Sample records for water processing kit

  1. Pilot Fullerton uses water dispenser kit gun to rehydrate food package

    NASA Image and Video Library

    1982-03-30

    STS003-26-254 (30 March 1982) --- Astronaut Gordon Fullerton, STS-3 pilot, wearing communications kit assembly (ASSY) mini-headset (HDST), inserts the JSC water dispenser kit water gun in rehydratable plastic food (cereal) package to fill it with hot water. Photo credit: NASA

  2. Low-cost field test kits for arsenic detection in water.

    PubMed

    Das, Joyati; Sarkar, Priyabrata; Panda, Jigisha; Pal, Priyabrata

    2014-01-01

    Arsenic, a common contaminant of groundwater, affects human health adversely. According to the World Health Organization (WHO), the maximum recommended contamination level of arsenic in drinking water is 10 μg/L. The purpose of this research was to develop user-friendly kits for detection of arsenic to measure at least up to 10 μg/L in drinking water, so that a preventive measure could be taken. Two different kits for detection of total arsenic in water are reported here. First, the arsenic in drinking water was converted to arsine gas by a strong reducing agent. The arsine produced was then detected by paper strips via generation of color due to reaction with either mercuric bromide (KIT-1) or silver nitrate (KIT-2). These were previously immobilized on the detector strip. The first one gave a yellow color and the second one grey. Both of these kits could detect arsenic contamination within a range of 10 μg/L-250 μg/L. The detection time for both the kits was only 7 min. The kits exhibited excellent performance compared to other kits available in the market with respect to detection time, ease of operation, cost and could be easily handled by a layman. The field trials with these kits gave very satisfactory results. A study on interference revealed that these kits could be used in the presence of 24 common ions present in the arsenic contaminated water. Though the kits were meant for qualitative assay, the results with unknown concentrations of real samples, when compared with atomic absorption spectrophotometer (AAS) were in good agreement as revealed by the t-test.

  3. UNESCO's HOPE Initiative—Providing Free and Open-Source Hydrologic Software for Effective and Sustainable Management of Africa's Water Resources Temporary Title

    NASA Astrophysics Data System (ADS)

    Barlow, P. M.; Filali-Meknassi, Y.; Sanford, W. E.; Winston, R. B.; Kuniansky, E.; Dawson, C.

    2015-12-01

    UNESCO's HOPE Initiative—the Hydro Free and (or) Open-source Platform of Experts—was launched in June 2013 as part of UNESCO's International Hydrological Programme. The Initiative arose in response to a recognized need to make free and (or) open-source water-resources software more widely accessible to Africa's water sector. A kit of software is being developed to provide African water authorities, teachers, university lecturers, and researchers with a set of programs that can be enhanced and (or) applied to the development of efficient and sustainable management strategies for Africa's water resources. The Initiative brings together experts from the many fields of water resources to identify software that might be included in the kit, to oversee an objective process for selecting software for the kit, and to engage in training and other modes of capacity building to enhance dissemination of the software. To date, teams of experts from the fields of wastewater treatment, groundwater hydrology, surface-water hydrology, and data management have been formed to identify relevant software from their respective fields. An initial version of the HOPE Software Kit was released in late August 2014 and consists of the STOAT model for wastewater treatment developed by the Water Research Center (United Kingdom) and the MODFLOW-2005 model for groundwater-flow simulation developed by the U.S. Geological Survey. The Kit is available on the UNESCO HOPE website (http://www.hope-initiative.net/).Training in the theory and use of MODFLOW-2005 is planned in southern Africa in conjunction with UNESCO's study of the Kalahari-Karoo/Stampriet Transboundary Aquifer, which extends over an area that includes parts of Botswana, Namibia, and South Africa, and in support of the European Commission's Horizon 2020 FREEWAT project (FREE and open source software tools for WATer resource management; see the UNESCO HOPE website).

  4. EVALUATION OF A FIELD TEST KIT FOR MONITORING LEAD IN DRINKING WATER.

    EPA Science Inventory

    This article describes a conceptual framework for designing evaluation studies of test kits for the analysis of significant drinking water constituents. A commercial test kit for the analysis of lead in tap waters was evaluated and compared with a standard graphite furnace atomic...

  5. Immunomagnetic Separation of Cryptosporidium parvum from Source Water Samples of Various Turbidities

    PubMed Central

    Bukhari, Z.; McCuin, R. M.; Fricker, C. R.; Clancy, J. L.

    1998-01-01

    Immunomagnetic separation (IMS) procedures which specifically capture Cryptosporidium oocysts and have the potential to isolate oocysts from debris have become commercially available. We compared two IMS kits (kit DB [Dynabeads anti-Cryptosporidium; product no. 730.01; Dynal A.S., Oslo, Norway] and kit IC1 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC, Portland, Maine]) and a modification of kit IC1 (kit IC2 [Crypto Scan IMS; product no. R10; Clearwater Diagnostics Company, LLC]) at three turbidity levels (50, 500, and 5,000 nephelometric turbidity units [ntu]) by using water matrices obtained from different geographical locations. In deionized water, kit DB yielded recoveries between 68 and 83%, whereas the recoveries obtained with kits IC1 and IC2 were more variable and ranged from 0.2 to 74.5%. In water matrices with turbidity levels up to 500 ntu, the oocyst recoveries were more variable with kit DB; however, the recoveries were similar to those obtained in deionized water. In contrast, there were notable reductions in oocyst recoveries in the turbid matrices with kits IC1 and IC2, and the highest recovery (8.3%) was obtained with a 50-ntu sample. An examination of the effects of age on oocyst recovery with kit DB revealed that oocysts up to 16 weeks old yielded recoveries similar to the recoveries observed with fresh oocysts. These data indicate that all IMS kits do not perform equally well, and it is important to conduct in-house quality assurance work before a commercially available IMS kit is selected to replace flotation procedures for recovery of Cryptosporidium oocysts. PMID:9797313

  6. Space processing applications payload equipment study. Volume 2D: SPA supplemental power and heat rejection kit

    NASA Technical Reports Server (NTRS)

    Hammel, R. L. (Editor); Smith, A. G. (Editor)

    1974-01-01

    The design and application of a supplementary power and heat rejection kit for the Spacelab are discussed. Two subsystems of electric power and thermal control were analyzed to define the requirements for the power and heat rejection kit (PHRK). Twelve exemplary experiments were defined and power timelines were developed. From these timeline, the experiment requirements for sustained power, peak power, and energy were determined. The electrical power subsystem of the PHRK will consist of two fuel cells, oxygen and hydrogen reactant tank assemblies, water storage tanks, plumbing, cabling, and inverters to convert the nominal 28 volt dc fuel cell output to ac power.

  7. AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) PERFORMANCE TESTING OF THE INDUSTRIAL TEST SYSTEM, INC. CYANIDE REAGENTSTRIP™ TEST KIT

    EPA Science Inventory

    Cyanide can be present in various forms in water. The cyanide test kit evaluated in this verification study (Industrial Test System, Inc. Cyanide Regent Strip ™ Test Kit) was designed to detect free cyanide in water. This is done by converting cyanide in water to cyanogen...

  8. Statistical modeling of dental unit water bacterial test kit performance.

    PubMed

    Cohen, Mark E; Harte, Jennifer A; Stone, Mark E; O'Connor, Karen H; Coen, Michael L; Cullum, Malford E

    2007-01-01

    While it is important to monitor dental water quality, it is unclear whether in-office test kits provide bacterial counts comparable to the gold standard method (R2A). Studies were conducted on specimens with known bacterial concentrations, and from dental units, to evaluate test kit accuracy across a range of bacterial types and loads. Colony forming units (CFU) were counted for samples from each source, using R2A and two types of test kits, and conformity to Poisson distribution expectations was evaluated. Poisson regression was used to test for effects of source and device, and to estimate rate ratios for kits relative to R2A. For all devices, distributions were Poisson for low CFU/mL when only beige-pigmented bacteria were considered. For higher counts, R2A remained Poisson, but kits exhibited over-dispersion. Both kits undercounted relative to R2A, but the degree of undercounting was reasonably stable. Kits did not grow pink-pigmented bacteria from dental-unit water identified as Methylobacterium rhodesianum. Only one of the test kits provided results with adequate reliability at higher bacterial concentrations. Undercount bias could be estimated for this device and used to adjust test kit results. Insensitivity to methylobacteria spp. is problematic.

  9. Portable field kit for determining uranium in water

    USGS Publications Warehouse

    McHugh, John B.

    1979-01-01

    The pressing need for on-site field analyses of the uranium content of surface and ground waters has promoted the development of a simple, light-weight, relatively cheap, portable kit to make such determinations in the field. Forty to sixty water samples per day can be analyzed for uranium to less than 0.2 parts per billion. The kit was tested in the field with excellent results.

  10. 46 CFR 160.058-1 - Applicable specification.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-1... the desalter kits are manufactured, forms a part of this subpart: (1) Military specification. MIL-D-5531D—Desalter Kit, Sea Water, Mark 2. (2) [Reserved] (b) Copies on file. A copy of the specification...

  11. 46 CFR 160.058-1 - Applicable specification.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-1... the desalter kits are manufactured, forms a part of this subpart: (1) Military specification. MIL-D-5531D—Desalter Kit, Sea Water, Mark 2. (2) [Reserved] (b) Copies on file. A copy of the specification...

  12. 46 CFR 160.058-1 - Applicable specification.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-1... the desalter kits are manufactured, forms a part of this subpart: (1) Military specification. MIL-D-5531D—Desalter Kit, Sea Water, Mark 2. (2) [Reserved] (b) Copies on file. A copy of the specification...

  13. 46 CFR 160.058-1 - Applicable specification.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-1... the desalter kits are manufactured, forms a part of this subpart: (1) Military specification. MIL-D-5531D—Desalter Kit, Sea Water, Mark 2. (2) [Reserved] (b) Copies on file. A copy of the specification...

  14. 46 CFR 160.058-1 - Applicable specification.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-1... the desalter kits are manufactured, forms a part of this subpart: (1) Military specification. MIL-D-5531D—Desalter Kit, Sea Water, Mark 2. (2) [Reserved] (b) Copies on file. A copy of the specification...

  15. Efforts to Reduce International Space Station Crew Maintenance Time in the Management of the Extravehicular Mobility Unit Transport Loop Water Quality

    NASA Technical Reports Server (NTRS)

    Etter,David; Rector, Tony; Boyle, robert; Zande, Chris Vande

    2012-01-01

    The EMU (Extravehicular Mobility Unit) contains a semi-closed-loop re-circulating water circuit (Transport Loop) to absorb heat into a LCVG (Liquid Coolant and Ventilation Garment) worn by the astronaut. A second, single-pass water circuit (Feed-water Loop) provides water to a cooling device (Sublimator) containing porous plates, and that water sublimates through the porous plates to space vacuum. The cooling effect from the sublimation of this water translates to a cooling of the LCVG water that circulates through the Sublimator. The quality of the EMU Transport Loop water is maintained through the use of a water processing kit (ALCLR - Airlock Cooling Loop Remediation) that is used to periodically clean and disinfect the water circuit. Opportunities to reduce crew time associated with ALCLR operations include a detailed review of the historical water quality data for evidence to support an extension to the implementation cycle. Furthermore, an EMU returned after 2-years of use on the ISS (International Space Station) is being used as a test bed to evaluate the results of extended and repeated ALCLR implementation cycles. Finally, design, use and on-orbit location enhancements to the ALCLR kit components are being considered to allow the implementation cycle to occur in parallel with other EMU maintenance and check-out activities, and to extend the life of the ALCLR kit components. These efforts are undertaken to reduce the crew-time and logistics burdens for the EMU, while ensuring the long-term health of the EMU water circuits for a post- Shuttle 6-year service life.

  16. Efforts to Reduce International Space Station Crew Maintenance for the Management of the Extravehicular Mobility Unit Transport Loop Water Quality

    NASA Technical Reports Server (NTRS)

    Steele, John W.; Etter, David; Rector, Tony; Boyle, Robert; Vandezande, Christopher

    2013-01-01

    The EMU (Extravehicular Mobility Unit) contains a semi-closed-loop re-circulating water circuit (Transport Loop) to absorb heat into a LCVG (Liquid Coolant and Ventilation Garment) worn by the astronaut. A second, single-pass water circuit (Feed-water Loop) provides water to a cooling device (Sublimator) containing porous plates, and that water sublimates through the porous plates to space vacuum. The cooling effect from the sublimation of this water translates to a cooling of the LCVG water that circulates through the Sublimator. The quality of the EMU Transport Loop water is maintained through the use of a water processing kit (ALCLR Airlock Cooling Loop Remediation) that is used to periodically clean and disinfect the water circuit. Opportunities to reduce crew time associated with on-orbit ALCLR operations include a detailed review of the historical water quality data for evidence to support an extension to the implementation cycle. Furthermore, an EMU returned after 2-years of use on the ISS (International Space Station) is being used as a test bed to evaluate the results of extended and repeated ALCLR implementation cycles. Finally, design, use and on-orbit location enhancements to the ALCLR kit components are being considered to allow the implementation cycle to occur in parallel with other EMU maintenance and check-out activities, and to extend the life of the ALCLR kit components. These efforts are undertaken to reduce the crew-time and logistics burdens for the EMU, while ensuring the long-term health of the EMU water circuits for a post-Shuttle 6-year service life.

  17. Water treatment and handwashing behaviors among non-pregnant friends and relatives of participants in an antenatal hygiene promotion program in Malawi.

    PubMed

    Russo, Elizabeth T; Sheth, Anandi; Menon, Manoj; Wannemuehler, Kathleen; Weinger, Merri; Kudzala, Amose C; Tauzie, Blessius; Masuku, Humphreys D; Msowoya, Tapona E; Quick, Robert

    2012-05-01

    Access to safe drinking water and improved hygiene are essential for preventing diarrheal diseases. To integrate hygiene improvement with antenatal care, free hygiene kits (water storage containers, water treatment solution, soap) and educational messages were distributed to pregnant women at antenatal clinics in Malawi. We assessed water treatment and hygiene practices of 275 non-pregnant friends and relatives of the hygiene kit recipients at baseline and follow-up nine months later to measure program impact on non-participants in the same communities. At follow-up, friends and relatives who did not receive kits or education were more likely than at baseline to purchase and use water treatment solution (25% versus 1%; P < 0.0001) and demonstrate correct handwashing practices (60% versus 18%; P < 0.0001). This antenatal clinic-based program resulted in improved water treatment and hygiene behaviors among non-pregnant friends and relatives living in the same communities as hygiene kit recipients, suggesting that program benefits extended beyond direct beneficiaries.

  18. A First Application of Enzyme-Linked Immunosorbent Assay for Screening Cyclodiene Insecticides in Ground Water

    USGS Publications Warehouse

    Dombrowski, T.R.; Thurman, E.M.; Mohrman, G.B.

    1996-01-01

    A commercially available enzyme-linked immunosorbent assay (ELISA) plate kit for screening of cyclodiene insecticides (aldrin, chlordane, dieldrin, endosulfan, endrin, and heptachlor) was evaluated for sensitivity, cross reactivity, and overall performance using groundwater samples from a contaminated site. Ground-water contaminants included several pesticide compounds and their manufacturing byproducts, as well as many other organic and inorganic compounds. Cross-reactivity studies were carried out for the cyclodiene compounds, and results were compared to those listed by the manufacturer. Data obtained were used to evaluate the sensitivity of the ELISA kit to the cyclodiene compounds in ground water samples with a contaminated matrix. The method quantitation limit for the ELISA kit was 15 ??g/L (as chlordane). Of the 56 ground-water samples analyzed using the ELISA plate kits, more than 85% showed cyclodiene insecticide contamination. The ELISA kit showed excellent potential as a screening tool for sites with suspected groundwater contamination by insecticides.

  19. Design, Certification, and Deployment of the Colorimetric Water Quality Monitoring Kit (CWQMK)

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Nolan, Daniel J.; Rutz, Jeff A.; Schultz, John R.; Siperko, Lorraine M.; Porter, Marc D.; Lipert, Robert J.; Flint, Stephanie M.; McCoy, J. Torin

    2010-01-01

    In August 2009, an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE) technology was delivered to the International Space Station (ISS) aboard STS-128/17A. The kit, called the Colorimetric Water Quality Monitoring Kit (CWQMK), was flown and deployed as a Station Development Test Objective (SDTO) experiment on the ISS. The goal of the SDTO experiment is to evaluate the acceptability of CSPE technology for routine water quality monitoring on the ISS. This paper provides an overview of the SDTO experiment, as well as a detailed description of the CWQMK hardware and a summary of the testing and analysis conducted to certify the CWQMK for use on the ISS. The initial results obtained from the SDTO experiment are also reported and discussed in detail

  20. DEVELOPMENT OF AN IDENTIFICATION KIT FOR SPILLED HAZARDOUS MATERIALS

    EPA Science Inventory

    The Chemical Systems Laboratory (CSL) has developed a field kit to identify spilled hazardous materials in inland waters and on the ground. The Hazardous Materials Spills Identification Kit is a two-component kit consisting of an inverter/shortwave UV lamp unit for photochemical ...

  1. DEMONSTRATION BULLETIN: HNU-HANBY PCP IMMUNOASSAY TEST KIT - HNU - SYSTEMS, INC.

    EPA Science Inventory

    The HNU-Hanby test kit rapidly analyzes for petroleum hydrocarbons in soil and water samples. The test kit can be used to estimate pentachlorophenol (PCP) concentrations in samples when the carrier solvent is a petroleum hydrocarbon. The test kit estimates PCP concentrations in ...

  2. Water Developments and Canids in Two North American Deserts: A Test of the Indirect Effect of Water Hypothesis

    PubMed Central

    Hall, Lucas K.; Larsen, Randy T.; Knight, Robert N.; Bunnell, Kevin D.; McMillan, Brock R.

    2013-01-01

    Anthropogenic modifications to landscapes intended to benefit wildlife may negatively influence wildlife communities. Anthropogenic provisioning of free water (water developments) to enhance abundance and distribution of wildlife is a common management practice in arid regions where water is limiting. Despite the long-term and widespread use of water developments, little is known about how they influence native species. Water developments may negatively influence arid-adapted species (e.g., kit fox, Vulpes macrotis) by enabling water-dependent competitors (e.g., coyote, Canis latrans) to expand distribution in arid landscapes (i.e., indirect effect of water hypothesis). We tested the two predictions of the indirect effect of water hypothesis (i.e., coyotes will visit areas with free water more frequently and kit foxes will spatially and temporally avoid coyotes) and evaluated relative use of free water by canids in the Great Basin and Mojave Deserts from 2010 to 2012. We established scent stations in areas with (wet) and without (dry) free water and monitored visitation by canids to these sites and visitation to water sources using infrared-triggered cameras. There was no difference in the proportions of visits to scent stations in wet or dry areas by coyotes or kit foxes at either study area. We did not detect spatial (no negative correlation between visits to scent stations) or temporal (no difference between times when stations were visited) segregation between coyotes and kit foxes. Visitation to water sources was not different for coyotes between study areas, but kit foxes visited water sources more in Mojave than Great Basin. Our results did not support the indirect effect of water hypothesis in the Great Basin or Mojave Deserts for these two canids. PMID:23844097

  3. Water Treatment and Handwashing Behaviors among Non-Pregnant Friends and Relatives of Participants in an Antenatal Hygiene Promotion Program in Malawi

    PubMed Central

    Russo, Elizabeth T.; Sheth, Anandi; Menon, Manoj; Wannemuehler, Kathleen; Weinger, Merri; Kudzala, Amose C.; Tauzie, Blessius; Masuku, Humphreys D.; Msowoya, Tapona E.; Quick, Robert

    2012-01-01

    Access to safe drinking water and improved hygiene are essential for preventing diarrheal diseases. To integrate hygiene improvement with antenatal care, free hygiene kits (water storage containers, water treatment solution, soap) and educational messages were distributed to pregnant women at antenatal clinics in Malawi. We assessed water treatment and hygiene practices of 275 non-pregnant friends and relatives of the hygiene kit recipients at baseline and follow-up nine months later to measure program impact on non-participants in the same communities. At follow-up, friends and relatives who did not receive kits or education were more likely than at baseline to purchase and use water treatment solution (25% versus 1%; P < 0.0001) and demonstrate correct handwashing practices (60% versus 18%; P < 0.0001). This antenatal clinic–based program resulted in improved water treatment and hygiene behaviors among non-pregnant friends and relatives living in the same communities as hygiene kit recipients, suggesting that program benefits extended beyond direct beneficiaries. PMID:22556088

  4. Arsenic exposure in drinking water: an unrecognized health threat in Peru.

    PubMed

    George, Christine Marie; Sima, Laura; Arias, M Helena Jahuira; Mihalic, Jana; Cabrera, Lilia Z; Danz, David; Checkley, William; Gilman, Robert H

    2014-08-01

    To assess the extent of arsenic contamination of groundwater and surface water in Peru and, to evaluate the accuracy of the Arsenic Econo-Quick(™) (EQ) kit for measuring water arsenic concentrations in the field. Water samples were collected from 151 water sources in 12 districts of Peru, and arsenic concentrations were measured in the laboratory using inductively-coupled plasma mass spectrometry. The EQ field kit was validated by comparing a subset of 139 water samples analysed by laboratory measurements and the EQ kit. In 86% (96/111) of the groundwater samples, arsenic exceeded the 10 µg/l arsenic concentration guideline given by the World Health Organization (WHO) for drinking water. In 56% (62/111) of the samples, it exceeded the Bangladeshi threshold of 50 µg/l; the mean concentration being 54.5 µg/l (range: 0.1-93.1). In the Juliaca and Caracoto districts, in 96% (27/28) of groundwater samples arsenic was above the WHO guideline; and in water samples collected from the section of the Rímac river running through Lima, all had arsenic concentrations exceeding the WHO limit. When validated against laboratory values, the EQ kit correctly identified arsenic contamination relative to the guideline in 95% (106/111) of groundwater and in 68% (19/28) of surface water samples. In several districts of Peru, drinking water shows widespread arsenic contamination, exceeding the WHO arsenic guideline. This poses a public health threat requiring further investigation and action. For groundwater samples, the EQ kit performed well relative to the WHO arsenic limit and therefore could provide a vital tool for water arsenic surveillance.

  5. DeePMD-kit: A deep learning package for many-body potential energy representation and molecular dynamics

    NASA Astrophysics Data System (ADS)

    Wang, Han; Zhang, Linfeng; Han, Jiequn; E, Weinan

    2018-07-01

    Recent developments in many-body potential energy representation via deep learning have brought new hopes to addressing the accuracy-versus-efficiency dilemma in molecular simulations. Here we describe DeePMD-kit, a package written in Python/C++ that has been designed to minimize the effort required to build deep learning based representation of potential energy and force field and to perform molecular dynamics. Potential applications of DeePMD-kit span from finite molecules to extended systems and from metallic systems to chemically bonded systems. DeePMD-kit is interfaced with TensorFlow, one of the most popular deep learning frameworks, making the training process highly automatic and efficient. On the other end, DeePMD-kit is interfaced with high-performance classical molecular dynamics and quantum (path-integral) molecular dynamics packages, i.e., LAMMPS and the i-PI, respectively. Thus, upon training, the potential energy and force field models can be used to perform efficient molecular simulations for different purposes. As an example of the many potential applications of the package, we use DeePMD-kit to learn the interatomic potential energy and forces of a water model using data obtained from density functional theory. We demonstrate that the resulted molecular dynamics model reproduces accurately the structural information contained in the original model.

  6. 33 CFR 149.323 - What are the requirements for first aid kits?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... first aid kits? 149.323 Section 149.323 Navigation and Navigable Waters COAST GUARD, DEPARTMENT OF... Lifesaving Equipment Manned Deepwater Port Requirements § 149.323 What are the requirements for first aid kits? (a) Each manned deepwater port must have an industrial first aid kit, approved by an appropriate...

  7. Impact and sustainability of low-head drip irrigation kits, in the semi-arid Gwanda and Beitbridge Districts, Mzingwane Catchment, Limpopo Basin, Zimbabwe

    NASA Astrophysics Data System (ADS)

    Moyo, Richard; Love, David; Mul, Marloes; Mupangwa, Walter; Twomlow, Steve

    Resource-poor smallholder farmers in the semi-arid Gwanda and Beitbridge districts face food insecurity on an annual basis due to a combination of poor and erratic rainfall (average 500 mm/a and 345 mm/a, respectively, for the period 1970-2003) and technologies inappropriate to their resource status. This impacts on both household livelihoods and food security. In an attempt to improve food security in the catchment a number of drip kit distribution programmes have been initiated since 2003 as part of an on-going global initiative aimed at 2 million poor households per year. A number of recent studies have assessed the technical performance of the drip kits in-lab and in-field. In early 2005 a study was undertaken to assess the impacts and sustainability of the drip kit programme. Representatives of the NGOs, local government, traditional leadership and agricultural extension officers were interviewed. Focus group discussions with beneficiaries and other villagers were held at village level. A survey of 114 households was then conducted in two districts, using a questionnaire developed from the output of the interviews and focus group discussions. The results from the study showed that the NGOs did not specifically target the distribution of the drip kits to poor members of the community (defined for the purpose of the study as those not owning cattle). Poor households made up 54% of the beneficiaries. This poor targeting of vulnerable households could have been a result of conditions set by some implementing NGOs that beneficiaries must have an assured water source. On the other hand, only 2% of the beneficiaries had used the kit to produce the expected 5 harvests over the 2 years, owing to problems related to water shortage, access to water and also pests and diseases. About 51% of the respondents had produced at least 3 harvests and 86% produced at least 2 harvests. Due to water shortages during the dry season 61% of production with the drip kit occurred during the wet season. This suggests that most households use the drip kits as supplementary irrigation. Conflicts between beneficiaries and water point committees or other water users developed in some areas especially during the dry season. The main finding from this study was that low cost drip kit programs can only be a sustainable intervention if implemented as an integral part of a long-term development program, not short-term relief programs and the programme should involve a broad range of stakeholders. A first step in any such program, especially in water scarce areas such as Gwanda and Beitbridge, is a detailed analysis of the existing water resources to assess availability and potential conflicts, prior to distribution of drip kits.

  8. Development and Certification of Station Development Test Objective (SDTO) Experiment # 15012-U, "Near RealTime Water Quality Monitoring Demonstration for ISS Biocides Using Colorimetric Solid Phase Extraction (CSPE)"

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Nolan, Daniel J.; Rutz, Jeffrey A.; Shcultz, John R.; Siperko, Lorraine M.; Porter, Marc D,; Lipert, Robert J.; Limardo, Jose G.; McCoy, J. Torin

    2009-01-01

    Scientists and engineers from the Wyle Integrated Science and Engineering Group are working with researchers at the University of Utah and Iowa State University to develop and certify an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE). The kit will be launched as a Station Development Test Objective (SDTO) experiment and evaluated on the International Space Station (ISS) to determine the acceptability of CSPE technology for routine inflight water quality monitoring. Iodine and silver, the biocides used in the US and Russian on-orbit water systems, will serve as test analytes for the technology evaluation. This manuscript provides an overview of the CSPE SDTO experiment and details the development and certification of the experimental water quality monitoring kit. Initial results from reagent and standard solution stability testing and environmental testing performed on the kit hardware are also reported.

  9. Gravity flow operated small electricity generator retrofit kit to flour mill industry.

    PubMed

    Shekara, Prithivi; Kumar V, Pavan; Hosamane, Gangadharappa Gundabhakthara

    2013-10-01

    Flour milling is a grinding process to produce flour from wheat through comprehensive stages of grinding and separation. The primary energy is required to provide power used in grinding of wheat. In wheat milling, tempering is the process of adding water to wheat before milling to toughen the bran and mellow the endosperm. Gravity flow of the wheat is utilized to rotate the dampener wheel with cups to add water. Low cost gravity flow operated small electricity generator retrofit kit for dampener was designed and developed to justify low cost energy production without expensive solutions. Results of statistical analysis indicated that there was significant difference in mean values for voltage, rpm and flow rate at the 95% probability level. The resulted maximum mechanical power and measured electrical power were 5.1 W and 4.9 W respectively at wheat flow rate of 1.6 Kg/s and dampener wheel rotational velocity of 4.4 rad/s.

  10. Analytical Bibliography for Water Supply and Conservation Techniques.

    DTIC Science & Technology

    1982-01-01

    effective due to rapidly increasing costs of water and wastewater services from centralized systems. The report may be used as a primary reference for...conservation kit. Each kit contained a toilet water dam, a plastic shower-head restrictor, and a packet of vegetable dye tablets to detect leaks from toilet...water in the 50 sub-basins of the North Atlantic Region (NAR) of the United States. The water-flow requirements (water demands) were disintegrated by

  11. 33 CFR 144.01-30 - First-aid kit.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 33 Navigation and Navigable Waters 2 2014-07-01 2014-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall be...

  12. 33 CFR 144.01-30 - First-aid kit.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 33 Navigation and Navigable Waters 2 2012-07-01 2012-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall be...

  13. 33 CFR 144.01-30 - First-aid kit.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 33 Navigation and Navigable Waters 2 2013-07-01 2013-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall be...

  14. 33 CFR 144.01-30 - First-aid kit.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 33 Navigation and Navigable Waters 2 2011-07-01 2011-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall be...

  15. 33 CFR 144.01-30 - First-aid kit.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 33 Navigation and Navigable Waters 2 2010-07-01 2010-07-01 false First-aid kit. 144.01-30 Section...) OUTER CONTINENTAL SHELF ACTIVITIES LIFESAVING APPLIANCES Manned Platforms § 144.01-30 First-aid kit. On each manned platform a first-aid kit approved by the Commandant or the U.S. Bureau of Mines shall be...

  16. Test/QA Plan for Verification of Microcystin Test Kits

    EPA Science Inventory

    Microcystin test kits are used to quantitatively measure total microcystin in recreational waters. These test kits are based on enzyme-linked immunosorbent assays (ELISA) with antibodies that bind specifically to microcystins or phosphate activity inhibition where the phosphatas...

  17. A "three-in-one" sample preparation method for simultaneous determination of B-group water-soluble vitamins in infant formula using VitaFast(®) kits.

    PubMed

    Zhang, Heng; Lan, Fang; Shi, Yupeng; Wan, Zhi-Gang; Yue, Zhen-Feng; Fan, Fang; Lin, Yan-Kui; Tang, Mu-Jin; Lv, Jing-Zhang; Xiao, Tan; Yi, Changqing

    2014-06-15

    VitaFast(®) test kits designed for the microbiological assay in microtiter plate format can be applied to quantitative determination of B-group water-soluble vitamins such as vitamin B12, folic acid and biotin, et al. Compared to traditional microbiological methods, VitaFast(®) kits significantly reduce sample processing time and provide greater reliability, higher productivity and better accuracy. Recently, simultaneous determination of vitamin B12, folic acid and biotin in one sample is urgently required when evaluating the quality of infant formulae in our practical work. However, the present sample preparation protocols which are developed for individual test systems, are incompatible with simultaneous determination of several analytes. To solve this problem, a novel "three-in-one" sample preparation method is herein developed for simultaneous determination of B-group water-soluble vitamins using VitaFast(®) kits. The performance of this novel "three-in-one" sample preparation method was systematically evaluated through comparing with individual sample preparation protocols. The experimental results of the assays which employed "three-in-one" sample preparation method were in good agreement with those obtained from conventional VitaFast(®) extraction methods, indicating that the proposed "three-in-one" sample preparation method is applicable to the present three VitaFast(®) vitamin test systems, thus offering a promising alternative for the three independent sample preparation methods. The proposed new sample preparation method will significantly improve the efficiency of infant formulae inspection. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Arsenic exposure in drinking water: an unrecognized health threat in Peru

    PubMed Central

    Sima, Laura; Arias, M Helena Jahuira; Mihalic, Jana; Cabrera, Lilia Z; Danz, David; Checkley, William; Gilman, Robert H

    2014-01-01

    Abstract Objective To assess the extent of arsenic contamination of groundwater and surface water in Peru and, to evaluate the accuracy of the Arsenic Econo-Quick™ (EQ) kit for measuring water arsenic concentrations in the field. Methods Water samples were collected from 151 water sources in 12 districts of Peru, and arsenic concentrations were measured in the laboratory using inductively-coupled plasma mass spectrometry. The EQ field kit was validated by comparing a subset of 139 water samples analysed by laboratory measurements and the EQ kit. Findings In 86% (96/111) of the groundwater samples, arsenic exceeded the 10 µg/l arsenic concentration guideline given by the World Health Organization (WHO) for drinking water. In 56% (62/111) of the samples, it exceeded the Bangladeshi threshold of 50 µg/l; the mean concentration being 54.5 µg/l (range: 0.1–93.1). In the Juliaca and Caracoto districts, in 96% (27/28) of groundwater samples arsenic was above the WHO guideline; and in water samples collected from the section of the Rímac river running through Lima, all had arsenic concentrations exceeding the WHO limit. When validated against laboratory values, the EQ kit correctly identified arsenic contamination relative to the guideline in 95% (106/111) of groundwater and in 68% (19/28) of surface water samples. Conclusion In several districts of Peru, drinking water shows widespread arsenic contamination, exceeding the WHO arsenic guideline. This poses a public health threat requiring further investigation and action. For groundwater samples, the EQ kit performed well relative to the WHO arsenic limit and therefore could provide a vital tool for water arsenic surveillance. PMID:25177071

  19. Preparation of acid-base bifunctional mesoporous KIT-6 (KIT: Korea Advanced Institute of Science and Technology) and its catalytic performance in Knoevenagel reaction

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

    Xu, Ling; Wang, Chunhua; Guan, Jingqi, E-mail: guanjq@jlu.edu.cn

    2014-05-01

    Acid-base bifunctional mesoporous catalysts Al-KIT-6-NH{sub 2} containing different aluminum content have been synthesized through post synthetic grafting method. The materials were characterized by X-ray diffraction (XRD), scanning electron micrographs (SEM), transmission electron micrographs (TEM), Fourier-transform infrared spectroscopy (FTIR), IR spectra of pyridine adsorption, NH{sub 3}-TPD and TG analysis. The characterization results indicated that the pore structure of KIT-6 was well kept after the addition of aluminum and grafting of aminopropyl groups. The acid amount of Al-KIT-6 increased with enhancing aluminum content. Catalytic results showed that weak acid and weak base favor the Knoevenagel reaction, while catalysts with strong acid andmore » weak base exhibited worse catalytic behavior. - Graphical abstract: The postulated steps of mechanism for the acid-base catalyzed process are as follows: (1) the aldehyde gets activated by the surface acidic sites which allow the amine undergoes nucleophilic to attack the carbonyl carbon of benzaldehyde. (2) Water is released in the formation of imine intermediate. (3) The ethyl cyanoacetate reacts with the intermediate. (4) The benzylidene ethyl cyanoacetate is formed and the amine is regenerated. - Highlights: • KIT-6 and Al-KIT-6-NH{sub 2} with different Si/Al ratios has been successfully prepared. • 79.4% Yield was obtained over 46-Al-KIT-6-NH{sub 2} within 20 min in Knoevenagel reaction. • Low Al-content Al-KIT-6-NH{sub 2} shows better catalytic stability than high Al-content catalysts. • There is acid-base synergistic effect in Knoevenagel reaction.« less

  20. SURVEY OF GROUND, SURFACE, AND POTABLE WATERS FOR THE PRESENCE OF LEGIONELLA SPECIES BY ENVIROAMP PCR LEGIONELLA KIT, CULTURE, AND IMMUNOFLUORESCENT STAINING

    EPA Science Inventory

    A total of 116 samples from numerous aquatic sources including water from faucets, showerheads, dental units, fire sprinklers, and surface waters were examined for the presence of Legionella by the EnviroAmp Legionella PCR kit, culture on BCYEx, or direct fluorescent antibody (DF...

