jatropha curcas irrigado com águas residuária e salina. Empregou-se o delineamento em blocos ao acaso, com quatro tratamentos e quatro repetições: irrigação plena com água de abastecimento a 0,6 dS m-1 (controle; irrigação plena com água salinizada a 2,4 dS m-1 (SAL; e irrigação com água residuária de esgoto, plena (R100 e a 50% da capacidade de campo (R50. O tratamento R50 reduziu fortemente as trocas gasosas e a área foliar, comparado ao controle, seguido pelos tratamentos SAL e R100. A redução na fotossíntese diminuiu o nível de sacarose nas folhas, nos quatro tratamentos. Os teores de açúcares solúveis aumentaram nos tratamentos R50, SAL e R100, enquanto o conteúdo de amido permaneceu praticamente inalterado. Os conteúdos de prolina e glicina betaína aumentaram nos três tratamentos, mas a última foi mais importante, em termos quantitativos, como protetor celular e osmótico. A irrigação plena com água residuária induz efeitos similares aos causados pela irrigação com água salina. A irrigação limitada com água residuária causa estresse agudo às plantas, provavelmente pela combinação de deficiência hídrica e acúmulo de solutos no solo.

Corrosividade Causada por Soluções Salinas Empregadas na Estabilização Dimensional da Madeira

Directory of Open Access Journals (Sweden)

Juarez Benigno Paes

2017-08-01

Full Text Available RESUMO Objetivou-se avaliar a corrosividade de soluções salinas antes e depois de sua impregnação na madeira. Soluções com 5% de concentração de cloreto de lítio, carbonato de sódio, sulfato de magnésio, sulfato de zinco e sulfato de cobre II foram mantidas durante 6 horas, a 25 °C, 45 °C e 100 °C em contato com amostras de 1,25 cm × 0,1 cm × 7,5 cm (largura × espessura × comprimento produzidas com chapas de aço carbono antes e depois de seu contato com amostras de madeira de Corymbia torelliana e Eucalyptus cloeziana. Não se observou efeito da temperatura na corrosão das soluções puras. Depois do contato com a madeira houve incremento da corrosividade para as soluções de sulfato de zinco e de cobre II. O sulfato de cobre II ocasionou a maior perda de massa nas chapas, não sendo recomendado seu contato com superfícies metálicas, como usinas de tratamento e câmaras de secagem de madeira.

Conversion of a Capture ELISA to a Luminex xMAP Assay using a Multiplex Antibody Screening Method

Science.gov (United States)

Baker, Harold N.; Murphy, Robin; Lopez, Erica; Garcia, Carlos

2012-01-01

The enzyme-linked immunosorbent assay (ELISA) has long been the primary tool for detection of analytes of interest in biological samples for both life science research and clinical diagnostics. However, ELISA has limitations. It is typically performed in a 96-well microplate, and the wells are coated with capture antibody, requiring a relatively large amount of sample to capture an antigen of interest . The large surface area of the wells and the hydrophobic binding of capture antibody can also lead to non-specific binding and increased background. Additionally, most ELISAs rely upon enzyme-mediated amplification of signal in order to achieve reasonable sensitivity. Such amplification is not always linear and can thus skew results. In the past 15 years, a new technology has emerged that offers the benefits of the ELISA, but also enables higher throughput, increased flexibility, reduced sample volume, and lower cost, with a similar workflow 1, 2. Luminex xMAP Technology is a microsphere (bead) array platform enabling both monoplex and multiplex assays that can be applied to both protein and nucleic acid applications 3-5. The beads have the capture antibody covalently immobilized on a smaller surface area, requiring less capture antibody and smaller sample volumes, compared to ELISA, and non-specific binding is significantly reduced. Smaller sample volumes are important when working with limiting samples such as cerebrospinal fluid, synovial fluid, etc. 6. Multiplexing the assay further reduces sample volume requirements, enabling multiple results from a single sample. Recent improvements by Luminex include: the new MAGPIX system, a smaller, less expensive, easier-to-use analyzer; Low-Concentration Magnetic MagPlex Microspheres which eliminate the need for expensive filter plates and come in a working concentration better suited for assay development and low-throughput applications; and the xMAP Antibody Coupling (AbC) Kit, which includes a protocol, reagents, and

Backbending in the N = 96 isotones

International Nuclear Information System (INIS)

Michel, C.; Vervier, J.

1981-01-01

The backbending in the even-even, N = 96 isotones can be quantitatively accounted for by the rotation-alignment of the spins of neutrons in i 13/2 orbits, as shown by comparing the aligned angular momentum and relative Routhian for the s-bands in these isotones and for the i 13/2 bands in the corresponding isotopes with N = 97. The influence of protons on this backbending situation is shown to be indirect, acting through a change of the nuclear deformation, which yields a change of the moment of inertia of the g.s. band and of the non-rigid character of the rotation. The experimental data on the N = 96 and 97 isotones are in reasonable agreement with cranking model calculations. Possible reasons for the inhibition of backbending in the h 9/2 proton bands in the odd-Z, N = 96 isotones, all related to a change of deformation, are presented. (orig.)

Combined fluorimetric caspase 3/7 assay and bradford protein determination for assessment of polycation-mediated cytotoxicity.

Science.gov (United States)

Larsen, Anna K; Hall, Arnaldur; Lundsgart, Henrik; Moghimi, S Moein

2013-01-01

Cationic polyplexes and lipoplexes are widely used as artificial systems for nucleic acid delivery into the cells, but they can also induce cell death. Mechanistic understanding of cell toxicity and biological side effects of these cationic entities is essential for optimization strategies and design of safe and efficient nucleic acid delivery systems. Numerous methods are presently available to detect and delineate cytotoxicity and cell death-mediated signals in cell cultures. Activation of caspases is part of the classical apoptosis program and increased caspase activity is therefore a well-established hallmark of programmed cell death. Additional methods to monitor cell death-related signals must, however, also be carried out to fully define the type of cell toxicity in play. These may include methods that detect plasma membrane damage, loss of mitochondrial membrane potential, phosphatidylserine exposure, and cell morphological changes (e.g., membrane blebbing, nuclear changes, cytoplasmic swelling, cell rounding). Here we describe a 96-well format protocol for detection of capsase-3/7 activity in cell lysates, based on a fluorescent caspase-3 assay, combined with a method to simultaneously determine relative protein contents in the individual wells.

A comparison between the efficiency of the Xpert MTB/RIF assay and nested PCR in identifying Mycobacterium tuberculosis during routine clinical practice.

Science.gov (United States)

Kim, Cheol-Hong; Woo, Heungjeong; Hyun, In Gyu; Kim, Changhwan; Choi, Jeong-Hee; Jang, Seung-Hun; Park, Sang Myeon; Kim, Dong-Gyu; Lee, Myung Goo; Jung, Ki-Suck; Hyun, Jeongwon; Kim, Hyun Soo

2014-06-01

Polymerase chain reaction (PCR) for the detection of Mycobacterium tuberculosis (MTB) is more sensitive, specific, and rapid than the conventional methods of acid-fast bacilli (AFB) smear and culture. The aim of this study was to determine if the Xpert MTB/rifampicin (RIF) assay had additional advantages over nested PCR for the detection of MTB in a geographical area with intermediate tuberculosis (TB) incidence. Between February and December 2013, the Xpert MTB/RIF assay and MTB nested PCR, as well as AFB smear and culture, were simultaneously performed on 198 clinical samples (160 pulmonary and 38 non-pulmonary specimens) collected from 171 patients hospitalized at Hallym University Medical Center for possible TB. The accuracy of the diagnosis of MTB culture-positive TB and the turnaround time of reporting laboratory results were calculated and compared. Rifampin resistance by the Xpert MTB/RIF assay was reviewed with that of conventional drug susceptibility testing (DST). The sensitivity, specificity, and positive and negative predictive values of the Xpert MTB/RIF assay and MTB nested PCR for diagnosis of MTB culture-positive pulmonary TB were 86.1% vs. 69.4% (P=0.1563), 97.8% vs. 94.1% (P=0.2173), 91.2% vs. 75.8% (P=0.1695), and 96.4% vs. 92.0% (P=0.2032), respectively. The median turnaround times of the Xpert MTB/RIF assay and MTB nested PCR were 0 [0-4] days and 4 [1-11] days, respectively (Pnested PCR for identifying MTB among clinically suspected TB patients, and the assay can be valuable in giving a timely identification of resistance to rifampin.

Variabilidade sazonal dos constituintes da própolis vermelha e bioatividade em Artermia salina

Directory of Open Access Journals (Sweden)

Lívio César Cunha Nunes

Full Text Available A própolis é uma substância resinosa coletada pelas abelhas de diversas partes das plantas. Sua composição depende da época, vegetação e local de coleta. Apresenta diversas atividades biológicas como antimicrobiana, antioxidante, antitumoral, dentre outras. Foi realizado estudo da variabilidade sazonal, nos meses de fevereiro, junho e outubro de 2006, dos constituintes voláteis da própolis vermelha de Pernambuco através da extração por headspace dinâmico e identificação por cromatografia gasosa acoplada com espectrometria de massas (CG-EM. Foram identificados 34 constituintes voláteis, sendo monoterpenos e monoterpenóides, sesquiterpenos e sesquiterpenóides, fenilpropanóides, aldeídos, cetonas e η-alcanos. Os constituintes majoritários foram o trans-anetol, α-copaeno e o metil cis-isoeugenol. Também foi realizado o perfil fitoquímico por cromatografia em camada delgada (CCD, através da qual os constituintes fenólicos foram identificados como majoritários. Com o extrato bruto metanólico da própolis, realizou-se o ensaio de letalidade em Artemia salina, que demonstrou DL50 de 18,9 µg/mL, sugerindo uma possível atividade antitumoral.

A high throughput colorimetric assay of β-1,3-D-glucans by Congo red dye.

Science.gov (United States)

Semedo, Magda C; Karmali, Amin; Fonseca, Luís

2015-02-01

Mushroom strains contain complex nutritional biomolecules with a wide spectrum of therapeutic and prophylactic properties. Among these compounds, β-d-glucans play an important role in immuno-modulating and anti-tumor activities. The present work involves a novel colorimetric assay method for β-1,3-d-glucans with a triple helix tertiary structure by using Congo red. The specific interaction that occurs between Congo red and β-1,3-d-glucan was detected by bathochromic shift from 488 to 516 nm (>20 nm) in UV-Vis spectrophotometer. A micro- and high throughput method based on a 96-well microtiter plate was devised which presents several advantages over the published methods since it requires only 1.51 μg of polysaccharides in samples, greater sensitivity, speed, assay of many samples and very cheap. β-D-Glucans of several mushrooms (i.e., Coriolus versicolor, Ganoderma lucidum, Pleurotus ostreatus, Ganoderma carnosum, Hericium erinaceus, Lentinula edodes, Inonotus obliquus, Auricularia auricular, Polyporus umbellatus, Cordyseps sinensis, Agaricus blazei, Poria cocos) were isolated by using a sequence of several extractions with cold and boiling water, acidic and alkaline conditions and quantified by this microtiter plate method. FTIR spectroscopy was used to study the structural features of β-1,3-D-glucans in these mushroom samples as well as the specific interaction of these polysaccharides with Congo red. The effect of NaOH on triple helix conformation of β-1,3-D-glucans was investigated in several mushroom species. Copyright © 2014 Elsevier B.V. All rights reserved.