  1. Buffer substitution in malaria rapid diagnostic tests causes false-positive results

    PubMed Central

    2010-01-01

    Background Malaria rapid diagnostic tests (RDTs) are kits that generally include 20 to 25 test strips or cassettes, but only a single buffer vial. In field settings, laboratory staff occasionally uses saline, distilled water (liquids for parenteral drugs dilution) or tap water as substitutes for the RDT kit's buffer to compensate for the loss of a diluent bottle. The present study assessed the effect of buffer substitution on the RDT results. Methods Twenty-seven RDT brands were run with EDTA-blood samples of five malaria-free subjects, who were negative for rheumatoid factor and antinuclear antibodies. Saline, distilled water and tap water were used as substitute liquids. RDTs were also run with distilled water, without adding blood. Results were compared to those obtained with the RDT kit's buffer and Plasmodium positive samples. Results Only eight cassettes (in four RDT brands) showed no control line and were considered invalid. Visible test lines occurred for at least one malaria-free sample and one of the substitutes in 20/27 (74%) RDT brands (saline: n = 16; distilled water: n = 17; and tap water: n = 20), and in 15 RDTs which were run with distilled water only. They occurred for all Plasmodium antigens and RDT formats (two-, three- and four-band RDTs). Clearance of the background of the strip was excellent except for saline. The aspects (colour, intensity and crispness) of the control and the false-positive test lines were similar to those obtained with the RDT kits' buffer and Plasmodium positive samples. Conclusion Replacement of the RDT kit's dedicated buffer by saline, distilled water and tap water can cause false-positive test results. PMID:20650003

  2. Water Microbiology Kit/Microbial Capture Devices (WMK MCD)

    NASA Image and Video Library

    2009-08-04

    ISS020-E-027318 (4 Aug. 2009) --- Canadian Space Agency astronaut Robert Thirsk, Expedition 20 flight engineer, performs a subsequent in-flight analysis with a Water Microbiology Kit/Microbial Capture Devices (WMK MCD) for microbial traces in the Destiny laboratory of the International Space Station.

  3. 46 CFR 160.058-4 - Inspections.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-4 Inspections. (a) Desalter kits specified by this subpart are not inspected at regularly scheduled factory inspections; however, the Commander of the Coast Guard District in which the desalter kits are manufactured...

  4. 46 CFR 160.058-4 - Inspections.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-4 Inspections. (a) Desalter kits specified by this subpart are not inspected at regularly scheduled factory inspections; however, the Commander of the Coast Guard District in which the desalter kits are manufactured...

  5. 46 CFR 160.058-4 - Inspections.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-4 Inspections. (a) Desalter kits specified by this subpart are not inspected at regularly scheduled factory inspections; however, the Commander of the Coast Guard District in which the desalter kits are manufactured...

  6. 46 CFR 160.058-4 - Inspections.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-4 Inspections. (a) Desalter kits specified by this subpart are not inspected at regularly scheduled factory inspections; however, the Commander of the Coast Guard District in which the desalter kits are manufactured...

  7. 46 CFR 160.058-4 - Inspections.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-4 Inspections. (a) Desalter kits specified by this subpart are not inspected at regularly scheduled factory inspections; however, the Commander of the Coast Guard District in which the desalter kits are manufactured...

  8. FlowGo: An Educational Kit for Fluid Dynamics and Heat Transfer

    NASA Astrophysics Data System (ADS)

    Guri, Dominic; Portsmore, Merredith; Kemmerling, Erica

    2015-11-01

    The authors have designed and prototyped an educational toolkit that will help middle-school-aged students learn fundamental fluid mechanics and heat transfer concepts in a hands-on play environment. The kit allows kids to build arbitrary flow rigs to solve fluid mechanics and heat transfer challenge problems. Similar kits for other engineering fields, such as structural and electrical engineering, have resulted in pedagogical improvements, particularly in early engineering education, where visual demonstrations have a significant impact. Using the FlowGo kit, students will be able to conduct experiments and develop new design ideas to solve challenge problems such as building plant watering systems or modeling water and sewage reticulation. The toolkit consists of components such as tubes, junctions, and reservoirs that easily snap together via a modular, universal connector. Designed with the Massachusetts K-12 science standards in mind, this kit is intended to be affordable and suitable for classroom use. Results and user feedback from students conducting preliminary tests of the kit will be presented.

  9. [Application of DNA extraction kit, 'GM quicker' for detection of genetically modified soybeans].

    PubMed

    Sato, Noriko; Sugiura, Yoshitsugu; Tanaka, Toshitsugu

    2012-01-01

    Several DNA extraction methods have been officially introduced to detect genetically modified soybeans, but the choice of DNA extraction kits depend on the nature of the samples, such as grains or processed foods. To overcome this disadvantage, we examined whether the GM quicker kit is available for both grains and processed foods. We compared GM quicker with four approved DNA extraction kits in respect of DNA purity, copy numbers of lectin gene, and working time. We found that the DNA quality of GM quicker was superior to that of the other kits for grains, and the procedure was faster. However, in the case of processed foods, GM quicker was not superior to the other kits. We therefore investigated an unapproved GM quicker 3 kit, which is available for DNA extraction from processed foods, such as tofu and boiled soybeans. The GM quicker 3 kit provided good DNA quality from both grains and processed foods, so we made a minor modification of the GM quicker-based protocol that was suitable for processed foods, using GM quicker and its reagents. The modified method enhanced the performance of GM quicker with processed foods. We believe that GM quicker with the modified protocol is an excellent tool to obtain high-quality DNA from grains and processed foods for detection of genetically modified soybeans.

  10. Performance of the Extravehicular Mobility Unit (EMU): Airlock Coolant Loop Recovery (A/L CLR) Hardware - Phase II

    NASA Technical Reports Server (NTRS)

    Steele, John; Rector, tony; Gazda, Daniel; Lewis, John

    2009-01-01

    An EMU water processing kit (Airlock Coolant Loop Recovery A/L CLR) was developed as a corrective action to Extravehicular Mobility Unit (EMU) coolant flow disruptions experienced on the International Space Station (ISS) in May of 2004 and thereafter. Conservative schedules for A/L CLR use and component life were initially developed and implemented based on prior analysis results and analytical modeling. The examination of postflight samples and EMU hardware in November of 2006 indicated that the A/L CLR kits were functioning well and had excess capacity that would allow a relaxation of the initially conservative schedules of use and component life. A relaxed use schedule and list of component lives was implemented thereafter. Since the adoption of the relaxed A/L CLR schedules of use and component lives, several A/L CLR kit components, transport loop water samples and sensitive EMU transport loop components have been examined to gage the impact of the relaxed requirements. The intent of this paper is to summarize the findings of that evaluation, and to outline updated schedules for A/L CLR use and component life.

  11. Commander Lousma adds water to a beverage container on middeck

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Commander Lousma, wearing communications kit assembly (assy) mini headset (HDST), fills beverage container using the JSC water dispenser kit water gun to prepare a juice drink. Lousma is wearing the trousers and shirt of a three-piece shuttle constant wear garment as he floats above the potable water tank on the middeck floor. The constant wear garment jacket is secured on a side hatch handle (background) to avoid zero gravity effect.

  12. UPA Fill Drain Valve Modification kit installation

    NASA Image and Video Library

    2016-01-25

    ISS046e023885 (01/25/2016) --- NASA astronaut Tim Kopra performs regular maintenance on the Urine Processing Assembly (UPA) aboard the International Space Station. The UPA is used by the crew to recycle water for use on the station. The image shows Tim replacing the brine filter from the UPA Fill Drain Valve enclosure.

  13. AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) OF FOUR TEST KITS FOR THE ANALYSIS OF ATRAZINE IN WATER: ABRAXIS LLC ATRAZINE ELISA KIT, BEACON ANALYTICAL SYSTEMS, INC. ATRAZINE TUBE KIT, SILVER LAKE RESEARCH CORP. WATERSAFE PESTICIDE TEST AND STRATEGIC DIAGNOSTICS, INC. RAPID ASSAY KIT

    EPA Science Inventory

    The Environmental Technology Verification (ETV) Program, beginning as an initiative of the U.S. Environmental Protection Agency (EPA) in 1995, verifies the performance of commercially available, innovative technologies that can be used to measure environmental quality. The ETV ...

  14. Evaluation of ELISA tests specific for Shiga toxin 1 and 2 in food and water samples

    USDA-ARS?s Scientific Manuscript database

    Two enzyme-linked immunosorbent assay (ELISA) kits were evaluated for their effectiveness in detecting and differentiating between Shiga toxin 1 and 2 (Stx1 and Stx2) produced by Shiga toxin-producing E. coli (STEC) inoculated into food and water samples. Each kit incorporated monoclonal antibodies ...

  15. Johnny Horizon Environmental Test Kit.

    ERIC Educational Resources Information Center

    Bentley, Richard; Bentley, William

    Derived from tests presently used by state and federal agencies involved with pollution detection, this Environmental Test Kit contains materials and instructions for ten experiments. Each experiment is designed to test a different aspect of air and water, to find out whether or not the air and water in the tester's immediate area has been…

  16. Design, Certification, and Deployment of the Colorimetric Water Quality Monitoring Kit (CWQMK)

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Nolan, Daniel J.; Rutz, Jeff A.; Schultz, John R.; Siperko, Lorraine M.; Porter, Marc D.; Lipert, Robert J.; Carrizales, Stephanie M.; McCoy, J. Torin

    2009-01-01

    In August 2009, an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE) technology was delivered to the International Space Station (ISS) aboard STS-128/17A. The kit, called the Colorimetric Water Quality Monitoring Kit (CWQMK), was developed by a team of scientists and engineers from NASA s Habitability and Environmental Factors Division in the Space Life Sciences Directorate at Johnson Space Center, the Wyle Integrated Science and Engineering Group in Houston, Texas, the University of Utah, and Iowa State University. The CWQMK was flown and deployed as a Station Development Test Objective (SDTO) experiment on ISS. The goal of the SDTO experiment was to evaluate the acceptability of CSPE technology for routine water quality monitoring on ISS. This paper provides an overview of the SDTO experiment, as well as a detailed description of the CWQMK hardware and a summary of the testing and analysis conducted to certify the CWQMK for use on ISS. The results obtained from the SDTO experiment are also reported and discussed in detail.

  17. Survival Kit - Food Kit

    NASA Image and Video Library

    1962-10-01

    S62-08743 (1962) --- Food kit used by Mercury astronauts. Some is dehydrated and needs water, other packets are ready to eat. Size is measured relative to a ruler. Included are packets of mushroom soup, orange-grapefruit juice, cocoa beverage, pineapple juice, chicken with gravy, pears, strawberries, beef and vegetables and other assorted food containers. Photo credit: NASA

  18. Survival Kit - Food Kit

    NASA Image and Video Library

    1962-10-01

    S62-08742 (1962) --- Food kit used by Mercury astronauts. Some is dehydrated and needs water, other packets are ready to eat. Size is measured relative to a ruler. Included are packets of mushroom soup, orange-grapefruit juice, cocoa beverage, pineapple juice, chicken with gravy, pears, strawberries, beef and vegetables and other assorted food containers. Photo credit: NASA

  19. 46 CFR 160.054-2 - Type and size.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Inflatable Liferafts § 160.054-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight type... special consideration. (b) Size. First-aid kits shall be of a size adequate for packing 12 standard single...

  20. 46 CFR 160.054-2 - Type and size.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Inflatable Liferafts § 160.054-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight type... special consideration. (b) Size. First-aid kits shall be of a size adequate for packing 12 standard single...

  1. 46 CFR 160.054-2 - Type and size.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Inflatable Liferafts § 160.054-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight type... special consideration. (b) Size. First-aid kits shall be of a size adequate for packing 12 standard single...

  2. 46 CFR 160.054-2 - Type and size.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Inflatable Liferafts § 160.054-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight type... special consideration. (b) Size. First-aid kits shall be of a size adequate for packing 12 standard single...

  3. 46 CFR 160.054-2 - Type and size.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Inflatable Liferafts § 160.054-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight type... special consideration. (b) Size. First-aid kits shall be of a size adequate for packing 12 standard single...

  4. [Examination of drinking water used in livestock production. Microbiological and physico-chemical methods. Ready to use test kits in field experiments].

    PubMed

    Sassen, J

    2000-08-01

    Livestock health care service is very much involved and interested in surveillance of the drinking water as well. However, in order to examine the water immediately "on the fly", test kits have to be provided, which offer results comparable to these obtained in the laboratories according to official prescription. The German Army was confronted with a similar situation during the secently performed mission in crisis regions. At the early state of a mission usually laboratory equipment is not yet established. Therefore a set of test kits was compiled suitable for mobile microbiological examination of drinking water. This set was excessively examined comparison with reference methods. In conclusion it is shown, that the mobile set gains equal or even better results compared to those obtained according to legally prescribed standard procedures.

  5. Assessment of RFID Read Accuracy for ISS Water Kit

    NASA Technical Reports Server (NTRS)

    Chu, Andrew

    2011-01-01

    The Space Life Sciences Directorate/Medical Informatics and Health Care Systems Branch (SD4) is assessing the benefits Radio Frequency Identification (RFID) technology for tracking items flown onboard the International Space Station (ISS). As an initial study, the Avionic Systems Division Electromagnetic Systems Branch (EV4) is collaborating with SD4 to affix RFID tags to a water kit supplied by SD4 and studying the read success rate of the tagged items. The tagged water kit inside a Cargo Transfer Bag (CTB) was inventoried using three different RFID technologies, including the Johnson Space Center Building 14 Wireless Habitat Test Bed RFID portal, an RFID hand-held reader being targeted for use on board the ISS, and an RFID enclosure designed and prototyped by EV4.

  6. 46 CFR 160.041-2 - Type and size.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant Vessels § 160.041-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight cabinet carrying... consideration. (b) Size. First-aid kits shall be of a size (approximately 9″×9″×21/2″ inside) adequate for...

  7. 46 CFR 160.041-2 - Type and size.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant Vessels § 160.041-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight cabinet carrying... consideration. (b) Size. First-aid kits shall be of a size (approximately 9″×9″×21/2″ inside) adequate for...

  8. 46 CFR 160.041-2 - Type and size.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant Vessels § 160.041-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight cabinet carrying... consideration. (b) Size. First-aid kits shall be of a size (approximately 9″ × 9″ × 21/2″ inside) adequate for...

  9. 46 CFR 160.041-2 - Type and size.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant Vessels § 160.041-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight cabinet carrying... consideration. (b) Size. First-aid kits shall be of a size (approximately 9″ × 9″ × 21/2″ inside) adequate for...

  10. 46 CFR 160.041-2 - Type and size.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant Vessels § 160.041-2 Type and size. (a) Type. First-aid kits covered by this specification shall be of the water-tight cabinet carrying... consideration. (b) Size. First-aid kits shall be of a size (approximately 9″×9″×21/2″ inside) adequate for...

  11. Management of the Post-Shuttle Extravehicular Mobility Unit (EMU) Water Circuits

    NASA Technical Reports Server (NTRS)

    Steele, John W.; Etter, David; Rector, Tony; Hill, Terry; Wells, Kevin

    2011-01-01

    The EMU incorporates two separate water circuits for the rejection of metabolic heat from the astronaut and the cooling of electrical components. The first (the Transport Water Loop) circulates in a semi-closed-loop manner and absorbs heat into a Liquid Coolant and Ventilation Garment (LCVG) warn by the astronaut. The second (the Feed Water Loop) provides water to a cooling device (Sublimator) with a porous plate, and that water subsequently sublimates to space vacuum. The cooling effect from the sublimation of this water translates to a cooling of the LCVG water that circulates through the Sublimator. Efforts are underway to streamline the use of a water processing kit (ALCLR) that is being used to periodically clean and disinfect the Transport Loop Water. Those efforts include a fine tuning of the duty cycle based on a review of prior performance data as well as an assessment of a fixed installation of this kit into the EMU backpack or within on-orbit EMU interface hardware. Furthermore, testing is being conducted to ensure compatibility between the International Space Station (ISS) Water Processor Assembly (WPA) effluent and the EMU Sublimator as a prelude to using the WPA effluent as influent to the EMU Feed Water loop. This work is undertaken to reduce the crew-time and logistics burdens for the EMU, while ensuring the long-term health of the EMU water circuits for a post-Shuttle 6-year service life.

  12. Management of the Post-Shuttle Extravehicular Mobility Unit (EMU) Water Circuits

    NASA Technical Reports Server (NTRS)

    Steele, John W.; Etter, David; Rector, Tony; Hill, Terry; Wells, Kevin

    2012-01-01

    The EMU incorporates two separate water circuits for the rejection of metabolic heat from the astronaut and the cooling of electrical components. The first (the Transport Water Loop) circulates in a semi-closed-loop manner and absorbs heat into a Liquid Coolant and Ventilation Garment (LCVG) worn by the astronaut. The second (the Feed-water Loop) provides water to a cooling device (Sublimator) with a porous plate, and that water subsequently sublimates to space vacuum. The cooling effect from the sublimation of this water translates to a cooling of the LCVG water that circulates through the Sublimator. Efforts are underway to streamline the use of a water processing kit (ALCLR) that is being used to periodically clean and disinfect the Transport Loop Water. Those efforts include a fine tuning of the duty cycle based on a review of prior performance data as well as an assessment of a fixed installation of this kit into the EMU backpack, within on-orbit EMU interface hardware or as a stand-alone unit. Furthermore, testing is being conducted to ensure compatibility between the International Space Station (ISS) Water Processor Assembly (WPA) effluent and the EMU Sublimator as a prelude to using the WPA effluent as influent to the EMU Feed Water loop. This work is undertaken to reduce the crewtime and logistics burdens for the EMU, while ensuring the long-term health of the EMU water circuits for a 6-year service life.

  13. Pilot Fullerton prepares meal on middeck

    NASA Image and Video Library

    1982-03-30

    STS003-26-253 (30 March 1982) --- Astronaut Gordon Fullerton, STS-3 pilot, wearing communications kit assembly (assy) mini-headset (HDST), prepares meal on middeck. Fullerton clips corner of rehydratable food (cereal) package with scissors. The opening will allow Fullerton to insert JSC water dispenser kit water gun in order to heat contents with hot water. Meal tray assembly is secured to forward middeck locker and holds additional food packages and beverage containers. Photo credit: NASA

  14. The challenges of lean manufacturing implementation in kitting assembly

    NASA Astrophysics Data System (ADS)

    Fansuri, A. F. H.; Rose, A. N. M.; Nik Mohamed, N. M. Z.; Ahmad, H.

    2017-10-01

    Literature studies shows that lean manufacturing goes way back with the original founder Eli Whitney in year 1799. The main purpose of lean manufacturing is to identify and eliminate waste in production. The application of lean manufacturing can be carried out in any industrial processes with regards to the understanding of lean principles, theories and practices. Kitting is one of the important aspects in a successful production. The continuous supply of materials from store to production has to be systematic and able to achieve lean standard for it to be successful. The objective of this paper is to review the implementation of lean manufacturing in kitting assembly. Previous papers show that, the implementation of lean manufacturing in kitting assembly may be beneficial to the organization such as reduce in space occupancy, part shortages, lead time and manpower. Based on previous research, some industries may tend to change between kitting and line stocking which are due to lack of understanding when implementing kitting and causes longer lead time and materials overflow in store. With a proper understanding on what to kit, where to kit, how to kit, why to kit and who kits the material with a standardised process flow may ensure the success of kitting.

  15. Technologies for safe births.

    PubMed

    1984-01-01

    The basic elements of a safe birth are proper prenatal care, adequate preparation of the mother, health worker, and site, awareness of the progress of labor and safe delivery, recognition of danger signs, and appropriate follow-up care. Technologies are differentiated by determining 1) the needs of rural birth attendants, 2) the nature of delivery kits, 3) proper cleanliness of the hands and equipment, and appropriate use of 5) disinfecting equipment, 6) drugs and medications, 7) the vertical position, 8) specialized instruments, and 9) records and support materials. Alternatives for measuring time are indicated. Customized kits available from UNICEF are described; some of the problems with these kits are reported. The logistics, referral procedures, and training and supervision needed for appropriate program managements are discussed. Adapting technologies to the local environment requires assessing the practices of traditional birth attendants (TBAs), the provision of kits (cost, ease of use and maintenance, replacement, durability, availability), the training required for proper use of equipment, the logistics of kit use, side effects of technologies, community attitudes, and evaluation. The advantages and disadvantages of including or not including particular supplies in the kit are discussed, i.e., the container for boiling water would either be a local pot or the aluminum carrying case. In lieu of a fingernail brush, a twig may be used for nail cleaning. Hand washing where water shortages exist might entail using a tin with a hole plugged with a stick to let water trickle as needed. Antiseptic solutions such a Dettol or Savlon can be used where a severe shortage exists. Basic equipment includes: soap and water, a container for boiling, other sterile containers, a protective cover of delivery area, towels, swabs, an optional apron, cord ties, a cutting instrument, gauze, a receiving blanket, records, and a carrying case.

  16. Differences in staining intensities affect reported occurrences and concentrations of Giardia spp. in surface drinking water sources.

    PubMed

    Alderisio, K A; Villegas, L F; Ware, M W; McDonald, L A; Xiao, L; Villegas, E N

    2017-12-01

    USEPA Method 1623, or its equivalent, is currently used to monitor for protozoan contamination of surface drinking water sources worldwide. At least three approved staining kits used for detecting Cryptosporidium and Giardia are commercially available. This study focuses on understanding the differences among staining kits used for Method 1623. Merifluor and EasyStain labelling kits were used to monitor Cryptosporidium oocyst and Giardia cyst densities in New York City's raw surface water sources. In the year following a change to the approved staining kits for use with Method 1623, an anomaly was noted in the occurrence of Giardia cysts in New York City's raw surface water. Specifically, Merifluor-stained samples had higher Giardia cyst densities as compared with those stained with EasyStain. Side by side comparison revealed significantly lower fluorescence intensities of Giardia muris as compared with Giardia duodenalis cysts when labelled with EasyStain. This study showed very poor fluorescence intensity signals by EasyStain on G. muris cysts resulting in lower cyst counts, while Merifluor, with its broader Giardia cyst staining specificity, resulted in higher cyst counts, when using Methods 1623. These results suggest that detected Giardia cyst concentrations are dependent on the staining kits used, which can result in a more or less conservative estimation of occurrences and densities of zoonotic Giardia cysts by detecting a broader range of Giardia species/Assemblages. Published 2017. This article is a U.S. Government work and is in the public domain in the USA.

  17. 46 CFR 160.058-2 - Type.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-2 Type. (a) Desalter kits specified by this subpart shall be of the type described in the specification listed in § 160.058...

  18. 46 CFR 160.058-2 - Type.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-2 Type. (a) Desalter kits specified by this subpart shall be of the type described in the specification listed in § 160.058...

  19. 46 CFR 160.058-2 - Type.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-2 Type. (a) Desalter kits specified by this subpart shall be of the type described in the specification listed in § 160.058...

  20. 46 CFR 160.058-2 - Type.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-2 Type. (a) Desalter kits specified by this subpart shall be of the type described in the specification listed in § 160.058...

  1. Village Infrastructure Kit-Alpha. Global Innovation and Strategy Center

    DTIC Science & Technology

    2009-05-01

    populations in the world. These kits could be capable of providing renewable energy, water, and telecommunication services, and therefore could impact...for development. The VIKA team adhered to the following five parameters during research efforts: Make available potable water, renewable energy...ENERGY Solar Energy Overview Solar energy is a renewable source that can provide an alternative to the electrical grid. It is an increasingly

  2. c-kit expression profile and regulatory factors during spermatogonial stem cell differentiation

    PubMed Central

    2013-01-01

    Background It has been proven that c-kit is crucial for proliferation, migration, survival and maturation of spermatogenic cells. A periodic expression of c-kit is observed from primordial germ cells (PGCs) to spermatogenetic stem cells (SSCs), However, the expression profile of c-kit during the entire spermatogenesis process is still unclear. This study aims to reveal and compare c-kit expression profiles in the SSCs before and after the anticipated differentiation, as well as to examine its relationship with retinoic acid (RA) stimulation. Results We have found that there are more than 4 transcripts of c-kit expressed in the cell lines and in the testes. The transcripts can be divided into short and long categories. The long transcripts include the full-length canonical c-kit transcript and the 3′ end short transcript. Short transcripts include the 3.4 kb short transcript and several truncated transcripts (1.9-3.2 kb). In addition, the 3.4 kb transcript (starting from intron 9 and covering exons 10 ~ 21) is discovered to be specifically expressed in the spermatogonia. The extracellular domain of Kit is obtained in the spermatogonia stage, but the intracellular domain (50 kDa) is constantly expressed in both SSCs and spermatogonia. The c-kit expression profiles in the testis and the spermatogonial stem cell lines vary after RA stimulation. The wave-like changes of the quantitative expression pattern of c-kit (increase initially and decrease afterwards) during the induction process are similar to that of the in vivo male germ cell development process. Conclusions There are dynamic transcription and translation changes of c-kit before and after SSCs’ anticipated differentiation and most importantly, RA is a significant upstream regulatory factor for c-kit expression. PMID:24161026

  3. In-Flight Water Quality Monitoring on the International Space Station (ISS): Measuring Biocide Concentrations with Colorimetric Solid Phase Extraction (CSPE)

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Schultz, John R.; Siperko, Lorraine M.; Porter, Marc D.; Lipert, Robert J.; Flint, Stephanie M.; McCoy, J. Torin

    2011-01-01

    The colorimetric water quality monitoring kit (CWQMK) was delivered to the International Space Station (ISS) on STS-128/17A and was initially deployed in September 2009. The kit was flown as a station development test objective (SDTO) experiment to evaluate the acceptability of colorimetric solid phase extraction (CSPE) technology for routine water quality monitoring on the ISS. During the SDTO experiment, water samples from the U.S. water processor assembly (WPA), the U.S. potable water dispenser (PWD), and the Russian system for dispensing ground-supplied water (SVO-ZV) were collected and analyzed with the CWQMK. Samples from the U.S. segment of the ISS were analyzed for molecular iodine, which is the biocide added to water in the WPA. Samples from the SVOZV system were analyzed for ionic silver, the biocide used on the Russian segment of the ISS. In all, thirteen in-flight analysis sessions were completed as part of the SDTO experiment. This paper provides an overview of the experiment and reports the results obtained with the CWQMK. The forward plan for certifying the CWQMK as operational hardware and expanding the capabilities of the kit are also discussed.

  4. Hydrogen-Rich Water Ameliorates Total Body Irradiation-Induced Hematopoietic Stem Cell Injury by Reducing Hydroxyl Radical

    PubMed Central

    Xue, Xiaolei; Han, Xiaodan; Li, Yuan; Lu, Lu; Li, Deguan

    2017-01-01

    We examined whether consumption of hydrogen-rich water (HW) could ameliorate hematopoietic stem cell (HSC) injury in mice with total body irradiation (TBI). The results indicated that HW alleviated TBI-induced HSC injury with respect to cell number alteration and to the self-renewal and differentiation of HSCs. HW specifically decreased hydroxyl radical (∙OH) levels in the c-kit+ cells of 4 Gy irradiated mice. Proliferative bone marrow cells (BMCs) increased and apoptotic c-kit+ cells decreased in irradiated mice uptaken with HW. In addition, the mean fluorescence intensity (MFI) of γ-H2AX and percentage of 8-oxoguanine positive cells significantly decreased in HW-treated c-kit+ cells, indicating that HW can alleviate TBI-induced DNA damage and oxidative DNA damage in c-kit+ cells. Finally, the cell cycle (P21), cell apoptosis (BCL-XL and BAK), and oxidative stress (NRF2, HO-1, NQO1, SOD, and GPX1) proteins were significantly altered by HW in irradiated mouse c-kit+ cells. Collectively, the present results suggest that HW protects against TBI-induced HSC injury. PMID:28243358

  5. Hydrogen-Rich Water Ameliorates Total Body Irradiation-Induced Hematopoietic Stem Cell Injury by Reducing Hydroxyl Radical.

    PubMed

    Zhang, Junling; Xue, Xiaolei; Han, Xiaodan; Li, Yuan; Lu, Lu; Li, Deguan; Fan, Saijun

    2017-01-01

    We examined whether consumption of hydrogen-rich water (HW) could ameliorate hematopoietic stem cell (HSC) injury in mice with total body irradiation (TBI). The results indicated that HW alleviated TBI-induced HSC injury with respect to cell number alteration and to the self-renewal and differentiation of HSCs. HW specifically decreased hydroxyl radical ( ∙ OH) levels in the c-kit + cells of 4 Gy irradiated mice. Proliferative bone marrow cells (BMCs) increased and apoptotic c-kit + cells decreased in irradiated mice uptaken with HW. In addition, the mean fluorescence intensity (MFI) of γ -H2AX and percentage of 8-oxoguanine positive cells significantly decreased in HW-treated c-kit + cells, indicating that HW can alleviate TBI-induced DNA damage and oxidative DNA damage in c-kit + cells. Finally, the cell cycle (P21), cell apoptosis (BCL-XL and BAK), and oxidative stress (NRF2, HO-1, NQO1, SOD, and GPX1) proteins were significantly altered by HW in irradiated mouse c-kit + cells. Collectively, the present results suggest that HW protects against TBI-induced HSC injury.

  6. My River My Home: Both Art and Science

    NASA Astrophysics Data System (ADS)

    Gillies, S. L.; Janmaat, A.; Marsh, S. J.; Peucker-Ehrenbrink, B.; Voss, B.; Holmes, R. M.; King, S.; Bertrand, K.

    2014-12-01

    The University of the Fraser Valley has been researching the water chemistry of the Fraser River since 2009 as a member of the Global Rivers Observatory coordinated by Woods Hole Oceanographic Institution and Woods Hole Research Center. The Global Rivers Observatory is advancing our understanding of how climate change, deforestation, and other disturbances are impacting river chemistry and land-ocean linkages. This knowledge is vital for tracking the health of Earth's watersheds and predicting how Earth's water and chemical cycles will change in the future. The Global Rivers Observatory also promotes the communication of science to the general public. In September 2013, the My River My Home art and science exhibit opened at the Fraser River Discovery Centre, New Westminster, BC. The exhibit is a global exchange of artwork created by children living along the rivers being studied by the Global Rivers Observatory scientists. The exhibit is intended to inspire young students to develop an awareness of the environment and the importance of rivers. Scientists from UFV, WHOI, and WHRC worked together with the Fraser River Discovery Centre on the science communication aspects of the display and to develop hands-on science activities looking at different aspects of river water quality. The exhibition has led to the creation of My River My Home, An Activity Kit for Educators about the sustainability of the Fraser River. The kit is being offered through the Fraser River Discovery Centre and deals with issues such as the importance of water, water quality, and encouraging a global perspective. The resource kit was classroom tested by several teachers, and four UBC teacher candidates worked on incorporating teacher suggestions into the kit. The resource kit will be available on-line at the end of September 2014 and contains inquiry based activities suitable for a variety of educational levels.

  7. Lysozyme adsorption onto mesoporous materials: effect of pore geometry and stability of adsorbents.

    PubMed

    Vinu, Ajayan; Miyahara, Masahiko; Hossain, Kazi Zakir; Takahashi, Motoi; Balasubramanian, Veerappan Vaithilingam; Mori, Toshiyuki; Ariga, Katsuhiko

    2007-03-01

    In this paper, adsorption of lysozyme onto two kinds of mesoporous adsorbents (KIT-5 and AISBA-15) has been investigated and the results on the effects of pore geometry and stability of the adsorbents are also discussed. The KIT-5 mesoporous silica materials possess cage-type pore geometry while the AISBA-15 adsorbent has mesopores of cylindrical type with rather large diameter (9.7 nm). Adsorption of lysozyme onto AISBA-15 aluminosilicate obeys a Langmuir isotherm, resulting in pore occupation of 25 to 30%. In contrast, the KIT-5 adsorbents showed very small adsorption capacities for the lysozyme adsorption, typically falling in 6 to 13% of pore occupation. The cage-type KIT-5 adsorbents have narrow channel (4 to 6 nm) where penetration of the lysozyme (3 x 3 x 4.5 nm) might be restricted. The KIT-5 adsorbent tends to collapse after long-time immersion in water, as indicated by XRD patterns, while the AISBA-15 adsorbent retains its regular structure even after immersion in basic water for 4 days. These facts confirm superiority of the AISBA-15 as an adsorbent as compared with the KIT-5 mesoporous silicates. This research strikingly demonstrates the selection of mesoporous materials is crucial to achieve efficient immobilization of biomaterials in aqueous environment.

  8. Immunochemical analytical methods for the determination of peanut proteins in foods.

    PubMed

    Whitaker, Thomas B; Williams, Kristina M; Trucksess, Mary W; Slate, Andrew B

    2005-01-01

    Peanut proteins can cause allergenic reactions that can result in respiratory and circulatory effects in the body sometimes leading to shock and death. The determination of peanut proteins in foods by analytical methods can reduce the risk of serious reactions in the highly sensitized individual by allowing for the detection of these proteins in a food at various stages of the manufacturing process. The method performance of 4 commercially available enzyme-linked immunosorbent assay (ELISA) kits was evaluated for the detection of peanut proteins in milk chocolate, ice cream, cookies, and breakfast cereals: ELISA-TEK Peanut Protein Assay, now known as "Bio-Kit" for peanut proteins, from ELISA Technologies Inc.; Veratox for Peanut Allergens from Neogen Corp.; RIDASCREEN Peanut Kit from R-Biopharm GmbH; and ProLisa from Canadian Food Technology Ltd. The 4 test kits were evaluated for accuracy (recovery) and precision using known concentrations of peanut or peanut proteins in the 4 food matrixes. Two different techniques, incurred and spiked, were used to prepare samples with 4 known concentrations of peanut protein. Defatted peanut flour was added in the incurred samples, and water-soluble peanut proteins were added in the spiked samples. The incurred levels were 0.0, 10, 20, and 100 microg whole peanut per g food; the spiked levels were 0.0, 5, 10, and 20 microg peanut protein per g food. Performance varied by test kit, protein concentration, and food matrix. The Veratox kit had the best accuracy or lowest percent difference between measured and incurred levels of 15.7% when averaged across all incurred levels and food matrixes. Recoveries associated with the Veratox kit varied from 93 to 115% for all food matrixes except cookies. Recoveries for all kits were about 50% for cookies. The analytical precision, as measured by the variance, increased with an increase in protein concentration. However, the coefficient of variation (CV) was stable across the 4 incurred protein levels and was 7.0% when averaged across the 4 food matrixes and analytical kits. The R-Biopharm test kit had the best precision or a CV of 4.2% when averaged across all incurred levels and food matrixes. Because measured protein values varied by test kit and food matrix, a method was developed to normalize or transform measured protein concentrations to an adjusted protein value that was equal to the known protein concentration. The normalization method adjusts measured protein values to equal the true protein value regardless of the type test kit or type food matrix.

  9. Loss of c-Kit function impairs arteriogenesis in a mouse model of hindlimb ischemia.

    PubMed

    Hernandez, Diana R; Artiles, Adriana; Duque, Juan C; Martinez, Laisel; Pinto, Mariana T; Webster, Keith A; Velazquez, Omaida C; Vazquez-Padron, Roberto I; Lassance-Soares, Roberta M

    2018-04-01

    Arteriogenesis is a process whereby collateral vessels remodel usually in response to increased blood flow and/or wall stress. Remodeling of collaterals can function as a natural bypass to alleviate ischemia during arterial occlusion. Here we used a genetic approach to investigate possible roles of tyrosine receptor c-Kit in arteriogenesis. Mutant mice with loss of c-Kit function (Kit W/W-v ), and controls were subjected to hindlimb ischemia. Blood flow recovery was evaluated pre-, post-, and weekly after ischemia. Foot ischemic damage and function were assessed between days 1 to 14 post-ischemia while collaterals remodeling were measured 28 days post-ischemia. Both groups of mice also were subjected to wild type bone marrow cells transplantation 3 weeks before hindlimb ischemia to evaluate possible contributions of defective bone marrow c-Kit expression on vascular recovery. Kit W/W-v mice displayed impaired blood flow recovery, greater ischemic damage and foot dysfunction after ischemia compared to controls. Kit W/W-v mice also demonstrated impaired collateral remodeling consistent with flow recovery findings. Because arteriogenesis is a biological process that involves bone marrow-derived cells, we investigated which source of c-Kit signaling (bone marrow or vascular) plays a major role in arteriogenesis. Kit W/W-v mice transplanted with bone marrow wild type cells exhibited similar phenotype of impaired blood flow recovery, greater tissue ischemic damage and foot dysfunction as nontransplanted Kit W/W-v mice. This study provides evidence that c-Kit signaling is required during arteriogenesis. Also, it strongly suggests a vascular role for c-Kit signaling because rescue of systemic c-Kit activity by bone marrow transplantation did not augment the functional recovery of Kit W/W-v mouse hindlimbs. Copyright © 2017 Elsevier Inc. All rights reserved.

  10. Ballast water compliance monitoring: A new application for ATP

    NASA Astrophysics Data System (ADS)

    Lo Curto, A.; Stehouwer, P.; Gianoli, C.; Schneider, G.; Raymond, M.; Bonamin, V.

    2018-03-01

    The coming into force of the USCG ballast water regulations and the IMO ballast water management convention resulted in the development of several technologies approved for the treatment of ballast water. To ensure compliance of these technologies, the development of rapid and robust analysis methods was necessary. In collaboration with the SGS Group (Switzerland) and LuminUltra (Canada), Aqua-tools (France) has developed an innovative Ballast Water Treatment Monitoring (BWTM) kit for rapid onboard testing. The affordable kit provides results in less than 1 h, is easy to use and durable ensuring that the ballast water treatment system on the ship is fully compliant with the discharge standards upon arrival in port. The core of this method is a combination of high-quality reagents (lysis solution and ATP 2G Luminase™ enzyme) not inhibited by salinity and a patented fast homogenizing method for ATP extraction developed for a higher ATP recovery from zooplankton and phytoplankton. Compared to traditional analysis methods, the BWTM Kit provides fast and accurate results for all three fractions of microorganisms (≥ 50 μm, ≥ 10 ÷ < 50 μm and bacteria). Preliminary tests carried out in cooperation with SGS showed that the proposed method was able to detect onboard the efficiency of the treatment systems used. Compliance limits were established for all size fractions and a correlation between the standard methods (microscopy, plate count, MPN) and ATP was evaluated. The BWTM kit can provide a fast indication of compliance or gross exceedance. The rare borderline cases, when encountered, of course require additional confirmation.

  11. Adaptation of commercial biomarker kits and proposal for 'drug development kits' to support bioanalysis: call for action.

    PubMed

    Islam, Rafiqul; Kar, Sumit; Islam, Clarinda; Farmen, Raymond

    2018-06-01

    There has been an increased use of commercial kits for biomarker measurement, commensurate with the increased demand for biomarkers in drug development. However, in most cases these kits do not meet the quality attributes for use in regulated environment. The process for adaptation of these kits can be frustrating, time consuming and resource intensive. In addition, a lack of harmonized guidance for the validation of biomarker poses a significant challenge in the adaptation of kits in a regulated environment. The purpose of this perspective is to propose a tiered approach to commercial drug development kits with clearly defined quality attributes and to demonstrate how these kits can be adapted to perform analytical validation in a regulated environment.

  12. The history and development of NASA survival equipment.

    NASA Technical Reports Server (NTRS)

    Radnofsky, M. I.

    1972-01-01

    A research and development program on survival equipment was begun in early 1960 with the Mercury Program. The Mercury survival kit is discussed together with Gemini survival equipment, and Apollo I survival equipment. A study program is conducted to assess potential survival problems that may be associated with future space flights landing in polar waters. Survival kit requirements for applications on the Skylab program are also considered. Other investigations are concerned with the design of a global survival kit in connection with Space Shuttle missions.

  13. Food kit used by Mercury astronauts

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Food kit used by Mercury astronauts. Some is dehydrated and needs water, other packets are ready to eat. Size is measured relative to a ruler. Included are packets of mushroom soup, orange-grapefruit juice, cocoa beverage, pineapple juice, chicken with gravy, pears, strawberries, beef and vegetables and other assorted food containers (08742-3); mechanism for connecting water dispensor to dehydrated food containers to facilitate rehydration (08744); Group packets of ready to eat space food, with size being measured by a ruler (8745).

  14. Reiter works with SWAB ASD Filter Kit in the U.S. Laboratory during Expedition 13

    NASA Image and Video Library

    2006-09-10

    ISS013-E-80066 (10 Sept. 2006) --- European Space Agency (ESA) astronaut Thomas Reiter, Expedition 13 flight engineer, works with the surface, water and air biocharacterization (SWAB) air sampling device (ASD) filter kit in the Destiny laboratory of the International Space Station.

  15. AN ENVIRONMENTAL TECHNOLOGY VERIFICATION (ETV) TESTING OF ENZYMATIC TEST KITS FOR WARFARE AGENTS AND PESTICIDES IN DRINKING WATER

    EPA Science Inventory

    Enzymatic test kits, generally designed to be handheld and portable, detect the presence of chemical agents, carbamate pesticides, and/or organophosphate pesticides by relying on the reaction of the cholinesterase enzyme. Under normal conditions, the enzyme reacts as expected wi...

  16. Automating Disk Forensic Processing with SleuthKit, XML and Python

    DTIC Science & Technology

    2009-05-01

    1 Automating Disk Forensic Processing with SleuthKit, XML and Python Simson L. Garfinkel Abstract We have developed a program called fiwalk which...files themselves. We show how it is relatively simple to create automated disk forensic applications using a Python module we have written that reads...software that the portable device may contain. Keywords: Computer Forensics; XML; Sleuth Kit; Python I. INTRODUCTION In recent years we have found many

  17. Validation of Two Portable Instruments to Measure Iron Concentration in Groundwater in Rural Bangladesh

    PubMed Central

    Merrill, Rebecca D.; Shamim, Abu Ahmed; Ali, Hasmot; Schulze, Kerry; Rashid, Mahbubur; Christian, Parul; West, Jr., Keith P.

    2009-01-01

    Iron is ubiquitous in natural water sources used around the world for drinking and cooking. The health impact of chronic exposure to iron through water, which in groundwater sources can reach well above the World Health Organization's defined aesthetic limit of 0.3 mg/L, is not currently understood. To quantify the impact of consumption of iron in groundwater on nutritional status, it is important to accurately assess naturally-occurring exposure levels among populations. In this study, the validity of iron quantification in water was evaluated using two portable instruments: the HACH DR/890 portable colorimeter (colorimeter) and HACH Iron test-kit, Model IR-18B (test-kit), by comparing field-based iron estimates for 25 tubewells located in northwestern Bangladesh with gold standard atomic absorption spectrophotometry analysis. Results of the study suggest that the HACH test-kit delivers more accurate point-of-use results across a wide range of iron concentrations under challenging field conditions. PMID:19507757

  18. Validation of two portable instruments to measure iron concentration in groundwater in rural Bangladesh.

    PubMed

    Merrill, Rebecca D; Shamim, Abu Ahmed; Labrique, Alain B; Ali, Hasmot; Schulze, Kerry; Rashid, Mahbubur; Christian, Parul; West, Keith P

    2009-06-01

    Iron is ubiquitous in natural water sources used around the world for drinking and cooking. The health impact of chronic exposure to iron through water, which in groundwater sources can reach well above the World Health Organization's defined aesthetic limit of 0.3 mg/L, is not currently understood. To quantify the impact of consumption of iron in groundwater on nutritional status, it is important to accurately assess naturally-occurring exposure levels among populations. In this study, the validity of iron quantification in water was evaluated using two portable instruments: the HACH DR/890 portable colorimeter (colorimeter) and HACH Iron test-kit, Model IR-18B (test-kit), by comparing field-based iron estimates for 25 tubewells located in northwestern Bangladesh with gold standard atomic absorption spectrophotometry analysis. Results of the study suggest that the HACH test-kit delivers more accurate point-of-use results across a wide range of iron concentrations under challenging field conditions.

  19. Preappointment testing for BRAF/KIT mutation in advanced melanoma: a model in molecular data delivery for individualized medicine.

    PubMed

    Mounajjed, Taofic; Brown, Char L; Stern, Therese K; Bjorheim, Annette M; Bridgeman, Andrew J; Rumilla, Kandelaria M; McWilliams, Robert R; Flotte, Thomas J

    2014-11-01

    The emergence of individualized medicine is driven by developments in precision diagnostics, epitomized by molecular testing. Because treatment decisions are being made based on such molecular data, data management is gaining major importance. Among data management challenges, creating workflow solutions for timely delivery of molecular data has become pivotal. This study aims to design and implement a scalable process that permits preappointment BRAF/KIT mutation analysis in melanoma patients, allowing molecular results necessary for treatment plans to be available before the patient's appointment. Process implementation aims to provide a model for efficient molecular data delivery for individualized medicine. We examined the existing process of BRAF/KIT testing in melanoma patients visiting our institution for oncology consultation. We created 5 working groups, each designing a specific segment of an alternative process that would allow preappointment BRAF/KIT testing and delivery of results. Data were captured and analyzed to evaluate the success of the alternative process. For 1 year, 35 (59%) of 55 patients had prior BRAF/KIT testing. The remaining 20 patients went through the new process of preappointment testing; results were available at the time of appointment for 12 patients (overall preappointment results availability, 85.5%). The overall process averaged 13.4 ± 4.7 days. In conclusion, we describe the successful implementation of a scalable workflow solution that permits preappointment BRAF/KIT mutation analysis and result delivery in melanoma patients. This sets the stage for further applications of this model to other conditions, answering an increasing demand for robust delivery of molecular data for individualized medicine. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Experiment kits for processing biological samples inflight on SLS-2

    NASA Technical Reports Server (NTRS)

    Savage, P. D.; Hinds, W. E.; Jaquez, R.; Evans, J.; Dubrovin, L.

    1995-01-01

    This paper describes development of an innovative, modular approach to packaging the instruments used to obtain and preserve the inflight rodent tissue and blood samples associated with hematology experiments on the Spacelab Life Sciences-2 (SLS-2) mission. The design approach organized the multitude of instruments into twelve 5- x 6- x l-in. kits which were each used for a particular experiment. Each kit contained the syringes, vials, microscope slides, etc., necessary for processing and storing blood and tissue samples for one rat on a particular day. A total of 1245 components, packaged into 128 kits and stowed in 17 Zero(registered trademark) boxes, were required. Crewmembers found the design easy to use and laid out in a logical, simple configuration which minimized chances for error during the complex procedures in flight. This paper also summarizes inflight performance of the kits on SLS-2.

  1. Method modification of the Legipid® Legionella fast detection test kit.

    PubMed

    Albalat, Guillermo Rodríguez; Broch, Begoña Bedrina; Bono, Marisa Jiménez

    2014-01-01

    Legipid(®) Legionella Fast Detection is a test based on combined magnetic immunocapture and enzyme-immunoassay (CEIA) for the detection of Legionella in water. The test is based on the use of anti-Legionella antibodies immobilized on magnetic microspheres. Target microorganism is preconcentrated by filtration. Immunomagnetic analysis is applied on these preconcentrated water samples in a final test portion of 9 mL. The test kit was certified by the AOAC Research Institute as Performance Tested Method(SM) (PTM) No. 111101 in a PTM validation which certifies the performance claims of the test method in comparison to the ISO reference method 11731-1998 and the revision 11731-2004 "Water Quality: Detection and Enumeration of Legionella pneumophila" in potable water, industrial water, and waste water. The modification of this test kit has been approved. The modification includes increasing the target analyte from L. pneumophila to Legionella species and adding an optical reader to the test method. In this study, 71 strains of Legionella spp. other than L. pneumophila were tested to determine its reactivity with the kit based on CEIA. All the strains of Legionella spp. tested by the CEIA test were confirmed positive by reference standard method ISO 11731. This test (PTM 111101) has been modified to include a final optical reading. A methods comparison study was conducted to demonstrate the equivalence of this modification to the reference culture method. Two water matrixes were analyzed. Results show no statistically detectable difference between the test method and the reference culture method for the enumeration of Legionella spp. The relative level of detection was 93 CFU/volume examined (LOD50). For optical reading, the LOD was 40 CFU/volume examined and the LOQ was 60 CFU/volume examined. Results showed that the test Legipid Legionella Fast Detection is equivalent to the reference culture method for the enumeration of Legionella spp.

  2. Decontamination formulations for disinfection and sterilization

    DOEpatents

    Tucker, Mark D.; Engler, Daniel E.

    2007-09-18

    Aqueous decontamination formulations that neutralize biological pathogens for disinfection and sterilization applications. Examples of suitable applications include disinfection of food processing equipment, disinfection of areas containing livestock, mold remediation, sterilization of medical instruments and direct disinfection of food surfaces, such as beef carcasses. The formulations include at least one reactive compound, bleaching activator, inorganic base, and water. The formulations can be packaged as a two-part kit system, and can have a pH value in the range of 7-8.

  3. Project W.A.T.E.R.

    ERIC Educational Resources Information Center

    EnviroTeach, 1992

    1992-01-01

    Introduces networking projects for studying rivers and water quality. Describes two projects in South Africa (Project W.A.T.E.R and SWAP) associated with the international network, Global Rivers Environmental Education Network. Discusses water test kits and educational material developed through Project W.A.T.E.R. (Water Awareness through…

  4. Healthy Water Healthy People Field Monitoring Guide

    ERIC Educational Resources Information Center

    Project WET Foundation, 2003

    2003-01-01

    This 100-page manual serves as a technical reference for the "Healthy Water, Healthy People Water Quality Educators Guide" and the "Healthy Water Healthy People Testing Kits". Yielding in-depth information about ten water quality parameters, it answers questions about water quality testing using technical overviews, data interpretation guidelines,…

  5. Synthesis and characterization of Pd-poly(N-vinyl-2-pyrrolidone)/KIT-5 nanocomposite as a polymer-inorganic hybrid catalyst for the Suzuki-Miyaura cross-coupling reaction

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

    Kalbasi, Roozbeh Javad, E-mail: rkalbasi@iaush.ac.ir; Mosaddegh, Neda

    2011-11-15

    Composite poly(N-vinyl-2-pyrrolidone)/KIT-5 (PVP/KIT-5) was prepared by in situ polymerization method and used as a support for palladium nanoparticles obtained through the reduction of Pd(OAc){sub 2} by hydrazine hydrate. The physical and chemical properties of the catalyst were investigated by XRD, FT-IR, UV-vis, TG, BET, SEM, and TEM techniques. The catalytic performance of this novel heterogeneous catalyst was determined for the Suzuki-Miyaura cross-coupling reaction between aryl halides and phenylboronic acid in the presence of water at room temperature. The stability of the nanocomposite catalyst was excellent and could be reused 8 times without much loss of activity in the Suzuki-Miyaura cross-couplingmore » reaction. - Graphical Abstract: Pd-poly(N-vinyl-2-pyrrolidone)/KIT-5 was prepared as an organic-inorganic hybrid catalyst for the Suzuki-Miyaura reaction. The stability of the catalyst was excellent and could be reused 8 times in the Suzuki-Miyaura reaction. Highlights: > Pd-poly(N-vinyl-2-pyrrolidone)/KIT-5 was prepared as a novel nanocomposite. > Nanocomposite was prepared based on a cage-type mesoporous system. > Catalyst showed excellent activity for Suzuki-Miyaura reaction in water. > Stability of the catalyst was excellent and could be reused 8 times.« less

  6. Development of Experiment Kits for Processing Biological Samples In-Flight on SLS-2

    NASA Technical Reports Server (NTRS)

    Jaquez, R.; Savage, P. D.; Hinds, W. E.; Evans, J.; Dubrovin, L.

    1994-01-01

    The design of the hematology experiment kits for SLS-2 has resulted in a modular, flexible configuration which maximizes crew efficiency and minimizes error and confusion when dealing with over 1200 different components over the course of the mission. The kit layouts proved to be very easy to use and their packaging design provided for positive, secure containment of the many small components. The secondary Zero(Tm) box enclosure also provided an effective means for transport of the kits within the Spacelab and for grouping individual kits by flight day usage. The kits are readily adaptable to use on future flights by simply replacing the inner components as required and changing the labelling scheme to match new mission requirements.

  7. Improved kit formulation for preparation of (99m)Tc-HYNIC-TOC: results of preliminary clinical evaluation in imaging patients with neuroendocrine tumors.

    PubMed

    Korde, Aruna; Mallia, Madhava; Shinto, Ajit; Sarma, H D; Samuel, Grace; Banerjee, Sharmila

    2014-11-01

    (99m)Tc-HYNIC-TOC is a cost-effective and logistically viable agent for scintigraphy of neuroendocrine tumors overexpressing somatostatin receptors as compared with [(111)In-DTPA-D-Phe(1)] Octreotide (Octreoscan(®)). Several studies have been reported, wherein the efficacy of this agent is demonstrated. In the present article, the authors report the preparation of a single-vial HYNIC-TOC kit suitable for the preparation of 4-5 patient doses (15 mCi/patient) of (99m)Tc-HYNIC-TOC. The kits were tested for sterility and bacterial endotoxins to assure safety of the product. A significant modification in this kit is the inclusion of buffer in the kit itself, unlike in commercially available kits where the buffer solution has to be added during preparation. (99m)Tc-HYNIC-TOC was prepared by adding 20-80 mCi (740-2960 MBq) of freshly eluted Na(99m)TcO4 in 1-3 mL of sterile saline directly into the kit vial and heating the vial in a water bath at 100°C for 20 minutes. The labeling yield and radiochemical purity of (99m)Tc-HYNIC-TOC, prepared using the lyophilized cold kit, were consistently >90%. The kits were evaluated over a period of 9 months and found to be stable when stored at -20°C. Limited clinical studies performed with the (99m)Tc-HYNIC-TOC, formulated using the kit, showed adequate sensitivity and specificity for the detection of gasteroenteropancreatic neuroendocrine tumors.

  8. Library Construction from Subnanogram DNA for Pelagic Sea Water and Deep-Sea Sediments

    PubMed Central

    Hirai, Miho; Nishi, Shinro; Tsuda, Miwako; Sunamura, Michinari; Takaki, Yoshihiro; Nunoura, Takuro

    2017-01-01

    Shotgun metagenomics is a low biased technology for assessing environmental microbial diversity and function. However, the requirement for a sufficient amount of DNA and the contamination of inhibitors in environmental DNA leads to difficulties in constructing a shotgun metagenomic library. We herein examined metagenomic library construction from subnanogram amounts of input environmental DNA from subarctic surface water and deep-sea sediments using two library construction kits: the KAPA Hyper Prep Kit and Nextera XT DNA Library Preparation Kit, with several modifications. The influence of chemical contaminants associated with these environmental DNA samples on library construction was also investigated. Overall, shotgun metagenomic libraries were constructed from 1 pg to 1 ng of input DNA using both kits without harsh library microbial contamination. However, the libraries constructed from 1 pg of input DNA exhibited larger biases in GC contents, k-mers, or small subunit (SSU) rRNA gene compositions than those constructed from 10 pg to 1 ng DNA. The lower limit of input DNA for low biased library construction in this study was 10 pg. Moreover, we revealed that technology-dependent biases (physical fragmentation and linker ligation vs. tagmentation) were larger than those due to the amount of input DNA. PMID:29187708

  9. Identification of a polymorphism in the transmembrane domain of the protooncogene c-kit in healthy subjects.

    PubMed

    Nagata, H; Worobec, A S; Metcalfe, D D

    1996-01-01

    c-Kit is the receptor for stem cell factor (SCF) and is found on hematopoietic stem cells, mast cells, melanocytes, and germ cells. Aggregation of c-Kit by SCF regulates cell proliferation, differentiation, and survival. In the process of examining c-Kit, a polymorphism in the transmembrane domain of the protooncogene c-Kit was identified. This polymorphism consisted of an A-to-C transversion at nucleotide (nt) 1642, and was deduced to substitute leucine for methionine at codon 541. The frequency of the allele with 'C' at nt 1642 was 0.09 in 64 unrelated subjects. Analysis of a two-generation family with the polymorphism suggested that this polymorphism did not result in disease. This is the first report of a polymorphism in the transmembrane domain of c-Kit, and may be of value in understanding and following the function of c-Kit in normal subjects and in those with other abnormalities of c-Kit.

  10. The Integration of Nonprint Media. SPEC Kit 33.

    ERIC Educational Resources Information Center

    Association of Research Libraries, Washington, DC. Office of Management Studies.

    This kit on the integration of nonprint media into Association of Research Libraries (ARL) institutions contains a concise summary of a 1977 phone survey with ARL members, eight descriptions of programs and services, and five acquisition and processing policies. To collect data on practices regarding the acquisition, processing, and use of…

  11. Ultrastructural study of relationships between c-kit immunoreactive interstitial cells and other cellular elements in the human colon.

    PubMed

    Mazzia, C; Porcher, C; Julé, Y; Christen, M O; Henry, M

    2000-05-01

    C-kit immunocytochemistry was performed on ultrathin sections of human distal colon. Our attention was focused on relationships between c-kit immunoreactive interstitial cells (c-kit ICs) and muscular cells and nervous elements located in the external muscular layers of the colonic wall. C-kit ICs established membrane apposition with both nerve fibers and smooth muscle cells of, respectively, the longitudinal and circular muscle layers, the myenteric area, and the extremus submucosus plexus. C-kit ICs also surrounded the external submucosus plexus and established membrane appositions with nerve elements located inside the myenteric ganglia. These membrane appositions were observed either at the level of the c-kit IC bodies or at that of their cytoplasmic processes. In some cases, membrane appositions were observed concomitantly between the c-kit ICs, nerve fibers, and smooth muscle cells. In all the regions studied, the c-kit ICs were also found to be located in the close vicinity of blood vessels and to have established close contacts with non-immunoreactive fibroblast-like cells. The results of the present study shed essential light on the relationships of c-kit ICs with the neighboring muscle cells and nerve elements, and confirm that the intercalated c-kit ICs well fit with the so-called "interstitial cells of Cajal".

  12. SRC-like adaptor protein 2 (SLAP2) is a negative regulator of KIT-D816V-mediated oncogenic transformation.

    PubMed

    Rupar, Kaja; Moharram, Sausan A; Kazi, Julhash U; Rönnstrand, Lars

    2018-04-23

    KIT is a receptor tyrosine kinase (RTK) involved in several cellular processes such as regulation of proliferation, survival and differentiation of early hematopoietic cells, germ cells and melanocytes. Activation of KIT results in phosphorylation of tyrosine residues in the receptor, and recruitment of proteins that mediate downstream signaling and also modulate receptor signaling. Here we show that the SRC-like adaptor protein 2 (SLAP2) binds to wild-type KIT in a ligand-dependent manner and is furthermore found constitutively associated with the oncogenic mutant KIT-D816V. Peptide fishing analysis mapped pY568 and pY570 as potential SLAP2 association sites in KIT, which overlaps with the SRC binding sites in KIT. Expression of SLAP2 in cells expressing the transforming mutant KIT-D816V led to reduced cell viability and reduced colony formation. SLAP2 also partially blocked phosphorylation of several signal transduction molecules downstream of KIT such as AKT, ERK, p38 and STAT3. Finally, SLAP2 expression enhanced ubiquitination of KIT and its subsequent degradation. Taken together, our data demonstrate that SLAP2 negatively modulates KIT-D816V-mediated transformation by enhancing degradation of the receptor.

  13. Transfer of adapted water supply technologies through a demonstration and teaching facility

    NASA Astrophysics Data System (ADS)

    Nestmann, F.; Oberle, P.; Ikhwan, M.; Stoffel, D.; Blaß, H. J.; Töws, D.; Schmidt, S.

    2016-09-01

    Water scarcity can be defined as a lack of sufficient water resources or as the limited or even missing access to a safe water supply. Latter can be classified as `economic water scarcity' which among others can commonly be met in tropical and subtropical karst regions of emerging and developing countries. Karst aquifers, mostly consisting of limestone and carbonate rock, show high infiltration rates which leads to a lack of above ground storage possibilities. Thus, the water will drain rapidly into the underground and evolve vast river networks. Considering the lack of appropriate infrastructure and limited human capacities in the affected areas, these underground water resources cannot be exploited adequately. Against this, background innovative and adapted technologies are required to utilize hard-to-access water resources in a sustainable way. In this context, the German-Indonesian joint R&D project "Integrated Water Resources Management (IWRM) Indonesia" dealt with the development of highly adaptable water technologies and management strategies. Under the aegis of the Karlsruhe Institute of Technology (KIT) and funded by the German Ministry of Education and Research (BMBF), these innovative technical concepts were exemplarily implemented to remedy this deficiency in the model region Gunung Sewu, a karst area situated on the southern coast of Java Island, Indonesia. The experiences gained through the interdisciplinary joint R&D activities clearly showed that even in the case of availability of appropriate technologies, a comprising transfer of knowhow and the buildup of capabilities (Capacity Development) is inevitable to sustainably implement and disseminate new methods. In this context, an adapted water supply facility was developed by KIT which hereafter shall serve for demonstration, teaching, and research purposes. The plant's functionality, its teaching and research concept, as well as the design process, which was accomplished in collaboration with the University Gadjah Mada (UGM), Yogyakarta, Indonesia, is the content of this present paper.

  14. The FOT tool kit concept

    NASA Technical Reports Server (NTRS)

    Fatig, Michael

    1993-01-01

    Flight operations and the preparation for it has become increasingly complex as mission complexities increase. Further, the mission model dictates that a significant increase in flight operations activities is upon us. Finally, there is a need for process improvement and economy in the operations arena. It is therefore time that we recognize flight operations as a complex process requiring a defined, structured, and life cycle approach vitally linked to space segment, ground segment, and science operations processes. With this recognition, an FOT Tool Kit consisting of six major components designed to provide tools to guide flight operations activities throughout the mission life cycle was developed. The major components of the FOT Tool Kit and the concepts behind the flight operations life cycle process as developed at NASA's GSFC for GSFC-based missions are addressed. The Tool Kit is therefore intended to increase productivity, quality, cost, and schedule performance of the flight operations tasks through the use of documented, structured methodologies; knowledge of past lessons learned and upcoming new technology; and through reuse and sharing of key products and special application programs made possible through the development of standardized key products and special program directories.

  15. Ease fabrication of PCR modular chip for portable DNA detection kit

    NASA Astrophysics Data System (ADS)

    Whulanza, Yudan; Aditya, Rifky; Arvialido, Reyhan; Utomo, Muhammad S.; Bachtiar, Boy M.

    2017-02-01

    Engineering a lab-on-a-chip (LoC) to perform the DNA polymerase chain reaction (PCR) for malaria detection is the ultimate goal of this study. This paper investigates the ability to fabricate an LoC kit using conventional method to achieve the lowest production cost by using existing fabrication process. It has been known that majority of LoC was made of polydimethylsiloxane (PDMS) which in this study was realized through a contact mold process. CNC milling process was utilized to create channel features in the range of 150-250 µm on the mold. Characterization on the milling process was done to understand the shrinkage/contraction between mold to product, roughness and also angle of contact of PDMS surface. Ultimately, this paper also includes analysis on flow measurement and heat distribution of an assembled LoC PCR kit. The results show that the achieved dimension of microchannel is 227 µm wide with a roughness of 0.01 µm. The flow measurement indicates a deviation with simulation in the range of 10%. A heat distribution through the kit is achieved following the three temperature zones as desired.

  16. FE-2 Stott analyzes Water Samples using the CQQMK in the US Lab

    NASA Image and Video Library

    2009-10-20

    ISS021-E-010311 (20 Oct. 2009) --- NASA astronaut Nicole Stott, Expedition 21 flight engineer, conducts a water quality analysis using the Colorimetric Water Quality Monitoring Kit (CWQMK) in the Destiny laboratory of the International Space Station.

  17. Reviews

    NASA Astrophysics Data System (ADS)

    2005-11-01

    WE RECOMMEND Joule and Watt Meter This versatile tool is aimed at low-voltage DC devices. Wireless datalogger Bluetooth enables data to be downloaded to your computer. WORTH A LOOK Amusement Park Physics This handbook contains teaching materials to excite students. Copper in the Curriculum Two free CDs cover aspects of Key Stages 3, 4 and 5. Launchpad A kit for making 25 devices that fire ping-pong balls. Solar water-heating kit This kit provides a good introduction to green energy. Solar voltaic cell The unit is ideal for Key Stages 1, 2 and 3. Wind turbine kit An impressive turbine that has many educational uses. HANDLE WITH CARE Hurdles and Strategies in the Teaching of Algebra A collection of tips that is really just for maths teachers. WEB WATCH A wealth of resources on the Internet support physics activities for students at amusement parks.

  18. Home bowel cancer tests and informed choice--is current information sufficient?

    PubMed

    Howard, K; Salkeld, G

    2003-10-01

    To evaluate the type of information that is available to purchasers of home-based bowel cancer test kits. Manufacturers, pharmacies and independent testing programs were contacted to obtain faecal occult blood test (FOBT) kits. State cancer organisations were contacted for information on bowel cancer screening. Information on bowel cancer, the FOBT kit, the testing process and potential benefits and harms of the screening process were assessed using guidelines provided by the UK General Medical Council (GMC). FOBT kits and cancer organisation information provided adequate information on the purpose of screening, the screening process itself and potential benefits, but provided no information concerning uncertainties of screening or potential harms. On the basis of both the UK GMC criteria and patient desires for information, the information available at present falls short of being considered adequate for an informed decision to purchase a home-based FOBT. We must ensure adequate and balanced information is available to redress the present information asymmetry to facilitate informed participation in a potentially valuable public health initiative.

  19. Factors for C-Kit Expression in Cardiac Outgrowth Cells and Human Heart Tissue.

    PubMed

    Matsushita, Satoshi; Minematsu, Kazuo; Yamamoto, Taira; Inaba, Hirotaka; Kuwaki, Kenji; Shimada, Akie; Yokoyama, Yasutaka; Amano, Atsushi

    2017-12-12

    We determined the factors associated with the expression of c-kit in the heart and the proliferation of c-kit-positive (c-kit pos ) cardiac stem cells among the outgrowth cells cultured from human cardiac explants.Samples of the right atrium (RA), left atrium (LA), and left ventricle obtained from patients during open-heart surgery were processed for cell culture of outgrowth cells and tissue analysis. The total number of growing cells and the population of c-kit pos cells were measured and compared with c-kit expression in native tissues and characteristics of the patients according to the region of the heart.We analyzed 452 samples from 334 patients. Atrial fibrillation (AF) in the patients reduced the number of outgrowth cells from the RA and LA, and aging was a co-factor for the LA. The c-kit pos population from the RA was associated with serum brain natriuretic peptide (BNP). C-kit expression in native tissue was also associated with BNP expression. However, we observed no relationship in expression between outgrowth cells and native tissue. In addition, the RA tissue provided the highest number of c-kit pos cells, and the left ventricle provided the lowest.C-kit was weakly expressed in response to damage. In addition, no correlation between outgrowth cells and native tissue was found for c-kit expression.

  20. Pilot Overmyer on middeck

    NASA Technical Reports Server (NTRS)

    1982-01-01

    On middeck port side, Pilot Overmyer, looks down at freefloating object. On his left, attached to port side wall, are a dessert package, a prepackaged meal, control panel ML86B, and water dispenser kit with water gun.

  1. 75 FR 64248 - Approval for Processing Authority Foreign-Trade Zones 73 and 74; The Belt's Corporation (Kitting...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-19

    ... DEPARTMENT OF COMMERCE Foreign-Trade Zones Board [Order No. 1713] Approval for Processing Authority Foreign-Trade Zones 73 and 74; The Belt's Corporation (Kitting of Liquor Gift Sets), Elkridge and Baltimore, MD Pursuant to its authority under the Foreign-Trade Zones Act of June 18, 1934, as amended (19 U...

  2. Safe Water and Hygiene Integration with Human Immunodeficiency Virus and Antenatal Services: Leveraging Opportunities for Public Health Interventions and Improved Service Uptake.

    PubMed

    Routh, Janell A; Loharikar, Anagha; Chemey, Elly; Msoma, Aulive; Ntambo, Maureen; Mvula, Richard; Ayers, Tracy; Gunda, Andrews; Russo, Elizabeth T; Barr, Beth Tippett; Wood, Siri; Quick, Robert

    2018-05-01

    Integrating public health interventions with antenatal clinic (ANC) visits may motivate women to attend ANC, thereby improving maternal and neonatal health, particularly for human immunodeficiency virus (HIV)-infected persons. In 2009, in an integrated ANC/Preventing Mother-to-Child Transmission program, we provided free hygiene kits (safe storage containers, WaterGuard water treatment solution, soap, and oral rehydration salts) to women at their first ANC visit and refills at subsequent visits. To increase fathers' participation, we required partners' presence for women to receive hygiene kits. We surveyed pregnant women at baseline and at 12-month follow-up to assess ANC service utilization, HIV counseling and testing (HCT), test drinking water for residual chlorine, and observe handwashing. We conducted in-depth interviews with pregnant women, partners, and health workers. We enrolled 106 participants; 97 (92%) were found at follow-up. During the program, 99% of pregnant women and their partners received HCT, and 99% mutually disclosed. Fifty-six percent of respondents had ≥ 4 ANC visits and 90% delivered at health facilities. From baseline to follow-up, the percentage of women who knew how to use WaterGuard (23% versus 80%, P < 0.0001), had residual chlorine in stored water (0% versus 73%, P < 0.0001), had confirmed WaterGuard use (0% versus 70%, P < 0.0003), and demonstrated proper handwashing technique (21% versus 64% P < 0.0001) increased. Program participants showed significant improvements in water treatment and hygiene, and high use of ANC services and HCT. This evaluation suggests that integration of hygiene kits, refills, and HIV testing during ANC is feasible and may help improve household hygiene and increase use of health services.

  3. Developing Save Your Food Kit (Sayofu Kit) to Support Inquiry, Improve Student Learning Outcomes at SMP Plus Hidayatul Mubtadiin and Public Awareness on Food Additives

    NASA Astrophysics Data System (ADS)

    Astutik, J.

    2017-02-01

    Food additives are materials that can not be separated from the lives of students and the community. Based on the preliminary questionnaire, it indicates the lack of kit supporting material additives in some schools and communities. The research objectives of this development are (1) to develop Kit experiment (SAYOFU KIT) and supplementary books to improve student learning outcomes in the classroom and public awareness on food additives (2) to describe the feasibility and potential effectiveness of SAYOFU KIT developed (3) to analyze the practice of SAYOFU KIT and benefits for students and the community. This development study uses 4-D models Thiagarajan, et al (1974). Through some stages, they are: defining, designing, developing and disseminating which involes the students and community. The developed SAYOFU KIT includes additives sample kit, borax test kit, curcumin test kit, formaldehyde test kit, modification heater to the identification of dyes and dye test paper. The study is conducted at SMP Plus Hidayatul Mubtadiin, and TKIT Al Uswah. The products are validated by experts and education practitioners. Qualitative data processing uses descriptive method, whereas quantitative data by using the N-gain. The average yield of expert validation of SAYOFU KIT with supplementary books 76.50% teacher’s book and 76.30% student’s book are eligible. The average yield of 96.81% validation of educational practitioners criteria, piloting a small group of 83.15%, and 82.89% field trials are very decent. The average yield on the student questionnaire responses SAYOFU kit and supplementary book is 87.6% with the criteria very well worth it. N-Gain 0:56 cognitive achievement with the criteria enough. The results of the public poll showed 95% feel the benefits SAYOFU kits for testing food. Based from description indicates that SAYOFU Kit developed feasible, practical, useful to support inquiry learning and improve student learning outcomes as well as public awareness of food additives.

  4. Incorporating ethical principles into clinical research protocols: a tool for protocol writers and ethics committees.

    PubMed

    Li, Rebecca H; Wacholtz, Mary C; Barnes, Mark; Boggs, Liam; Callery-D'Amico, Susan; Davis, Amy; Digilova, Alla; Forster, David; Heffernan, Kate; Luthin, Maeve; Lynch, Holly Fernandez; McNair, Lindsay; Miller, Jennifer E; Murphy, Jacquelyn; Van Campen, Luann; Wilenzick, Mark; Wolf, Delia; Woolston, Cris; Aldinger, Carmen; Bierer, Barbara E

    2016-04-01

    A novel Protocol Ethics Tool Kit ('Ethics Tool Kit') has been developed by a multi-stakeholder group of the Multi-Regional Clinical Trials Center of Brigham and Women's Hospital and Harvard. The purpose of the Ethics Tool Kit is to facilitate effective recognition, consideration and deliberation of critical ethical issues in clinical trial protocols. The Ethics Tool Kit may be used by investigators and sponsors to develop a dedicated Ethics Section within a protocol to improve the consistency and transparency between clinical trial protocols and research ethics committee reviews. It may also streamline ethics review and may facilitate and expedite the review process by anticipating the concerns of ethics committee reviewers. Specific attention was given to issues arising in multinational settings. With the use of this Tool Kit, researchers have the opportunity to address critical research ethics issues proactively, potentially speeding the time and easing the process to final protocol approval. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

  5. LEM-CF Premixed Tool Kit

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

    2015-01-19

    The purpose of LEM-CF Premixed Tool Kit is to process premixed flame simulation data from the LEM-CF solver (https://fileshare.craft-tech.com/clusters/view/lem-cf) into a large-eddy simulation (LES) subgrid model database. These databases may be used with a user-defined-function (UDF) that is included in the Tool Kit. The subgrid model UDF may be used with the ANSYS FLUENT flow solver or other commercial flow solvers.