A Soluble Fluorescent Binding Assay Reveals PIP2 Antagonism of TREK-1 Channels

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Cerrone Cabanos

2017-08-01

Full Text Available Lipid regulation of ion channels by low-abundance signaling lipids phosphatidylinositol 4,5-bisphosphate (PIP2 and phosphatidic acid (PA has emerged as a central cellular mechanism for controlling ion channels and the excitability of nerves. A lack of robust assays suitable for facile detection of a lipid bound to a channel has hampered the probing of the lipid binding sites and measuring the pharmacology of putative lipid agonists for ion channels. Here, we show a fluorescent PIP2 competition assay for detergent-purified potassium channels, including TWIK-1-related K+-channel (TREK-1. Anionic lipids PA and phosphatidylglycerol (PG bind dose dependently (9.1 and 96 μM, respectively and agonize the channel. Our assay shows PIP2 binds with high affinity (0.87 μM but surprisingly can directly antagonize TREK-1 in liposomes. We propose a model for TREK-1 lipid regulation where PIP2 can compete with PA and PG agonism based on the affinity of the lipid for a site within the channel.

Real-Time PCR Assay To Detect Smallpox Virus

Science.gov (United States)

Sofi Ibrahim, M.; Kulesh, David A.; Saleh, Sharron S.; Damon, Inger K.; Esposito, Joseph J.; Schmaljohn, Alan L.; Jahrling, Peter B.

2003-01-01

We developed a highly sensitive and specific assay for the rapid detection of smallpox virus DNA on both the Smart Cycler and LightCycler platforms. The assay is based on TaqMan chemistry with the orthopoxvirus hemagglutinin gene used as the target sequence. With genomic DNA purified from variola virus Bangladesh 1975, the limit of detection was estimated to be approximately 25 copies on both machines. The assay was evaluated in a blinded study with 322 coded samples that included genomic DNA from 48 different isolates of variola virus; 25 different strains and isolates of camelpox, cowpox, ectromelia, gerbilpox, herpes, monkeypox, myxoma, rabbitpox, raccoonpox, skunkpox, vaccinia, and varicella-zoster viruses; and two rickettsial species at concentrations mostly ranging from 100 fg/μl to 1 ng/μl. Contained within those 322 samples were variola virus DNA, obtained from purified viral preparations, at concentrations of 1 fg/μl to 1 ng/μl. On the Smart Cycler platform, 2 samples with false-positive results were detected among the 116 samples not containing variola virus tested; i.e., the overall specificity of the assay was 98.3%. On the LightCycler platform, five samples with false-positive results were detected (overall specificity, 95.7%). Of the 206 samples that contained variola virus DNA ranging in concentrations from 100 fg/μl to 1 ng/μl, 8 samples were considered negative on the Smart Cycler platform and 1 sample was considered negative on the LightCycler platform. Thus, the clinical sensitivities were 96.1% for the Smart Cycler instrument and 99.5% for the LightCycler instrument. The vast majority of these samples were derived from virus-infected cell cultures and variola virus-infected tissues; thus, the DNA material contained both viral DNA and cellular DNA. Of the 43 samples that contained purified variola virus DNA ranging in concentration from 1 fg/μl to 1 ng/μl, the assay correctly detected the virus in all 43 samples on both the Smart Cycler

Proteomic Analyses Reveal the Mechanism of Dunaliella salina Ds-26-16 Gene Enhancing Salt Tolerance in Escherichia coli.

Directory of Open Access Journals (Sweden)

Yanlong Wang

Full Text Available We previously screened the novel gene Ds-26-16 from a 4 M salt-stressed Dunaliella salina cDNA library and discovered that this gene conferred salt tolerance to broad-spectrum organisms, including E. coli (Escherichia coli, Haematococcus pluvialis and tobacco. To determine the mechanism of this gene conferring salt tolerance, we studied the proteome of E. coli overexpressing the full-length cDNA of Ds-26-16 using the iTRAQ (isobaric tags for relative and absolute quantification approach. A total of 1,610 proteins were identified, which comprised 39.4% of the whole proteome. Of the 559 differential proteins, 259 were up-regulated and 300 were down-regulated. GO (gene ontology and KEGG (Kyoto encyclopedia of genes and genomes enrichment analyses identified 202 major proteins, including those involved in amino acid and organic acid metabolism, energy metabolism, carbon metabolism, ROS (reactive oxygen species scavenging, membrane proteins and ABC (ATP binding cassette transporters, and peptidoglycan synthesis, as well as 5 up-regulated transcription factors. Our iTRAQ data suggest that Ds-26-16 up-regulates the transcription factors in E. coli to enhance salt resistance through osmotic balance, energy metabolism, and oxidative stress protection. Changes in the proteome were also observed in E. coli overexpressing the ORF (open reading frame of Ds-26-16. Furthermore, pH, nitric oxide and glycerol content analyses indicated that Ds-26-16 overexpression increases nitric oxide content but has no effect on glycerol content, thus confirming that enhanced nitric oxide synthesis via lower intercellular pH was one of the mechanisms by which Ds-26-16 confers salt tolerance to E. coli.

Multiplex Real-Time PCR Assay Targeting Eight Parasites Customized to the Korean Population: Potential Use for Detection in Diarrheal Stool Samples from Gastroenteritis Patients.

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Eun Jeong Won

Full Text Available Intestinal parasitic diseases occur worldwide and can cause diarrhea or gastroenteritis; however, their diagnosis is quite difficult, especially in low-endemism countries. We developed a multiplex real-time PCR assay for detection of eight intestinal parasites and prospectively evaluated it for patients with gastroenteritis. The assay targeted Cryptosporidium parvum, Giardia lamblia, Entamoeba histolytica, Blastocystis hominis, Dientamoeba fragilis, Clonorchis sinensis, Metagonimus yokogawai, and Gymnophalloides seoi. Performance characteristics were evaluated based on recovery after DNA extraction, analytical sensitivity, specificity, reproducibility, cross-reactivity, and interference characteristics. Clinical performance was validated against microscopy on 123 diarrheal samples. The assay demonstrated strong correlations between DNA concentrations and Ct values (R2, 0.9924-0.9998, and had a high PCR efficiency (83.3%-109.5%. Polymerase chain reactions detected as few as 10-30 copies of genomic DNA, and coefficient of variance was 0-7%. There was no cross-reactivity to the other 54 microorganisms tested. Interference occurred only in presence of high concentrations of erythrocytes or leukocytes. This assay had a higher correct identification rate (100.0% vs. 90.2% and lower incorrect ID rate (0.0% vs. 9.8% when compared to microscopy. Overall, this assay showed a higher sensitivity (100.0%; 95% confidence interval [CI] of 80.5-100.0 than microscopy (29.4%; 95% CI 10.31-55.96, and the specificity levels were comparable for both methods (100.0%; 95% CI 96.58-100.0. This newly developed multiplex real-time PCR assay offers a potential use for detecting intestinal parasitic pathogens customized to the Korean population.

Development of a solid-phase assay for measurement of proteolytic enzyme activity

International Nuclear Information System (INIS)

Varani, J.; Johnson, K.; Kaplan, J.

1980-01-01

A solid-phase, plate assay was developed for the measurement of proteolytic enzyme activity. In this assay procedure, radiolabeled substrates were dried onto the surface of microtiter wells. Following drying, the wells were washed two times with saline to remove the nonadherent substrate. When proteolytic enzymes were added to the wells, protein hydrolysis occurred, releasing radioactivity into the supernatant fluid. The amount of protein hydrolysis that occurred was reflected by the amount of radioactivity in the supernatant fluid. When 125 I-hemoglobin was used as the substrate, it was as susceptible to hydrolysis by trypsin in the solid-phase assay as it was in solution in a standard assay procedure. Protease activity from a variety of sources (including from viable cells as well as from extracellular sources) were also able to hydrolyze the hemoglobin on the plate. 125 I-Labeled serum albumen, fibrinogen, and rat pulmonary basement membrane were also susceptible to hydrolysis by trypsin in the solid phase. When [ 14 C]elastin was dried onto the plate, it behaved in a similar manner to elastin in solution. It was resistant to hydrolysis by nonspecific proteases such as trypsin and chymotrypsin but was highly susceptible to hydrolysis by elastase. The solid-phase plate assay has several features which recommended it for routine use. It is as sensitive as standard tube assays (and much more sensitive than routinely used colormetric assays). It is quick and convenient; there are no precipitation, centrifugation, or filtration steps. In addition, very small volumes of radioactive wastes are generated. Another advantage of the solid-phase plate assay is the resistance of the dried substrates to spontaneous breakdown and to microbial contamination. Finally, this assay is suitable for use with viable cells as well as for extracellular proteases

Transfer plate radioassay using cell monolayers to detect anti-cell surface antibodies synthesized by lymphocyte hybridomas

International Nuclear Information System (INIS)

Schneider, M.D.; Eisenbarth, G.S.

1979-01-01

A solid phase [ 125 I] Protein A radioassay for anti-cell surface antibodies is described, which employs target cell monolayers cultured on fenestrated polyvinyl chloride 96-well plates ('transfer plates'). The calibrated aperture in the bottom of each well is small enough to retain fluid contents by surface tension during monolayer growth, but also permits fluid to enter the wells when transfer plate are lowered into receptacles containing washing buffer on test sera. To assay for antibodies directed against target cell surface antigens, transfer plates bearing monolayers are inserted into microculture plates with corresponding 96-well geometry, thereby simultaneously sampling 96 wells. This assay allows rapid screening of hundreds of hybrid cell colonies for production of antibodies with desired tissue specificity. (Auth.)

Effect of temperature and viscosity on swimming velocity of the copepod Acartia tonsa, brine shrimp Artemia salina and rotifer Brachionus plicatilis

DEFF Research Database (Denmark)

Larsen, Poul Scheel; Madsen, C.V.; Riisgard, H.U.

2008-01-01

Beating cilia are important organelles for swimming in many zooplanktonic aquatic organisms, including many invertebrate larvae, rotifers and ciliates, but other planktonic organisms, such as copepods and brine shrimps, use muscle-powered swimming appendages. In recent studies we found...... of swimming velocity for a 10 degrees C temperature reduction) that is found to be largest for the brine shrimp Artemia salina nauplius (37 %) and the rotifer Brachionus plicatilis (26%), but negligible for the copepod Acartia tonsa (4%). We suggest that experimental data on change in swimming velocity (V......) due to change in kinematic viscosity (v) be correlated in terms of a power law, V proportional to v(-m). The present data on swimming velocity of copepods, brine shrimps and rotifers show values of exponent m approximate to 1.5 to 3, with a trend of decreasing values for increasing size of species...