  6. Colorimetric Solid Phase Extraction for the Measurement of Total I (Iodine, Iodide, and Triiodide) in Spacecraft Drinking Water

    NASA Technical Reports Server (NTRS)

    Lipert, Robert J.; Porter, Marc D.; Siperko, Lorraine M.; Gazda, Daniel B.; Rutz, Jeff A.; Schultz, John R.; Carrizales, Stephanie M.; McCoy, J. Torin

    2009-01-01

    An experimental drinking water monitoring kit for the measurement of iodine and silver(I) was recently delivered to the International Space Station (ISS). The kit is based on Colorimetric Solid Phase Extraction (CSPE) technology, which measures the change in diffuse reflectance of indicator disks following exposure to a water sample. To satisfy additional spacecraft water monitoring requirements, CSPE has now been extended to encompass the measurement of total I (iodine, iodide, and triiodide) through the introduction of an oxidizing agent, which converts iodide and triiodide to iodine, for measurement using the same indicator disks currently being tested on ISS. These disks detect iodine, but are insensitive to iodide and triiodide. We report here the operational considerations, design, and ground-based performance of the CSPE method for total I. The results demonstrate that CSPE technology is poised to meet NASA's total I monitoring requirements.

  7. Preliminary characterization of digestive enzymes in freshwater mussels

    USGS Publications Warehouse

    Sauey, Blake W.; Amberg, Jon J.; Cooper, Scott T.; Grunwald, Sandra K.; Newton, Teresa J.; Haro, Roger J.

    2015-01-01

    Resource managers lack an effective chemical tool to control the invasive zebra mussel Dreissena polymorpha. Zebra mussels clog water intakes for hydroelectric companies, harm unionid mussel species, and are believed to be a reservoir of avian botulism. Little is known about the digestive physiology of zebra mussels and unionid mussels. The enzymatic profile of the digestive glands of zebra mussels and native threeridge (Amblema plicata) and plain pocketbook mussels (Lampsilis cardium) are characterized using a commercial enzyme kit, api ZYM, and validated the kit with reagent-grade enzymes. A linear correlation was shown for only one of nineteen enzymes, tested between the api ZYM kit and a specific enzyme kit. Thus, the api ZYM kit should only be used to make general comparisons of enzyme presence and to observe trends in enzyme activities. Enzymatic trends were seen in the unionid mussel species, but not in zebra mussels sampled 32 days apart from the same location. Enzymatic classes, based on substrate, showed different trends, with proteolytic and phospholytic enzymes having the most change in relative enzyme activity.

  8. The AskA Starter Kit: How To Build and Maintain Digital Reference Services.

    ERIC Educational Resources Information Center

    Lankes, R. David; Kasowitz, Abby S.

    This Starter Kit is designed to help organizations and individuals who wish to offer human-mediated information services via the Internet to users in the K-12 community. A six-step process is proposed for organizations to follow in creating an "AskA" service. This process addresses all aspects involved in building and maintaining an AskA…

  9. Sowing the Seeds of Environmental Education.

    ERIC Educational Resources Information Center

    Moloi, Dudley

    1994-01-01

    This article examines Umgeni Water's External Education Unit (EEU) to explore the potential of one approach to environmental education. The EEU uses a simple water quality monitoring kit to educate communities on the state of their rivers and catchment areas and warn them of potential dangers facing users of untreated water. (LZ)

  10. The stem cell factor (SCF)/c-KIT system in carcinogenesis of reproductive tissues: What does the hormonal regulation tell us?

    PubMed

    Figueira, Marília I; Cardoso, Henrique J; Correia, Sara; Maia, Cláudio J; Socorro, Sílvia

    2017-10-01

    The tyrosine kinase receptor c-KIT and its ligand, the stem cell factor (SCF) are expressed in several tissues of male and female reproductive tract, playing an important role in the regulation of basic biological processes. The activation of c-KIT by SCF controls, cell survival and death, cell differentiation and migration. Also, the SCF/c-KIT system has been implicated in carcinogenesis of reproductive tissues due to its altered expression pattern or overactivation in consequence of gain-of-functions mutations. Over the years, it has also been shown that hormones, the primary regulators of reproductive function and causative agents in the case of hormone-dependent cancers, are also able to control the SCF/c-KIT tissue levels. Therefore, it is liable to suppose that disturbed SCF/c-KIT expression driven by (de)regulated hormone actions can be a relevant step towards carcinogenesis. The present review describes the SCF and c-KIT expression in cancers of reproductive tissues, discussing the implications of the hormonal regulation of the SCF/c-KIT system in cancer development. Understanding the relationship between hormonal imbalance and the SCF/c-KIT expression and activity would be relevant in the context of novel therapeutic approaches in reproductive cancers. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Optimising the quantification of cytokines present at low concentrations in small human mucosal tissue samples using Luminex assays☆

    PubMed Central

    Staples, Emily; Ingram, Richard James Michael; Atherton, John Christopher; Robinson, Karen

    2013-01-01

    Sensitive measurement of multiple cytokine profiles from small mucosal tissue biopsies, for example human gastric biopsies obtained through an endoscope, is technically challenging. Multiplex methods such as Luminex assays offer an attractive solution but standard protocols are not available for tissue samples. We assessed the utility of three commercial Luminex kits (VersaMAP, Bio-Plex and MILLIPLEX) to measure interleukin-17A (IL-17) and interferon-gamma (IFNγ) concentrations in human gastric biopsies and we optimised preparation of mucosal samples for this application. First, we assessed the technical performance, limits of sensitivity and linear dynamic ranges for each kit. Next we spiked human gastric biopsies with recombinant IL-17 and IFNγ at a range of concentrations (1.5 to 1000 pg/mL) and assessed kit accuracy for spiked cytokine recovery and intra-assay precision. We also evaluated the impact of different tissue processing methods and extraction buffers on our results. Finally we assessed recovery of endogenous cytokines in unspiked samples. In terms of sensitivity, all of the kits performed well within the manufacturers' recommended standard curve ranges but the MILLIPLEX kit provided most consistent sensitivity for low cytokine concentrations. In the spiking experiments, the MILLIPLEX kit performed most consistently over the widest range of concentrations. For tissue processing, manual disruption provided significantly improved cytokine recovery over automated methods. Our selected kit and optimised protocol were further validated by measurement of relative cytokine levels in inflamed and uninflamed gastric mucosa using Luminex and real-time polymerase chain reaction. In summary, with proper optimisation Luminex kits (and for IL-17 and IFNγ the MILLIPLEX kit in particular) can be used for the sensitive detection of cytokines in mucosal biopsies. Our results should help other researchers seeking to quantify multiple low concentration cytokines in small tissue samples. PMID:23644159

  12. Using Art to Enhance the Learning of Math and Science: Developing an Educational Art-Science Kit about Fractal Patterns in Nature

    NASA Astrophysics Data System (ADS)

    Rao, Deepa

    This study documents the development of an educational art-science kit about natural fractals, whose aim is to unite artistic and scientific inquiry in the informal learning of science and math. Throughout this research, I argue that having an arts-integrated approach can enhance the learner of science and math concepts. A guiding metaphor in this thesis is the Enlightenment-era cabinet of curiosities that represents a time when art and science were unified in the process of inquiry about the natural world. Over time, increased specialization in the practice of arts and science led to a growing divergence between the disciplines in the educational system. Recently, initiatives like STEAM are underway at the national level to integrate "Arts and Design" into the Science, Technology, Engineering, and Math (STEM) formal education agenda. Learning artifacts like science kits present an opportunity to unite artistic and scientific inquiry in informal settings. Although science kits have been introduced to promote informal learning, presently, many science kits have a gap in their design, whereby the activities consist of recipe-like instructions that do not encourage further inquiry-based learning. In the spirit of the cabinet of curiosities, this study seeks to unify visual arts and science in the process of inquiry. Drawing from educational theories of Dewey, Piaget, and Papert, I developed a novel, prototype "art-science kit" that promotes experiential, hands-on, and active learning, and encourages inquiry, exploration, creativity, and reflection through a series of art-based activities to help users learn science and math concepts. In this study, I provide an overview of the design and development process of the arts-based educational activities. Furthermore, I present the results of a pilot usability study (n=10) conducted to receive user feedback on the designed materials for use in improving future iterations of the art-science fractal kit. The fractal kit booklet that I designed can be found in the supplemental materials to this thesis.

  13. The stem cell growth factor receptor KIT is not expressed on interstitial cells in bladder.

    PubMed

    Gevaert, Thomas; Ridder, Dirk De; Vanstreels, Els; Daelemans, Dirk; Everaerts, Wouter; Aa, Frank Van Der; Pintelon, Isabel; Timmermans, Jean-Pierre; Roskams, Tania; Steiner, Clara; Neuhaus, Jochen

    2017-06-01

    The mast/stem cell growth factor receptor KIT has long been assumed to be a specific marker for interstitial cells of Cajal (ICC) in the bladder, with possible druggable perspectives. However, several authors have challenged the presence of KIT + ICC in recent years. The aim of this study was therefore to attempt to clarify the conflicting reports on KIT expression in the bladder of human beings, rat, mouse and guinea pig and to elucidate the possible role of antibody-related issues and interspecies differences in this matter. Fresh samples were obtained from human, rat, mouse and guinea pig cystectomies and processed for single/double immunohistochemistry/immunofluorescence. Specific antibodies against KIT, mast cell tryptase (MCT), anoctamin-1 (ANO1) and vimentin were used to characterize the cell types expressing KIT. Gut (jejunum) tissue was used as an external antibody control. Our results revealed KIT expression on mast cells but not on ICC in human, rat, mouse and guinea pig bladder. Parallel immunohistochemistry showed KIT expression on ICC in human, rat, mouse and guinea pig gut, which confirmed the selectivity of the KIT antibody clones. In conclusion, we have shown that KIT + cells in human, rat, mouse and guinea pig bladder are mast cells and not ICC. The present report is important as it opposes the idea that KIT + ICC are present in bladder. In this perspective, functional concepts of KIT + ICC being involved in sensory and/or motor aspects of bladder physiology should be revised. © 2016 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

  14. Molecular Alterations of KIT Oncogene in Gliomas

    PubMed Central

    Gomes, Ana L.; Reis-Filho, Jorge S.; Lopes, José M.; Martinho, Olga; Lambros, Maryou B. K.; Martins, Albino; Schmitt, Fernando; Pardal, Fernando; Reis, Rui M.

    2007-01-01

    Gliomas are the most common and devastating primary brain tumours. Despite therapeutic advances, the majority of gliomas do not respond either to chemo or radiotherapy. KIT, a class III receptor tyrosine kinase (RTK), is frequently involved in tumourigenic processes. Currently, KIT constitutes an attractive therapeutic target. In the present study we assessed the frequency of KIT overexpression in gliomas and investigated the genetic mechanisms underlying KIT overexpression. KIT (CD117) immunohistochemistry was performed in a series of 179 gliomas of various grades. KIT activating gene mutations (exons 9, 11, 13 and 17) and gene amplification analysis, as defined by chromogenic in situ hybridization (CISH) and quantitative real-time PCR (qRT-PCR) were performed in CD117 positive cases. Tumour cell immunopositivity was detected in 15.6% (28/179) of cases, namely in 25% (1/4) of pilocytic astrocytomas, 25% (5/20) of diffuse astrocytomas, 20% (1/5) of anaplastic astrocytomas, 19.5% (15/77) of glioblastomas and one third (3/9) of anaplastic oligoastrocytomas. Only 5.7% (2/35) of anaplastic oligodendrogliomas showed CD117 immunoreactivity. No association was found between tumour CD117 overexpression and patient survival. In addition, we also observed CD117 overexpression in endothelial cells, which varied from 0–22.2% of cases, being more frequent in high-grade lesions. No KIT activating mutations were identified. Interestingly, CISH and/or qRT-PCR analysis revealed the presence of KIT gene amplification in 6 glioblastomas and 2 anaplastic oligoastrocytomas, corresponding to 33% (8/24) of CD117 positive cases. In conclusion, our results demonstrate that KIT gene amplification rather than gene mutation is a common genetic mechanism underlying KIT expression in subset of malignant gliomas. Further studies are warranted to determine whether glioma patients exhibiting KIT overexpression and KIT gene amplification may benefit from therapy with anti-KIT RTK inhibitors. PMID:17726262

  15. Quantification of Parvovirus B19 DNA Using COBAS AmpliPrep Automated Sample Preparation and LightCycler Real-Time PCR

    PubMed Central

    Schorling, Stefan; Schalasta, Gunnar; Enders, Gisela; Zauke, Michael

    2004-01-01

    The COBAS AmpliPrep instrument (Roche Diagnostics GmbH, D-68305 Mannheim, Germany) automates the entire sample preparation process of nucleic acid isolation from serum or plasma for polymerase chain reaction analysis. We report the analytical performance of the LightCycler Parvovirus B19 Quantification Kit (Roche Diagnostics) using nucleic acids isolated with the COBAS AmpliPrep instrument. Nucleic acids were extracted using the Total Nucleic Acid Isolation Kit (Roche Diagnostics) and amplified with the LightCycler Parvovirus B19 Quantification Kit. The kit combination processes 72 samples per 8-hour shift. The lower detection limit is 234 IU/ml at a 95% hit-rate, linear range approximately 104-1010 IU/ml, and overall precision 16 to 40%. Relative sensitivity and specificity in routine samples from pregnant women are 100% and 93%, respectively. Identification of a persistent parvovirus B19-infected individual by the polymerase chain reaction among 51 anti-parvovirus B19 IgM-negative samples underlines the importance of additional nucleic acid testing in pregnancy and its superiority to serology in identifying the risk of parvovirus B19 transmission via blood or blood products. Combination of the Total Nucleic Acid Isolation Kit on the COBAS AmpliPrep instrument with the LightCycler Parvovirus B19 Quantification Kit provides a reliable and time-saving tool for sensitive and accurate detection of parvovirus B19 DNA. PMID:14736825

  16. The Evaluation of Water Conservation for Municipal and Industrial Water Supply: Illustrative Examples. Water Conservation and Supply Information Transfer and Analysis Program. Revision.

    DTIC Science & Technology

    1981-02-01

    108 NED Project Plan........................................ 108 EQ Project Plan ......................................... 115 Performance of Water...Conservation Proposal: ED Objective ............. 115 50. Atlanta ED Water Conservation Proposal: EQasObjective............. 115 52. Atlanta EQ Water... differential inflation of energy prices, this would provide an annualized energy-related advantageous effect of $655,000/year. Measure A2--Moderate Kit

  17. Teacher's Guide to Water Play: The MATCH Box Project; Prototype Edition.

    ERIC Educational Resources Information Center

    Olson, Nancy; And Others

    The multimedia kit on water was designed for use by nursery school and first grade children. Water is an intriguing part of a child's world; for this reason, water was chosen as the medium for this set of instructional materials. The child's natural way of exploring his environment is through play; he is really learning ways to discover and…

  18. Bowersox works with the WMK in Destiny during Expedition Six

    NASA Image and Video Library

    2003-02-17

    ISS006-E-27226 (17 February 2003) --- Astronaut Kenneth D. Bowersox, Expedition Six mission commander, uses the water microbiology kit (WMK) to collect water samples for in-flight chemistry/microbiology analysis in the Destiny laboratory on the International Space Station (ISS).

  19. Evaluation of two platelet-rich plasma processing methods and two platelet-activation techniques for use in llamas and alpacas.

    PubMed

    Semevolos, Stacy A; Youngblood, Cori D; Grissom, Stephanie K; Gorman, M Elena; Larson, Maureen K

    2016-11-01

    OBJECTIVE To evaluate 2 processing methods (commercial kit vs conical tube centrifugation) for preparing platelet rich plasma (PRP) for use in llamas and alpacas. SAMPLES Blood samples (30 mL each) aseptically collected from 6 healthy llamas and 6 healthy alpacas. PROCEDURES PRP was prepared from blood samples by use of a commercial kit and by double-step conical tube centrifugation. A CBC was performed for blood and PRP samples. Platelets in PRP samples were activated by means of a freeze-thaw method with or without 23mM CaCl 2 , and concentrations of platelet-derived growth factor-BB and transforming growth factor-β 1 were measured. Values were compared between processing methods and camelid species. RESULTS Blood CBC values for llamas and alpacas were similar. The commercial kit yielded a significantly greater degree of platelet enrichment (mean increase, 8.5 fold vs 2.8 fold) and WBC enrichment (mean increase, 3.7 fold vs 1.9 fold) than did conical tube centrifugation. Llamas had a significantly greater degree of platelet enrichment than alpacas by either processing method. No difference in WBC enrichment was identified between species. Concentrations of both growth factors were significantly greater in PRP samples obtained by use of the commercial kit versus those obtained by conical tube centrifugation. CONCLUSIONS AND CLINICAL RELEVANCE For blood samples from camelids, the commercial kit yielded a PRP product with a higher platelet and WBC concentration than achieved by conical tube centrifugation. Optimal PRP platelet and WBC concentrations for various applications need to be determined for llamas and alpacas.

  20. Effect of blanching treatments on antioxidant activity of frozen green capsicum (Capsicum annuum L. var bell pepper) using radical scavenging activity (DPPH) assay

    NASA Astrophysics Data System (ADS)

    Azizzuddin, Norafida; Abdullah, Aminah

    2016-11-01

    Blanching treatments are needed to deactivate enzymes in frozen vegetables. Antioxidant activity using DPPH radical scavenging activity assay were evaluated in steaming, boiling water, and microwave blanching at different temperature, time and microwave power level on frozen green capsicum. Green capsicum was chosen for frozen treatment compared to other capsicum with different maturity index because of the firm texture. The objective of this study was to compare the antioxidant activity of frozen green capsicum between conventional and Oxi Count Kit® assay for DPPH radical scavenging activity. Results showed frozen green capsicum blanched using microwave at high level/90 seconds (sample J) contained higher level of DPPH in both conventional method and Oxi Count Kit® compared to other treatments. However, there were no significant differences between sample J and fresh sample (sample A). Overall, the sequences from highest to lowest in blanching treatments for both DPPH conventional method, and DPPH Oxi Count Kit® were J (microwave high level/90 seconds) > A (Fresh) > H (Microwave Medium Level/120 seconds) > D (Boiling Water 80°C/150 seconds) > K (Microwave High Level/120 seconds) > I (Microwave Medium Level/150 seconds) > F (Microwave Low Level/150 seconds)> B (Steam 100°C/150 seconds) > E (Boiling Water 100°C /120 seconds) > G (Microwave Low Level /180 seconds)> C (Steam 100°C/180 seconds). Almost all frozen green capsicum samples showed no significant differences for comparison between test using DPPH conventional method and Oxi Count Kit®. Frozen storage for 0, and 3rd months showed no significant differences which indicate no changes on antioxidant activity during frozen storage at -18°C.

  1. A framework for measurement and harmonization of pediatric multiple sclerosis etiologic research studies: The Pediatric MS Tool-Kit.

    PubMed

    Magalhaes, Sandra; Banwell, Brenda; Bar-Or, Amit; Fortier, Isabel; Hanwell, Heather E; Lim, Ming; Matt, Georg E; Neuteboom, Rinze F; O'Riordan, David L; Schneider, Paul K; Pugliatti, Maura; Shatenstein, Bryna; Tansey, Catherine M; Wassmer, Evangeline; Wolfson, Christina

    2018-06-01

    While studying the etiology of multiple sclerosis (MS) in children has several methodological advantages over studying etiology in adults, studies are limited by small sample sizes. Using a rigorous methodological process, we developed the Pediatric MS Tool-Kit, a measurement framework that includes a minimal set of core variables to assess etiological risk factors. We solicited input from the International Pediatric MS Study Group to select three risk factors: environmental tobacco smoke (ETS) exposure, sun exposure, and vitamin D intake. To develop the Tool-Kit, we used a Delphi study involving a working group of epidemiologists, neurologists, and content experts from North America and Europe. The Tool-Kit includes six core variables to measure ETS, six to measure sun exposure, and six to measure vitamin D intake. The Tool-Kit can be accessed online ( www.maelstrom-research.org/mica/network/tool-kit ). The goals of the Tool-Kit are to enhance exposure measurement in newly designed pediatric MS studies and comparability of results across studies, and in the longer term to facilitate harmonization of studies, a methodological approach that can be used to circumvent issues of small sample sizes. We believe the Tool-Kit will prove to be a valuable resource to guide pediatric MS researchers in developing study-specific questionnaire.

  2. Comparison of commercial DNA extraction kits for isolation and purification of bacterial and eukaryotic DNA from PAH-contaminated soils.

    PubMed

    Mahmoudi, Nagissa; Slater, Greg F; Fulthorpe, Roberta R

    2011-08-01

    Molecular characterization of the microbial populations of soils and sediments contaminated with polycyclic aromatic hydrocarbons (PAHs) is often a first step in assessing intrinsic biodegradation potential. However, soils are problematic for molecular analysis owing to the presence of organic matter, such as humic acids. Furthermore, the presence of contaminants, such as PAHs, can cause further challenges to DNA extraction, quantification, and amplification. The goal of our study was to compare the effectiveness of four commercial soil DNA extraction kits (UltraClean Soil DNA Isolation kit, PowerSoil DNA Isolation kit, PowerMax Soil DNA Isolation kit, and FastDNA SPIN kit) to extract pure, high-quality bacterial and eukaryotic DNA from PAH-contaminated soils. Six different contaminated soils were used to determine if there were any biases among the kits due to soil properties or level of contamination. Extracted DNA was used as a template for bacterial 16S rDNA and eukaryotic 18S rDNA amplifications, and PCR products were subsequently analyzed using denaturing gel gradient electrophoresis (DGGE). We found that the FastDNA SPIN kit provided significantly higher DNA yields for all soils; however, it also resulted in the highest levels of humic acid contamination. Soil texture and organic carbon content of the soil did not affect the DNA yield of any kit. Moreover, a liquid-liquid extraction of the DNA extracts found no residual PAHs, indicating that all kits were effective at removing contaminants in the extraction process. Although the PowerSoil DNA Isolation kit gave relatively low DNA yields, it provided the highest quality DNA based on successful amplification of both bacterial and eukaryotic DNA for all six soils. DGGE fingerprints among the kits were dramatically different for both bacterial and eukaryotic DNA. The PowerSoil DNA Isolation kit revealed multiple bands for each soil and provided the most consistent DGGE profiles among replicates for both bacterial and eukaryotic DNA.

  3. [Preparation of freeze - drying control materials of IgG antibody against Schistosoma japonicum for immunodetection kits].

    PubMed

    Jin, Huang; Chun-Lian, Tang; Zu-Wu, Tu; Li, Tang; Ke-Hui, Zhang; Qian, Li; Jun, Ye

    2018-04-18

    To prepare freeze-drying control materials of IgG antibody against Schistosoma japonicum for detection kits. The serum samples of schistosomiasis patients from endemic areas and normal people without history of schistosome infection or contact with infested water in Hubei Province were collected. All the sera were detected by the method approved by China Food and Drug Administration and selected for preparation of quality control samples. Totally twelve positive quality control materials, ten negative quality control materials, and one sensitive and one precision quality control materials were screened. According to the positive serum level, the positive degrees of quality control materials were divided into strong, medium and weak levels. The stability could be valid for one year. The freeze-drying quality control materials of IgG antibody against S. japonicum for detection kits are prepared. They are easy to use and have good stability, and therefore, they may meet the requirement of quality control for the detection of schistosomiasis diagnostics kits.

  4. Incorporating ethical principles into clinical research protocols: a tool for protocol writers and ethics committees

    PubMed Central

    Li, Rebecca H; Wacholtz, Mary C; Barnes, Mark; Boggs, Liam; Callery-D'Amico, Susan; Davis, Amy; Digilova, Alla; Forster, David; Heffernan, Kate; Luthin, Maeve; Lynch, Holly Fernandez; McNair, Lindsay; Miller, Jennifer E; Murphy, Jacquelyn; Van Campen, Luann; Wilenzick, Mark; Wolf, Delia; Woolston, Cris; Aldinger, Carmen; Bierer, Barbara E

    2016-01-01

    A novel Protocol Ethics Tool Kit (‘Ethics Tool Kit’) has been developed by a multi-stakeholder group of the Multi-Regional Clinical Trials Center of Brigham and Women's Hospital and Harvard. The purpose of the Ethics Tool Kit is to facilitate effective recognition, consideration and deliberation of critical ethical issues in clinical trial protocols. The Ethics Tool Kit may be used by investigators and sponsors to develop a dedicated Ethics Section within a protocol to improve the consistency and transparency between clinical trial protocols and research ethics committee reviews. It may also streamline ethics review and may facilitate and expedite the review process by anticipating the concerns of ethics committee reviewers. Specific attention was given to issues arising in multinational settings. With the use of this Tool Kit, researchers have the opportunity to address critical research ethics issues proactively, potentially speeding the time and easing the process to final protocol approval. PMID:26811365

  5. Tyrosine Kinase Inhibitors Induce Down-Regulation of c-Kit by Targeting the ATP Pocket

    PubMed Central

    Descarpentries, Clotilde; Frisan, Emilie; Adam, Kevin; Verdier, Frederique; Floquet, Célia; Dubreuil, Patrice; Lacombe, Catherine; Fontenay, Michaela; Mayeux, Patrick; Kosmider, Olivier

    2013-01-01

    The stem cell factor receptor (SCF) c-Kit plays a pivotal role in regulating cell proliferation and survival in many cell types. In particular, c-Kit is required for early amplification of erythroid progenitors, while it must disappear from cell surface for the cell entering the final steps of maturation in an erythropoietin-dependent manner. We initially observed that imatinib (IM), an inhibitor targeting the tyrosine kinase activity of c-Kit concomitantly down-regulated the expression of c-Kit and accelerated the Epo-driven differentiation of erythroblasts in the absence of SCF. We investigated the mechanism by which IM or related masitinib (MA) induce c-Kit down-regulation in the human UT-7/Epo cell line. We found that the down-regulation of c-Kit in the presence of IM or MA was inhibited by a pre-incubation with methyl-β-cyclodextrin suggesting that c-Kit was internalized in the absence of ligand. By contrast to SCF, the internalization induced by TKI was independent of the E3 ubiquitin ligase c-Cbl. Furthermore, c-Kit was degraded through lysosomal, but not proteasomal pathway. In pulse-chase experiments, IM did not modulate c-Kit synthesis or maturation. Analysis of phosphotyrosine peptides in UT-7/Epo cells treated or not with IM show that IM did not modify overall tyrosine phosphorylation in these cells. Furthermore, we showed that a T670I mutation preventing the full access of IM to the ATP binding pocket, did not allow the internalization process in the presence of IM. Altogether these data show that TKI-induced internalization of c-Kit is linked to a modification of the integrity of ATP binding pocket. PMID:23637779

  6. An Evaluation of the NIDS (registered trademark) ACE (trademark) Test

    DTIC Science & Technology

    2014-06-30

    included chemicals commonly used for drinking water disinfection (chlorine and chloramine), byproducts of cyanobacteria blooms (geosmin and 2...duration field toxicity test for Army drinking water . One component of the ESB the ACE Test) is an enzymatic assay designed to detect neurotoxicants...adverse health effects is the presence of toxic industrial chemicals (TICs) in drinking water . The current field water test kit – the water quality

  7. A look at the purchase and use of home pregnancy-test kits.

    PubMed

    Coons, S J

    1989-04-01

    A study was conducted to obtain information regarding the purchase and use of home pregnancy-test kits. Questionnaires were distributed to 438 women entering a family-planning clinic in the fall of 1987. A total of 153 questionnaires were completed and returned, providing a response rate of 34.9%. Results indicated that nearly 40% of the respondents had used a home pregnancy-test kit at least once. Of those who had used a home pregnancy-test kit, the majority did so because of "the speed of obtaining results" or "convenience." Although nearly 87% of the pregnancy-test kits had been purchased in a pharmacy, pharmacists played only a minor role in the decision-making process concerning purchase and use. Some three-fourths of the subjects listed "information on the side of the package," "price," or "advertisements" as the most important factor in the selection of a specific brand of test kit. Only about 7% of the subjects selected "recommendation of the pharmacist" as the most important factor. The results suggest that pharmacists could be doing more to promote the appropriate use of self-testing products, specifically home pregnancy-test kits.

  8. Development and testing of monoclonal antibody-based rapid immunodiagnostic test kits for direct detection of Vibrio cholerae O139 synonym Bengal.

    PubMed

    Hasan, J A; Huq, A; Nair, G B; Garg, S; Mukhopadhyay, A K; Loomis, L; Bernstein, D; Colwell, R R

    1995-11-01

    We report on the development and testing of two monoclonal antibody-based rapid immunodiagnostic test kits, BengalScreen, a coagglutination test, and Bengal DFA, a direct fluorescent-antibody test, for direct detection of Vibrio cholerae O139 synonym Bengal in clinical and environmental specimens. The BengalScreen test requires less than 5 min to complete and can be used in the field. Bengal DFA, being more sensitive than BengalScreen, requires only one reagent and less than 20 min for detection and enumeration of V. cholerae O139 synonym Bengal. In tests for specificity, all 40 strains of V. cholerae O139 reacted with both test kits, whereas 157 strains of heterologous species examined did not, yielding 100% specificity in this study. A field trial was conducted in with both BengalScreen and Bengal DFA, and the results were compared with those obtained by conventional culture methods. BengalScreen demonstrated a sensitivity of 95%, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 94%. Results obtained by Bengal DFA, on the other hand, were 100% sensitive and 100% specific and yielded 100% positive and negative predictive values compared with culture methods. In a second evaluation, 93 stool specimens from Mexico that were negative for V. cholerae O139 by culture were also tested with both the BengalScreen and Bengal DFA kits. None of the 93 specimens were positive for V. cholerae O139 by both tests. A concentration method was optimized for screening of environmental water samples for V. cholerae O139 synonym Bengal with rapid test kits. BengalScreen results were unequivocally positive when water samples contained at least 2.0 x 10(3) CFU/ml, whereas Bengal DFA demonstrated an unequivocally positive reaction when the water sample contained at least 1.5 x 10(2) CFU/ml. When Bengal DFA was compared with conventional culture methods for enumeration of V. cholerae O139 synonym Bengal organisms, no difference was observed.

  9. Phosphorylation of the Activation Loop Tyrosine 823 in c-Kit Is Crucial for Cell Survival and Proliferation*

    PubMed Central

    Agarwal, Shruti; Kazi, Julhash U.; Rönnstrand, Lars

    2013-01-01

    The receptor tyrosine kinase c-Kit, also known as the stem cell factor receptor, plays a key role in several developmental processes. Activating mutations in c-Kit lead to alteration of these cellular processes and have been implicated in many human cancers such as gastrointestinal stromal tumors, acute myeloid leukemia, testicular seminomas and mastocytosis. Regulation of the catalytic activity of several kinases is known to be governed by phosphorylation of tyrosine residues in the activation loop of the kinase domain. However, in the case of c-Kit phosphorylation of Tyr-823 has been demonstrated to be a late event that is not required for kinase activation. However, because phosphorylation of Tyr-823 is a ligand-activated event, we sought to investigate the functional consequences of Tyr-823 phosphorylation. By using a tyrosine-to-phenylalanine mutant of tyrosine 823, we investigated the impact of Tyr-823 on c-Kit signaling. We demonstrate here that Tyr-823 is crucial for cell survival and proliferation and that mutation of Tyr-823 to phenylalanine leads to decreased sustained phosphorylation and ubiquitination of c-Kit as compared with the wild-type receptor. Furthermore, the mutated receptor was, upon ligand-stimulation, quickly internalized and degraded. Phosphorylation of the E3 ubiquitin ligase Cbl was transient, followed by a substantial reduction in phosphorylation of downstream signaling molecules such as Akt, Erk, p38, Shc, and Gab2. Thus, we propose that activation loop tyrosine 823 is crucial for activation of both the MAPK and PI3K pathways and that its disruption leads to a destabilization of the c-Kit receptor and decreased survival of cells. PMID:23803604

  10. Comparison of DNA extraction protocols for microbial communities from soil treated with biochar

    PubMed Central

    Leite, D.C.A.; Balieiro, F.C.; Pires, C.A.; Madari, B.E.; Rosado, A.S.; Coutinho, H.L.C.; Peixoto, R.S.

    2014-01-01

    Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit Miniprep™ (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples. PMID:24948928

  11. Comparison of DNA extraction protocols for microbial communities from soil treated with biochar.

    PubMed

    Leite, D C A; Balieiro, F C; Pires, C A; Madari, B E; Rosado, A S; Coutinho, H L C; Peixoto, R S

    2014-01-01

    Many studies have evaluated the effects of biochar application on soil structure and plant growth. However, there are very few studies describing the effect of biochar on native soil microbial communities. Microbial analysis of environmental samples requires accurate and reproducible methods for the extraction of DNA from samples. Because of the variety among microbial species and the strong adsorption of the phosphate backbone of the DNA molecule to biochar, extracting and purifying high quality microbial DNA from biochar-amended soil is not a trivial process and can be considerably more difficult than the extraction of DNA from other environmental samples. The aim of this study was to compare the relative efficacies of three commercial DNA extraction kits, the FastDNA® SPIN Kit for Soil (FD kit), the PowerSoil® DNA Isolation Kit (PS kit) and the ZR Soil Microbe DNA Kit Miniprep™ (ZR kit), for extracting microbial genomic DNA from sand treated with different types of biochar. The methods were evaluated by comparing the DNA yields and purity and by analysing the bacterial and fungal community profiles generated by PCR-DGGE. Our results showed that the PCR-DGGE profiles for bacterial and fungal communities were highly affected by the purity and yield of the different DNA extracts. Among the tested kits, the PS kit was the most efficient with respect to the amount and purity of recovered DNA and considering the complexity of the generated DGGE microbial fingerprint from the sand-biochar samples.

  12. Effects of grain species and cultivar, thermal processing, and enzymatic hydrolysis on gluten quantitation.

    PubMed

    Pahlavan, Autusa; Sharma, Girdhari M; Pereira, Marion; Williams, Kristina M

    2016-10-01

    Gluten from wheat, rye, and barley can trigger IgE-mediated allergy or Celiac disease in sensitive individuals. Gluten-free labeled foods are available as a safe alternative. Immunoassays such as the enzyme-linked immunosorbent assay (ELISA) are commonly used to quantify gluten in foods. However, various non-assay related factors can affect gluten quantitation. The effect of gluten-containing grain cultivars, thermal processing, and enzymatic hydrolysis on gluten quantitation by various ELISA kits was evaluated. The ELISA kits exhibited variations in gluten quantitation depending on the gluten-containing grain and their cultivars. Acceptable gluten recoveries were obtained in 200mg/kg wheat, rye, and barley-spiked corn flour thermally processed at various conditions. However, depending on the enzyme, gluten grain source, and ELISA kit used, measured gluten content was significantly reduced in corn flour spiked with 200mg/kg hydrolyzed wheat, rye, and barley flour. Thus, the gluten grain source and processing conditions should be considered for accurate gluten analysis. Published by Elsevier Ltd.

  13. Suicide Prevention with Diverse College Students

    ERIC Educational Resources Information Center

    Shadick, Richard; Akhter, Sarah

    2014-01-01

    Because of a dearth of experience in preventing suicide in diverse student populations, Pace University developed a multicultural suicide prevention kit. This article details the process used to develop the kit. The rationale for approaching suicide prevention in a culturally competent manner is presented, and methods used to gain culture-specific…

  14. Menstrual Education and Personal Hygiene Supplies to Empower Young Women in Haiti.

    PubMed

    Coker-Bolt, Patricia; Jansson, Annika; Bigg, Sherridan; Hammond, Elizabeth; Hudson, Harmony; Hunkler, Sarah; Kitch, Jana; Richardson, Heather; Tiedemann, Erica; O'Flynn, Janet; Laurent, Marie Dominque

    2017-10-01

    Recent studies report that Haitian women are concerned about unmet women's health issues. The Days for Girls (DfG) International program features women's health education and personal hygiene kits to ensure women understand the process of menstruation and sanitary hygiene practices. The aim was to train Haitian seamstresses to produce the DfG kits during an in-country workshop and investigate the perceived benefit of the DfG program in young women who used the DfG kits. Posttest only design was used to measure the effectiveness of DfG workshop and postsurvey to study perception of women using DfG kits. It was found that the workshop participants demonstrated an understanding of the DfG program (90% average). Forty-four young women (89.8%) who used the DfG hygiene kits for 2 months agreed that there is a need for feminine hygiene programs in Haiti and that the kits were easy to use and clean (97.1% and 92.1%, respectively). The DfG program could provide a cost-effective feminine hygiene program for Haiti and decrease waste from traditional hygiene products.

  15. Cross-compartment evaluation of a fully-coupled hydrometeorological modeling system using comprehensive observation data

    NASA Astrophysics Data System (ADS)

    Fersch, Benjamin; Senatore, Alfonso; Kunstmann, Harald

    2017-04-01

    Fully-coupled hydrometeorological modeling enables investigations about the complex and often non-linear exchange mechanisms among subsurface, land, and atmosphere with respect to water and energy fluxes. The consideration of lateral redistribution of surface and subsurface water in such modeling systems is a crucial enhancement, allowing for a better representation of surface spatial patterns and providing also channel discharge predictions. However, the evaluation of fully-coupled simulations is difficult since the amount of physical detail along with feedback mechanisms leads to high degrees of freedom. Therefore, comprehensive observation data is required to obtain meaningful model configurations. We present a case study for a medium-sized river catchment in southern Germany that includes the calibration of the stand-alone and the evaluation of the fully-coupled WRF-Hydro modeling system with a horizontal resolution of 1 x 1 km2, for the period June to August 2015. ECMWF ERA-Interim reanalysis is used for model driving. Land-surface processes are represented by the Noah-MP land surface model. Land-cover is described by the EU CORINE data set. Observations for model evaluation are obtained from the TERENO Pre-Alpine observatory (http://www.imk-ifu.kit.edu/tereno.php) and are complemented by further measurements from the ScaleX campaign (http://scalex.imk-ifu.kit.edu) such as atmospheric profiles obtained from radiometer sounding and airborne systems as well as soil moisture and -temperature networks. We show how well water budgets and heat-fluxes are being reproduced by the stand-alone WRF, the stand-alone WRF-Hydro and the fully-coupled WRF-Hydro model.

  16. Defense Science Board 2006 Summer Study on 21st Century Strategic Technology Vectors. Volume 4. Accelerating the Transition of Technologies into U.S. Capabilities

    DTIC Science & Technology

    2007-04-01

    perform more research on future defense technology, the DOD should invest in companies that are leaders in the development of innovative sources of next...well. In fact, the kit from one vender out-performed the standard up-armor kits being produced for the Army’s acquisition team. That Army company ...subsequently purchased the company that had built the improved performance kit. As part of the process to look at alternatives, the Army Material

  17. 78 FR 79391 - Foreign-Trade Zone (FTZ) 22-Chicago, Illinois, Notification of Proposed Production Activity...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-30

    ...; cable clamps; locks; belts; O-rings; sealing gaskets; support packages; filter bags; disposable bags; assembly bags; dust bags; maintenance packs; paper bags; dirt tube kits; paper adaptor bags; filters...; wire racks; bake pans; pizza pans; coffee water filters; water filters; base assemblies; dust cup...

  18. Occurrence of ground waters of low hardness and of high chloride content in Lyon County, Minnesota

    USGS Publications Warehouse

    Rodis, Harry G.; Schneider, Robert

    1960-01-01

    Hardness and chloride determinations were made with field-testing kits at the time data were obtained on most of the farm wells in the county. Tests were made on wells that were reported to yield relatively soft or "salty" water.

  19. Differences in staining intensities affect reported occurrences and concentrations of Giardia spp. in surface drinking water sources

    EPA Science Inventory

    Aim USEPA Method 1623, or its equivalent, is currently used to monitor for protozoan contamination of surface drinking water sources worldwide. At least three approved staining kits used for detecting Cryptosporidium and Giardia are commercially available. This study focuses on ...

  20. Reduced weight decontamination formulation for neutralization of chemical and biological warfare agents

    DOEpatents

    Tucker, Mark D.

    2014-06-03

    A reduced weight DF-200 decontamination formulation that is stable under high temperature storage conditions. The formulation can be pre-packed as an all-dry (i.e., no water) or nearly-dry (i.e., minimal water) three-part kit, with make-up water (the fourth part) being added later in the field at the point of use.

  1. Water Quality Monitoring of an Urban Estuary and a Coastal Aquifer Using Field Kits and Meters: A Community-Based Environmental Research Project

    ERIC Educational Resources Information Center

    Jung, Hun Bok; Zamora, Felix; Duzgoren-Aydin, Nurdan S.

    2017-01-01

    Water quality is an important interdisciplinary environmental topic for project-based learning. An undergraduate summer research internship program at a public minority serving institution engaged environmental science majors in community-based research experiences. The research focused on the field monitoring of water quality for surface water…

  2. Mixed Alcohol Dehydration over Bronsted and Lewis Acidic Catalysts

    DOE PAGES

    Nash, Connor P.; Ramanathan, Anand; Ruddy, Daniel A.; ...

    2015-12-01

    Mixed alcohols are attractive oxygenated products of biomass-derived syngas because they may be catalytically converted to a range of hydrocarbon products, including liquid hydrocarbon fuels. Catalytic dehydration to form olefins is a potential first step in the conversion of C 2–C 4 alcohols into longer-chain hydrocarbons. Here, we describe the physical and chemical characterization along with catalytic activity and selectivity of 4 Brønsted and Lewis acidic catalysts for the dehydration of two mixed alcohol feed streams that are representative of products from syngas conversion over K-CoMoS type catalysts (i.e., ethanol, 1-propanol, 1-butanol and 2-methyl-1-propanol). Specifically, a Lewis acidic Zr-incorporated mesoporousmore » silicate (Zr-KIT-6), a commercial Al-containing mesoporous silicate (Al-MCM-41), a commercial microporous aluminosilicate (HZSM-5), and a commercial microporous silicoaluminophosphate (SAPO-34) were tested for mixed alcohol dehydration at 250, 300 and 350 °C. The zeolite materials exhibited high activity (>98% ethanol conversion) at all temperatures while the mesoporous materials only displayed significant activity (>10% ethanol conversion) at or above 300 °C. The turnover frequencies for ethanol dehydration at 300 °C decreased in the following order: HZSM-5 > SAPO-34 > Al-MCM-41 > Zr-KIT-6, suggesting that Brønsted acidic sites are more active than Lewis acidic sites for alcohol dehydration. At 300 °C, SAPO-34 produced the highest yield of olefin products from both a water-free ethanol rich feed stream and a C 3+-alcohol rich feed stream containing water. Post-reaction characterization indicated changes in the Brønsted-to-Lewis acidic site ratios for Zr-KIT-6, Al-MCM-41 and HZSM-5. Ammonia temperature programmed desorption indicated that the acid sites of post-reaction samples could be regenerated following treatment in air. The post-reaction SAPO-34 catalyst contained more aromatic, methylated aromatic and polyaromatic compounds than its zeolite counterpart HZSM-5, while no aromatic compounds were observed on post-reaction Al-MCM-41 or Zr-KIT-6 catalysts. Olefin yield at 300 °C over SAPO-34 (>95%) was comparable to published values for the methanol-to-olefins process, indicating the potential industrial application of mixed alcohol dehydration. Furthermore, the olefin product distribution over SAPO-34 was tunable by the composition of the alcohol feed mixture.« less

  3. Mixed Alcohol Dehydration over Bronsted and Lewis Acidic Catalysts

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

    Nash, Connor P.; Ramanathan, Anand; Ruddy, Daniel A.

    Mixed alcohols are attractive oxygenated products of biomass-derived syngas because they may be catalytically converted to a range of hydrocarbon products, including liquid hydrocarbon fuels. Catalytic dehydration to form olefins is a potential first step in the conversion of C 2–C 4 alcohols into longer-chain hydrocarbons. Here, we describe the physical and chemical characterization along with catalytic activity and selectivity of 4 Brønsted and Lewis acidic catalysts for the dehydration of two mixed alcohol feed streams that are representative of products from syngas conversion over K-CoMoS type catalysts (i.e., ethanol, 1-propanol, 1-butanol and 2-methyl-1-propanol). Specifically, a Lewis acidic Zr-incorporated mesoporousmore » silicate (Zr-KIT-6), a commercial Al-containing mesoporous silicate (Al-MCM-41), a commercial microporous aluminosilicate (HZSM-5), and a commercial microporous silicoaluminophosphate (SAPO-34) were tested for mixed alcohol dehydration at 250, 300 and 350 °C. The zeolite materials exhibited high activity (>98% ethanol conversion) at all temperatures while the mesoporous materials only displayed significant activity (>10% ethanol conversion) at or above 300 °C. The turnover frequencies for ethanol dehydration at 300 °C decreased in the following order: HZSM-5 > SAPO-34 > Al-MCM-41 > Zr-KIT-6, suggesting that Brønsted acidic sites are more active than Lewis acidic sites for alcohol dehydration. At 300 °C, SAPO-34 produced the highest yield of olefin products from both a water-free ethanol rich feed stream and a C 3+-alcohol rich feed stream containing water. Post-reaction characterization indicated changes in the Brønsted-to-Lewis acidic site ratios for Zr-KIT-6, Al-MCM-41 and HZSM-5. Ammonia temperature programmed desorption indicated that the acid sites of post-reaction samples could be regenerated following treatment in air. The post-reaction SAPO-34 catalyst contained more aromatic, methylated aromatic and polyaromatic compounds than its zeolite counterpart HZSM-5, while no aromatic compounds were observed on post-reaction Al-MCM-41 or Zr-KIT-6 catalysts. Olefin yield at 300 °C over SAPO-34 (>95%) was comparable to published values for the methanol-to-olefins process, indicating the potential industrial application of mixed alcohol dehydration. Furthermore, the olefin product distribution over SAPO-34 was tunable by the composition of the alcohol feed mixture.« less

  4. The ability of two commercially available quick test kits to detect drug-facilitated sexual assault drugs in beverages.

    PubMed

    Beynon, C M; Sumnall, H R; McVeigh, J; Cole, J C; Bellis, M A

    2006-10-01

    Assessment of the sensitivity and specificity of two commercially available 'drug-facilitated sexual assault' drug detector kits, Drink Guard and Drink Detective. Experimental. Laboratory. Gamma hydroxybutyrate (GHB) sodium salt, ketamine hydrochloride, temazepam, flunitrazepam and diazepam were dissolved (Tween added to benzodiazepine solutions) as separate stock solutions and added to 330 ml samples of cola (Pepsi Max), beer (Stella Artois), 'alcopop' (Bacardi Breezer) and placebo (distilled water). The doses used are reported to be common in cases of intoxication. Each kit was tested 10 times for each drink/drug combination. Two blind, independent observers scored each test (presence/absence of drug) in accordance with kit instructions; chi 2 was used to compare the proportion of times raters scored tests correctly and incorrectly. Sensitivity and specificity were calculated overall, for each drink, and sensitivity was calculated for each drug. Inter-observer agreement was evaluated using the kappa statistic. While both raters were able to score significantly more tests correctly than incorrectly using Drink Detective, and one rater scored similarly using Drink Guard, the overall sensitivity of Drink Detective and Drink Guard was 69.0% (95% CI 64.2-73.5%) and 37.5% (95% CI 30.1-45.5%), respectively. Sensitivity was drink-dependent. Drink Detective was unable to detect our dose of GHB in water, with all tests scored negatively by both raters for this drink/drug combination (n = 20 negative scores). Overall, specificity was 76.6% (95% CI 71.5-81.0%) and 87.9% (95% CI 83.0-91.6%) for Drink Guard and Drink Detective, respectively, but was affected by the beverage. Inter-rater agreement was poor for Drink Guard (kappa = 0.278 +/- 0.069) but excellent for Drink Detective (kappa = 0.894 +/- 0.245). Inter-observer agreement was drug-dependent. Use of drug detector kits by the public in the night-time environment needs further investigation and may create a false sense of security (false negatives) and undue concern (false positives) among kit users.

  5. Gloss and surface roughness produced by polishing kits on resin composites.

    PubMed

    Sadidzadeh, Ramtin; Cakir, Deniz; Ramp, Lance C; Burgess, John O

    2010-08-01

    To compare in vitro the surface roughness (Ra) and gloss (G) produced by three conventional and one experimental polishing kits on four resin composites. 24 discs were prepared (d = 12 mm, t = 4 mm) for each resin composite: Filtek Supreme Plus Body/A2 (FSB), Yellow Translucent (FST), Heliomolar/A2 (HM), and EsthetX/A2 (EX) following the manufacturers' instructions. They were finished with 320 grit silicon carbide paper for 80 seconds each. Polishing systems: Sof-Lex, Enhance-Pogo, Astropol and Experimental Discs/EXL-695, were applied following manufacturers' instructions. Each specimen was ultrasonically cleaned with distilled water and dried. Gloss and Ra were measured with a small area glossmeter (Novo-curve) and non-contact profilometer (Proscan 2000) following ISO 4288, respectively. The results were evaluated by two-way ANOVA followed by separate one-way ANOVA and Tukey/Kramer test (P = 0.05). There was a significant interaction of surface roughness and gloss between the composites and polishing systems (P < 0.05). The lowest surface roughness was recorded for FST polished with the Experimental kit. The highest gloss was obtained for FSB composite polished with the Experimental kit. The experimental polishing system produced smoothest surfaces (P < 0.05). The Enhance-Pogo and the experimental polishing kit produced highest gloss (P < 0.05).

  6. The human tyrosine kinase Kit and its gatekeeper mutant T670I, show different kinetic properties: Implications for drug design.

    PubMed

    Kissova, Miroslava; Maga, Giovanni; Crespan, Emmanuele

    2016-10-01

    The tyrosine kinase Kit, a receptor for Stem Cell Factor, is involved, among others, in processes associated to cell survival, proliferation and migration. Upon physiological conditions, the activity of Kit is tightly regulated. However, primary mutations that lead to its constitutive activation are the causal oncogenic driver of gastrointestinal stromal tumours (GISTs). GISTs are known to be refractory to conventional therapies but the introduction of Imatinib, a selective inhibitor of tyrosine kinases Abl and Kit, significantly ameliorated the treatment options of GISTs patients. However, the acquisition of secondary mutations renders Kit resistant towards all available drugs. Mutation involving gatekeeper residues (such as V654a and T670I) influence both the structure and the catalytic activity of the enzyme. Therefore, detailed knowledge of the enzymatic properties of the mutant forms, in comparison with the wild type enzyme, is an important pre-requisite for the rational development of specific inhibitors. In this paper we report a thorough kinetic analysis of the reaction catalyzed by the Kit kinase and its gatekeeper mutated form T670I. Our results revealed the different mechanisms of action of these two enzymes and may open a new avenue for the future design of specific Kit inhibitors. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. A change in structural integrity of c-Kit mutant D816V causes constitutive signaling.

    PubMed

    Raghav, Pawan Kumar; Singh, Ajay Kumar; Gangenahalli, Gurudutta

    2018-03-01

    Several signaling pathways, ligands, and genes that regulate proliferative and self-renewal properties of the Hematopoietic Stem Cells (HSCs) have been studied meticulously. One of the signaling pathways that play a crucial role in the process of hematopoiesis is the Stem Cell Factor (SCF) mediated c-Kit pathway. The c-Kit is a Receptor Tyrosine Kinase (RTK), which is expressed in the cells including HSCs. It undergoes dimerization upon binding with its cognate ligand SCF. As a result, phosphorylation of the Juxtamembrane (JM) domain of c-Kit takes place at Tyr568 and Tyr570 residues. These phosphorylated residues become the docking sites for protein tyrosine phosphatases (PTPs) namely SHP-1 and SHP-2, which in turn cause dephosphorylation and negative regulation of the downstream signaling responsible for the cell proliferation. Interestingly, it has been reported that the mutation of c-Kit at D816V makes it independent of SCF stimulation and SHP-1/SHP-2 inhibition, thereby, causing its constitutive activation. The present study was commenced to elucidate the structural behavior of this mutation in the JM and A-loop region of c-Kit using Molecular Dynamics (MD) simulations of the wild-type and mutant c-Kit in unphosphorylated and phosphorylated states. The energy difference computed between the wild type and mutant (D816V) c-Kit, and protein-protein docking and complex analysis revealed the impact of this single residue mutation on the integrity dynamics of c-Kit that makes it independent of SHP-1/SHP-2 negative regulation. Copyright © 2018 Elsevier B.V. All rights reserved.

  8. Resource Kit Tips for Teaching Textiles and Clothing.

    ERIC Educational Resources Information Center

    New York State Education Dept., Albany. Bureau of Continuing Education Curriculum Development.

    This kit has been designed to acquaint the instructor of adult textiles and clothing programs with some of the teaching aids that might be used to improve the learning process. The main parts of the publication include: Preparing and Using Transparencies; Developing a Learning Experience Using a Transparency; A Master Transparency with Overlays;…

  9. World War II Spy Kit: "The Great Nazi Intelligence Coup."

    ERIC Educational Resources Information Center

    Haight, David

    This instructional packet is designed to introduce students to primary source material by having them participate in an historical "what might have been." Students engage in critical thinking and document analysis, and through the process learn about Operation OVERLORD and World War II in general. This spy kit centers on Operation…

  10. An improved kit for culturing clean nematode, larvae from faeces.

    PubMed

    Ndalahwa, J B; Lwambo, N J

    1989-03-01

    A kit for rapid recovery of clean hookworm and Strongyloides stercoralis larvae from faeces was devised. It consists of a 2.0dl perspex cup shaped container 35 x 30 x 24mm in size. A horse shoe stirrup aluminium sheet 54mm long and 20mm wide is inserted into the container so that the free ends stand at the bottom of the apparatus. The back of the stirrup carries a plastic disc 30mm in diameter through which two adjacent crescent shaped openings are cut. A strip of filter paper 40 x 20mm in size passes through the crescent openings astride the stirrup and its loose ends are bound together by means of a stapler. When the kit is assembled, the disk fits firmly in the middle of the container dividing it into lower and upper chambers. The staples free ends of the filter paper hang immersed into water in the lower chamber of the apparatus. A faecal sample is smeared on the upper part of the filter paper. Nematode larvae, if present, crawl from the faeces down the damp filter paper into the water at the bottom of the container.

  11. Technology assessment and strategy for development of a Rapid Field Water Microbiology Test Kit. Technical report

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

    Preston, D.R.; Schaub, S.A.

    1991-09-01

    A literature and market search of existing technology for the detection, identification, and quantification of microorganisms in water was conducted. Based upon the availability of technologies and their configurations, an assessment of the appropriate strategies to pursue for the near and long term development plans in development of the Rapid Field Bacteriology Test Kit was performed. Near term technologies to improve the Army's capability to detect microorganisms would appear to be essentially improvements in versatility and measurement of coliform indicator organisms. New chromogenic and fluorogenic indicator substances associated with new substrates appear to be best suited for test kit developmentmore » either for quantitative membrane filter tests or presence/absence and multiple fermentation tests. Test times, incubator requirements, and operator involvement appear to be similar to older technologies. Long term development would appear to favor such technologies as genetic probes with amplification of the hydridized nucleic acid materials of positive samples, and some immunological based systems such as enzyme linked, immuno-sorbent assays. In both cases, the major problems would appear to be sample preparation and development of signal strengths from the reactions which would allow the user to see results in 1 hour.« less

  12. Field demonstration of on-site analytical methods for TNT and RDX in ground water

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

    Craig, H.; Ferguson, G.; Markos, A.

    1996-12-31

    A field demonstration was conducted to assess the performance of eight commercially-available and emerging colorimetric, immunoassay, and biosensor on-site analytical methods for explosives 2,4,6-trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) in ground water and leachate at the Umatilla Army Depot Activity, Hermiston, Oregon and US Naval Submarine Base, Bangor, Washington, Superfund sites. Ground water samples were analyzed by each of the on-site methods and results compared to laboratory analysis using high performance liquid chromatography (HPLC) with EPA SW-846 Method 8330. The commercial methods evaluated include the EnSys, Inc., TNT and RDX colorimetric test kits (EPA SW-846 Methods 8515 and 8510) with amore » solid phase extraction (SPE) step, the DTECH/EM Science TNT and RDX immunoassay test kits (EPA SW-846 Methods 4050 and 4051), and the Ohmicron TNT immunoassay test kit. The emerging methods tested include the antibody-based Naval Research Laboratory (NRL) Continuous Flow Immunosensor (CFI) for TNT and RDX, and the Fiber Optic Biosensor (FOB) for TNT. Accuracy of the on-site methods were evaluated using linear regression analysis and relative percent difference (RPD) comparison criteria. Over the range of conditions tested, the colorimetric methods for TNT and RDX showed the highest accuracy of the emerging methods for TNT and RDX. The colorimetric method was selected for routine ground water monitoring at the Umatilla site, and further field testing on the NRL CFI and FOB biosensors will continue at both Superfund sites.« less

  13. Comparison of DNA extraction methods used to detect bacterial and yeast DNA from spiked whole blood by real-time PCR.

    PubMed

    Dalla-Costa, Libera M; Morello, Luis G; Conte, Danieli; Pereira, Luciane A; Palmeiro, Jussara K; Ambrosio, Altair; Cardozo, Dayane; Krieger, Marco A; Raboni, Sonia M

    2017-09-01

    Sepsis is the leading cause of death in intensive care units (ICUs) worldwide and its diagnosis remains a challenge. Blood culturing is the gold standard technique for blood stream infection (BSI) identification. Molecular tests to detect pathogens in whole blood enable early use of antimicrobials and affect clinical outcomes. Here, using real-time PCR, we evaluated DNA extraction using seven manual and three automated commercially available systems with whole blood samples artificially contaminated with Escherichia coli, Staphylococcus aureus, and Candida albicans, microorganisms commonly associated with BSI. Overall, the commercial kits evaluated presented several technical limitations including long turnaround time and low DNA yield and purity. The performance of the kits was comparable for detection of high microorganism loads (10 6 CFU/mL). However, the detection of lower concentrations was variable, despite the addition of pre-processing treatment to kits without such steps. Of the evaluated kits, the UMD-Universal CE IVD kit generated a higher quantity of DNA with greater nucleic acid purity and afforded the detection of the lowest microbial load in the samples. The inclusion of pre-processing steps with the kit seems to be critical for the detection of microorganism DNA directly from whole blood. In conclusion, future application of molecular techniques will require overcoming major challenges such as the detection of low levels of microorganism nucleic acids amidst the large quantity of human DNA present in samples or differences in the cellular structures of etiological agents that can also prevent high-quality DNA yields. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Evaluation of Immunomagnetic Separation for Recovery of Infectious Cryptosporidium parvum Oocysts from Environmental Samples

    PubMed Central

    Rochelle, Paul A.; De Leon, Ricardo; Johnson, Anne; Stewart, Mic H.; Wolfe, Roy L.

    1999-01-01

    Two commercial immunomagnetic separation (IMS) kits for Cryptosporidium were compared for recovery of oocysts from environmental samples. Oocyst recovery efficiencies with the Dynal and Crypto-Scan kits ranged from 62 to 100% and 34 to 74%, respectively, for seeded environmental water concentrates (turbidity of 210 to 11,480 nephelometric turbidity units). Recovery efficiencies were dependent on the mechanism of agitation during the magnetic capture procedure. An assay combining in vitro cell culture and reverse transcriptase PCR demonstrated that oocysts recovered by IMS retained their infectivity. PMID:9925626

  15. Commander Lousma records PGU data on middeck

    NASA Image and Video Library

    1982-03-30

    STS003-22-122 (30 March 1982) --- STS-3 Commander Lousma, wearing communications kit assembly (ASSY) mini-headset (HDST), records Plant Growth Unit (PGU) data for the Influence of Weightlessness on Plant Lignification Experiment at forward middeck locker MF14K. The experiment is designed to demonstrate the effect of weightlessness on the quantity and rate of lignin formation in different plant species during early stages of development. Port side bulkhead with window shade and filter kit appears behind Lousma and potable water tank below him. Trash bag also appears in view. Photo credit: NASA

  16. FUELS IN SOIL TEST KIT: FIELD USE OF DIESEL DOG SOIL TEST KITS

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

    Unknown

    2001-05-31

    Western Research Institute (WRI) is commercializing Diesel Dog Portable Soil Test Kits for performing analysis of fuel-contaminated soils in the field. The technology consists of a method developed by WRI (U.S. Patents 5,561,065 and 5,976,883) and hardware developed by WRI that allows the method to be performed in the field (patent pending). The method is very simple and does not require the use of highly toxic reagents. The aromatic components in a soil extract are measured by absorption at 254 nm with a field-portable photometer. WRI added significant value to the technology by taking the method through the American Societymore » for Testing and Materials (ASTM) approval and validation processes. The method is designated ASTM Method D-5831-96, Standard Test Method for Screening Fuels in Soils. This ASTM designation allows the method to be used for federal compliance activities. In FY 99, twenty-five preproduction kits were successfully constructed in cooperation with CF Electronics, Inc., of Laramie, Wyoming. The kit components work well and the kits are fully operational. In the calendar year 2000, kits were provided to the following entities who agreed to participate as FY 99 and FY 00 JSR (Jointly Sponsored Research) cosponsors and use the kits as opportunities arose for field site work: Wyoming Department of Environmental Quality (DEQ) (3 units), F.E. Warren Air Force Base, Gradient Corporation, The Johnson Company (2 units), IT Corporation (2 units), TRC Environmental Corporation, Stone Environmental, ENSR, Action Environmental, Laco Associates, Barenco, Brown and Caldwell, Dames and Moore Lebron LLP, Phillips Petroleum, GeoSyntek, and the State of New Mexico. By early 2001, ten kits had been returned to WRI following the six-month evaluation period. On return, the components of all ten kits were fully functional. The kits were upgraded with circuit modifications, new polyethylene foam inserts, and updated instruction manuals.« less

  17. C-MORE Science Kits: Putting Technology in the Hands of K-12 Teachers and Students

    NASA Astrophysics Data System (ADS)

    Achilles, K.; Weersing, K.; Daniels, C.; Puniwai, N.; Matsuzaki, J.; Bruno, B. C.

    2008-12-01

    The Center for Microbial Oceanography: Research and Education (C-MORE) is a NSF Science and Technology Center based at the University of Hawaii. The C-MORE education and outreach program offers a variety of resources and professional development opportunities for science educators, including online resources, participation in oceanography research cruises, teacher-training workshops, mini-grants to incorporate microbial oceanography-related content and activities into their classroom and, most recently, C- MORE science kits. C-MORE science kits provide hands-on classroom, field, and laboratory activities related to microbial oceanography for K-12 students. Each kit comes with complete materials and instructions, and is available free of charge to Hawaii's public school teachers. Several kits are available nationwide. C-MORE science kits cover a range of topics and technologies and are targeted at various grade levels. Here is a sampling of some available kits: 1) Marine Murder Mystery: The Case of the Missing Zooxanthellae. Students learn about the effect of climate change and other environmental threats on coral reef destruction through a murder-mystery experience. Participants also learn how to use DNA to identify a suspect. Grades levels: 3-8. 2) Statistical sampling. Students learn basic statistics through an exercise in random sampling, with applications to microbial oceanography. The laptops provided with this kit enable students to enter, analyze, and graph their data using EXCEL. Grades levels: 6-12. 3) Chlorophyll Lab. A research-quality fluorometer is used to measure the chlorophyll content in marine and freshwater systems. This enables students to compare biomass concentrations in samples collected from various locations. Grades levels: 9-12. 4) Conductivity-Temperature-Depth (CTD). Students predict how certain variables (e.g., temperature, pressure, chlorophyll, oxygen) vary with depth. A CTD, attached to a laptop computer, is deployed into deep water off a dock or a ship to collect real-time data and test their hypotheses. Grades levels: 9-12.

  18. The Central Auditory Processing Kit[TM]. Book 1: Auditory Memory [and] Book 2: Auditory Discrimination, Auditory Closure, and Auditory Synthesis [and] Book 3: Auditory Figure-Ground, Auditory Cohesion, Auditory Binaural Integration, and Compensatory Strategies.

    ERIC Educational Resources Information Center

    Mokhemar, Mary Ann

    This kit for assessing central auditory processing disorders (CAPD), in children in grades 1 through 8 includes 3 books, 14 full-color cards with picture scenes, and a card depicting a phone key pad, all contained in a sturdy carrying case. The units in each of the three books correspond with auditory skill areas most commonly addressed in…

  19. The Moon Topography Model as an Astronomy Educational Kit for Visual Impaired Student

    NASA Astrophysics Data System (ADS)

    Pramudya, Y.; Hikmah, F. N.; Muchlas

    2016-08-01

    The visual impaired students need science educational kit at the school to assist their learning process in science. However, there are lack of the educational kit especially on the topic of astronomy. To introduce the structure of the moon, the moon topography model has been made in circular shape only shown the near side of the moon. The moon topography module are easy to be made since it was made based on low cost material. The expertise on astronomy and visual impaired media marked the 76.67% and 94% ideal percentage, respectively. The visual impaired students were able to study the moon crater and mare by using the kit and the braille printed learning book. They also showed the improvement in the material understanding skill.

  20. The Tapwater Tour. The Hands-On Test Kit & Mini Curriculum for Exploring Drinking Water.

    ERIC Educational Resources Information Center

    LaMotte Co., Chestertown, MD.

    The goal of this mini-curriculum is for students to discover what drinking water really is. Five units are designed to be presented as a lecture and to allow 30 to 40 students to perform every procedure, each testing their own tap water sample. Games are provided in each section to reinforce the concepts presented in each unit. Unit 1 introduces…

  1. Sterile technique

    MedlinePlus

    ... the crook of your elbow. Getting Your Supplies Ready To open a sterile pad or kit: Wash your hands with soap and running water for at least 1 minute. Wash the backs, palms, fingers, and between ...

  2. Hot Weather Tips

    MedlinePlus

    ... Caregiver Stories Connections e-Newsletter Research Registry FCA+(plus) Services Press Media Kit What’s New FCA Blog ... person plenty of water and fruit or vegetable juice even if they say they’re not thirsty. ...

  3. Effect of processing on recovery and variability associated with immunochemical analytical methods for multiple allergens in a single matrix: sugar cookies.

    PubMed

    Khuda, Sefat; Slate, Andrew; Pereira, Marion; Al-Taher, Fadwa; Jackson, Lauren; Diaz-Amigo, Carmen; Bigley, Elmer C; Whitaker, Thomas; Williams, Kristina M

    2012-05-02

    Among the major food allergies, peanut, egg, and milk are the most common. The immunochemical detection of food allergens depends on various factors, such as the food matrix and processing method, which can affect allergen conformation and extractability. This study aimed to (1) develop matrix-specific incurred reference materials for allergen testing, (2) determine whether multiple allergens in the same model food can be simultaneously detected, and (3) establish the effect of processing on reference material stability and allergen detection. Defatted peanut flour, whole egg powder, and spray-dried milk were added to cookie dough at seven incurred levels before baking. Allergens were measured using five commercial enzyme-linked immunosorbent assay (ELISA) kits. All kits showed decreased recovery of all allergens after baking. Analytical coefficients of variation for most kits increased with baking time, but decreased with incurred allergen level. Thus, food processing negatively affects the recovery and variability of peanut, egg, and milk detection in a sugar cookie matrix when using immunochemical methods.

  4. The Munchsters Talk about Food: Preschool Nutrition Program.

    ERIC Educational Resources Information Center

    National Live Stock and Meat Board, Chicago, IL.

    This nutrition education kit is designed for use with 3- to 5-year old children. It introduces children to new foods, involves them in the process of food preparation, and creates an awareness of the importance of eating foods from all food groups. The kit has been designed to provide a variety of opportunities to develop and practice language…

  5. A Guide to Delegate Preparation: UNA-USA Model UN Survival Kit. 1988-89 Edition.

    ERIC Educational Resources Information Center

    Muldoon, James P., Jr., Ed.; Diehl, John, Ed.

    Written by students for students, the purpose of the book, one of four components of the Model UN Survival kit, is to help prepare secondary and college student delegates for participation in Model United Nations (UN) Conferences. Model UN programs help students discover the difficulties of world political processes and understand other nations…

  6. Intermediate SCDC Spanish Curricula Units. Science/Health, Unit 1, Kits 1-4, Teacher's Guide.

    ERIC Educational Resources Information Center

    Spanish Curricula Development Center, Miami Beach, FL.

    Unified by the theme "our community", this unit, part of nine basic instructional units for intermediate level, reflects the observations of Mexican Americans, Puerto Ricans, and Cubans in various regions of the United States. Comprised of Kits 1-4, the unit extends the following basic and interpreted science processes: observing, communicating,…

  7. Assessment, Referral and Placement Kit for Adult Literacy & Basic Education Programs in Victoria.

    ERIC Educational Resources Information Center

    Purdey, Margaret

    This kit is an aid to the assessment, referral, placement, and recognition of achievement of adult literacy and basic education students across Victoria (Australia). It is designed as a guide to the integration of current assessment with new placement processes within the context of the new Adult Basic Education Accreditation Framework and the…

  8. Microbial community structures in high rate algae ponds for bioconversion of agricultural wastes from livestock industry for feed production.

    PubMed

    Mark Ibekwe, A; Murinda, Shelton E; Murry, Marcia A; Schwartz, Gregory; Lundquist, Trygve

    2017-02-15

    Dynamics of seasonal microbial community compositions in algae cultivation ponds are complex. However, there is very limited knowledge on bacterial communities that may play significant roles with algae in the bioconversion of manure nutrients to animal feed. In this study, water samples were collected during winter, spring, summer, and fall from the dairy lagoon effluent (DLE), high rate algae ponds (HRAP) that were fed with diluted DLE, and municipal waste water treatment plant (WWTP) effluent which was included as a comparison system for the analysis of total bacteria, Cyanobacteria, and microalgae communities using MiSeq Illumina sequencing targeting the 16S V4 rDNA region. The main objective was to examine dynamics in microbial community composition in the HRAP used for the production of algal biomass. DNA was extracted from the different sample types using three commercially available DNA extraction kits; MoBio Power water extraction kit, Zymo fungi/bacterial extraction kit, and MP Biomedicals FastDNA SPIN Kit. Permutational analysis of variance (PERMANOVA) using distance matrices on each variable showed significant differences (P=0.001) in beta-diversity based on sample source. Environmental variables such as hydraulic retention time (HRT; P<0.031), total N (P<0.002), total inorganic N (P<0.002), total P (P<0.002), alkalinity (P<0.002), pH (P<0.022), total suspended solid (TSS; P<0.003), and volatile suspended solids (VSS; P<0.002) significantly affected microbial communities in DLE, HRAP, and WWTP. Of the operational taxonomic units (OTUs) identified to phyla level, the dominant classes of bacteria identified were: Cyanobacteria, Alpha-, Beta-, Gamma-, Epsilon-, and Delta-proteobacteria, Bacteroidetes, Firmicutes, and Planctomycetes. Our data suggest that microbial communities were significantly affected in HRAP by different environmental variables, and care must be taken in extraction procedures when evaluating specific groups of microbial communities for specific functions. Published by Elsevier B.V.

  9. c-Kit mediates chemoresistance and tumor-initiating capacity of ovarian cancer cells through activation of Wnt/β-catenin-ATP-binding cassette G2 signaling.

    PubMed

    Chau, W K; Ip, C K; Mak, A S C; Lai, H-C; Wong, A S T

    2013-05-30

    Cisplatin and paclitaxel are standard chemotherapy for metastatic ovarian cancer, but with limited efficacy. Cancer stem/progenitor cells (or tumor-initiating cells, TICs) are hypothesized to be chemoresistant, and the existence of TICs in ovarian cancer has been previously demonstrated. However, the key signals and molecular events regulating the formation and expansion of ovarian tumor-initiating cells (OTICs) remain elusive. Here, we show that c-Kit is not just a marker of OTICs, but also a critical mediator of the phenotype that can be a viable target for the treatment of ovarian cancer. In contrast to non-OICs, c-Kit was overexpressed in OTICs. Moreover, the use of small interfering RNA to inhibit c-Kit expression markedly attenuated the number and size of OTIC subpopulations, inhibited the expression of stem cell markers and decreased the tumorigenic capabilities of OTICs. Imatinib (Gleevec), a clinical drug that blocks c-Kit kinase activity, also demonstrated its inhibition potency on OTICs. In addition, cisplatin/paclitaxel, which killed non-OTICs, with c-Kit knockdown or imatinib revealed that this was critically required for intervening ovarian cancer progression and recurrence in vitro and in xenograft tumors in vivo. Similar results were obtained with OTICs derived from ovarian carcinoma patients. Studies into the mechanisms suggest an important role for the activation of Wnt/β-catenin and ATP-binding cassette G2 downstream of c-Kit. The tumor-promoting microenvironment, such as hypoxia, could promote OTICs via upregulation of c-Kit expression. These results unravel an integral role for c-Kit in ovarian neoplastic processes and shed light on its mechanisms of action.