Use of GenoType® MTBDRplus assay to assess drug resistance and mutation patterns of multidrug-resistant tuberculosis isolates in northern India

Directory of Open Access Journals (Sweden)

A K Maurya

2013-01-01

Full Text Available Purpose: The emergence and spread of multidrug-resistant tuberculosis (MDR-TB is a major public health problem. The diagnosis of MDR-TB is of paramount importance in establishing appropriate clinical management and infection control measures. The aim of this study was to evaluate drug resistance and mutational patterns in clinical isolates MDR-TB by GenoType® MTBDRplus assay. Material and Methods: A total of 350 non-repeated sputum specimens were collected from highly suspected drug-resistant pulmonary tuberculosis (PTB cases; which were processed by microscopy, culture, differentiation and first line drug susceptibility testing (DST using BacT/ALERT 3D system. Results: Among a total of 125 mycobacterium tuberculosis complex (MTBC strains, readable results were obtained from 120 (96% strains by GenoType® MTBDRplus assay. Only 45 MDR-TB isolates were analysed for the performance, frequency and mutational patterns by GenoType® MTBDRplus assay. The sensitivity of the GenoType® MDRTBplus assay for detecting individual resistance to rifampicin (RIF, isoniazid (INH and multidrug resistance was found to be 95.8%, 96.3% and 97.7%, respectively. Mutation in codon S531L of the rpoB gene and codon S315T1 of katG genes were dominated in MDR-TB strains, respectively (P < 0.05. Conclusions: The GenoType® MTBDRplus assay is highly sensitive with short turnaround times and a rapid test for the detection of the most common mutations conferring resistance in MDR-TB strains that can readily be included in a routine laboratory workflow.

Pentobarbital quantitation using EMIT serum barbiturate assay reagents: application to monitoring of high-dose pentobarbital therapy.

Science.gov (United States)

Pape, B E; Cary, P L; Clay, L C; Godolphin, W

1983-01-01

Pentobarbital serum concentrations associated with a high-dose therapeutic regimen were determined using EMIT immunoassay reagents. Replicate analyses of serum controls resulted in a within-assay coefficient of variation of 5.0% and a between-assay coefficient of variation of 10%. Regression analysis of 44 serum samples analyzed by this technique (y) and a reference procedure (x) were y = 0.98x + 3.6 (r = 0.98; x = ultraviolet spectroscopy) and y = 1.04x + 2.4 (r = 0.96; x = high-performance liquid chromatography). Clinical evaluation of the results indicates the immunoassay is sufficiently sensitive and selective for pentobarbital to allow accurate quantitation within the therapeutic range associated with high-dose therapy.

40 CFR 96.3 - Measurements, abbreviations, and acronyms.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false Measurements, abbreviations, and acronyms. 96.3 Section 96.3 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR... pounds. CO2—carbon dioxide. NOX—nitrogen oxides. O2—oxygen. ...

A novel method for radioimmunoassay and its application to the assay of digoxin

International Nuclear Information System (INIS)

Lader, S.R.

1977-01-01

A novel radioimmunoassay method has been developed for digoxin which eliminates the need for centrifugation while retaining a short incubation time at ambient temperature. The apparatus used consists of a plastic reaction chamber containing a pad of absorbent material impregnated with solid phase antibody. A mixture of tracer and standard (or tracer and sample) is pipetted onto the antibody-containing pad and at the end of the thirty minute incubation period radioactivity remaining unbound to antibody is drawn through the pad into an absorbent filter by capillary attraction. The specificity, sensitivity, precision and accuracy of the method are described. Clinical evaluation of the method has been carried out in two laboratories in the United Kingdom and four in the United States of America. The consistency of performance of the new digoxin method as judged by precision and accuracy were quite striking compared with the wide variation in performance of routine in-house methods. Thus, results for between assay reproducibility show that C.V.'s for the novel method ranged from 9.6% to 15.5% while those for the routine assays ranged from 7% to 27,8%. The recovery of digoxin added to serum varied from 85% to 117% for the new method and from 88% to 164% with in-house methods. In a total of 640 clinical samples the results compared well with those obtained with established in-house procedures. (orig.) [de

Radioimmunoassay of type D oncovirus from continuous J-96 cells

International Nuclear Information System (INIS)

Vlasenkova, N.K.; Altshtejn, A.D.; Zhdanov, V.M.; Kitsak, V.Ya.

1978-01-01

The radioimmunoassay of the J-96 virus and an extract of J-96 cells in the homologous and heterologous systems aimed at detecting antigenic determinants of p25 of Mason-Pfizer virus and group-specific and interspecies antigenic determinants p30 of Rauscher leukaemia virus demonstrated that (1) J-96 virus contains a major internal protein immunologically identical with p25 protein of Mason-Pfizer virus based on the antigenic determinants detectable by the radioimmunoassay used; and (2) no interspecies antigenic determinants characteristic of the major internal protein of mammalian type C viruses were detectable in the J-96 virus or the J-96 cell extract. (author)

DOE Handbook: Supplementary guidance and design experience for the fusion safety standards DOE-STD-6002-96 and DOE-STD-6003-96

International Nuclear Information System (INIS)

1999-01-01

Two standards have been developed that pertain to the safety of fusion facilities. These are DOE- STD-6002-96, Safety of Magnetic Fusion Facilities: Requirements, and DOE-STD-6003-96, Safety of Magnetic Fusion Facilities: Guidance. The first of these standards identifies requirements that subscribers to that standard must meet to achieve safety in fusion facilities. The second standard contains guidance to assist in meeting the requirements identified inthefirst This handbook provides additional documentation on good operations and design practices as well as lessons learned from the experiences of designers and operators of previous fusion facilities and related systems. It is intended to capture the experience gained in the various fields and pass it on to designers of future fusion facilities as a means of enhancing success and safeiy. The sections of this document are presented according to the physical location of the major systems of a t%sion facility, beginning with the vacuum vessel and proceeding to those systems and components outside the vacuum vessel (the ''Ex-vessel Systems''). The last section describes administrative procedures that cannot be localized to specific components. It has been tacitly assumed that the general structure of the fusion facilities addressed is that of a tokamak though the same principles would apply to other magnetic confinement options

DOE Handbook: Supplementary guidance and design experience for the fusion safety standards DOE-STD-6002-96 and DOE-STD-6003-96

Energy Technology Data Exchange (ETDEWEB)

None

1999-01-01

Two standards have been developed that pertain to the safety of fusion facilities. These are DOE- STD-6002-96, Safety of Magnetic Fusion Facilities: Requirements, and DOE-STD-6003-96, Safety of Magnetic Fusion Facilities: Guidance. The first of these standards identifies requirements that subscribers to that standard must meet to achieve safety in fusion facilities. The second standard contains guidance to assist in meeting the requirements identified in the first This handbook provides additional documentation on good operations and design practices as well as lessons learned from the experiences of designers and operators of previous fusion facilities and related systems. It is intended to capture the experience gained in the various fields and pass it on to designers of future fusion facilities as a means of enhancing success and safety. The sections of this document are presented according to the physical location of the major systems of a fusion facility, beginning with the vacuum vessel and proceeding to those systems and components outside the vacuum vessel (the "Ex-vessel Systems"). The last section describes administrative procedures that cannot be localized to specific components. It has been tacitly assumed that the general structure of the fusion facilities addressed is that of a tokamak though the same principles would apply to other magnetic confinement options.

New limits for the 2 νββ decay of 96Zr to excited nuclear states of 96Mo

Science.gov (United States)

Finch, Sean; Tornow, Werner

2015-10-01

The final results from our search for the 2 νββ decay of 96Zr to excited 0+ and 2+ states of 96Mo are presented. Such measurements provide valuable test cases for 2 νββ -decay nuclear matrix element calculations, which in turn are used to tune 0 νββ -decay nuclear matrix element calculations. After undergoing double- β decay to an excited state, the excited daughter nucleus decays to the ground state, emitting two coincident γ rays. These two γ rays are detected in coincidence by two HPGe detectors sandwiching the 96Zr sample, with a NaI veto in anti-coincidence. This experimental apparatus, located at the Kimballton Underground Research Facility (KURF), has previously measured the 2 νββ decay of 100Mo and 150Nd to excited nuclear states. Experimental limits on the T1 / 2 and corresponding nuclear matrix element are presented for each of these decays. As a byproduct of this experiment, limits were also set on the single- β decay of 96Zr. Supported by DOE Grant: DE-FG02-97ER41033.

Developing high throughput quantitative PCR assays for diagnosing Ikeda and other Theileria orientalis types common to New Zealand in bovine blood samples.

Science.gov (United States)

Pulford, D J; Gias, E; Bueno, I M; McFadden, Amj

2016-01-01

To develop rapid, quantitative PCR (qPCR) assays using high resolution melt (HRM) analysis and type-specific TaqMan assays for identifying the prevalent types of Theileria orientalis found in New Zealand cattle; and to evaluate their analytical and diagnostic characteristics compared with other assays for T. orientalis. Nucleotide sequences aligned with T. orientalis Buffeli, Chitose and Ikeda types, obtained from DNA extracted from blood samples from infected cattle, were used to design HRM and type-specific probe-based qPCR assays. The three type-specific assays were also incorporated into a single-tube multiplex qPCR assay. These assays were validated using DNA extracted from blood samples from cattle in herds with or without clinical signs of T. orientalis infection, other veterinary laboratory samples, as well as plasmids containing T. orientalis type-specific sequences. Diagnostic specificity (DSp) and sensitivity (DSe) estimates for the qPCR assays were compared to blood smear piroplasm results, and other PCR assays for T. orientalis. Copy number estimates of Ikeda DNA in blood were determined from cattle exhibiting anaemia using the Ikeda-specific qPCR assay. The T. orientalis type-specific and the HRM qPCR assays displayed 100% analytical specificity. The Ikeda-specific qPCR assay exhibited linearity (R(2) = 0.997) with an efficiency of 94.3%. Intra-assay CV were ≤0.08 and inter-assay CV were ≤0.095. For blood samples from cows with signs of infection with T. orientalis, the DSp and DSe of the multiplex probe qPCR assay were 93 and 96%, respectively compared with blood smears, and 97 and 100%, respectively compared with conventional PCR assays. For the Ikeda-specific qPCR assay, the number of positive samples (n=66) was slightly higher than a conventional PCR assay (n=64). The concentration of Ikeda genomes in blood samples from 41 dairy cows with signs of infection with T. orientalis ranged between 5.6 × 10(4) and 3.3 × 10(6) genomes per

Growth of mono- and mixed cultures of Nannochloropsis salina and Phaeodactylum tricornutum on struvite as a nutrient source

Energy Technology Data Exchange (ETDEWEB)

Davis, Ryan W. [Sandia National Lab. (SNL-CA), Livermore, CA (United States); Siccardi, Anthony J. [Texas AgriLife Research Mariculture Lab., Corpus Christi, TX (United States); Huysman, Nathan D. [Texas AgriLife Research Mariculture Lab., Corpus Christi, TX (United States); Wyatt, Nicholas B. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Hewson, John C. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Lane, Todd W. [Sandia National Lab. (SNL-CA), Livermore, CA (United States)

2015-09-26

In this paper, the suitability of crude and purified struvite (MgNH₄PO₄), a major precipitate in wastewater streams, was investigated for renewable replacement of conventional nitrogen and phosphate resources for cultivation of microalgae. Bovine effluent wastewater stone, the source of crude struvite, was characterized for soluble N/P, trace metals, and biochemical components and compared to the purified mineral. Cultivation trials using struvite as a major nutrient source were conducted using two microalgae production strains, Nannochloropsis salina and Phaeodactylum tricornutum, in both lab and outdoor pilot-scale raceways in a variety of seasonal conditions. Both crude and purified struvite-based media were found to result in biomass productivities at least as high as established media formulations (maximum outdoor co-culture yield ~20 ± 4 g AFDW/m²/day). Finally, analysis of nutrient uptake by the alga suggest that struvite provides increased nutrient utilization efficiency, and that crude struvite satisfies the trace metals requirement and results in increased pigment productivity for both microalgae strains.