  10. KIT D816V-mutated bone marrow mesenchymal stem cells in indolent systemic mastocytosis are associated with disease progression.

    PubMed

    Garcia-Montero, Andres C; Jara-Acevedo, Maria; Alvarez-Twose, Ivan; Teodosio, Cristina; Sanchez-Muñoz, Laura; Muñiz, Carmen; Muñoz-Gonzalez, Javier I; Mayado, Andrea; Matito, Almudena; Caldas, Carolina; Morgado, Jose M; Escribano, Luis; Orfao, Alberto

    2016-02-11

    Multilineage involvement of bone marrow (BM) hematopoiesis by the somatic KIT D816V mutation is present in a subset of adult indolent systemic mastocytosis (ISM) patients in association with a poorer prognosis. Here, we investigated the potential involvement of BM mesenchymal stem cells (MSCs) from ISM patients by the KIT D816V mutation and its potential impact on disease progression and outcome. This mutation was investigated in highly purified BM MSCs and other BM cell populations from 83 ISM patients followed for a median of 116 months. KIT D816V-mutated MSCs were detected in 22 of 83 cases. All MSC-mutated patients had multilineage KIT mutation (100% vs 30%, P = .0001) and they more frequently showed involvement of lymphoid plus myeloid BM cells (59% vs 22%; P = .03) and a polyclonal pattern of inactivation of the X-chromosome of KIT-mutated BM mast cells (64% vs 0%; P = .01) vs other multilineage ISM cases. Moreover, presence of KIT-mutated MSCs was associated with more advanced disease features, a greater rate of disease progression (50% vs 17%; P = .04), and a shorter progression-free survival (P ≤ .003). Overall, these results support the notion that ISM patients with mutated MSCs may have acquired the KIT mutation in a common pluripotent progenitor cell, prior to differentiation into MSCs and hematopoietic precursor cells, before the X-chromosome inactivation process occurs. From a clinical point of view, acquisition of the KIT mutation in an earlier BM precursor cell confers a significantly greater risk for disease progression and a poorer outcome. © 2016 by The American Society of Hematology.

  11. A Frameshift Mutation in KIT is Associated with White Spotting in the Arabian Camel

    PubMed Central

    Holl, Heather; Isaza, Ramiro; Mohamoud, Yasmin; Ahmed, Ayeda; Almathen, Faisal; Youcef, Cherifi; Gaouar, Semir; Antczak, Douglas F.; Brooks, Samantha

    2017-01-01

    While the typical Arabian camel is characterized by a single colored coat, there are rare populations with white spotting patterns. White spotting coat patterns are found in virtually all domesticated species, but are rare in wild species. Theories suggest that white spotting is linked to the domestication process, and is occasionally associated with health disorders. Though mutations have been found in a diverse array of species, fewer than 30 genes have been associated with spotting patterns, thus providing a key set of candidate genes for the Arabian camel. We obtained 26 spotted camels and 24 solid controls for candidate gene analysis. One spotted and eight solid camels were whole genome sequenced as part of a separate project. The spotted camel was heterozygous for a frameshift deletion in KIT (c.1842delG, named KITW1 for White spotting 1), whereas all other camels were wild-type (KIT+/KIT+). No additional mutations unique to the spotted camel were detected in the EDNRB, EDN3, SOX10, KITLG, PDGFRA, MITF, and PAX3 candidate white spotting genes. Sanger sequencing of the study population identified an additional five KITW1/KIT+ spotted camels. The frameshift results in a premature stop codon five amino acids downstream, thus terminating KIT at the tyrosine kinase domain. An additional 13 spotted camels tested KIT+/KIT+, but due to phenotypic differences when compared to the KITW1/KIT+ camels, they likely represent an independent mutation. Our study suggests that there are at least two causes of white spotting in the Arabian camel, the newly described KITW1 allele and an uncharacterized mutation. PMID:28282952

  12. Evaluation of commercial kit based on loop-mediated isothermal amplification for rapid detection of low levels of uninjured and injured Salmonella on duck meat, bean sprouts, and fishballs in Singapore.

    PubMed

    Lim, Hazel Sin Yue; Zheng, Qianwang; Miks-Krajnik, Marta; Turner, Matthew; Yuk, Hyun-Gyun

    2015-06-01

    The objective of this study was to evaluate performance of the commercial kit based on loop-mediated isothermal amplification (LAMP) in comparison with the International Organization for Standardization method for detecting uninjured and sublethally injured Salmonella cells artificially inoculated at levels of 10(0) and 10(1) CFU/25 g on raw duck wing, raw mung bean sprouts, and processed fishballs. Injured cells were prepared by a heat treatment for duck wings and fishball samples and a chlorine treatment for bean sprout samples. Additionally, a validation study was performed on naturally contaminated food samples sold in Singapore. A total of 110 samples of each commodity were analyzed in this study. Regardless of inoculum levels, the detection by the commercial LAMP kit showed 100% sensitivity and specificity for both inoculated and uninoculated samples compared with the International Organization for Standardization method, with the exception of bean sprout samples. Only 20% of bean sprout samples inoculated with 10(0) CFU/25 g injured Salmonella cells were positive by using the commercial LAMP-based kit. However, all negative samples became positive following a secondary enrichment in Rappaport-Vassiliadis medium with soy broth or after concentration by centrifugation. These results suggest that secondary enrichment or centrifugation should be considered as an additional step to increase the sensitivity of the commercial LAMP-based kit with low numbers of injured target cells in samples with high background microflora (such as mung bean sprouts). The validation study also showed that the commercial LAMP-based kit provided 91% sensitivity and 95% specificity for naturally contaminated samples. Thus, this study demonstrates that the commercial LAMP-based kit might be a cost-effective method, as this system could provide rapid, accurate detection of both uninjured and injured Salmonella cells on raw duck wings, raw mung bean sprouts, and processed fishballs in less than 26 h.

  13. Inventions leading to the development of the diagnostic test kit industry--from the modern pregnancy test to the sandwich assays.

    PubMed

    Wide, Leif

    2005-01-01

    The universities are encouraged by the government nowadays to stimulate innovations and also to provide the proper machinery for assisting the protection and commercialisation of innovations. A better understanding of the innovation process may help to create an atmosphere suitable for inventions at the university. Examples can be taken from successful innovations previously made at the university. During the 1960's I made a series of inventions, which ultimately led to the development of the diagnostic test kit industry. The first, which I made as an undergraduate, was a simple and reliable test kit for diagnosis of pregnancy. This was followed by the solid phase radioimmunoassay and a solid phase assay for vitamin B12; next, the dual specific non-competitive sandwich assay and the in-vitro test for diagnosis of allergy, called RAST (Radioallergosorbent test). Organon in Holland with the pregnancy test kit, and Pharmacia in Sweden with test kits for radioimmunoassay, became pioneers among the diagnostic test kit industries. Pharmacia Diagnostics later became one of the leading diagnostic test kit companies in the world and has continued to be so in the field of allergy diagnosis. Each one of these inventions started with a few unique observations leading to a technical development. The pregnancy test as well as the allergy test emerged from the development of assay methods with unique qualities with the subsequent search for appropriate applications. The foreseeing of a commercial value on a future market was a very important step. This was followed by the search for a suitable industry interested to exploit the invention with its new business opportunity i.e. apply for a patent, produce and market the products, which in my case consisted of the necessary reagents and equipments for particular diagnostic tests. Finally, an agreement had to be settled between the entrepreneur and the inventors. This report describes these inventions and particularly discusses some crucial steps of the innovation processes.

  14. Engineering CHO cells with an oncogenic KIT improves cells growth, resilience to stress, and productivity.

    PubMed

    Mahameed, Mohamed; Tirosh, Boaz

    2017-11-01

    An optimized biomanufacturing process in mammalian cells is contingent on the ability of the producing cells to reach high viable cell densities. In addition, at the peak of growth, cells need to continue producing the biological entity at a consistent quality. Thus, engineering cells with robust growth performance and resilience to variable stress conditions is highly desirable. The tyrosine kinase receptor, KIT, plays a key role in cell differentiation and the survival of several immune cell types. Its oncogenic mutant, D816V, endows cells with high proliferation capacity, and resistance to kinase inhibitors. Importantly, this onco-KIT mutant when introduced into various cell types is arrested in the endoplasmic reticulum in a constitutively active form. Here, we investigated the effect of oncogenic D816V KIT on the performance of CHO-K1 cells under conventional tissue culture growth settings and when adapted, to shaking conditions. The onco-KIT promoted global protein synthesis, elevated the expression of a secretable transgene, enhanced proliferation, and improved the overall titers of a model glycoprotein. Moreover, the expression of the onco-KIT endowed the cells with a remarkable resistance to various stress conditions. Our data suggest that the introduction of onco-KIT can serve as a strategy for improving glycoprotein biomanufacturing. Biotechnol. Bioeng. 2017;114: 2560-2570. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

  15. Optimising the diagnostic strategy for onychomycosis from sample collection to FUNGAL identification evaluation of a diagnostic kit for real-time PCR.

    PubMed

    Petinataud, Dimitri; Berger, Sibel; Ferdynus, Cyril; Debourgogne, Anne; Contet-Audonneau, Nelly; Machouart, Marie

    2016-05-01

    Onychomycosis is a common nail disorder mainly due to dermatophytes for which the conventional diagnosis requires direct microscopic observation and culture of a biological sample. Nevertheless, antifungal treatments are commonly prescribed without a mycological examination having been performed, partly because of the slow growth of dermatophytes. Therefore, molecular biology has been applied to this pathology, to support a quick and accurate distinction between onychomycosis and other nail damage. Commercial kits are now available from several companies for improving traditional microbiological diagnosis. In this paper, we present the first evaluation of the real-time PCR kit marketed by Bio Evolution for the diagnosis of dermatophytosis. Secondly, we compare the efficacy of the kit on optimal and non-optimal samples. This study was conducted on 180 nails samples, processed by conventional methods and retrospectively analysed using this kit. According to our results, this molecular kit has shown high specificity and sensitivity in detecting dermatophytes, regardless of sample quality. On the other hand, and as expected, optimal samples allowed the identification of a higher number of dermatophytes by conventional mycological diagnosis, compared to non-optimal samples. Finally, we have suggested several strategies for the practical use of such a kit in a medical laboratory for quick pathogen detection. © 2016 Blackwell Verlag GmbH.

  16. Detection of genetically modified soybean in crude soybean oil.

    PubMed

    Nikolić, Zorica; Vasiljević, Ivana; Zdjelar, Gordana; Ðorđević, Vuk; Ignjatov, Maja; Jovičić, Dušica; Milošević, Dragana

    2014-02-15

    In order to detect presence and quantity of Roundup Ready (RR) soybean in crude oil extracted from soybean seed with a different percentage of GMO seed two extraction methods were used, CTAB and DNeasy Plant Mini Kit. The amplifications of lectin gene, used to check the presence of soybean DNA, were not achieved in all CTAB extracts of DNA, while commercial kit gave satisfactory results. Comparing actual and estimated GMO content between two extraction methods, root mean square deviation for kit is 0.208 and for CTAB is 2.127, clearly demonstrated superiority of kit over CTAB extraction. The results of quantification evidently showed that if the oil samples originate from soybean seed with varying percentage of RR, it is possible to monitor the GMO content at the first stage of processing crude oil. Copyright © 2013 Elsevier Ltd. All rights reserved.

  17. Space Science

    NASA Image and Video Library

    2003-06-07

    The heart of a colorimetric solid phase extractor (CSPE) test kit quickly measures the concentration of the biocides silver or iodine in astronauts’ drinking water to determine whether concentrations are safe. When 10 milliliters (ml) of water is drawn through the disk, the disk will turn color (yellow in this picture for iodine) indicating the presence of the biocides. The device could someday be used to test water safety at reservoirs and water treatment plants on Earth. (photo credit: Microanalytical Instrumentation Center, Iowa State University).

  18. Operational Analysis of Time-Optimal Maneuvering for Imaging Spacecraft

    DTIC Science & Technology

    2013-03-01

    imaging spacecraft. The analysis is facilitated through the use of AGI’s Systems Tool Kit ( STK ) software. An Analytic Hierarchy Process (AHP)-based...the Singapore-developed X-SAT imaging spacecraft. The analysis is facilitated through the use of AGI’s Systems Tool Kit ( STK ) software. An Analytic...89  B.  FUTURE WORK................................................................................. 90  APPENDIX A. STK DATA AND BENEFIT

  19. IMMUNOASSAY TEST KITS FOR ATRAZINE

    EPA Science Inventory

    The U.S. Environmental Protection Agency (EPA) Environmental Technology Verification (ETV) program evaluates the performance of innovative air, water, pollution prevention and monitoring technologies that have the potential to improve human health and the environment. This techn...

  20. Treatment of mining acidic leachates with indigenous limestone, Zimapan Mexico.

    PubMed

    Labastida, I; Armienta, M A; Lara-Castro, R H; Aguayo, A; Cruz, O; Ceniceros, N

    2013-11-15

    An experimental study to evaluate the potential of using indigenous limestones in a passive system to treat acid mine drainage, at a mining zone of Mexico was carried out. Chemical and mineralogical characteristics of four types of native rocks (KIT1, KIT2, KSS, QZ) showed distinct CaCO3 contents. Synthetic aqueous leachates from an old tailings impoundment had a pH of 2.18, 34 mg/L As, 705 mg/L Fetotal, and 3975 mg/L SO4(2-). To evaluate dissolution behavior of rocks, kinetic batch experiments with an acid Fe-rich solution were performed. Decaying kinetic constants adjusting H(+) concentration to a first order exponential process were: KIT1 (k = 2.89), KIT2 (k = 0.89) and KSS (k = 0.47). Infrared spectrum and XRD of precipitates showed schwertmannite formation. To determine As and heavy metals (Fe, Cd, Zn, Al) removal from the synthetic leachates, batch experiments using KIT1 were developed. Arsenic decreased from 34.00 mg/L to 0.04 mg/L, Fe and Al were totally removed, and concentrations of Zn and Cd decreased 88% and 91% respectively. Analyses by IR and SEM-EDS indicate that co-precipitation with Fe-Hydroxides formed upon leachate interaction with limestone is the main As removal process. Chamosite, identified by XRD may participate in the removal of Al, SiO2 and a fraction of Fe. Copyright © 2012 Elsevier B.V. All rights reserved.

  1. Nucleic acid stains as indicators of Giardia muris viability following cyst inactivation.

    PubMed

    Taghi-Kilani, R; Gyürék, L L; Millard, P J; Finch, G R; Belosevic, M

    1996-06-01

    A reliable viability assay for Giardia is required for the development of disinfection process design criteria and pathogen monitoring by water treatment utilities. Surveys of single-staining nucleic acid dyes (stain dead parasites only), and double-staining vital dye kits from Molecular Probes (stain live and dead parasites) were conducted to assess the viability of untreated, heat-killed, and chemically inactivated Giardia muris cysts. Nucleic acid staining results were compared to those of in vitro excystation and animal infectivity. Nucleic acid stain, designated as SYTO-9, was considered the best among the single-staining dyes for its ability to stain dead cysts brightly and its relatively slow decay rate of visible light emission following DNA binding. SYTO-9 staining was correlated to animal infectivity. A Live/Dead BacLight was found to be the better of 2 double-staining viability kits tested. Logarithmic survival ratios based on SYTO-9 and Live/Dead BacLight were compared to excystation and infectivity results for G. muris cysts exposed to ozone or free chlorine. The results indicate that SYTO-9 and Live/Dead BacLight staining is stable following treatment of cysts with chemical disinfectants.

  2. Resveratrol Improves Cell Cycle Arrest in Chronic Prostatitis Rats, by C-kit/SCF Suppression.

    PubMed

    He, Yi; Zeng, Huizhi; Yu, Yang; Zhang, Jiashu; Zeng, Xiaona; Gong, Fengtao; Liu, Qi; Yang, Bo

    2017-08-01

    Chronic prostatitis (CP) with complex pathogenesis is difficult for treatment. c-kit has been associated with the control of cell proliferation of prostate cells. This study aims to evaluate the role of resveratrol, an activator of Sirt1, in regulating the expression of c-kit in CP and investigate the consequent effects on cell cycle. Rat model of CP was established through subcutaneous injections of diphtheria-pertussis-tetanus vaccine and subsequently treated with resveratrol. Hematoxylin and eosin staining was performed to identify the histopathological changes in prostates. Western blotting and immunohistochemical staining examined the expression level of c-kit, stem cell factor (SCF), Sirt1, and cell cycle-associated proteins. The model group exhibited severe diffuse chronic inflammation, characterized by leukocyte infiltration and papillary frond protrusion into the gland cavities, and a notable increase in prostatic epithelial height. Gland lumen diameter was also significantly smaller; the activity of c-kit/SCF in the CP rats was increased significantly compared to the control group. Meanwhile, the cell cycle proteins are dysregulated significantly in CP rats. Resveratrol treatment significantly improved these factors by Sirt1 activation. Dysregulation of cell cycle was involved in the pathological processes of CP, which was improved after resveratrol treatment by the downregulation of c-kit/SCF by activating Sirt1.

  3. Multiplex method for initial complex testing of antibodies to blood transmitted diseases agents.

    PubMed

    Poltavchenko, Alexander G; Nechitaylo, Oleg V; Filatov, Pavel V; Ersh, Anna V; Gureyev, Vadim N

    2016-10-01

    Initial screening of donors and population at high risk of infection with blood transmitted diseases involves a number of analyses using monospesific diagnostic systems, and therefore is expensive labor- and time-consuming process. The goal of this work is to construct a multiplex test enabling to carry out rapid initial complex testing at a low price. The paper describes a kit making it possible to detect simultaneously antibodies to six agents of the most significant blood transmitted diseases: HIV virus, hepatitis B and C viruses, cytomegalovirus, T. pallidum and T. gondii in blood products. The kit comprises multiplex dot-immunoassay based on plane protein arrays (immune chips) using colloidal gold conjugates and silver development. It provides an opportunity to carry out complex analysis within 70min at room temperature, and there is no need of well-qualified personnel. We compared laboratory findings of the kit with monospecific kits for ELISA produced by two Russian commercial companies. Dot-assay results correlate well with data obtained using commercial kits for ELISA. Furthermore, multiplex analysis is quicker and cheaper in comparison with ELISA and can be carried out in non-laboratory conditions. The kit for multiplex dot-immunoassay of antibodies to blood transmitted agents can significantly simplify initial complex testing. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Analytical framework and tool kit for SEA follow-up

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

    Nilsson, Mans; Wiklund, Hans; Finnveden, Goeran

    2009-04-15

    Most Strategic Environmental Assessment (SEA) research and applications have so far neglected the ex post stages of the process, also called SEA follow-up. Tool kits and methodological frameworks for engaging effectively with SEA follow-up have been conspicuously missing. In particular, little has so far been learned from the much more mature evaluation literature although many aspects are similar. This paper provides an analytical framework and tool kit for SEA follow-up. It is based on insights and tools developed within programme evaluation and environmental systems analysis. It is also grounded in empirical studies into real planning and programming practices at themore » regional level, but should have relevance for SEA processes at all levels. The purpose of the framework is to promote a learning-oriented and integrated use of SEA follow-up in strategic decision making. It helps to identify appropriate tools and their use in the process, and to systematise the use of available data and knowledge across the planning organization and process. It distinguishes three stages in follow-up: scoping, analysis and learning, identifies the key functions and demonstrates the informational linkages to the strategic decision-making process. The associated tool kit includes specific analytical and deliberative tools. Many of these are applicable also ex ante, but are then used in a predictive mode rather than on the basis of real data. The analytical element of the framework is organized on the basis of programme theory and 'DPSIR' tools. The paper discusses three issues in the application of the framework: understanding the integration of organizations and knowledge; understanding planners' questions and analytical requirements; and understanding interests, incentives and reluctance to evaluate.« less

  5. Disposable collection kit for rapid and reliable collection of saliva.

    PubMed

    Yamaguchi, Masaki; Tezuka, Yuki; Takeda, Kazunori; Shetty, Vivek

    2015-01-01

    To describe and evaluate disposable saliva collection kit for rapid, reliable, and reproducible collection of saliva samples. The saliva collection kit comprised of a saliva absorbent swab and an extractor unit was used to retrieve whole saliva samples from 10 subjects. The accuracy and precision of the extracted volumes (3, 10, and 30 μl) were compared to similar volumes drawn from control samples obtained by passive drool. Additionally, the impact of kit collection method on subsequent immunoassay results was verified by assessing salivary cortisol levels in the samples and comparing them to controls. The recovered volumes for the whole saliva samples were 3.85 ± 0.28, 10.79 ± 0.95, and 31.18 ± 1.72 μl, respectively (CV = 8.76%) and 2.91 ± 0.19, 9.75 ± 0.43, and 29.64 ± 0.91 μl, respectively, (CV = 6.36%) for the controls. There was a close correspondence between the salivary cortisol levels from the saliva samples obtained by the collection kit and the controls (R(2)  > 0.96). The disposable saliva collection kit allows accurate and repeatable collection of fixed amounts of whole saliva and does not interfere with subsequent measurements of salivary cortisol. The simple collection process, lack of elaborate specimen recovery steps, and the short turnaround time (<3 min) should render the kit attractive to test subjects and researchers alike. © 2015 Wiley Periodicals, Inc.

  6. Decontamination of breast pump milk collection kits and related items at home and in hospital: guidance from a Joint Working Group of the Healthcare Infection Society & Infection Prevention Society.

    PubMed

    Price, E; Weaver, G; Hoffman, P; Jones, M; Gilks, J; O'Brien, V; Ridgway, G

    2016-03-01

    A variety of methods are in use for decontaminating breast pump milk collection kits and related items associated with infant feeding. This paper aims to provide best practice guidance for decontamination of this equipment at home and in hospital. It has been compiled by a joint Working Group of the Healthcare Infection Society and the Infection Prevention Society. The guidance has been informed by a search of the literature in Medline, the British Nursing Index, the Cumulative Index to Nursing & Allied Health Literature, Midwifery & Infant Care and the results of two surveys of UK neonatal units in 2002/3 and 2006, and of members of the Infection Prevention Society in 2014. Since limited good quality evidence was available from these sources much of the guidance represents good practice based on the consensus view of the Working Group. Breast pump milk collection kits should not be reused by different mothers unless they have been sterilized in a Sterile Services Department between these different users.When used by the same mother, a detergent wash followed by thorough rinsing and drying after each use gives acceptable decontamination for most circumstances, as long as it is performed correctly.Additional decontamination precautions to washing, rinsing and drying may be used if indicated by local risk assessments and on advice from the departmental clinicians and Infection Prevention and Control Teams. The microbiological quality of the rinse water is an important consideration, particularly for infants on neonatal units.If bottle brushes or breast/nipple shields are used, they should be for use by one mother only. Decontamination should be by the processes used for breast pump milk collection kits.Dummies (soothers, pacifiers or comforters) needed for non-nutritive sucking by infants on neonatal units, should be for single infant use. Manufacturers should provide these dummies ready-to-use and individually packaged. They must be discarded at least every 24 hours or immediately if soiled with anything other than the baby's saliva. No attempt should be made to decontaminate the dummies, either before or during use. This guidance provides practical recommendations to support the safe decontamination of breast pump milk collection kits for healthcare professionals to use and communicate to other groups such as parents and carers.

  7. Decontamination of breast pump milk collection kits and related items at home and in hospital: guidance from a Joint Working Group of the Healthcare Infection Society & Infection Prevention Society*

    PubMed Central

    Price, E; Weaver, G; Hoffman, P; Jones, M; Gilks, J; O’Brien, V; Ridgway, G

    2015-01-01

    Introduction: A variety of methods are in use for decontaminating breast pump milk collection kits and related items associated with infant feeding. This paper aims to provide best practice guidance for decontamination of this equipment at home and in hospital. It has been compiled by a joint Working Group of the Healthcare Infection Society and the Infection Prevention Society. Methods: The guidance has been informed by a search of the literature in Medline, the British Nursing Index, the Cumulative Index to Nursing & Allied Health Literature, Midwifery & Infant Care and the results of two surveys of UK neonatal units in 2002/3 and 2006, and of members of the Infection Prevention Society in 2014. Since limited good quality evidence was available from these sources much of the guidance represents good practice based on the consensus view of the Working Group. Key recommendations: Breast pump milk collection kits should not be reused by different mothers unless they have been sterilized in a Sterile Services Department between these different users. When used by the same mother, a detergent wash followed by thorough rinsing and drying after each use gives acceptable decontamination for most circumstances, as long as it is performed correctly. Additional decontamination precautions to washing, rinsing and drying may be used if indicated by local risk assessments and on advice from the departmental clinicians and Infection Prevention and Control Teams. The microbiological quality of the rinse water is an important consideration, particularly for infants on neonatal units. If bottle brushes or breast/nipple shields are used, they should be for use by one mother only. Decontamination should be by the processes used for breast pump milk collection kits. Dummies (soothers, pacifiers or comforters) needed for non-nutritive sucking by infants on neonatal units, should be for single infant use. Manufacturers should provide these dummies ready-to-use and individually packaged. They must be discarded at least every 24 hours or immediately if soiled with anything other than the baby’s saliva. No attempt should be made to decontaminate the dummies, either before or during use. Conclusion: This guidance provides practical recommendations to support the safe decontamination of breast pump milk collection kits for healthcare professionals to use and communicate to other groups such as parents and carers. PMID:28989455

  8. 78 FR 32535 - National Hurricane Preparedness Week, 2013

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-05-30

    ... supply kit with food, water, and medicine; taking time now to learn evacuation routes, and how workplaces... with the tide, families whose worlds were torn apart with the loss of a loved one. But in the aftermath...

  9. PCP IMMUMOASSAY TECHNOLOGIES - INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    Three enzyme-linked immunosorbent assay technologies for pentachlorophenol (PCP) testing in soil and water were evaluated. Penta RISc Test Systems (formerly ENSYS, Inc.), EnviroGard™ PCP Immunoassay Test Kit (Millipore Corp.), and Pentachlorophenol RaPID Assay (formerly Ohmicron ...

  10. 77 FR 64311 - Foreign-Trade Zone 176-Rockford, IL, Notification of Proposed Production Activity, AndersonBrecon...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-19

    ... procedures that applies to SURGIFLO[supreg] Hemostatic Matrix Kits and SURGIFLO[supreg] Hemostatic Matrix... from abroad include: Sterile water in vials/non-needled syringes, hemostatic gelatin matrix pouches...

  11. Chemical and isotopic database of water and gas from hydrothermal systems with an emphasis for the western United States

    USGS Publications Warehouse

    Mariner, R.H.; Venezky, D.Y.; Hurwitz, S.

    2006-01-01

    Chemical and isotope data accumulated by two USGS Projects (led by I. Barnes and R. Mariner) over a time period of about 40 years can now be found using a basic web search or through an image search (left). The data are primarily chemical and isotopic analyses of waters (thermal, mineral, or fresh) and associated gas (free and/or dissolved) collected from hot springs, mineral springs, cold springs, geothermal wells, fumaroles, and gas seeps. Additional information is available about the collection methods and analysis procedures.The chemical and isotope data are stored in a MySQL database and accessed using PHP from a basic search form below. Data can also be accessed using an Open Source GIS called WorldKit by clicking on the image to the left. Additional information is available about WorldKit including the files used to set up the site.

  12. Optimizing techniques to capture and extract environmental DNA for detection and quantification of fish.

    PubMed

    Eichmiller, Jessica J; Miller, Loren M; Sorensen, Peter W

    2016-01-01

    Few studies have examined capture and extraction methods for environmental DNA (eDNA) to identify techniques optimal for detection and quantification. In this study, precipitation, centrifugation and filtration eDNA capture methods and six commercially available DNA extraction kits were evaluated for their ability to detect and quantify common carp (Cyprinus carpio) mitochondrial DNA using quantitative PCR in a series of laboratory experiments. Filtration methods yielded the most carp eDNA, and a glass fibre (GF) filter performed better than a similar pore size polycarbonate (PC) filter. Smaller pore sized filters had higher regression slopes of biomass to eDNA, indicating that they were potentially more sensitive to changes in biomass. Comparison of DNA extraction kits showed that the MP Biomedicals FastDNA SPIN Kit yielded the most carp eDNA and was the most sensitive for detection purposes, despite minor inhibition. The MoBio PowerSoil DNA Isolation Kit had the lowest coefficient of variation in extraction efficiency between lake and well water and had no detectable inhibition, making it most suitable for comparisons across aquatic environments. Of the methods tested, we recommend using a 1.5 μm GF filter, followed by extraction with the MP Biomedicals FastDNA SPIN Kit for detection. For quantification of eDNA, filtration through a 0.2-0.6 μm pore size PC filter, followed by extraction with MoBio PowerSoil DNA Isolation Kit was optimal. These results are broadly applicable for laboratory studies on carps and potentially other cyprinids. The recommendations can also be used to inform choice of methodology for field studies. © 2015 John Wiley & Sons Ltd.

  13. Multi-allergen Quantitation and the Impact of Thermal Treatment in Industry-Processed Baked Goods by ELISA and Liquid Chromatography-Tandem Mass Spectrometry.

    PubMed

    Parker, Christine H; Khuda, Sefat E; Pereira, Marion; Ross, Mark M; Fu, Tong-Jen; Fan, Xuebin; Wu, Yan; Williams, Kristina M; DeVries, Jonathan; Pulvermacher, Brian; Bedford, Binaifer; Zhang, Xi; Jackson, Lauren S

    2015-12-16

    Undeclared food allergens account for 30-40% of food recalls in the United States. Compliance with ingredient labeling regulations and the implementation of effective manufacturing allergen control plans require the use of reliable methods for allergen detection and quantitation in complex food products. The objectives of this work were to (1) produce industry-processed model foods incurred with egg, milk, and peanut allergens, (2) compare analytical method performance for allergen quantitation in thermally processed bakery products, and (3) determine the effects of thermal treatment on allergen detection. Control and allergen-incurred cereal bars and muffins were formulated in a pilot-scale industry processing facility. Quantitation of egg, milk, and peanut in incurred baked goods was compared at various processing stages using commercial enzyme-linked immunosorbent assay (ELISA) kits and a novel multi-allergen liquid chromatography (LC)-tandem mass spectrometry (MS/MS) multiple-reaction monitoring (MRM) method. Thermal processing was determined to negatively affect the recovery and quantitation of egg, milk, and peanut to different extents depending on the allergen, matrix, and analytical test method. The Morinaga ELISA and LC-MS/MS quantitative methods reported the highest recovery across all monitored allergens, whereas the ELISA Systems, Neogen BioKits, Neogen Veratox, and R-Biopharm ELISA Kits underperformed in the determination of allergen content of industry-processed bakery products.

  14. Expression profiling of c-kit and its impact after esiRNA silencing during gonadal development in catfish.

    PubMed

    Laldinsangi, C; Senthilkumaran, B

    2018-04-03

    C-kit receptor is a member of a family of growth factor receptors that have tyrosine kinase activity, and are involved in the transduction of growth regulatory signals across plasma membrane by activation of its ligand, kitl/scf. The present study analysed mRNA and protein expression profiles of c-kit in the gonads of catfish, Clarias gariepinus, using real time PCR, in situ hybridization and immunohistochemistry. Tissue distribution analysis revealed higher expression mainly in the catfish gonads. Ontogeny studies showed minimal expression during early developmental stages and highest during 50-75 days post hatch, and the dimorphic expression in gonads decreased gradually till adulthood, which might suggest an important role for this gene around later stages of sex differentiation and gonadal development. Expression of C-kit was analysed at various phases of gonadal cycle in both male and female, which showed minimal expression during the resting phase, and higher expression in male compared to females during the pre-spawning phase. In vitro and in vivo induction using human chorionic gonadotropin elevated the expression of c-kit indicating the regulatory influence of hypothalamo-hypophyseal axis. In vivo transient gene silencing using c-kit-esiRNA in adult catfish during gonadal recrudescence showed a decrease in c-kit expression, which affected the expression level of germ cell meiotic marker sycp3, as well as several factors and steroidogenic enzyme genes involved in germ cell development. Decrease in the levels of serum 11-KT and T were also observed after esiRNA silencing. The findings of this study suggest that c-kit has an important role in the process of germ cell proliferation, development and maturation during gonadal development and recrudescence in catfish. Copyright © 2018. Published by Elsevier Inc.

  15. AMD3100 treatment attenuates pulmonary angiogenesis by reducing the c-kit (+) cells and its pro-angiogenic activity in CBDL rat lungs.

    PubMed

    Shen, Cheng-Cheng; Chen, Bing; Gu, Jian-Teng; Ning, Jiao-Lin; Zeng, Jing; Yi, Bin; Lu, Kai-Zhi

    2018-03-01

    Recent studies have shown that pulmonary angiogenesis is an important pathological process in the development of hepatopulmonary syndrome (HPS), and growing evidence has indicated that Stromal cell-derived factor 1/C-X-C chemokine receptor type 4 (SDF-1/CXCR4) axis is involved in pulmonary vascular disease by mediating the accumulation of c-kit+ cells. This study aimed to test the effect of AMD3100, an antagonist of CXCR4, in HPS pulmonary angiogenesis. Common bile duct ligation (CBDL) rats were used as experimental HPS model and were treated with AMD3100 (1.25mg/kg/day, i.p.) or 0.9% saline for 3weeks. The sham rats underwent common bile duct exposure without ligation. The c-kit+ cells accounts and its angiogenic-related functions, prosurvival signals, pulmonary angiogenesis and arterial oxygenation were analysed in these groups. Our results showed that pulmonary SDF-1/CXCR4, Akt, Erk and VEGF/VEGFR2 were significantly activated in CBDL rats, and the numbers of circulating and pulmonary c-kit+ cells were increased in CBDL rats compared with control rats. Additionally, the angiogenic-related functions of c-kit+ cells and pulmonary microvessel counts were also elevated in CBDL rats. CXCR4 inhibition reduced pulmonary c-kit+ cells and microvessel counts and improved arterial oxygenation within 3weeks in CBDL rats. The pulmonary prosurvival signals and pro-angiogenic activity of c-kit+ cells were also down-regulated in AMD3100-treated rats. In conclusion, AMD3100 treatment attenuated pulmonary angiogenesis in CBDL rats and prevented the development of HPS via reductions in pulmonary c-kit+ cells and inhibition of the prosurvival signals. Our study provides new insights in HPS treatment. Copyright © 2017. Published by Elsevier B.V.

  16. A Frameshift Mutation in KIT is Associated with  White Spotting in the Arabian Camel.

    PubMed

    Holl, Heather; Isaza, Ramiro; Mohamoud, Yasmin; Ahmed, Ayeda; Almathen, Faisal; Youcef, Cherifi; Gaouar, Semir; Antczak, Douglas F; Brooks, Samantha

    2017-03-09

    While the typical Arabian camel is characterized by a single colored coat, there are rare populations with white spotting patterns. White spotting coat patterns are found in virtually all domesticated species, but are rare in wild species. Theories suggest that white spotting is linked to the domestication process, and is occasionally associated with health disorders. Though mutations have been found in a diverse array of species, fewer than 30 genes have been associated with spotting patterns, thus providing a key set of candidate genes for the Arabian camel. We obtained 26 spotted camels and 24 solid controls for candidate gene analysis. One spotted and eight solid camels were whole genome sequenced as part of a separate project. The spotted camel was heterozygous for a frameshift deletion in KIT (c.1842delG, named KITW1 for White spotting 1), whereas all other camels were wild-type (KIT+/KIT+). No additional mutations unique to the spotted camel were detected in the EDNRB, EDN3, SOX10, KITLG, PDGFRA, MITF, and PAX3 candidate white spotting genes. Sanger sequencing of the study population identified an additional five kITW1/KIT+ spotted camels. The frameshift results in a premature stop codon five amino acids downstream, thus terminating KIT at the tyrosine kinase domain. An additional 13 spotted camels tested KIT+/KIT+, but due to phenotypic differences when compared to the KITW1/KIT+ camels, they likely represent an independent mutation. Our study suggests that there are at least two causes of white spotting in the Arabian camel, the newly described KITW1 allele and an uncharacterized mutation.

  17. Levels of use of an elementary school inquiry-based instructional innovation among a selected group of teacher participants in the Delaware Elementary Science Initiative

    NASA Astrophysics Data System (ADS)

    Bouchelle, Henry Ellsworth Wirt, III

    Science education in Delaware's public elementary and middle schools has experienced much change in recent years as a result of the adoption of state standards and, in particular, the adoption by school districts of the Smithsonian/National Science Resources Council-sponsored inquiry-based instruction modules as part of the "Elementary Science Initiative." As part of this adoption process, each participating elementary teacher and middle school science teacher receives extensive training in the use of several discrete science kits. The trainings include reinforcement and development of content knowledge, in addition to the modeling of and practice with complementary pedagogy. One measure of the effectiveness of the science kit training process (and perhaps the Initiative itself) is the teachers' levels of use of the Initiative. The purpose of this study was to determine the participating teachers' use of the science kit innovation through the use of the Concerns-based Adoption Model Levels of Use Questionnaire. Eight K--5 elementary classroom teachers who had completed at least three science kit trainings participated. The results of this study indicate that on the Overall Level of Use Rating Scale, teachers who had completed training in at least three science kits generally scored at the Routine (IVA) level. All of the teachers, regardless of the wide range in the number of years of experience, had achieved the Mechanical Use level in Overall (III) LoU, and 6 of the 8 participants (75%) were operating at no less than the Refinement (IVA) Overall LoU level.

  18. Therapeutic drug monitoring of infliximab: performance evaluation of three commercial ELISA kits.

    PubMed

    Schmitz, Ellen M H; van de Kerkhof, Daan; Hamann, Dörte; van Dongen, Joost L J; Kuijper, Philip H M; Brunsveld, Luc; Scharnhorst, Volkher; Broeren, Maarten A C

    2016-07-01

    Therapeutic drug monitoring (TDM) of infliximab (IFX, Remicade®) can aid to optimize therapy efficacy. Many assays are available for this purpose. However, a reference standard is lacking. Therefore, we evaluated the analytical performance, agreement and clinically relevant differences of three commercially available IFX ELISA kits on an automated processing system. The kits of Theradiag (Lisa Tracker Infliximab), Progenika (Promonitor IFX) and apDia (Infliximab ELISA) were implemented on an automated processing system. Imprecision was determined by triplicate measurements of patient samples on five days. Agreement was evaluated by analysis of 30 patient samples and four spiked samples by the selected ELISA kits and the in-house IFX ELISA of Sanquin Diagnostics (Amsterdam, The Netherlands). Therapeutic consequences were evaluated by dividing patients into four treatment groups using cut-off levels of 1, 3 and 7 μg/mL and determining assay concordance. Within-run and between-run imprecision were acceptable (≤12% and ≤17%, respectively) within the quantification range of the selected ELISA kits. The apDia assay had the best precision and agreement to target values. Statistically significant differences were found between all assays except between Sanquin Diagnostics and the Lisa Tracker assay. The Promonitor assay measured the lowest IFX concentrations, the apDia assay the highest. When patients were classified in four treatment categories, 70% concordance was achieved. Although all assays are suitable for TDM, significant differences were observed in both imprecision and agreement. Therapeutic consequences were acceptable when patients were divided in treatment categories, but this could be improved by assay standardization.

  19. Generation of Functional Blood Vessels from a Single c-kit+ Adult Vascular Endothelial Stem Cell

    PubMed Central

    Fang, Shentong; Wei, Jing; Pentinmikko, Nalle; Leinonen, Hannele; Salven, Petri

    2012-01-01

    In adults, the growth of blood vessels, a process known as angiogenesis, is essential for organ growth and repair. In many disorders including cancer, angiogenesis becomes excessive. The cellular origin of new vascular endothelial cells (ECs) during blood vessel growth in angiogenic situations has remained unknown. Here, we provide evidence for adult vascular endothelial stem cells (VESCs) that reside in the blood vessel wall endothelium. VESCs constitute a small subpopulation within CD117+ (c-kit+) ECs capable of undergoing clonal expansion while other ECs have a very limited proliferative capacity. Isolated VESCs can produce tens of millions of endothelial daughter cells in vitro. A single transplanted c-kit-expressing VESC by the phenotype lin−CD31+CD105+Sca1+CD117+ can generate in vivo functional blood vessels that connect to host circulation. VESCs also have long-term self-renewal capacity, a defining functional property of adult stem cells. To provide functional verification on the role of c-kit in VESCs, we show that a genetic deficit in endothelial c-kit expression markedly decreases total colony-forming VESCs. In vivo, c-kit expression deficit resulted in impaired EC proliferation and angiogenesis and retardation of tumor growth. Isolated VESCs could be used in cell-based therapies for cardiovascular repair to restore tissue vascularization after ischemic events. VESCs also provide a novel cellular target to block pathological angiogenesis and cancer growth. PMID:23091420

  20. Development of an incurred cornbread model for gluten detection by immunoassays.

    PubMed

    Sharma, Girdhari M; Khuda, Sefat E; Pereira, Marion; Slate, Andrew; Jackson, Lauren S; Pardo, Christopher; Williams, Kristina M; Whitaker, Thomas B

    2013-12-11

    Gluten that is present in food as a result of cross-contact or misbranding can cause severe health concerns to wheat-allergic and celiac patients. Immunoassays, such as enzyme-linked immunosorbent assay (ELISA) and lateral flow device (LFD), are commonly used to detect gluten traces in foods. However, the performance of immunoassays can be affected by non-assay-related factors, such as food matrix and processing conditions. Gluten (0-500 ppm) and wheat flour (20-1000 ppm) incurred cornbread was prepared at different incurred levels and baking conditions (204.4 °C for 20, 27, and 34 min) to study the accuracy and precision of gluten measurement by seven immunoassay kits (three LFD and four ELISA kits). The stability and immunoreactivity of gluten proteins, as measured by western blot using three different antibodies, were not adversely affected by the baking conditions. However, the gluten recovery varied depending upon the ELISA kit and the gluten source used to make the incurred cornbread, affecting the accuracy of gluten quantification (BioKits, 9-77%; Morinaga, 91-137%; R-Biopharm, 61-108%; and Romer Labs, 113-190%). Gluten recovery was reduced with increased baking time for most ELISA kits analyzed. Both the sampling and analytical variance increased with an increase in the gluten incurred level. The predicted analytical coefficient of variation associated with all ELISA kits was below 12% for all incurred levels, indicative of good analytical precision.

  1. Tumor Necrosis Factor-α Is Required for Mast Cell-Mediated Host Immunity Against Cutaneous Staphylococcus aureus Infection.

    PubMed

    Liu, Chao; Ouyang, Wei; Xia, Jingyan; Sun, Xiaoru; Zhao, Liying; Xu, Feng

    2018-06-05

    Mast cells (MCs) play a key role in immune process response to invading pathogens. This study assessed the involvement of MCs in controlling Staphylococcus aureus infection in a cutaneous infection model of MC-deficient (KitW-sh/W-sh) mice. KitW-sh/W-sh mice developed significantly larger skin lesions after the cutaneous S. aureus challenge, when compared to wild-type (WT) mice, while MC dysfunction reduced the inflammation response to S. aureus. The levels of tumor necrosis factor (TNF)-α in skin tissues were significantly decreased in KitW-sh/W-sh mice upon infection. Moreover, the exogenous administration of MCs or recombinant TNF-α effectively restored the immune response against S. aureus in KitW-sh/W-sh mice via the recruitment of neutrophils to the infected site. These results indicate that the effects of MC deficiency are largely attributed to the decrease in production of TNF-α in cutaneous S. aureus infection. In addition, S. aureus-induced MC activation was dependent on the c-kit receptor-activated phosphoinositide 3-kinase (PI3K)/AKT/P65-nuclear factor (NF-κB) pathway, which was confirmed by treatment with Masitinib (a c-kit receptor inhibitor), Wortmannin (a PI3K inhibitor), and pyrrolidine dithiocarbamate (a NF-κB inhibitor), respectively. The present study identifies the critical role of MCs in the host defense against S. aureus infection.

  2. Disposable Collection Kit for Rapid and Reliable Collection of Saliva

    PubMed Central

    Yamaguchi, Masaki; Tezuka, Yuki; Takeda, Kazunori; Shetty, Vivek

    2015-01-01

    Objectives To describe and evaluate disposable saliva collection kit for rapid, reliable, and reproducible collection of saliva samples. Methods The saliva collection kit comprised of a saliva absorbent swab and an extractor unit was used to retrieve whole saliva samples from 10 subjects. The accuracy and precision of the extracted volumes (3, 10, and 30 μl) were compared to similar volumes drawn from control samples obtained by passive drool. Additionally, the impact of kit collection method on subsequent immunoassay results was verified by assessing salivary cortisol levels in the samples and comparing them to controls. Results The recovered volumes for the whole saliva samples were 3.85 ± 0.28, 10.79 ± 0.95, and 31.18 ± 1.72 μl, respectively (CV = 8.76%) and 2.91 ± 0.19, 9.75 ± 0.43, and 29.64 ± 0.91 μl, respectively, (CV = 6.36%) for the controls. There was a close correspondence between the salivary cortisol levels from the saliva samples obtained by the collection kit and the controls (R2 > 0.96). Conclusions The disposable saliva collection kit allows accurate and repeatable collection of fixed amounts of whole saliva and does not interfere with subsequent measurements of salivary cortisol. The simple collection process, lack of elaborate specimen recovery steps, and the short turnaround time (<3 min) should render the kit attractive to test subjects and researchers alike. Am. J. Hum. Biol. 27:720–723, 2015. © 2015 The Authors American Journal of Human Biology Published by Wiley Periodicals, Inc. PMID:25754371

  3. MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore.

    PubMed

    Ren, Jian; Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

    2010-01-01

    During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0).

  4. MiCroKit 3.0: an integrated database of midbody, centrosome and kinetochore

    PubMed Central

    Liu, Zexian; Gao, Xinjiao; Jin, Changjiang; Ye, Mingliang; Zou, Hanfa; Wen, Longping; Zhang, Zhaolei; Xue, Yu; Yao, Xuebiao

    2010-01-01

    During cell division/mitosis, a specific subset of proteins is spatially and temporally assembled into protein super complexes in three distinct regions, i.e. centrosome/spindle pole, kinetochore/centromere and midbody/cleavage furrow/phragmoplast/bud neck, and modulates cell division process faithfully. Although many experimental efforts have been carried out to investigate the characteristics of these proteins, no integrated database was available. Here, we present the MiCroKit database (http://microkit.biocuckoo.org) of proteins that localize in midbody, centrosome and/or kinetochore. We collected into the MiCroKit database experimentally verified microkit proteins from the scientific literature that have unambiguous supportive evidence for subcellular localization under fluorescent microscope. The current version of MiCroKit 3.0 provides detailed information for 1489 microkit proteins from seven model organisms, including Saccharomyces cerevisiae, Schizasaccharomyces pombe, Caenorhabditis elegans, Drosophila melanogaster, Xenopus laevis, Mus musculus and Homo sapiens. Moreover, the orthologous information was provided for these microkit proteins, and could be a useful resource for further experimental identification. The online service of MiCroKit database was implemented in PHP + MySQL + JavaScript, while the local packages were developed in JAVA 1.5 (J2SE 5.0). PMID:19783819

  5. 46 CFR 160.151-21 - Equipment required for SOLAS A and SOLAS B inflatable liferafts.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ...). (g) Tin-opener (Regulation -III/38.5.1.7). Each sharp part of a tin-opener described by Regulation... fishing tackle must be in a kit approved by the Commandant under approval series 160.061. (r) Food rations.... (s) Drinking water (Regulation -III/38.5.1.19). The fresh water required by Regulation III/38.5.1.19...

  6. 46 CFR 160.151-21 - Equipment required for SOLAS A and SOLAS B inflatable liferafts.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ...). (g) Tin-opener (Regulation -III/38.5.1.7). Each sharp part of a tin-opener described by Regulation... fishing tackle must be in a kit approved by the Commandant under approval series 160.061. (r) Food rations.... (s) Drinking water (Regulation -III/38.5.1.19). The fresh water required by Regulation III/38.5.1.19...

  7. Encapsulation of ultrafine metal-oxide nanoparticles within mesopores for biomass-derived catalytic applications† †Electronic supplementary information (ESI) available: Experimental section, additional characterization and reaction results. See DOI: 10.1039/c7sc04724j

    PubMed Central

    Fang, Ruiqi; Tian, Panliang; Yang, Xianfeng

    2018-01-01

    The development of efficient encapsulation strategies has attracted intense interest for preparing highly active and stable heterogeneous metal catalysts. However, issues related to low loadings, costly precursors and complex synthesis processes restrict their potential applications. Herein, we report a novel and general strategy to encapsulate various ultrafine metal-oxides nanoparticles (NPs) into the mesoporous KIT-6. The synthesis is facile, which only involves self-assembly of a metal–organic framework (MOF) precursor in the silica mesopores and a subsequent calcination process to transform the MOF into metal-oxide NPs. After the controlled calcination, the metal-oxide NPs produced from MOF decomposition are exclusively confined and uniformly distributed in the mesopores of KIT-6 with high metal loadings. Benefitting from the encapsulation effects, as-synthesized Co@KIT-6 materials exhibit superior catalytic activity and recycling stability in biomass-derived HMF oxidation under mild reaction conditions. PMID:29675231

  8. Fast and simple DNA extraction from saliva and sperm cells obtained from the skin or isolated from swabs.

    PubMed

    von Wurmb-Schwark, Nicole; Mályusz, Victoria; Fremdt, Heike; Koch, Christine; Simeoni, Eva; Schwark, Thorsten

    2006-05-01

    The forensic scientist often has to cope with problematic samples from the crime scene due to their minute size and thus the low amount of extractable DNA. The retrieval of DNA from swabs taken from the surface of the skin, for example, in cases of strangulation, can be especially difficult. We systematically investigated swabs taken from the skin (to obtain a genetic profile from the victim and also from a possible offender) and from sperm cell containing swabs using two extraction kits: the Invisorb forensic and the Invisorb spin swab kit (both Invitek, Germany). DNA quality and quantity were tested on ethidium bromide containing agarose gels and in a highly sensitive duplex-PCR, which amplifies fragments specific for mitochondrial and nuclear DNA. Absolute quantification was done using real time PCR. Samples, which were positive in the duplex-PCR, were also employed to genetic fingerprinting using the Powerplex ES and the AmpFlSTRIdentifiler(TM) kits. Our study shows that the easy-to-use Invisorb spin swab kit is very suitable for DNA isolation from swabs taken from the skin and also from sperm cells. Retrieval of cells from the skin with swabs moistened in extraction buffer, not in distilled water, led to a significant higher DNA yield.

  9. Deployment of the DosiKit System Under Operational Conditions: Experience From a French Defense National Nuclear Exercise.

    PubMed

    Entine, F; Bensimon Etzol, J; Bettencourt, C; Dondey, M; Michel, X; Gagna, G; Gellie, G; Corre, Y; Ugolin, N; Chevillard, S; Amabile, J-C

    2018-07-01

    Estimation of the dose received by accidentally irradiated victims is based on a tripod: clinical, biological, and physical dosimetry. The DosiKit system is an operational and mobile biodosimetry device allowing the measurement of external irradiation directly on the site of a radiological accident. This tool is based on capillary blood sample and hair follicle collection. The aim is to obtain a whole-body and local-surface dose assessment. This paper is about the technical evaluation of the DosiKit; the analytical process and scientific validation are briefly described. The Toulon exercise scenario was based on a major accident involving the reactor of a nuclear attack submarine. The design of the scenario made it impossible for several players (firefighters, medical team) to leave the area for a long time, and they were potentially exposed to high dose rates. The DosiKit system was fully integrated into a deployable radiological emergency laboratory, and the response to operational needs was very satisfactory.

  10. Logistics Study -- Shelter Stockage.

    DTIC Science & Technology

    1979-11-01

    Supplies such as paper cups, toilet tissue, sanitary napkins , commode 4 chemicals, etc., must be provided. 4 WATER REQUIREMENTS Potable water and its...PROPOSED SANITATION KITS ITEM UI SK-III SK-IV SK-V Container ea 1 1 1 * Toilet Tissue ro 5 10 10 Can Opener ea 1 1 - Sanitary Napkins ea 36 60 60 Water...34 3.3.5 Unit No. 5. -- One set of assembly instructions as delineated in Figure 4. -he instructions shall be printed in black or blue ink on a sheet

  11. 49 CFR 173.161 - Chemical kits and first aid kits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 2 2010-10-01 2010-10-01 false Chemical kits and first aid kits. 173.161 Section... Class 7 § 173.161 Chemical kits and first aid kits. (a) Chemical kits and First aid kits must conform to... 10 kg. (b) Chemical kits and First aid kits are excepted from the specification packaging...

  12. Optimizing Medical Kits for Space Flight

    NASA Technical Reports Server (NTRS)

    Minard, Charles G.; FreiredeCarvalho, Mary H.; Iyengar, M. Sriram

    2010-01-01

    The Integrated Medical Model (IMM) uses Monte Carlo methodologies to predict the occurrence of medical events, their mitigation, and the resources required during space flight. The model includes two modules that utilize output from a single model simulation to identify an optimized medical kit for a specified mission scenario. This poster describes two flexible optimization routines built into SAS 9.1. The first routine utilizes a systematic process of elimination to maximize (or minimize) outcomes subject to attribute constraints. The second routine uses a search and mutate approach to minimize medical kit attributes given a set of outcome constraints. There are currently 273 unique resources identified that are used to treat at least one of 83 medical conditions currently in the model.

  13. Lessons from Jerusalem.

    PubMed

    Daniell, Z

    1993-01-01

    The Gihon spring can no longer meet Jerusalem's water needs due to population growth. It provides just a small amount of water to an Arab community near Jerusalem called Silwan. 3 pipelines carry water to Jerusalem from coastal and mountain aquifers whose water tables are falling. A 4th pipeline is being built. Nevertheless, Jerusalem has been able to reduce water use, e.g. water use was lower in 1991 than in 1983, even though its population grew 25%. An engineer from the Department of Water attributed the reduced water use to conservation. The 1987 drought reduced the water in one of the city's main water sources, the Sea of Galilee, to very low levels. City authorities first responded by physically improving the water system, e.g., repairing system blow out valves and reducing water pressure where it was more than 60 m pressure. Next, the department distributed pamphlets on how to adjust household toilets to use less water per flush. The schools replaced all toilets with the new dual system. The city advertised water conservation in newspapers and buses. The department clamped down on industries and businesses using a disproportionate 23% of all water. In 1987. these efforts reduced water consumption 8-10%. A temporary law banned irrigation during the day when evaporation is greatest. The city charged residents more money for water used for gardening. Some residents have switched from grass laws to less thirsty local vegetation. The newness and efficiency of Jerusalem's pumping system and the commitment to replacing leaky pipes also contributes to the city's success. Jerusalem loses only about 10% of its water to leaks while other cities lose 40-60% of its water to leaks. Currently, the department partially subsidizes water-saving kits to be distributed to all households at a cost less than $4. 20% of households now use these kits. The department hopes to add another 40% of households. This holy city could be a water conservation model for all people.

  14. Global 2000 Countdown Kit.

    ERIC Educational Resources Information Center

    Zero Population Growth, Inc., Washington, DC.

    Fourteen units for high school global education classes are based on "The Global 2000 Report to the President," which examines the relationships between worldwide population growth and resource and environmental consequences. Topics of the units are population; income; food; fisheries; forests; water; nonfuel minerals; energy; impacts on…

  15. 49 CFR 173.161 - Chemical kits and first aid kits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 2 2012-10-01 2012-10-01 false Chemical kits and first aid kits. 173.161 Section... Class 7 § 173.161 Chemical kits and first aid kits. (a) Applicability. Chemical kits and first aid kits... assigned to the chemical kit and first aid kit as a whole must be the most stringent packing group assigned...

  16. DNA extraction from benthic Cyanobacteria: comparative assessment and optimization.

    PubMed

    Gaget, V; Keulen, A; Lau, M; Monis, P; Brookes, J D

    2017-01-01

    Benthic Cyanobacteria produce toxic and odorous compounds similar to their planktonic counterparts, challenging the quality of drinking water supplies. The biofilm that benthic algae and other micro-organisms produce is a complex and protective matrix. Monitoring to determine the abundance and identification of Cyanobacteria, therefore, relies on molecular techniques, with the choice of DNA isolation technique critical. This study investigated which DNA extraction method is optimal for DNA recovery in order to guarantee the best DNA yield for PCR-based analysis of benthic Cyanobacteria. The conventional phenol-chloroform extraction method was compared with five commercial kits, with the addition of chemical and physical cell-lysis steps also trialled. The efficacy of the various methods was evaluated by measuring the quantity and quality of DNA by UV spectrophotometry and by quantitative PCR (qPCR) using Cyanobacteria-specific primers. The yield and quality of DNA retrieved with the commercial kits was significantly higher than that of DNA obtained with the phenol-chloroform protocol. Kits including a physical cell-lysis step, such as the MO BIO Power Soil and Biofilm kits, were the most efficient for DNA isolation from benthic Cyanobacteria. These commercial kits allow greater recovery and the elimination of dangerous chemicals for DNA extraction, making them the method of choice for the isolation of DNA from benthic mats. They also facilitate the extraction of DNA from benthic Cyanobacteria, which can help to improve the characterization of Cyanobacteria in environmental studies using qPCRs or population composition analysis using next-generation sequencing. © 2016 The Society for Applied Microbiology.

  17. Tool kit development to refine and visualize essential climate data and information for marine protected areas

    NASA Astrophysics Data System (ADS)

    Cecil, L.; Stachniewicz, J.; Shein, K. A.; Ansari, S.; Jarvis, C.

    2013-05-01

    Marine ecosystem responses to climate variability and change such as changing water temperature, water chemistry (e.g., pH, salinity), water level, or storminess may result in adverse impacts including mass mortality, loss of habitat, increased disease susceptibility, and trophic cascade feedbacks. Unfortunately, while marine ecosystem resource managers are aware of these threats, they often lack sufficient expertise with identifying, accessing and using the many large and complex climate data products that would inform ecosystem-scale climate impact assessments. NOAA's National Climatic Data Center (NCDC) has been working with the Gulf of the Farallones National Marine Sanctuary Ocean Climate Center to enhance and expand the functionality of NCDC's Weather and Climate Toolkit (WCT) to begin to address this limitation. The WCT is a freely available, Java-based user interface (http://www.ncdc.noaa.gov/oa/wct/) designed to access, analyze, and display a variety of NCDC's georeferenced climate data products (e.g., satellite data, radar, reanalysis datasets, in-situ observations). However, the WCT requires the user to have already identified a data set of interest and gained access to it. This can limit its utility by users who are not knowledgeable about which data sets are relevant to their needs and where those data sets can be found. The Integrated Marine Protected Area Climate Tools (IMPACT) prototype modification to the WCT addresses those requirements through an iterative process between climate scientists and resource managers. The WCT-IMPACT prototype couples a user query approach with a quasi-expert system that determines, retrieves, and loads the appropriate data products for visualization and analysis by the user. Relevant data products are identified based on the environmental variables in which ecosystem managers have indicated an importance to their ecosystems. To improve response time, the user, through the WCT-IMPACT interface, crops (or subsets) the larger gridded data products, such as NOAA's satellite Climate Data Records to the geographic boundaries of each included marine protected area (MPA). These clipped data sets are processed to produce MPA-specific analytics (e.g., files for averages, extremes, peaks over threshold, etc). Once a specific MPA has been selected, the associated data may be visualized, analyzed, and exported to other formats (e.g., netCDF, KML) from within the tool. The WCT-IMPACT tool kit will provide marine ecosystem managers with the capacity to answer such questions as what was the climate like during periods of optimal ecological health, or have climate conditions changed equally across an ecosystem's domain? The WCT-IMPACT extension is being developed specifically to address the needs of marine ecosystem managers to have access to relevant climate data and information for developing ecosystem-scale climate assessments, while retaining the ability for a WCT user to identify and access the full suite of georeferenced climate data provided by NCDC. In this tool kit development scheme, the need to coordinate with the resource managers is paramount and end user participation in an iterative process with the climate scientists is essential.

  18. Effect of using different U/S probe Standoff materials in image geometry for interventional procedures: the example of prostate

    PubMed Central

    Diamantopoulos, Stefanos; Butt, Saeed; Katsilieri, Zaira; Kefala, Vasiliki; Zogal, Pawel; Sakas, George; Baltas, Dimos

    2011-01-01

    Purpose This study investigates the distortion of geometry of catheters and anatomy in acquired U/S images, caused by utilizing various stand-off materials for covering a transrectal bi-planar ultrasound probe in HDR and LDR prostate brachytherapy, biopsy and other interventional procedures. Furthermore, an evaluation of currently established water-bath based quality assurance (QA) procedures is presented. Material and methods Image acquisitions of an ultrasound QA setup were carried out at 5 MHz and 7 MHz. The U/S probe was covered by EA 4015 Silicone Standoff kit, or UA0059 Endocavity balloon filled either with water or one of the following: 40 ml of Endosgel®, Instillagel®, Ultraschall gel or Space OAR™ gel. The differences between images were recorded. Consequently, the dosimetric impact of the observed image distortion was investigated, using a tissue equivalent ultrasound prostate phantom – Model number 053 (CIRS Inc., Norfolk, VA, USA). Results By using the EA 4015 Silicone Standoff kit in normal water with sound speed of 1525 m/s, a 3 mm needle shift was observed. The expansion of objects appeared in radial direction. The shift deforms also the PTV (prostate in our case) and other organs at risk (OARs) in the same way leading to overestimation of volume and underestimation of the dose. On the other hand, Instillagel® and Space OAR™ “shrinks” objects in an ultrasound image for 0.65 mm and 0.40 mm, respectively. Conclusions The use of EA 4015 Silicone Standoff kit for image acquisition, leads to erroneous contouring of PTV and OARs and reconstruction and placement of catheters, which results to incorrect dose calculation during prostate brachytherapy. Moreover, the reliability of QA procedures lies mostly in the right temperature of the water used for accurate simulation of real conditions of transrectal ultrasound imaging. PMID:23346130

  19. Comparison of Quantifiler(®) Trio and InnoQuant™ human DNA quantification kits for detection of DNA degradation in developed and aged fingerprints.

    PubMed

    Goecker, Zachary C; Swiontek, Stephen E; Lakhtakia, Akhlesh; Roy, Reena

    2016-06-01

    The development techniques employed to visualize fingerprints collected from crime scenes as well as post-development ageing may result in the degradation of the DNA present in low quantities in such evidence samples. Amplification of the DNA samples with short tandem repeat (STR) amplification kits may result in partial DNA profiles. A comparative study of two commercially available quantification kits, Quantifiler(®) Trio and InnoQuant™, was performed on latent fingerprint samples that were either (i) developed using one of three different techniques and then aged in ambient conditions or (ii) undeveloped and then aged in ambient conditions. The three fingerprint development techniques used were: cyanoacrylate fuming, dusting with black powder, and the columnar-thin-film (CTF) technique. In order to determine the differences between the expected quantities and actual quantities of DNA, manually degraded samples generated by controlled exposure of DNA standards to ultraviolet radiation were also analyzed. A total of 144 fingerprint and 42 manually degraded DNA samples were processed in this study. The results indicate that the InnoQuant™ kit is capable of producing higher degradation ratios compared to the Quantifiler(®) Trio kit. This was an expected result since the degradation ratio is a relative value specific for a kit based on the length and extent of amplification of the two amplicons that vary from one kit to the other. Additionally, samples with lower concentrations of DNA yielded non-linear relationships of degradation ratio with the duration of aging, whereas samples with higher concentrations of DNA yielded quasi-linear relationships. None of the three development techniques produced a noticeably different degradation pattern when compared to undeveloped fingerprints, and therefore do not impede downstream DNA analysis. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  20. Internal validation of the GlobalFiler™ Express PCR Amplification Kit for the direct amplification of reference DNA samples on a high-throughput automated workflow.

    PubMed

    Flores, Shahida; Sun, Jie; King, Jonathan; Budowle, Bruce

    2014-05-01

    The GlobalFiler™ Express PCR Amplification Kit uses 6-dye fluorescent chemistry to enable multiplexing of 21 autosomal STRs, 1 Y-STR, 1 Y-indel and the sex-determining marker amelogenin. The kit is specifically designed for processing reference DNA samples in a high throughput manner. Validation studies were conducted to assess the performance and define the limitations of this direct amplification kit for typing blood and buccal reference DNA samples on various punchable collection media. Studies included thermal cycling sensitivity, reproducibility, precision, sensitivity of detection, minimum detection threshold, system contamination, stochastic threshold and concordance. Results showed that optimal amplification and injection parameters for a 1.2mm punch from blood and buccal samples were 27 and 28 cycles, respectively, combined with a 12s injection on an ABI 3500xL Genetic Analyzer. Minimum detection thresholds were set at 100 and 120RFUs for 27 and 28 cycles, respectively, and it was suggested that data from positive amplification controls provided a better threshold representation. Stochastic thresholds were set at 250 and 400RFUs for 27 and 28 cycles, respectively, as stochastic effects increased with cycle number. The minimum amount of input DNA resulting in a full profile was 0.5ng, however, the optimum range determined was 2.5-10ng. Profile quality from the GlobalFiler™ Express Kit and the previously validated AmpFlSTR(®) Identifiler(®) Direct Kit was comparable. The validation data support that reliable DNA typing results from reference DNA samples can be obtained using the GlobalFiler™ Express PCR Amplification Kit. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  1. Oct4-GFP expression during transformation of gonocytes into spermatogonial stem cells in the perinatal mouse testis.

    PubMed

    Li, Ruili; Vannitamby, Amanda; Zhang, Jian-Guo; Fehmel, Emma L; Southwell, Bridget R; Hutson, John M

    2015-12-01

    In cryptorchidism perinatal failure to switch off Oct4, a germ cell (GC) marker, may lead to carcinoma in situ. We aimed to analyze Oct4 expression during mouse gonocyte transformation into spermatogonial stem cells (SSC). Testes from OG2 (Oct4-promoter driven eGFP) mice at embryonic day (E) 17 and postnatal day P0-10 underwent immunohistochemistry and immunoblotting. Antibodies against MVH, AMH, Ki67, and c-Kit were visualized by confocal microscopy. Numbers of Oct4-GFP(+) GC and Oct4-GFP(-) GC/tubule were counted using ImageJ. Data were analyzed using nonparametric one-way ANOVA. GC from E17-P4 were Oct4-GFP(+). Numbers of Oct4-GFP(-) GC/tubule increased from P6-10, whereas Oct4-GFP(+) GC/tubule numbers remained similar between P6 and P10. Sertoli cells proliferated from E17-P10, whereas GC only proliferated from P2. Gonocytes (Oct4-GFP(+)/c-Kit(-)) central in tubules migrated to the basement membrane to become prospermatogonia (Oct4-GFP(+)/c-Kit(-)) and then SSC (Oct4-GFP(+)/c-Kit(+)) from day 4 and further developed into Oct4-GFP(-)/c-Kit(+) at P6. In Oct4-GFP mice both centrally located gonocytes and prospermatogonia located at the tubular basement membrane were Oct4-GFP(+)/c-Kit(-) before further developing into SSC (Oct4-GFP(+)/c-Kit(+)). This indicates that Oct4 is important in gonocyte transformation into SSC. Understanding this process will aid GC tumor diagnostics and fertility potential in boys with UDT undergoing orchidopexy. Copyright © 2015 Elsevier Inc. All rights reserved.

  2. Standardization and optimization of fluorescence in situ hybridization (FISH) for HER-2 assessment in breast cancer: A single center experience.

    PubMed

    Bogdanovska-Todorovska, Magdalena; Petrushevska, Gordana; Janevska, Vesna; Spasevska, Liljana; Kostadinova-Kunovska, Slavica

    2018-05-20

    Accurate assessment of human epidermal growth factor receptor 2 (HER-2) is crucial in selecting patients for targeted therapy. Commonly used methods for HER-2 testing are immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Here we presented the implementation, optimization and standardization of two FISH protocols using breast cancer samples and assessed the impact of pre-analytical and analytical factors on HER-2 testing. Formalin fixed paraffin embedded (FFPE) tissue samples from 70 breast cancer patients were tested for HER-2 using PathVysion™ HER-2 DNA Probe Kit and two different paraffin pretreatment kits, Vysis/Abbott Paraffin Pretreatment Reagent Kit (40 samples) and DAKO Histology FISH Accessory Kit (30 samples). The concordance between FISH and IHC results was determined. Pre-analytical and analytical factors (i.e., fixation, baking, digestion, and post-hybridization washing) affected the efficiency and quality of hybridization. The overall hybridization success in our study was 98.6% (69/70); the failure rate was 1.4%. The DAKO pretreatment kit was more time-efficient and resulted in more uniform signals that were easier to interpret, compared to the Vysis/Abbott kit. The overall concordance between IHC and FISH was 84.06%, kappa coefficient 0.5976 (p < 0.0001). The greatest discordance (82%) between IHC and FISH was observed in IHC 2+ group. A standardized FISH protocol for HER-2 assessment, with high hybridization efficiency, is necessary due to variability in tissue processing and individual tissue characteristics. Differences in the pre-analytical and analytical steps can affect the hybridization quality and efficiency. The use of DAKO pretreatment kit is time-saving and cost-effective.

  3. Gastroschisis in the rat model is associated with a delayed maturation of intestinal pacemaker cells and smooth muscle cells.

    PubMed

    Midrio, P; Faussone-Pellegrini, M S; Vannucchi, M G; Flake, A W

    2004-10-01

    A pacemaker system is required for peristalsis generation. The interstitial cells of Cajal (ICC) are considered the intestinal pacemaker, and are identified by expression of the c-kit gene--encoded protein. Gastroschisis is characterized by a severe gastrointestinal dysmotility in newborns. In spite of this clinical picture, few studies have focused on smooth muscle cells (SMC) morphology and none on ICC. Therefore, their morphology has been studied in fetuses at term in the rat model of gastroschisis. At 18.5 day's gestation (E18.5), 10 rat fetuses were killed, 10 underwent surgical creation of gastroschisis, and 10 underwent manipulation only. The small intestine of the latter 2 groups was harvested at E21.5. Specimens were processed for H&E, c-kit and actin (alpha smooth muscle antibody [alpha-SMA]) immunohistochemistry, and transmission electron microscopy (TEM). In the controls, SMC were c-kit+ and alpha-SMA+, with labeling intensity increasing by age. At E21.5, some cells around the Auerbach's plexus were more intensely c-kit+, and differentiating ICC were seen under TEM at this level. Gastroschisis fetuses had no c-kit+ cells referable to ICC. In the more damaged loops, SMC were very faintly c-kit+ and alpha-SMA+. Under TEM, there were few differentiated SMC and no presumptive ICC. In the less-damaged loops, SMC were faintly c-kit+ and alpha-SMA+ and had ultrastructural features intermediate between those of E18.5 and E21.5 controls; ICC were very immature. ICC and SMC differentiation is delayed in gastroschisis with the most damaged loops showing the most incomplete picture. These findings might help in understanding the delayed onset of peristalsis and the variable time-course of the recover seen in babies affected by gastroschisis.

  4. Development of an in-house fast real-time PCR method for detection of fish allergen in foods and comparison with a commercial kit.

    PubMed

    Herrero, Beatriz; Vieites, Juan M; Espiñeira, Montserrat

    2014-05-15

    Food allergy is recognised as an important human health problem. Fish represent one of the most important causes of food hypersensitivity reaction. Small amounts of the allergen can cause severe reactions in sensitive individuals, so correct labelling is essential to ensure the protection of consumers. The objective of the present work was to develop a reliable, sensitive and specific real-time PCR method for the detection of fish and traces of fish in all kind of products included those that have undergone aggressive treatments such as high temperature or pressure. This methodology was validated simulating products likely to contain this allergen and spiking them with fish cooking water. In addition, a comparison between the performance of in-house methodology and a commercial kit, both of them based on real-time PCR, was carried out. This work is relevant because it is the first, rapid real-time PCR method developed to date for the detection of fish in processed food products. The results obtained confirm the present assay is a useful tool in detecting fish and, therefore, minimising exposure and reducing incidences of allergic reaction to fish in contaminated products. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Development of a qualitative, multiplex real-time PCR kit for screening of genetically modified organisms (GMOs).