EL PAN VS. SALINAS O EL DILEMA DEL PRISIONERO: REFORMA INSTITUCIONAL Y COOPERACIÓN COMO ESTRATEGIA POLÍTICA

Directory of Open Access Journals (Sweden)

Alicia Gómez López

2002-01-01

Full Text Available Este artículo analiza, desde el punto de vista de la teoría de juegos, la estrategia que el Partido Acción Nacional desarrolló frente al gobierno de Carlos Salinas de Gortari luego de la turbulenta elección presidencial de 1988. El objetivo es demostrar que entre estos adversarios se estableció un tipo de juego cuyo modelo a menudo se utiliza para analizar procesos de transición: el Dilema del prisionero, con un desenlace, al menos en este caso, cooperativo. Como en todo dilema, en este juego la posibilidad de la cooperación implica la existencia de un escenario aceptable para ambos adversarios, pese a que en primer lugar se busquen resultados opuestos. En este caso, dicho escenario se relaciona con el hecho de que el objetivo central en la estrategia panista era lograr reformas institucionales que hicieran avanzar la transición mexicana, de manera que le fuera posible llegar al poder; un objetivo cuya materia —las reformas institucionales— resultaba perfectamente negociable para su adversario presidencial.

Study Bioprospecting of Medicinal Plant Extracts of the Semiarid Northeast: Contribution to the Control of Oral Microorganisms

Directory of Open Access Journals (Sweden)

Maria Suênia P. Silva

2012-01-01

Full Text Available Dental pathologies can be caused by plaque-forming bacteria and yeast, which reside in the oral cavity. The bacteria growing in dental plaque, a naturally occurring biofilm, display increased resistance to antimicrobial agents. The objective was the evaluation of a preclinical assay of medicinal plants of the semiarid region from the northeast against oral pathogenic microorganism, aiming at bioprospecting a new product. The selection of plant material for this study was based on the ethnobotanical data on the traditional use of plants from the semiarid region. The thirty extracts were subjected to the determination of antibiofilm activity against gram-positive, gram-negative bacteria and yeast. The hydroalcoholic extract which showed positive antibiofilm activity against most of the microorganisms tested in agar diffusion assay was further tested for the determination of minimum inhibitory concentration (MIC and Bioassay with Artemia salina. Plant samples tested in this study exhibited good antibiofilm activity for the treatment of oral problems. The Schinopsis brasiliensis showed greater activity for Pseudomonas aeruginosa and Staphylococcus aureus, but toxicity against Artemia salina.

Clinical Evaluation of Fully Automated Elecsys® Syphilis Assay for the Detection of Antibodies of Treponema pallidum.

Science.gov (United States)

Li, Dongdong; An, Jingna; Wang, Tingting; Tao, Chuanmin; Wang, Lanlan

2016-11-01

The resurgence of syphilis in recent years has become a serious threat to the public health worldwide, and the serological detection of specific antibodies against Treponema pallidum (TP) remains the most reliable method for laboratory diagnosis of syphilis. The performance of the Elecsys ® Syphilis assay, a brand new electrochemiluminescene immunoassay (ECLIA), was assessed by large amounts of samples in this study. In comparison with InTec assay, the Elecsys ® Syphilis assay was evaluated in 146 preselected samples from patients with syphilis, 1803 clinical routine samples, and 175 preselected samples from specific populations with reportedly increased rates of false-positive syphilis test results. Discrepancy samples must be investigated by Mikrogen Syphilis recomline assay. There was an overall agreement of 99.58% between two assays (Kappa = 0.975). The sensitivity and specificity of the Elecsys ® Syphilis assay were 100.0% (95% CI, 96.8-100.0%) and 99.8% (95% CI, 99.5-100.0%), respectively. The Elecsys syphilis assay displays better sensitivity (100%), specificity (99.8%), PPV (98.7%), and NPV (100%) in 2124 samples enrolled, compared with the InTec assay. Considering the excellent ease of use and automation, high throughput, and its superior sensitivity, especially in primary syphilis, the Elecsys ® Syphilis assay could represent an outstanding choice for screening of syphilis in high-volume laboratories. However, more attention was still needed, or the results must be confirmed by other treponemal immunoassays. The new Elecsys ® Syphilis assay is applied to patients with malignant neoplasm or HIV infection. © 2016 Wiley Periodicals, Inc.

Saliva Polymerase-Chain-Reaction Assay for Cytomegalovirus Screening in Newborns

Science.gov (United States)

Boppana, Suresh B.; Ross, Shannon A.; Shimamura, Masako; Palmer, April L.; Ahmed, Amina; Michaels, Marian G.; Sánchez, Pablo J.; Bernstein, David I.; Tolan, Robert W.; Novak, Zdenek; Chowdhury, Nazma; Britt, William J.; Fowler, Karen B.

2011-01-01

BACKGROUND Congenital cytomegalovirus (CMV) infection is an important cause of hearing loss, and most infants at risk for CMV-associated hearing loss are not identified early in life because of failure to test for the infection. The standard assay for newborn CMV screening is rapid culture performed on saliva specimens obtained at birth, but this assay cannot be automated. Two alternatives — real-time polymerase-chain-reaction (PCR)–based testing of a liquid-saliva or dried-saliva specimen obtained at birth — have been developed. METHODS In our prospective, multicenter screening study of newborns, we compared real-time PCR assays of liquid-saliva and dried-saliva specimens with rapid culture of saliva specimens obtained at birth. RESULTS A total of 177 of 34,989 infants (0.5%; 95% confidence interval [CI], 0.4 to 0.6) were positive for CMV, according to at least one of the three methods. Of 17,662 newborns screened with the use of the liquid-saliva PCR assay, 17,569 were negative for CMV, and the remaining 85 infants (0.5%; 95% CI, 0.4 to 0.6) had positive results on both culture and PCR assay. The sensitivity and specificity of the liquid-saliva PCR assay were 100% (95% CI, 95.8 to 100) and 99.9% (95% CI, 99.9 to 100), respectively, and the positive and negative predictive values were 91.4% (95% CI, 83.8 to 96.2) and 100% (95% CI, 99.9 to 100), respectively. Of 17,327 newborns screened by means of the dried-saliva PCR assay, 74 were positive for CMV, whereas 76 (0.4%; 95% CI, 0.3 to 0.5) were found to be CMV-positive on rapid culture. Sensitivity and specificity of the dried-saliva PCR assay were 97.4% (95% CI, 90.8 to 99.7) and 99.9% (95% CI, 99.9 to 100), respectively. The positive and negative predictive values were 90.2% (95% CI, 81.7 to 95.7) and 99.9% (95% CI, 99.9 to 100), respectively. CONCLUSIONS Real-time PCR assays of both liquid- and dried-saliva specimens showed high sensitivity and specificity for detecting CMV infection and should be

Absorbance and fluorometric sensing with capillary wells microplates

International Nuclear Information System (INIS)

Tan, Han Yen; Cheong, Brandon Huey-Ping; Neild, Adrian; Wah Ng, Tuck; Liew, Oi Wah

2010-01-01

Detection and readout from small volume assays in microplates are a challenge. The capillary wells microplate approach [Ng et al., Appl. Phys. Lett. 93, 174105 (2008)] offers strong advantages in small liquid volume management. An adapted design is described and shown here to be able to detect, in a nonimaging manner, fluorescence and absorbance assays minus the error often associated with meniscus forming at the air-liquid interface. The presence of bubbles in liquid samples residing in microplate wells can cause inaccuracies. Pipetting errors, if not adequately managed, can result in misleading data and wrong interpretations of assay results; particularly in the context of high throughput screening. We show that the adapted design is also able to detect for bubbles and pipetting errors during actual assay runs to ensure accuracy in screening.

Absorbance and fluorometric sensing with capillary wells microplates

Energy Technology Data Exchange (ETDEWEB)

Tan, Han Yen; Cheong, Brandon Huey-Ping; Neild, Adrian; Wah Ng, Tuck [Laboratory for Optics, Acoustics, and Mechanics, Department of Mechanical and Aerospace Engineering, Monash University, Clayton, Victoria 3800 (Australia); Liew, Oi Wah [Cardiovascular Biomarkers Laboratory, Cardiovascular Research Institute, 30 Medical Drive, Singapore 117609 (Singapore)

2010-12-15

Detection and readout from small volume assays in microplates are a challenge. The capillary wells microplate approach [Ng et al., Appl. Phys. Lett. 93, 174105 (2008)] offers strong advantages in small liquid volume management. An adapted design is described and shown here to be able to detect, in a nonimaging manner, fluorescence and absorbance assays minus the error often associated with meniscus forming at the air-liquid interface. The presence of bubbles in liquid samples residing in microplate wells can cause inaccuracies. Pipetting errors, if not adequately managed, can result in misleading data and wrong interpretations of assay results; particularly in the context of high throughput screening. We show that the adapted design is also able to detect for bubbles and pipetting errors during actual assay runs to ensure accuracy in screening.

Radioreceptor assay: theory and applications to pharmacology

International Nuclear Information System (INIS)

Perret, G.; Simon, P.

1984-01-01

The aim of the first part of this work is to present the theory of the radioreceptor assay and to compare it to the other techniques of radioanalysis (radioimmunoassay, competitive protein binding assays). The technology of the radioreceptor assay is then presented and its components (preparation of the receptors, radioligand, incubation medium) are described. The analytical characteristics of the radioreceptor assay (specificity, sensitivity, reproductibility, accuracy) and the pharmacological significance of the results are discussed. The second part is devoted to the description of the radioreceptor assays of some pharmacological classes (neuroleptics, tricyclic antidepressants, benzodiazepines, β-blockers, anticholinergic drugs) and to their use in therapeutic drug monitoring. In conclusion, by their nature, radioreceptor assays are highly sensitive, reliable, precise, accurate and simple to perform. Their chief disadvantage relates to specificity, since any substance having an appreciable affinity to the receptor site will displace the specifically bound radioligand. Paradoxically in some cases, this lack of specificity may be advantageous in that it allows for the detection of not only the apparent compound but of active metabolites and endogenous receptor agonists as well and in that radioreceptors assays can be devised for a whole pharmacological class and not only for one drug as it is the case for classical physico-chemical techniques. For all these reasons future of radioreceptor assay in pharmacology appears promising [fr

25-hydroxy-Vitamin D status: limitations in comparison and clinical interpretation of serum-levels across different assay methods.

Science.gov (United States)

Enko, Dietmar; Fridrich, Leo; Rezanka, Erwin; Stolba, Robert; Ernst, Juliane; Wendler, Iris; Fabian, Daniel; Hauptlorenz, Susanne; Halwachs-Baumann, Gabriele

2014-01-01

Background: Over the last decade, clinical interest to evaluate human 25-hydroxy-vitamin D (25[OH]D) serum levels has increased exponentially. In the present study, four chemiluminescence immunoassays (CLIA), one radioimmunoassy (RIA), and one high performance liquid chromatography (HPLC) method were compared and also with the liquid chromatography-tandem mass spectrometry (LC-MS/MS) method in view of 25(OH)D serum level determination. Methods: For the method comparison, blood samples from 133 consecutive patients were prospectively collected. All participants gave written informed consent for their blood samples to be used in this study. They came to the Department of Nuclear Medicine of the Central Hospital Steyr (Austria) for osteodensidometric measurement as part of their preventive medical check-up. Pearson's correlation coefficients, Bland-Altman plots, and paired t-tests were calculated. Assay-specific reference ranges were considered using blood samples from persons with normal parathormone, calcium, and total-protein values (n = 97). Results: The highest correlation was between the HPLC and the LC-MS/MS method (r = 0.96). The lowest correlation was between the cobas Vitamin D3 assay (Roche) and any of the evaluated assays (r = 0.46 - 0.63). Bland-Altman plots revealed a big negative mean bias in three assays (cobas Vitamin D3 assay [Roche]: -22.8; DiaSorin LIAISON [25[OH]D total CLIA [Diasorin]: -18.4; Diasorin 25[OH]D125 I RIA [Diasorin]: -23.8 [nmol/L]) and a much smaller positive mean bias in the other assays (ClinRep complete 25[OH]D2/D3 HPLC kit [Recipe]: 2.7; ADVIA Centaur Vitamin D total assay [Siemens]: 8.2; IDS total vitamin D assay [Immunodiagnostic Systems]: 12.1 [nmol/L]) compared to the LC-MS/MS method. Meanwhile, the manufacturer has withdrawn the cobas Vitamin D3 assay from the market. Conclusions: Poor antibody specificity with cross-reactivity to other vitamin D metabolites, incomplete extraction of the 25(OH)D analyte from the vitamin D

Application of neutron multiplicity counting to waste assay

Energy Technology Data Exchange (ETDEWEB)

Pickrell, M.M.; Ensslin, N. [Los Alamos National Lab., NM (United States); Sharpe, T.J. [North Carolina State Univ., Raleigh, NC (United States)

1997-11-01

This paper describes the use of a new figure of merit code that calculates both bias and precision for coincidence and multiplicity counting, and determines the optimum regions for each in waste assay applications. A {open_quotes}tunable multiplicity{close_quotes} approach is developed that uses a combination of coincidence and multiplicity counting to minimize the total assay error. An example is shown where multiplicity analysis is used to solve for mass, alpha, and multiplication and tunable multiplicity is shown to work well. The approach provides a method for selecting coincidence, multiplicity, or tunable multiplicity counting to give the best assay with the lowest total error over a broad spectrum of assay conditions. 9 refs., 6 figs.

Automation of a Nile red staining assay enables high throughput quantification of microalgal lipid production.

Science.gov (United States)

Morschett, Holger; Wiechert, Wolfgang; Oldiges, Marco

2016-02-09

Within the context of microalgal lipid production for biofuels and bulk chemical applications, specialized higher throughput devices for small scale parallelized cultivation are expected to boost the time efficiency of phototrophic bioprocess development. However, the increasing number of possible experiments is directly coupled to the demand for lipid quantification protocols that enable reliably measuring large sets of samples within short time and that can deal with the reduced sample volume typically generated at screening scale. To meet these demands, a dye based assay was established using a liquid handling robot to provide reproducible high throughput quantification of lipids with minimized hands-on-time. Lipid production was monitored using the fluorescent dye Nile red with dimethyl sulfoxide as solvent facilitating dye permeation. The staining kinetics of cells at different concentrations and physiological states were investigated to successfully down-scale the assay to 96 well microtiter plates. Gravimetric calibration against a well-established extractive protocol enabled absolute quantification of intracellular lipids improving precision from ±8 to ±2 % on average. Implementation into an automated liquid handling platform allows for measuring up to 48 samples within 6.5 h, reducing hands-on-time to a third compared to manual operation. Moreover, it was shown that automation enhances accuracy and precision compared to manual preparation. It was revealed that established protocols relying on optical density or cell number for biomass adjustion prior to staining may suffer from errors due to significant changes of the cells' optical and physiological properties during cultivation. Alternatively, the biovolume was used as a measure for biomass concentration so that errors from morphological changes can be excluded. The newly established assay proved to be applicable for absolute quantification of algal lipids avoiding limitations of currently established

A gene encoding an abscisic acid biosynthetic enzyme (LsNCED4) collocates with the high temperature germination locus Htg6.1 in lettuce (Lactuca sp.).

Science.gov (United States)

Argyris, Jason; Truco, María José; Ochoa, Oswaldo; McHale, Leah; Dahal, Peetambar; Van Deynze, Allen; Michelmore, Richard W; Bradford, Kent J

2011-01-01

Thermoinhibition, or failure of seeds to germinate when imbibed at warm temperatures, can be a significant problem in lettuce (Lactuca sativa L.) production. The reliability of stand establishment would be improved by increasing the ability of lettuce seeds to germinate at high temperatures. Genes encoding germination- or dormancy-related proteins were mapped in a recombinant inbred line population derived from a cross between L. sativa cv. Salinas and L. serriola accession UC96US23. This revealed several candidate genes that are located in the genomic regions containing quantitative trait loci (QTLs) associated with temperature and light requirements for germination. In particular, LsNCED4, a temperature-regulated gene in the biosynthetic pathway for abscisic acid (ABA), a germination inhibitor, mapped to the center of a previously detected QTL for high temperature germination (Htg6.1) from UC96US23. Three sets of sister BC(3)S(2) near-isogenic lines (NILs) that were homozygous for the UC96US23 allele of LsNCED4 at Htg6.1 were developed by backcrossing to cv. Salinas and marker-assisted selection followed by selfing. The maximum temperature for germination of NIL seed lots with the UC96US23 allele at LsNCED4 was increased by 2-3°C when compared with sister NIL seed lots lacking the introgression. In addition, the expression of LsNCED4 was two- to threefold lower in the former NIL lines as compared to expression in the latter. Together, these data strongly implicate LsNCED4 as the candidate gene responsible for the Htg6.1 phenotype and indicate that decreased ABA biosynthesis at high imbibition temperatures is a major factor responsible for the increased germination thermotolerance of UC96US23 seeds.

45 CFR 96.102 - Carryover of unobligated funds.

Science.gov (United States)

2010-10-01

... reason or if the State has determined that program objectives would be better served by deferring... 45 Public Welfare 1 2010-10-01 2010-10-01 false Carryover of unobligated funds. 96.102 Section 96.102 Public Welfare DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL ADMINISTRATION BLOCK GRANTS Primary...

Topography and stoichiometry of acidic proteins in large ribosomal subunits from Artemia salina as determined by crosslinking

International Nuclear Information System (INIS)

Uchiumi, T.; Wahba, A.J.; Traut, R.R.

1987-01-01

The 60S subunits isolated from Artemia salina ribosomes were treated with the crosslinking reagent 2-iminothiolane under mild conditions. Proteins were extracted and fractions containing crosslinked acidic proteins were obtained by stepwise elution from CM-cellulose. Each fraction was analyzed by diagonal (two-dimensional nonreducing-reducing) NaDodSO 4 /polyacrylamide gel electrophoresis. Crosslinked proteins below the diagonal were radioiodinated and identified by two-dimensional acidic urea-NaDodSO 4 gel electrophoresis. Each of the acidic proteins P1 and P2 was crosslinked individually to the same third protein, PO. The fractions containing acidic proteins were also analyzed by two-dimensional nonequilibrium isoelectric focusing-NaDodSO 4 /polyacrylamide gel electrophoresis. Two crosslinked complexes were observed that coincide in isoelectric positions with monomeric P1 and P2, respectively. Both P1 and P2 appear to form crosslinked homodimers. These results suggest the presence in the 60S subunit of (P1) 2 and (P2) 2 dimers, each of which is anchored to PO. Protein PO appears to play the same role as L10 in Escherichia coli ribosomes and may form a pentameric complex with the two dimers in the 60S subunits

Development of a Sensitive and Specific Serological Assay Based on Luminex Technology for Detection of Antibodies to Zaire Ebola Virus.

Science.gov (United States)

Ayouba, Ahidjo; Touré, Abdoulaye; Butel, Christelle; Keita, Alpha Kabinet; Binetruy, Florian; Sow, Mamadou S; Foulongne, Vincent; Delaporte, Eric; Peeters, Martine

2017-01-01

The recent Zaire Ebola virus (EBOV) outbreak in West Africa illustrates clearly the need for additional studies with humans and animals to elucidate the ecology of Ebola viruses (EBVs). In this study, we developed a serological assay based on the Luminex technology. Nine recombinant proteins representing different viral regions (nucleoprotein [NP], 40-kDa viral protein [VP40], and glycoprotein [GP]) from four of the five EBV lineages were used. Samples from 94 survivors of the EBOV outbreak in Guinea and negative samples from 108 patients in France were used to calculate test performance for EBOV detection and cross-reaction with other Ebola virus lineages. For EBOV antibody detection, sensitivities of 95.7%, 96.8%, and 92.5% and specificities of 94.4%, 95.4%, and 96.3% for NP, GP, and VP40, respectively, were observed. All EBOV-negative samples that presented a reaction, except for one, interacted with a single antigen, whereas almost all samples from EBOV survivors were simultaneously reactive with NP and GP (90/94) or with NP, GP, and VP40 (87/94). Considering as positive for past EBOV infection only samples that reacted with EBOV NP and GP, sensitivity was 95.7% and specificity increased to 99.1%. Comparing results with commercial EBOV NP and GP enzyme-linked immunosorbent assays (ELISAs; Alpha Diagnostic, San Antonio, TX), lower sensitivity (92.5%) and high specificity (100%) were observed with the same positivity criteria. Samples from EBOV survivors cross-reacted with GP from Sudan Ebola virus (GP-SUDV) (81.9%), GP from Bundibugyo Ebola virus (GP-BDBV) (51.1%), GP from Reston Ebola virus (GP-RESTV) (9.6%), VP40-SUDV (76.6%), and VP40-BDBV (38.3%). Overall, we developed a sensitive and specific high-throughput serological assay, and defined an algorithm, for epidemiological surveys with humans. Copyright © 2016 American Society for Microbiology.

Detection of knockdown resistance (kdr mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

Directory of Open Access Journals (Sweden)

Ball Amanda

2007-08-01

Full Text Available Abstract Background Knockdown resistance (kdr is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1 TaqMan probes and 2 high resolution melt (HRM analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR, Heated Oligonucleotide Ligation Assay (HOLA, Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost, and safety (requirement for hazardous chemicals. Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions and the most specific (with the lowest number of incorrect scores. Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS

Detection of knockdown resistance (kdr) mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

Science.gov (United States)

Bass, Chris; Nikou, Dimitra; Donnelly, Martin J; Williamson, Martin S; Ranson, Hilary; Ball, Amanda; Vontas, John; Field, Linda M

2007-01-01

Background Knockdown resistance (kdr) is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1) TaqMan probes and 2) high resolution melt (HRM) analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR), Heated Oligonucleotide Ligation Assay (HOLA), Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA) and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost), and safety (requirement for hazardous chemicals). Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions) and the most specific (with the lowest number of incorrect scores). Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS-PCR, SSOP-ELISA, PCR Dot

Low sensitivity of type VII collagen enzyme-linked immunosorbent assay in epidermolysis bullosa acquisita : serration pattern analysis on skin biopsy is required for diagnosis

NARCIS (Netherlands)

Terra, J. B.; Jonkman, M. F.; Diercks, G. F. H.; Pas, H. H.

BackgroundThe type VII collagen (coll VII) enzyme-linked immunosorbent assay (ELISA) has been reported to have high sensitivity (>93%) and specificity (>96%) for diagnosing epidermolysis bullosa acquisita (EBA) in patients who are seropositive on indirect immunofluorescence on salt-split skin (SSS).

40 CFR 96.40 - State trading program budget.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 20 2010-07-01 2010-07-01 false State trading program budget. 96.40... (CONTINUED) NOX BUDGET TRADING PROGRAM AND CAIR NOX AND SO2 TRADING PROGRAMS FOR STATE IMPLEMENTATION PLANS NOX Allowance Allocations § 96.40 State trading program budget. The State trading program budget...

40 CFR 86.134-96 - Running loss test.

Science.gov (United States)

2010-07-01

... 40 Protection of Environment 18 2010-07-01 2010-07-01 false Running loss test. 86.134-96 Section... Heavy-Duty Vehicles; Test Procedures § 86.134-96 Running loss test. (a) Overview. Gasoline- and methanol-fueled vehicles are to be tested for running loss emissions during simulated high-temperature urban...

Comparison of antibody responses to human papillomavirus vaccination as measured by three assays

Directory of Open Access Journals (Sweden)

Hilary Ann Robbins

2014-01-01

Full Text Available Background: Different assays, including the competitive Luminex immunoassay (cLIA, secreted alkaline phosphatase neutralization assay (SEAP-NA, and virus-like particle-based ELISA, are commonly used to measure antibody responses after human papillomavirus (HPV vaccination. Direct assay comparisons aid interpretation of immunogenicity data evaluated by different assays. Methods: We compared cLIA to SEAP-NA and ELISA among 51 HPV16/18-vaccinated women enrolled in the Costa Rica Vaccine Trial. We tested replicate serum samples collected at months 0, 1, and 12 by HPV16/18 cLIA, SEAP-NA, and ELISA. For a subset (N=10, we further tested month 24 and 36 samples. We calculated seroprevalence estimates and Spearman rank correlation coefficients comparing cLIA to SEAP-NA and ELISA.Results: After one vaccine dose, seroprevalence by SEAP-NA and ELISA was 100% (both HPV16 and HPV18, and by cLIA was 96% (95% CI 87%-100% for HPV16 and 71% (95% CI 56%-83% for HPV18. Seroprevalence was 100% by all assays after 3 doses. Correlation between assays was high after one vaccine dose (cLIA/SEAP-NA ρ=0.91 (HPV16 and ρ=0.86 (HPV18; cLIA/ELISA ρ=0.84 (HPV16 and ρ=0.74 (HPV18; all p<0.001 and remained high through month 36. Ratios of mean antibody levels to seropositivity cutoffs at month 36 were lower for cLIA than for SEAP-NA or ELISA, particularly for HPV18 (HPV18 ratio for cLIA 1.9, SEAP-NA 3.5, ELISA 3.4.Conclusion: Though correlation between cLIA and SEAP-NA/ELISA is high and stable after vaccination, the assays differ in scale and sensitivity, with notable differences after 1 vaccine dose and for HPV18. Our results demonstrate that comparisons of antibody responses to HPV vaccination measured by different assays are approximate, and must consider biological and technical differences between assays.

Alaska Geoid Heights (GEOID96)

Data.gov (United States)

National Oceanic and Atmospheric Administration, Department of Commerce — This 2' x 4' geoid height grid for Alaska is distributed as a GEOID96 model. The computation used 1.1 million terrestrial and marine gravity data held in the...

A 252Cf based nondestructive assay system for fissile material

International Nuclear Information System (INIS)

Menlove, H.O.; Crane, T.W.

1978-01-01

A modulated 252 Cf source assay system 'Shuffler' based on fast-or-thermal-neutron interrogation combined with delayed-neutron counting has been developed for the assay of fissile material. The 252 Cf neutron source is repetitively transferred from the interrogation position to a shielded position while the delayed neutrons are counted in a high efficiency 3 He neutron well-counter. For samples containing plutonium, this well-counter is also used in the passive coincidence mode to assay the effective 240 Pu content. The design of an optimized neutron tailoring assembly for fast-neutron interrogation using a Monte Carlo Neutron Computer Code is described. The Shuffler system has been applied to the assay of fuel pellets, inventory samples, irradiated fuel and plutonium mixed-oxide fuel. The system can assay samples with fissile contents from a few milligrams up to several kilograms using thermal-neutron interrogation for the low mass samples and fast-neutron interrogation for the high mass samples. Samples containing 235 U- 238 U, or 233 U-Th, or UO 2 -PuO 2 fuel mixtures have been assayed with the Shuffler system. (Auth.)

29 CFR 96.54 - Responsibility for subrecipient audits.

Science.gov (United States)

2010-07-01

... fiscal year or $500,000 for fiscal years ending after December 31, 2003 are audited and that any audit... 29 Labor 1 2010-07-01 2010-07-01 true Responsibility for subrecipient audits. 96.54 Section 96.54 Labor Office of the Secretary of Labor AUDIT REQUIREMENTS FOR GRANTS, CONTRACTS, AND OTHER AGREEMENTS...

32 CFR 552.96 - Violations.

Science.gov (United States)

2010-07-01

.... Lewis Area Access Section or the Military Police as soon as possible. ... Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY MILITARY RESERVATIONS AND NATIONAL CEMETERIES REGULATIONS AFFECTING MILITARY RESERVATIONS Fort Lewis Land Use Policy § 552.96 Violations. Anyone...

27 CFR 26.96 - Prepayment of tax-release of wine.

Science.gov (United States)

2010-04-01

... of wine. 26.96 Section 26.96 Alcohol, Tobacco Products and Firearms ALCOHOL AND TOBACCO TAX AND TRADE... Taxpayment of Liquors and Articles in Puerto Rico Wine § 26.96 Prepayment of tax—release of wine. (a) Action by proprietor. Where the wine is to be withdrawn from bonded storage after payment of the computed...

Si96: A New Silicon Allotrope with Interesting Physical Properties

Directory of Open Access Journals (Sweden)

Qingyang Fan

2016-04-01

Full Text Available The structural mechanical properties and electronic properties of a new silicon allotrope Si96 are investigated at ambient pressure by using a first-principles calculation method with the ultrasoft pseudopotential scheme in the framework of generalized gradient approximation. The elastic constants and phonon calculations reveal that Si96 is mechanically and dynamically stable at ambient pressure. The conduction band minimum and valence band maximum of Si96 are at the R and G point, which indicates that Si96 is an indirect band gap semiconductor. The anisotropic calculations show that Si96 exhibits a smaller anisotropy than diamond Si in terms of Young’s modulus, the percentage of elastic anisotropy for bulk modulus and shear modulus, and the universal anisotropic index AU. Interestingly, most silicon allotropes exhibit brittle behavior, in contrast to the previously proposed ductile behavior. The void framework, low density, and nanotube structure make Si96 quite attractive for applications such as hydrogen storage and electronic devices that work at extreme conditions, and there are potential applications in Li-battery anode materials.

27 CFR 71.96 - Disqualification.

Science.gov (United States)

2010-04-01

... before another administrative law judge. If the Administrator should decide against the disqualification... Administrative Law Judges § 71.96 Disqualification. An administrative law judge shall, at any time, withdraw from... personal bias or otherwise warranting the disqualification of any administrative law judge, the...

Chromosome aberration assays in barley (Hordeum vulgare)

Energy Technology Data Exchange (ETDEWEB)

Constantin, M J [Univ. of Tennessee, Knoxville; Nilan, R A

1982-01-01

Barley is an exceellent organism for studies of induced chromosome aberrations because of its few (2n = 2x = 14) relatively large chromosomes. Root-tip and shoot-tip cells have been used extensively for the study of ionizing radiation-induced chromosome aberrations. The general procedures are well known, the technology is simple and easy to learn, and the assays are relatively quick and inexpensive. Both root tips and shoot tips can be used for the study of chemical mutagens as well as ionizing radiations. Pollen mother cells are well suited for studying the effects of mutagens on meiotic chromosomes. The literature review for the Gene-Tox Program reported on 61 chemicals tested for their effects on barley chromosomes. Of these, 90% were reported to be either positive or positive dose-related, while 7% were negative and 3% were questionable. Barley assays based on chromosomal aberrations are useful to detect the clastogenic potency of chemicals under laboratory conditions. Indications are that the data from barley can be used to corroborate data obtained from other organisms. Among the classes of chemicals assayed were: alcohols and phenols; alkaloids; epoxides; alkyl sulfates; amides and sulfonamides; aromatic amines; aryl halides; aziridines; alkenes; carbamates; hydroazides; nitroaromatics; nitrosamides; nitrosources; phenothiazines; and polycyclic aromatic hydrocarbons.

A validated HPLC-MS/MS assay for quantifying unstable pharmacologically active metabolites of clopidogrel in human plasma: application to a clinical pharmacokinetic study.

Science.gov (United States)

Furlong, Michael T; Savant, Ishani; Yuan, Moucun; Scott, Laura; Mylott, William; Mariannino, Thomas; Kadiyala, Pathanjali; Roongta, Vikram; Arnold, Mark E

2013-05-01

Clopidogrel is prescribed for the treatment of Acute Coronary Syndrome and recent myocardial infarction, recent stroke, or established peripheral arterial disease. A sensitive and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay was developed and validated to enable reliable quantification of four diastereomeric and chemically reactive thiol metabolites, two of which are pharmacologically active, in human plasma. The metabolites were stabilized by alkylation of their reactive thiol moieties with 2-bromo-3'-methoxyacetophenone (MPB). Following organic solvent mediated-protein precipitation in a 96-well plate format, chromatographic separation was achieved by gradient elution on an Ascentis Express RP-amide column. Chromatographic conditions were optimized to ensure separation of the four derivatized active metabolites. Derivatized metabolites and stable isotope-labeled internal standards were detected by positive ion electrospray tandem mass spectrometry. The HPLC-MS/MS assay was validated over concentration ranges of 0.125-125 ng/mL for metabolites H1-H3 and 0.101-101 ng/mL for H4. Intra- and inter-assay precision values for replicate quality control samples were within 14.3% for all analytes during the assay validation. Mean quality control accuracy values were within ±6.3% of nominal values for all analytes. Assay recoveries were high (>79%). The four derivatized analytes were stable in human blood for at least 2 h at room temperature and on ice. The analytes were also stable in human plasma for at least 25 h at room temperature, 372 days at -20 °C and -70 °C, and following at least five freeze-thaw cycles. The validated assay was successfully applied to the quantification of all four thiol metabolites in human plasma in support of a human pharmacokinetic study. Copyright © 2013 Elsevier B.V. All rights reserved.

A plasmacytoid dendritic cell (CD123+/CD11c-) based assay system to predict contact allergenicity of chemicals

Science.gov (United States)

Ayehunie, Seyoum; Snell, Maureen; Child, Matthew; Klausner, Mitchell

2009-01-01

A predictive allergenicity test system for assessing the contact allergenicity of chemicals is needed by the cosmetic and pharmaceutical industry to monitor product safety in the marketplace. Development of such non-animal alternative assay systems for skin sensitization and hazard identification has been pursued by policy makers and regulatory agencies. We investigated whether phenotypic and functional changes to a subset of dendritic cells (DC), plasmacytoid DC (pDC), could be used to identify contact allergens. To achieve this goal, normal human DC were generated from CD34+ progenitor cells and cryopreserved. Frozen DC were thawed and the pDC fraction (CD123+/CD11c-) was harvested using FACS sorting. The pDC were cultured, expanded, and exposed to chemical allergens (N=26) or non-allergens (N=22). Concentrations of each chemical that resulted in >50% viability was determined using FACS analysis of propidium iodide stained cells using pDC from 2-5 donors. Expression of the surface marker, CD86, which has been implicated in dendritic cell maturation, was used as a marker of allergenicity. CD86 expression increased (≥ 1.5 fold) for 25 of 26 allergens (sensitivity = 96%) but did not increase for 19 of 22 non-allergens (specificity = 86%). In a direct comparison to historical data for the regulatory approved, mouse local lymph node assay (LLNA) for 23 allergens and 22 non-allergens, the pDC method had sensitivity and specificity of 96% and 86%, respectively, while the sensitivity and specificity of the LLNA assay was 83% and 82%, respectively. In conclusion, CD86 expression in pDC appears to be a sensitive and specific indicator to identify contact allergenicity. Such an assay method utilizing normal human cells will be useful for high throughput screening of chemicals for allergenicity. PMID:19665512

A plasmacytoid dendritic cell (CD123+/CD11c-) based assay system to predict contact allergenicity of chemicals

International Nuclear Information System (INIS)

Ayehunie, Seyoum; Snell, Maureen; Child, Matthew; Klausner, Mitchell

2009-01-01

A predictive allergenicity test system for assessing the contact allergenicity of chemicals is needed by the cosmetic and pharmaceutical industry to monitor product safety in the marketplace. Development of such non-animal alternative assay systems for skin sensitization and hazard identification has been pursued by policy makers and regulatory agencies. We investigated whether phenotypic and functional changes to a subset of dendritic cells (DC), plasmacytoid DC (pDC), could be used to identify contact allergens. To achieve this goal, normal human DC were generated from CD34+ progenitor cells and cryopreserved. Frozen DC were thawed and the pDC fraction (CD123+/CD11c-) was harvested using FACS sorting. The pDC were cultured, expanded, and exposed to chemical allergens (N = 26) or non-allergens (N = 22). Concentrations of each chemical that resulted in >50% viability was determined using FACS analysis of propidium iodide stained cells using pDC from 2 to 5 donors. Expression of the surface marker, CD86, which has been implicated in dendritic cell maturation, was used as a marker of allergenicity. CD86 expression increased (≥1.5-fold) for 25 of 26 allergens (sensitivity = 96%) but did not increase for 19 of 22 non-allergens (specificity = 86%). In a direct comparison to historical data for the regulatory approved, mouse local lymph node assay (LLNA) for 23 allergens and 22 non-allergens, the pDC method had sensitivity and specificity of 96% and 86%, respectively, while the sensitivity and specificity of the LLNA assay was 83% and 82%, respectively. In conclusion, CD86 expression in pDC appears to be a sensitive and specific indicator to identify contact allergenicity. Such an assay method utilizing normal human cells will be useful for high throughput screening of chemicals for allergenicity.

Novel Risk Stratification Assays for Acute Coronary Syndrome.

Science.gov (United States)

Ahmed, Haitham M; Hazen, Stanley L

2017-08-01

Since identification of aspartate aminotransferase as the first cardiac biomarker in the 1950s, there have been a number of new markers used for myocardial damage detection over the decades. There have also been several generations of troponin assays, each with progressively increasing sensitivity for troponin detection. Accordingly, the "standard of care" for myocardial damage detection continues to change. The purpose of this paper is to review the clinical utility, biological mechanisms, and predictive value of these various biomarkers in contemporary clinical studies. As of this writing, a fifth "next" generation troponin assay has now been cleared by the US Food and Drug Administration for clinical use in the USA for subjects presenting with suspected acute coronary syndromes. Use of these high-sensitivity assays has allowed for earlier detection of myocardial damage as well as greater negative predictive value for infarction after only one or two serial measurements. Recent algorithms utilizing these assays have allowed for more rapid rule-out of myocardial infarction in emergency department settings. In this review, we discuss novel assays available for the risk assessment of subjects presenting with chest pain, including both the "next generation" cardiac troponin assays as well as other novel biomarkers. We review the biological mechanisms for these markers, and explore the positive and negative predictive value of the assays in clinical studies, where reported. We also discuss the potential use of these new markers within the context of future clinical care in the modern era of higher sensitivity troponin testing. Finally, we discuss advances in new platforms (e.g., mass spectrometry) that historically have not been considered for rapid in vitro diagnostic capabilities, but that are taking a larger role in clinical diagnostics, and whose prognostic value and power promise to usher in new markers with potential for future clinical utility in acute coronary

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

International Nuclear Information System (INIS)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro; Onodera, Takeshi; Toko, Kiyoshi

2014-01-01

The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

Energy Technology Data Exchange (ETDEWEB)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Onodera, Takeshi [Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan); Toko, Kiyoshi, E-mail: toko@ed.kyushu-u.ac.jp [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan)

2014-05-30

The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Arqueología de un espacio habitado, trabajado y defendido. El sistema fortificado de Salinas de Añana (Álava

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Plata Montero, Alberto

2009-12-01

Full Text Available The stratigraphic analysis of the wall of the village of Salinas de Añana (Álava together with the excavations within its fortified area have offered a lot of information about the evolution of its built environment from the Early Medieval period until now. This research paper consists of two parts. Main recorded phases obtained thanks to the archaeological work are firstly described. Comprehension of spatial order resulted after the foundation of a village at the beginnings of the 12th century and the gathering of several hamlets that since the Late Antiquity populated, worked and defended the Valle Salado, is exposed in the second part. In order to understand this process, space syntax analysis, methodology developed within English scientific context, has been applied, showing an special attention to those instruments which enable to determine a global pattern of settlements (Alpha analysis, because this pattern links the social logic of space with the spatial logic of the society that created it.La lectura estratigráfica de la muralla de la villa alavesa de Salinas de Añana, así como la ejecución de excavaciones en su recinto fortificado, nos han proporcionado abundante información sobre la evolución de su paisaje construido desde la Alta Edad Media hasta la actualidad. La investigación que presentamos se divide en dos partes. En la primera, describimos las principales fases documentadas durante la intervención arqueológica. En la segunda, nos hemos centrado en comprender el ordenamiento espacial resultante tras el proceso de fundación de una villa creada a principios del siglo XII en la que, además, se agruparon las distintas comunidades aldeanas que desde la Tardoantigüedad habitaban, trabajaban y defendían el Valle Salado. Para ello, hemos empleado la metodología del «Análisis Sintáctico del Espacio» desarrollada en el mundo anglosajón, prestando una especial atención a las herramientas dirigidas a determinar el patr

33 CFR 96.120 - Definitions.

Science.gov (United States)

2010-07-01

....120 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF HOMELAND SECURITY VESSEL OPERATING REGULATIONS RULES FOR THE SAFE OPERATION OF VESSELS AND SAFETY MANAGEMENT SYSTEMS General § 96.120 Definitions... management operate in accordance with the approved safety management system. Safety Management System means a...

Estudo fitoquímico e bioensaio toxicológico frente a larvas de Artemia salina Leach. de três espécies medicinais do gênero Phyllanthus (Phyllanthaceae

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J. E. Nascimento

2009-02-01

Full Text Available

Avaliou-se o perfil fitoquímico e a toxicidade de três espécies do gênero Phyllanthus (P. niruri, P. amarus e P. tenellus, coletadas em diferentes localidades do estado de Pernambuco, nordeste do Brasil. Os extratos brutos das três espécies foram submetidos a testes convencionais por cromatografia em camada delgada analítica para verificação das classes de metabólitos presentes, e testados frente a larvas de Artemia salina para obtenção das concentrações letais médias (CL50. Os testes fitoquímicos demonstraram a presença de flavonóides, saponinas, terpenos, naftoquinonas, alcalóides, antraquinonas, lignanas e taninos. As três espécies de Phyllanthus apresentaram variações na composição fitoquímica e na toxicidade frente a A. salina. Dependendo do local de coleta, os valores de CL50 variaram de 404,43 ± 49,64 µg/mL a 770,84 ± 51,78 µg/mL para P. niruri, 837,65 ± 61,45 µg/mL a 1075,89 ± 70,72 µg/mL para P. amarus e 534,60 ± 46,83 µg/mL a 1003,62 ± 65,15 µg/mL para P. tenellus. Palavras-chave: Phyllanthus niruri; Phyllanthus amarus,/i>; Phyllanthus tenellus; análise fitoquímica preliminar; bioensaio toxicológico; CL50

Assaying Cellular Viability Using the Neutral Red Uptake Assay.

Science.gov (United States)

Ates, Gamze; Vanhaecke, Tamara; Rogiers, Vera; Rodrigues, Robim M

2017-01-01

The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

Microbiological assay for the analysis of certain macrolides in pharmaceutical dosage forms.

Science.gov (United States)

Mahmoudi, A; Fourar, R E-A; Boukhechem, M S; Zarkout, S

2015-08-01

Clarithromycin (CLA) and roxithromycin (ROX) are macrolide antibiotics with an expanded spectrum of activity that are commercially available as tablets. A microbiological assay, applying the cylinder-plate method and using a strain of Micrococcus luteus ATCC 9341 as test organism, has been used and validated for the quantification of two macrolide drugs; CLA and ROX in pure and pharmaceutical formulations. The validation of the proposed method was carried out for linearity, precision, accuracy and specificity. The linear dynamic ranges were from 0.1 to 0.5μg/mL for both compounds. Logarithmic calibration curve was obtained for each macrolide (r>0.989) with statistically equal slopes varying from 3.275 to 4.038, and a percentage relative standard deviation in the range of 0.24-0.92%. Moreover, the method was applied successfully for the assay of the studied drugs in pharmaceutical tablet dosage forms. Recovery from standard addition experiments in commercial products was 94.71-96.91% regarding clarithromycin and 93.94-98.12% regarding roxithromycin, with a precision (%RSD) 1.32-2.11%. Accordingly, this microbiological assay can be used for routine quality control analysis of titled drugs in tablet formulations. Copyright © 2015 Elsevier B.V. All rights reserved.

Verification of the harmonization of human epididymis protein 4 assays.

Science.gov (United States)

Ferraro, Simona; Borille, Simona; Carnevale, Assunta; Frusciante, Erika; Bassani, Niccolò; Panteghini, Mauro

2016-10-01

Serum human epididymis protein 4 (HE4) has gained relevance as an ovarian cancer (OC) biomarker and new automated methods have replaced the first released manual EIA by tracing results to it. We verified agreement and bias of automated methods vs. EIA as well as possible effects on patients' management. One hundred and fifteen serum samples were measured by Abbott Architect i2000, Fujirebio Lumipulse G1200, Roche Modular E170, and Fujirebio EIA. Passing-Bablok regression was used to compare automated assays to EIA and agreement between methods was estimated by Lin's concordance correlation coefficient (CCC). The bias vs. EIA was estimated and compared to specifications derived from HE4 biological variation. Median (25th-75th percentiles) HE4 concentrations (pmol/L) were 84.5 (60.1-148.8) for EIA, 82.7 (50.3-153.9) for Abbott, 89.1 (55.2-154.9) for Roche, and 112.2 (67.8-194.2) for Fujirebio. Estimated regressions and agreements (95% confidence interval) were: Abbott=1.01(0.98-1.03) EIA-4.8(-7.5/-2.6), CCC=0.99(0.99-1.00); Roche=0.91(0.89-0.93) EIA+5.7(4.2/8.0), CCC=0.98(0.98-0.99); Fujirebio=1.20(1.17-1.24) EIA+ 2.4(-0.6/4.9), CCC=0.97(0.96-0.98). The average bias vs. EIA resulted within the desirable goal for Abbott [-3.3% (-6.1/-0.5)] and Roche [-0.2% (-3.0/2.5)]. However, while for Abbott the bias was constant and acceptable along the measurement concentration range, Roche bias increased up to -28% for HE4 values >250 pmol/L. Lumipulse showed a markedly positive bias [25.3% (21.8/28.8)]. Abbott and Roche assays exhibited a good comparability in the range of HE4 values around the previously recommended 140 pmol/L cut-off. For patient monitoring, however, the assay used for determining serial HE4 must not be changed as results from different systems in lower and higher concentration ranges can markedly differ.

Produção e indicadores fisiológicos de alface sob hidroponia com água salina Production and physiologic indicators of lettuce grown in hydroponics with saline water

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Dalva Paulus

2010-03-01

Full Text Available O uso de água salina na produção de hortaliças constitui no momento atividade essencial, tendo em vista o aumento da demanda de água doce, tanto pela atividade agrícola quanto pelo abastecimento urbano e industrial. O objetivo do trabalho foi avaliar a produção e os indicadores fisiológicos de alface cultivada em hidroponia com a utilização de água salina. O experimento foi conduzido em ambiente protegido, em Piracicaba-SP. O delineamento experimental foi de blocos ao acaso, sendo estudados os efeitos de cinco níveis de salinidade da água de irrigação, utilizando-se NaCl [Condutividade elétrica (Cea: 0,42, 1,53, 3,52, 5,55 e 7,43 dS m-1] em duas cultivares de alface (Verônica e Pira Roxa, em esquema fatorial. Foram determinadas massa fresca e seca de folhas, caule, raízes e da parte aérea; teor de nitrato, prolina e clorofila. O aumento da salinidade da água reduziu linearmente as massas fresca e seca das folhas, caule, raízes e da parte aérea. As massas fresca e seca foram 36% e 57% superiores na cultivar Verônica, respectivamente. A cultivar Pira Roxa apresentou maior teor de nitrato 25% (3008 mg L-1, clorofila total 50% (1,46 mg g-1massa fresca e prolina, 71,43% (0,21 µM g-1massa fresca-1 em relação à Verônica, o que pode ser um mecanismo de adaptação daquela cultivar ao estresse salino. Com relação à produtividade comercial, obteve-se uma perda de 69 e 64% para as cultivares Pira Roxa e Verônica, quando se utilizou água mais salina (7,43 dS m-1. Em relação à produção de massa seca, a perda pelo uso dessa água foi de 53% e 44%, respectivamente. Os resultados obtidos em sistema de cultivo hidropônico podem indicar a possibilidade do uso da água salina como alternativa para produção de hortaliças para produtores que têm disponibilidade de água salina e restrita disponibilidade de água doce, embora com redução na produtividade.The use of saline water in the production of vegetables constitutes

Population screening for glucose-6-phosphate dehydrogenase deficiencies in Isabel Province, Solomon Islands, using a modified enzyme assay on filter paper dried bloodspots

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Landry Losi

2010-08-01

Full Text Available Abstract Background Glucose-6-phosphate dehydrogenase deficiency poses a significant impediment to primaquine use for the elimination of liver stage infection with Plasmodium vivax and for gametocyte clearance, because of the risk of life-threatening haemolytic anaemia that can occur in G6PD deficient patients. Although a range of methods for screening G6PD deficiency have been described, almost all require skilled personnel, expensive laboratory equipment, freshly collected blood, and are time consuming; factors that render them unsuitable for mass-screening purposes. Methods A published WST8/1-methoxy PMS method was adapted to assay G6PD activity in a 96-well format using dried blood spots, and used it to undertake population screening within a malaria survey undertaken in Isabel Province, Solomon Islands. The assay results were compared to a biochemical test and a recently marketed rapid diagnostic test. Results Comparative testing with biochemical and rapid diagnostic test indicated that results obtained by filter paper assay were accurate providing that blood spots were assayed within 5 days when stored at ambient temperature and 10 days when stored at 4 degrees. Screening of 8541 people from 41 villages in Isabel Province, Solomon Islands revealed the prevalence of G6PD deficiency as defined by enzyme activity Conclusions The assay enabled simple and quick semi-quantitative population screening in a malaria-endemic region. The study indicated a high prevalence of G6PD deficiency in Isabel Province and highlights the critical need to consider G6PD deficiency in the context of P. vivax malaria elimination strategies in Solomon Islands, particularly in light of the potential role of primaquine mass drug administration.

Fluorescence lifetime assays: current advances and applications in drug discovery.

Science.gov (United States)

Pritz, Stephan; Doering, Klaus; Woelcke, Julian; Hassiepen, Ulrich

2011-06-01

Fluorescence lifetime assays complement the portfolio of established assay formats available in drug discovery, particularly with the recent advances in microplate readers and the commercial availability of novel fluorescent labels. Fluorescence lifetime assists in lowering complexity of compound screening assays, affording a modular, toolbox-like approach to assay development and yielding robust homogeneous assays. To date, materials and procedures have been reported for biochemical assays on proteases, as well as on protein kinases and phosphatases. This article gives an overview of two assay families, distinguished by the origin of the fluorescence signal modulation. The pharmaceutical industry demands techniques with a robust, integrated compound profiling process and short turnaround times. Fluorescence lifetime assays have already helped the drug discovery field, in this sense, by enhancing productivity during the hit-to-lead and lead optimization phases. Future work will focus on covering other biochemical molecular modifications by investigating the detailed photo-physical mechanisms underlying the fluorescence signal.

Comet Assay in Cancer Chemoprevention.

Science.gov (United States)

Santoro, Raffaela; Ferraiuolo, Maria; Morgano, Gian Paolo; Muti, Paola; Strano, Sabrina

2016-01-01

The comet assay can be useful in monitoring DNA damage in single cells caused by exposure to genotoxic agents, such as those causing air, water, and soil pollution (e.g., pesticides, dioxins, electromagnetic fields) and chemo- and radiotherapy in cancer patients, or in the assessment of genoprotective effects of chemopreventive molecules. Therefore, it has particular importance in the fields of pharmacology and toxicology, and in both environmental and human biomonitoring. It allows the detection of single strand breaks as well as double-strand breaks and can be used in both normal and cancer cells. Here we describe the alkali method for comet assay, which allows to detect both single- and double-strand DNA breaks.

Simplified Casing Program for Development Wells in Mahu Well Block

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Lu Zongyu

2017-01-01

Full Text Available In the Mahu well block of Junggar basin, the complex formation has many sets of pressure system. Especially, the formation with microcracks in the middle layer is loose and the pressure bearing capacity is low. Lost circulation is prone to occur in this layer. At present, high investment and long drilling period were the main problems in the exploration and development process. The geostress 3D model of Mahu well block was established by means of logging and drilling data. The model provided the three-pressure profiles of Mahu well block for casing program optimization and safety drilling. Each well could be optimized the intermediate casing setting position. The intermediate casing was saved 160 meters long. The total of drilling speed was improved 5 times compared with the past drilling process. Slim hole drilling technology raised ROP 51.96% higher, and the average drilling period is shorten to 24.83 days.

32 CFR 96.1 - Purpose.