    PubMed

    Dörries, Hans-Henno; Remus, Ivonne; Grönewald, Astrid; Grönewald, Cordt; Berghof-Jäger, Kornelia

    2010-03-01

    The number of commercially available genetically modified organisms (GMOs) and therefore the diversity of possible target sequences for molecular detection techniques are constantly increasing. As a result, GMO laboratories and the food production industry currently are forced to apply many different methods to reliably test raw material and complex processed food products. Screening methods have become more and more relevant to minimize the analytical effort and to make a preselection for further analysis (e.g., specific identification or quantification of the GMO). A multiplex real-time PCR kit was developed to detect the 35S promoter of the cauliflower mosaic virus, the terminator of the nopaline synthase gene of Agrobacterium tumefaciens, the 35S promoter from the figwort mosaic virus, and the bar gene of the soil bacterium Streptomyces hygroscopicus as the most widely used sequences in GMOs. The kit contains a second assay for the detection of plant-derived DNA to control the quality of the often processed and refined sample material. Additionally, the plant-specific assay comprises a homologous internal amplification control for inhibition control. The determined limits of detection for the five assays were 10 target copies/reaction. No amplification products were observed with DNAs of 26 bacterial species, 25 yeasts, 13 molds, and 41 not genetically modified plants. The specificity of the assays was further demonstrated to be 100% by the specific amplification of DNA derived from reference material from 22 genetically modified crops. The applicability of the kit in routine laboratory use was verified by testing of 50 spiked and unspiked food products. The herein described kit represents a simple and sensitive GMO screening method for the reliable detection of multiple GMO-specific target sequences in a multiplex real-time PCR reaction.

  6. FUELS IN SOIL TEST KIT: FIELD USE OF DIESEL DOG SOIL TEST KITS

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

    Susan S. Sorini; John F. Schabron; Joseph F. Rovani, Jr.

    Western Research Institute (WRI) has developed a new commercial product ready for technology transfer, the Diesel Dog{reg_sign} Portable Soil Test Kit, for performing analysis of fuel-contaminated soils in the field. The technology consists of a method developed by WRI (U.S. Patents 5,561,065 and 5,976,883) and hardware developed by WRI that allows the method to be performed in the field (patent pending). The method is very simple and does not require the use of highly toxic reagents. The aromatic components in a soil extract are measured by absorption at 254 nm with a field-portable photometer. WRI added significant value to themore » technology by taking the method through the American Society for Testing and Materials (ASTM) approval and validation processes. The method is designated as ASTM Method D 5831-96, Standard Test Method for Screening Fuels in Soils. This ASTM designation allows the method to be used for federal compliance activities. In June 2001, the Diesel Dog technology won an American Chemical Society Regional Industrial Innovations Award. To gain field experience with the new technology, Diesel Dog kits have been used for a variety of site evaluation and cleanup activities. Information gained from these activities has led to improvements in hardware configurations and additional insight into correlating Diesel Dog results with results from laboratory methods. The Wyoming Department of Environmental Quality (DEQ) used Diesel Dog Soil Test Kits to guide cleanups at a variety of sites throughout the state. ENSR, of Acton, Massachusetts, used a Diesel Dog Portable Soil Test Kit to evaluate sites in the Virgin Islands and Georgia. ChemTrack and the U.S. Army Corps of Engineers successfully used a test kit to guide excavation at an abandoned FAA fuel-contaminated site near Fairbanks, Alaska. Barenco, Inc. is using a Diesel Dog Portable Soil Test Kit for site evaluations in Canada. A small spill of diesel fuel was cleaned up in Laramie, Wyoming using a Diesel Dog Soil Test Kit.« less

  7. Interest of the Ergo-Kit(®) for the clinical practice of the occupational physician. A study of 149 patients recruited in a rehabilitation program.

    PubMed

    Caron, J; Ronzi, Y; Bodin, J; Richard, I; Bontoux, L; Roquelaure, Y; Petit, A

    2015-10-01

    Functional capacity evaluation is commonly used to assess the abilities of patients to perform some tasks. Ergo-Kit(®) is a validated tool assessing both functional capacities of patients and workplace demands. The objective of this study was to evaluate the relevance of the Ergo-Kit(®) data for occupational physicians during the return-to-work process. A retrospective and monocenter study was conducted on all patients included in a rehabilitation program and assessed with the Ergo-Kit(®) tool between 2005 and 2014. Workplace demands and patients' functional capacities were evaluated and confronted. Self-beliefs and perceived disability were also assessed and compared to the functional capacity evaluation. One hundred and forty-nine working-age patients (85 men, 64 women; 39±12 years) suffering from musculoskeletal disorders or other diseases were included. Main causes of mismatch between workplace demands and functional capacities were manual handling of loads, postures with arms away from the body and repetitive motions at work; sitting posture was correlated with a lesser physical workload; and Oswestry score was correlated with functional capacities evaluated by the Ergo-Kit(®). Ergo-Kit(®) is a relevant tool to assess the multidimensional aspects of workplace demands and functional capacities. It could be very helpful for occupational physicians to manage return-to-work. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

  8. Developing intra-curricular photonics educational material for secondary schools in Europe

    NASA Astrophysics Data System (ADS)

    Prasad, Amrita; Debaes, Nathalie; Fischer, Robert; Thienpont, Hugo

    2014-07-01

    There is an imminent shortage of skilled workforce facing Europe's hi-tech industries mainly due to the declining interest of young people in science and engineering careers. To avert this trend the European Union funded the development of the `Photonics Explorer' - an intra-curricular educational kit designed to engage, excite and educate students about the fascination of working with optics hands-on, in their own classrooms! Each kit equips teachers with class sets of experimental components provided within a supporting didactic framework based on guided inquiry based learning techniques. The material has been specifically designed to integrate into the curriculum and enhance and complement the teaching and learning of science in the classroom. The kits are provided free of charge to teachers, in conjunction with teacher training courses. The main challenge of this program was the development of educational material that seamlessly integrates into the various national curricula across Europe. To achieve this, the development process included a preparatory EU wide curricula survey and a special `Review and Revise' process bringing together the expertise of over 35 teachers and pedagogic experts. This paper reports on the results of the preparatory study which identified two specific age groups at secondary schools for photonics educational material, the didactic content of the Photonics Explorer kit resulting from a pan-European collaboration of key stakeholders, EU wide dissemination and sustainability of the program.

  9. Performance of the Extravehicular Mobility Unit (EMU) Airlock Coolant Loop Remediation (A/L CLR) Hardware - Final

    NASA Technical Reports Server (NTRS)

    Steele, John W.; Rector, Tony; Gazda, Daniel; Lewis, John

    2011-01-01

    An EMU water processing kit (Airlock Coolant Loop Recovery -- A/L CLR) was developed as a corrective action to Extravehicular Mobility Unit (EMU) coolant flow disruptions experienced on the International Space Station (ISS) in May of 2004 and thereafter. A conservative duty cycle and set of use parameters for A/L CLR use and component life were initially developed and implemented based on prior analysis results and analytical modeling. Several initiatives were undertaken to optimize the duty cycle and use parameters of the hardware. Examination of post-flight samples and EMU Coolant Loop hardware provided invaluable information on the performance of the A/L CLR and has allowed for an optimization of the process. The intent of this paper is to detail the evolution of the A/L CLR hardware, efforts to optimize the duty cycle and use parameters, and the final recommendations for implementation in the post-Shuttle retirement era.

  10. Energy Audit . . . Here's How.

    ERIC Educational Resources Information Center

    American School and University, 1983

    1983-01-01

    After establishing building use patterns and complaints, a consulting engineer's walkthrough energy audit begins with the exterior. Then heating/cooling system efficiency is checked with a flue gases kit. Efficient use of water heaters, lighting, teacher lounges, and food preparation and eating areas saves energy. Most effective conservation…

  11. A novel honeycomb cell assay kit designed for evaluating horizontal cell migration in response to functionalized self-assembling peptide hydrogels

    NASA Astrophysics Data System (ADS)

    Guan, Fengyi; Lu, Jiaju; Wang, Xiumei

    2017-03-01

    A clear understanding on cell migration behaviors contributes to designing novel biomaterials in tissue engineering and elucidating related tissue regeneration processes. Many traditional evaluation methods on cell migration including scratch assay and transwell migration assay possess all kinds of limitations. In this study, a novel honeycomb cell assay kit was designed and made of photosensitive resin by 3D printing. This kit has seven hexagonal culture chambers so that it can evaluate the horizontal cell migration behavior in response to six surrounding environments simultaneously, eliminating the effect of gravity on cells. Here this cell assay kit was successfully applied to evaluate endothelial cell migration cultured on self-assembling peptide (SAP) RADA (AcN-RADARADARADARADA-CONH2) nanofiber hydrogel toward different functionalized SAP hydrogels. Our results indicated that the functionalized RADA hydrogels with different concentration of bioactive motifs of KLT or PRG could induce cell migration in a dose-dependent manner. The total number and migration distance of endothelial cells on functionalized SAP hydrogels significantly increased with increasing concentration of bioactive motif PRG or KLT. Therefore, the honeycomb cell assay kit provides a simple, efficient and convenient tool to investigate cell migration behavior in response to multi-environments simultaneously.

  12. Airborne hygrometer calibration inter-comparison against a metrological water vapour standard

    NASA Astrophysics Data System (ADS)

    Smorgon, Denis; Boese, Norbert; Ebert, Volker

    2014-05-01

    Water vapour is the most important atmospheric greenhouse gas, which causes a major feedback to warming and other changes in the climate system. Knowledge of the distribution of water vapour and its climate induced changes is especially important in the upper troposphere and lower stratosphere (UT/LS) where vapour plays a critical role in atmospheric radiative balance, cirrus cloud formation, and photochemistry. But, our understanding of water in the UT/LS is limited by significant uncertainties in current UT/LS water measurements. One of the most comprehensive inter-comparison campaigns for airborne hygrometers, termed AQUAVIT (AV1) [1], took place in 2007 at the AIDA chamber at the Karlsruhe Institute of Technology (KIT) in Germany. AV1 was a well-defined, referred, blind inter-comparison of 22 airborne field instruments from 17 international research groups. One major metrological deficit of AV1, however, was, that no traceable reference instrument participated in the inter-comparison experiments and that the calibration procedures of the participating instruments were not monitored or interrogated. Consequently a follow-up inter-comparison was organized in April 2013, which for the first time also provides a traceable link to the international humidity scale. This AQUAVIT2 (AV2) campaign (details see: http://www.imk-aaf.kit.edu/aquavit/index.php/Main_Page) was again located at KIT/AIDA and organised by an international organizing committee including KIT, PTB, FZJ and others. Generally AV2 is divided in two parallel comparisons: 1) AV2-A uses the AIDA chamber for a simultaneous comparison of all instruments (incl. sampling and in-situ instruments) over a broad range of conditions characteristic for the UT/LS; 2) AV2-B, about which this paper is reporting, is a sequential comparison of selected hygrometers and (when possible) their reference calibration infrastructures by means of a chilled mirror hygrometer traced back to the primary National humidity standard of PTB and a validated, two-pressure generator acting as a highly stable and reproducible source of water vapour. The aim of AV2-B was to perform an absolute, metrological comparison of the field instruments/calibration infrastructures to the metrological humidity scale, and to collect essential information about methods and procedures used by the atmospheric community for instrument calibration and validation, in order to investigate e.g. the necessity and possible comparability advantage by a standardized calibration procedure. The work will give an overview over the concept of the AV2-B inter-comparison, the various general measurement and calibration principles, and discuss the outcome and consequences of the comparison effort. The AQUAVIT effort is linked to the EMRP project METEOMET (ENV07) and partially supported by the EMRP and ENV07. The EMRP is jointly funded by the EMRP participating countries within EURAMET and the European Union. [1] H. Saathoff, C. Schiller, V. Ebert, D. W. Fahey, R.-S. Gao, O. Möhler, and the aquavit team, The AQUAVIT formal intercomparison of atmospheric water measurement methods, 5th General Assembly of the European Geosciences Union, 13-18 April 2008, Vienna, Austria Keywords: humidity, water vapour, inter-comparison, airborne instruments.

  13. On-farm biosecurity practices and causes of preweaning mortality in Canadian commercial mink kits.

    PubMed

    Compo, Nicole; Pearl, David L; Tapscott, Brian; Storer, Amanda; Hammermueller, Jutta; Brash, Marina; Turner, Patricia V

    2017-09-08

    Mink are an important animal commodity group in Canada and excessive kit mortality represents a significant loss to production. National biosecurity standards have been developed for Canadian mink farms, but it is unclear how well these standards have been implemented as there are no studies correlating management practices of mink producers with causes of death in mink kits. To that end, we surveyed Ontario mink producers on their biosecurity and management practices and conducted almost 5660 post mortem examinations on found-dead, preweaned kits to characterize mink farm biosecurity practices and causes of death in preweaned kits. We found that very few biosecurity and management practices were uniformly used by producers, despite good awareness of appropriate practices. Use of personal protective equipment was implemented by fewer than 50% of respondents, while control of mink shed access, disinfection of feed containers after use, and use of a rodent control program were the only practices implemented by greater than 70% of respondents. Only 18% of producers reported regular use of antimicrobials in feed or water, although 91% stated they used antimicrobials for treatment of bacterial diseases on a regular basis. On post mortem examination, no gross abnormalities were noted in 71% of the kits, 45% were thought to be stillborn or aborted, 27% had some form of abnormal fluid distribution in the body, and 2% had a congenital malformation. A subset of 69 gastrointestinal tract samples was submitted for bacterial culture, of which 45 samples yielded sufficient growth. Most interesting was the identification of Salmonella enterica serovar Heidelberg in 11% of samples. The results of this study will provide a benchmark for Canadian mink producers and their veterinarians, defining the areas to which greater attention should be given to ensure more rigorous biosecurity practices are in place. Ultimately, these improvements in practices may contribute to increased mink production and animal well-being.

  14. Crystal structure of a c-kit promoter quadruplex reveals the structural role of metal ions and water molecules in maintaining loop conformation.

    PubMed

    Wei, Dengguo; Parkinson, Gary N; Reszka, Anthony P; Neidle, Stephen

    2012-05-01

    We report here the 1.62 Å crystal structure of an intramolecular quadruplex DNA formed from a sequence in the promoter region of the c-kit gene. This is the first reported crystal structure of a promoter quadruplex and the first observation of localized magnesium ions in a quadruplex structure. The structure reveals that potassium and magnesium ions have an unexpected yet significant structural role in stabilizing particular quadruplex loops and grooves that is distinct from but in addition to the role of potassium ions in the ion channel at the centre of all quadruplex structures. The analysis also shows how ions cluster together with structured water molecules to stabilize the quadruplex arrangement. This particular quadruplex has been previously studied by NMR methods, and the present X-ray structure is in accord with the earlier topology assignment. However, as well as the observations of potassium and magnesium ions, the crystal structure has revealed a highly significant difference in the dimensions of the large cleft in the structure, which is a plausible target for small molecules. This difference can be understood by the stabilizing role of structured water networks.

  15. Comparison of two blanket surveys of arsenic in tubewells conducted 12 years apart in a 25 km(2) area of Bangladesh.

    PubMed

    van Geen, Alexander; Ahmed, Ershad B; Pitcher, Lynnette; Mey, Jacob L; Ahsan, Habibul; Graziano, Joseph H; Ahmed, Kazi Matin

    2014-08-01

    The arsenic (As) content of groundwater pumped from all tubewells within 61 contiguous villages of Araihazar, Bangladesh, was determined a first time in 2000-01 with laboratory measurements and a second time in 2012-13 using the ITS Arsenic Econo-Quick kit. The two surveys indicate that the total number of tubewells within the area almost doubled from 5560 to 10,879 over 12 years. The evolution of the distribution of well ages between the two surveys is consistent with a simple model that combines an annual increase of 42 wells/year in the rate of installations within the 61 villages starting in 1980 and a 7%/year rate of abandonment of wells as a function of well age. Colored placards were posted on each pumphead in 2012-13 on the basis of the kit results relative to the WHO guideline for As and the Bangladesh standard for As in drinking water: blue for As≤10 μg/L, green>10-50 μg/L, and red: >50 μg/L. According to quality-control samples collected from 502 tubewells for comparing the kit results with laboratory measurements, not a single well labeled blue in 2012-13 should have been labeled red and vice-versa. Field-kit testing in 2012-13 did not change the status of wells relative to the Bangladesh standard of 876 (87%) out of 1007 wells with a placard based on laboratory measurements in 2000-01 still attached to the pumphead. The high proportion of tubewells believed by households to be unsafe (66% out of 2041) that were still used for drinking and cooking in 2012-13 underlines the need for more widespread testing to identify low-As wells as an alternative source of drinking water. Copyright © 2013. Published by Elsevier B.V.

  16. Comparison of two blanket surveys of arsenic in tubewells conducted 12 years apart in a 25 km2 area of Bangladesh

    PubMed Central

    van Geen, Alexander; Sumon, Ershad B. A.; Pitcher, Lynnette; Mey, Jacob L.; Ahsan, Habibul; Graziano, Joseph H.; Ahmed, Kazi Matin

    2014-01-01

    The arsenic (As) content of groundwater pumped from all tubewells within 61 contiguous villages of Araihazar, Bangladesh, was determined a first time in 2000–01 with laboratory measurements and a second time in 2012–13 using the ITS Arsenic Econo-Quick kit. The two surveys indicate that the total number of tubewells within the area almost doubled from 5,560 to 10,879 over 12 years. The evolution of the distribution of well ages between the two surveys is consistent with a simple model that combines an annual increase of 42 wells/year in the rate of installations within the 61 villages starting in 1980 and a 7%/year rate of abandonment of wells as a function of well age. Colored placards were posted on each pumphead in 2012–2013 on the basis of the kit results relative to the WHO guideline for As and the Bangladesh standard for As in drinking water: blue for As ≤10 µg/L, green >10–50 µg/L, and red: >50 µg/L. According to quality-control samples collected from 502 tubewells for comparing the kit results with laboratory measurements, not a single well labeled blue in 2012–13 should have been labeled red and vice-versa. Field-kit testing in 2012–13 did not change the status of wells relative to the Bangladesh standard of 876 (87%) out of 1,007 wells with a placard based on laboratory measurements in 2000–01 still attached to the pumphead. The high proportion of tubewells believed by households to be unsafe (66% out of 2,041) that were still used for drinking and cooking in 2012–13 underlines the need for more widespread testing to identify low-As wells as an alternative source of drinking water. PMID:24438870

  17. 10 CFR 429.12 - General requirements applicable to certification reports.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... ceiling fan light kits, Residential showerheads, Residential faucets, Residential water closets, and.... 429.12 Section 429.12 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION CERTIFICATION, COMPLIANCE, AND... conservation standard set forth in parts 430 or 431, and annually thereafter on or before the dates provided in...

  18. Middle School Science Notes.

    ERIC Educational Resources Information Center

    School Science Review, 1983

    1983-01-01

    Describes a soap bubble motor, house insulation models, using hot-water bottles as heat sources, solar mobile, surface tension boat, evaporative cooling experiments, six activities on heat, and a magic trick based on friction. Also discusses using the Cambion Electronics Kit to introduce junior high students to the subject. (JM)

  19. 46 CFR 160.058-2 - Type.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 6 2010-10-01 2010-10-01 false Type. 160.058-2 Section 160.058-2 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-2 Type. (a) Desalter...

  20. Pilot Fullerton plans menu as packaged food and beverages float around him

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Pilot C. Gordon Fullerton, wearing the communications carrier assembly (ASSY) mini headset (HDST), beings food preparation on the middeck. Canned goods, sealed packages, beverage containers, etc are attached with velcro to meal tray assemblies (secured on middeck forward lockers) and freefloat around Fullerton. JSC water dispenser kit and portrait of G.W.S. Abbey appears behind Fullerton on port side bulkhead and potable water tank appears below him.

  1. Impact on arsenic exposure of a growing proportion of untested wells in Bangladesh

    PubMed Central

    2012-01-01

    Background In many areas of Bangladesh, it has been more than six years since a national campaign to test tubewells for arsenic (As) was conducted. Many households therefore draw their water for drinking and cooking from untested wells. Methods A household drinking water survey of 6646 households was conducted in Singair upazilla of Bangladesh. A subset of 795 untested wells used by 1000 randomly selected households was tested in the field by trained village workers with the Hach EZ kit, using an extended reaction time of 40 min, and in the laboratory by high-resolution inductively-coupled plasma-mass spectrometry (HR ICP-MS). Results The household survey shows that more than 80% of the wells installed since the national testing campaign in this area were untested. Less than 13% of the households with untested wells knew where a low-As well was located near their home. Village workers using the Hach EZ kit underestimated the As content of only 4 out of 795 wells relative to the Bangladesh standard. However, the As content of 168 wells was overestimated relative to the same threshold. Conclusion There is a growing need for testing tubewells in areas of Bangladesh where As concentrations in groundwater are elevated. This could be achieved by village workers trained to use a reliable field kit. Such an effort would result in a considerable drop in As exposure as it increases the opportunities for well switching by households. PMID:22353180

  2. MALDI-TOF identification of Gram-negative bacteria directly from blood culture bottles containing charcoal: Sepsityper® kits versus centrifugation-filtration method.

    PubMed

    Riederer, Kathleen; Cruz, Kristian; Shemes, Stephen; Szpunar, Susan; Fishbain, Joel T

    2015-06-01

    Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry has dramatically altered the way microbiology laboratories identify clinical isolates. Direct blood culture (BC) detection may be hampered, however, by the presence of charcoal in BC bottles currently in clinical use. This study evaluates an in-house process for extraction and MALDI-TOF identification of Gram-negative bacteria directly from BC bottles containing charcoal. Three hundred BC aliquots were extracted by a centrifugation-filtration method developed in our research laboratory with the first 96 samples processed in parallel using Sepsityper® kits. Controls were colonies from solid media with standard phenotypic and MALDI-TOF identification. The identification of Gram-negative bacteria was successful more often via the in-house method compared to Sepsityper® kits (94.7% versus 78.1%, P≤0.0001). Our in-house centrifugation-filtration method was further validated for isolation and identification of Gram-negative bacteria (95%; n=300) directly from BC bottles containing charcoal. Copyright © 2015 Elsevier Inc. All rights reserved.

  3. Radotinib induces high cytotoxicity in c-KIT positive acute myeloid leukemia cells.

    PubMed

    Heo, Sook-Kyoung; Noh, Eui-Kyu; Kim, Jeong Yi; Jo, Jae-Cheol; Choi, Yunsuk; Koh, SuJin; Baek, Jin Ho; Min, Young Joo; Kim, Hawk

    2017-06-05

    Previously, we reported that radotinib, a BCR-ABL1 tyrosine kinase inhibitor, induced cytotoxicity in acute myeloid leukemia (AML) cells. However, the effects of radotinib in the subpopulation of c-KIT-positive AML cells were unclear. We observed that low-concentration radotinib had more potent cytotoxicity in c-KIT-positive cells than c-KIT-negative cells from AML patients. To address this issue, cell lines with high c-KIT expression, HEL92.1.7, and moderate c-KIT expression, H209, were selected. HEL92.1.7 cells were grouped into intermediate and high c-KIT expression populations. The cytotoxicity of radotinib against the HEL92.1.7 cell population with intermediate c-KIT expression was not different from that of the population with high c-KIT expression. When H209 cells were grouped into c-KIT expression-negative and c-KIT expression-positive populations, radotinib induced cytotoxicity in the c-KIT-positive population, but not the c-KIT-negative population. Thus, radotinib induces cytotoxicity in c-KIT-positive cells, regardless of the c-KIT expression intensity. Therefore, radotinib induces significant cytotoxicity in c-KIT-positive AML cells, suggesting that radotinib is a potential target agent for the treatment of c-KIT-positive malignancies including AML. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Techni-kits and Techni-kit Building Systems

    NASA Technical Reports Server (NTRS)

    Callender, E. D.; Hartsough, C.; Morris, R. V.; Yamamoto, Y.

    1985-01-01

    Techni-kits consists of theories, methods, standards and computer based tools that assist in design of information-intensive systems. Techni-kit "building system" is techni-kit that builds techni-kits.

  5. International Space Station Common Cabin Air Assembly Water Separator On-Orbit Operation, Failure, and Redesign

    NASA Technical Reports Server (NTRS)

    Balistreri, Steven F., Jr.; Shaw, Laura A.; Laliberte, Yvon

    2010-01-01

    The ability to control the temperature and humidity of an environment or habitat is critical for human survival. These factors are important to maintaining human health and comfort, as well as maintaining mechanical and electrical equipment in good working order to support the human and to accomplish mission objectives. The temperature and humidity of the International Space Station (ISS) United States On-orbit Segment (USOS) cabin air is controlled by the Common Cabin Air Assembly (CCAA). The CCAA consists of a fan, a condensing heat exchanger (CHX), an air/water separator, temperature and liquid sensors, and electrical controlling hardware and software. The Water Separator (WS) pulls in air and water from the CHX, and centrifugally separates the mixture, sending the water to the condensate bus and the air back into the CHX outlet airstream. Two distinct early failures of the CCAA Water Separator in the Quest Airlock forced operational changes and brought about the re-design of the Water Separator to improve the useful life via modification kits. The on-orbit operational environment of the Airlock presented challenges that were not foreseen with the original design of the Water Separator. Operational changes were instituted to prolong the life of the third installed WS, while waiting for newly designed Water Separators to be delivered on-orbit. The modification kit design involved several different components of the Water Separator, including the innovative use of a fabrication technique to build the impellers used in Water Separators out of titanium instead of aluminum. The technique allowed for the cost effective production of the low quantity build. This paper will describe the failures of the Water Separators in the Quest Airlock, the operational constraints that were implemented to prolong the life of the installed Water Separators throughout the USOS, and the innovative re-design of the CCAA Water Separator.

  6. Direct Determination of ECD in ECD Kit: A Solid Sample Quantitation Method for Active Pharmaceutical Ingredient in Drug Product

    PubMed Central

    Chao, Ming-Yu; Liu, Kung-Tien; Hsia, Yi-Chih; Liao, Mei-Hsiu; Shen, Lie-Hang

    2011-01-01

    Technetium-99m ethyl cysteinate dimer (Tc-99m-ECD) is an essential imaging agent used in evaluating the regional cerebral blood flow in patients with cerebrovascular diseases. Determination of active pharmaceutical ingredient, that is, L-Cysteine, N, N′-1,2-ethanediylbis-, diethyl ester, dihydrochloride (ECD) in ECD Kit is a relevant requirement for the pharmaceutical quality control in processes of mass fabrication. We here presented a direct solid sample determination method of ECD in ECD Kit without sample dissolution to avoid the rapid degradation of ECD. An elemental analyzer equipped with a nondispersive infrared detector and a calibration curve of coal standard was used for the quantitation of sulfur in ECD Kit. No significant matrix effect was found. The peak area of coal standard against the amount of sulfur was linear over the range of 0.03–0.10 mg, with a correlation coefficient (r) of 0.9993. Method validation parameters were achieved to demonstrate the potential of this method. PMID:21687539

  7. Using technology to support HIV self-testing among MSM.

    PubMed

    LeGrand, Sara; Muessig, Kathryn E; Horvath, Keith J; Rosengren, Anna L; Hightow-Weidman, Lisa B

    2017-09-01

    Technology-based HIV self-testing (HST) interventions have the potential to improve access to HIV testing among gay, bisexual, and other MSM, as well as address concerns about HST use, including challenges with linkage to appropriate follow-up services. This review examines studies that use technology-based platforms to increase or improve the experience of HST among MSM. Seven published studies and eight funded studies were included in this review. Comprehensive prevention interventions with free HST kit distribution and interventions that provide free HST kits and support the HST process address a greater number of barriers (e.g., access, correct use of testing kits, and correct interpretation of results) than studies that only distribute free HST kits through technology-based platforms. By addressing HIV-testing barriers and specific HST concerns, these interventions address a critical need to improve first time and repeat testing rates among MSM. Additional research is needed to determine the efficacy of recent formative HST interventions. If proven efficacious, scale-up of these strategies have the potential to increase HIV testing among MSM via expanded HST uptake.

  8. JSC Pharmacy Services for Remote Operations

    NASA Technical Reports Server (NTRS)

    Stoner, Paul S.; Bayuse, Tina

    2005-01-01

    The Johnson Space Center Pharmacy began operating in March of 2003. The pharmacy serves in two main capacities: to directly provide medications and services in support of the medical clinics at the Johnson Space Center, physician travel kits for NASA flight surgeon staff, and remote operations, such as the clinics in Devon Island, Star City and Moscow; and indirectly provide medications and services for the International Space Station and Space Shuttle medical kits. Process changes that occurred and continued to evolve in the advent of the installation of the new JSC Pharmacy, and the process of stocking medications for each of these aforementioned areas will be discussed. Methods: The incorporation of pharmacy involvement to provide services for remote operations and supplying medical kits was evaluated. The first step was to review the current processes and work the JSC Pharmacy into the existing system. The second step was to provide medications to these areas. Considerations for the timeline of expiring medications for shipment are reviewed with each request. The third step was the development of a process to provide accountability for the medications. Results: The JSC Pharmacy utilizes a pharmacy management system to document all medications leaving the pharmacy. Challenges inherent to providing medications to remote areas were encountered. A process has been designed to incorporate usage into the electronic medical record upon return of the information from these remote areas. This is an evolving program and several areas have been identified for further improvement.

  9. Portable kit for the assessment of gait parameters in daily telerehabilitation.

    PubMed

    Giansanti, Daniele; Morelli, Sandra; Maccioni, Giovanni; Grigioni, Mauro

    2013-03-01

    When designing a complete process of daily telerehabilitation, it should be borne in mind that patients should be furnished with properly designed methodologies for executing specific motion tasks and the assessment of the relevant parameters. In general, such a process should comprehend three basic elements in both the hospital and the home: (a) instrumented walkways, (b) walking aids or supports, and (c) equipment for the assessment of parameters. The objective, with gait being the focus, of this study was thus to design a simple, portable kit-as an alternative to the complex and expensive instruments currently used-to be easily interfaced or integrated with the instrumented walkways and aids/supports both for self-monitoring while patients are exercising with their own aids and for clinical reporting. The proposed system is a portable kit that furnishes useful parameters with feedback to both the patient and the trainer/therapist. Capable of being integrated with the most common mechanical tools used in motion rehabilitation (handrail, scales, walkways, etc.), it constantly monitors and quantitatively assesses progress in rehabilitation care. It is composed of one step counter, photo-emitter detectors, one central unit for collecting and processing the telemetrically transmitted data, and a software interface. The system has been successfully validated on 16 subjects at the second level of the Tinetti test in a clinical application for both home and the hospital. The portable kit can be used with different rehabilitation tools and on varying ground rugosity. Advantages include (a) very low cost, when compared with optoelectronic solutions or other portable devices, (b) very high accuracy, also for subjects with imbalance problems, compared with other commercial solutions, and (c) integration (compatibility) with any rehabilitative tool.

  10. An economic analysis of adherence engineering to improve use of best practices during central line maintenance procedures.

    PubMed

    Nelson, Richard E; Angelovic, Aaron W; Nelson, Scott D; Gleed, Jeremy R; Drews, Frank A

    2015-05-01

    Adherence engineering applies human factors principles to examine non-adherence within a specific task and to guide the development of materials or equipment to increase protocol adherence and reduce human error. Central line maintenance (CLM) for intensive care unit (ICU) patients is a task through which error or non-adherence to protocols can cause central line-associated bloodstream infections (CLABSIs). We conducted an economic analysis of an adherence engineering CLM kit designed to improve the CLM task and reduce the risk of CLABSI. We constructed a Markov model to compare the cost-effectiveness of the CLM kit, which contains each of the 27 items necessary for performing the CLM procedure, compared with the standard care procedure for CLM, in which each item for dressing maintenance is gathered separately. We estimated the model using the cost of CLABSI overall ($45,685) as well as the excess LOS (6.9 excess ICU days, 3.5 excess general ward days). Assuming the CLM kit reduces the risk of CLABSI by 100% and 50%, this strategy was less costly (cost savings between $306 and $860) and more effective (between 0.05 and 0.13 more quality-adjusted life-years) compared with not using the pre-packaged kit. We identified threshold values for the effectiveness of the kit in reducing CLABSI for which the kit strategy was no longer less costly. An adherence engineering-based intervention to streamline the CLM process can improve patient outcomes and lower costs. Patient safety can be improved by adopting new approaches that are based on human factors principles.

  11. Evaluating the Medical Kit System for the International Space Station(ISS) - A Paradigm Revisited

    NASA Technical Reports Server (NTRS)

    Hailey, Melinda J.; Urbina, Michelle C.; Hughlett, Jessica L.; Gilmore, Stevan; Locke, James; Reyna, Baraquiel; Smith, Gwyn E.

    2010-01-01

    Medical capabilities aboard the International Space Station (ISS) have been packaged to help astronaut crew medical officers (CMO) mitigate both urgent and non-urgent medical issues during their 6-month expeditions. Two ISS crewmembers are designated as CMOs for each 3-crewmember mission and are typically not physicians. In addition, the ISS may have communication gaps of up to 45 minutes during each orbit, necessitating medical equipment that can be reliably operated autonomously during flight. The retirement of the space shuttle combined with ten years of manned ISS expeditions led the Space Medicine Division at the NASA Johnson Space Center to reassess the current ISS Medical Kit System. This reassessment led to the system being streamlined to meet future logistical considerations with current Russian space vehicles and future NASA/commercial space vehicle systems. Methods The JSC Space Medicine Division coordinated the development of requirements, fabrication of prototypes, and conducted usability testing for the new ISS Medical Kit System in concert with implementing updated versions of the ISS Medical Check List and associated in-flight software applications. The teams constructed a medical kit system with the flexibility for use on the ISS, and resupply on the Russian Progress space vehicle and future NASA/commercial space vehicles. Results Prototype systems were developed, reviewed, and tested for implementation. Completion of Preliminary and Critical Design Reviews resulted in a streamlined ISS Medical Kit System that is being used for training by ISS crews starting with Expedition 27 (June 2011). Conclusions The team will present the process for designing, developing, , implementing, and training with this new ISS Medical Kit System.

  12. Waterless Condensers for the Teaching Laboratory: An Adaptation of Traditional Glassware

    ERIC Educational Resources Information Center

    Baum, Erich W.; Esteb, John J.; Wilson, Anne M.

    2014-01-01

    A simple adaptation of traditional "chemistry kit" condensers for the organic chemistry teaching laboratory is described. These waterless condensers have been employed safely with most solvents. They can be easily fabricated, stored, and used in the same manner as water-cooled condensers. These condensers were utilized in several…

  13. 78 FR 34335 - Foreign-Trade Zone (FTZ) 141-Monroe County, New York; Notification of Proposed Production...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-06-07

    ... steel, fire and water resistant storage chests, gun safes, security safes, portable security safes, cash..., touchpads, plastic gun racks, keypad assemblies, panel lock assemblies, door backs, plastic trays, drawers..., wood gun shelf racks, cable assemblies, communication cables, gasket kits, door springs, metal handles...

  14. EQ Index, 1971.

    ERIC Educational Resources Information Center

    National Wildlife Federation, Washington, DC.

    To assess the status of our environmental quality and determine if we are winning or losing the campaign to save our environment, the National Wildlife Federation has developed this "EQ" kit. Multi-media materials help measure one's attitudes and activities in the areas of air, water, forests, wildlife, minerals, and soil conservation. Living…

  15. Literature Reference for Shiga and Shiga-like Toxin (Journal of Clinical Microbiology. 2007. 45(10): 3377–3380)

    EPA Pesticide Factsheets

    This procedure describes a rapid optical immunoassay for the detection of Stx-1 and Stx-2 using a commercially available kit for presumptive analysis of aerosol, solid, particulate, liquid and water samples contaminated with Shiga and Shiga-like toxin.

  16. Verification of ELISA Test Kits to Determine EDCs in Animal Feedlot Runoff

    EPA Science Inventory

    The key points discussed in this presentation are: (1) Climate change will very likely exacerbate the already stressed water supply in the U.S. Southeast, the only region that has experienced precipitation decline over >100 years; (2) The effect in precipitation change is locati...

  17. 46 CFR 160.058-5 - Labeling and marking.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 6 2013-10-01 2013-10-01 false Labeling and marking. 160.058-5 Section 160.058-5 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-5...

  18. 46 CFR 160.058-5 - Labeling and marking.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 6 2012-10-01 2012-10-01 false Labeling and marking. 160.058-5 Section 160.058-5 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-5...

  19. 46 CFR 160.058-5 - Labeling and marking.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 6 2014-10-01 2014-10-01 false Labeling and marking. 160.058-5 Section 160.058-5 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-5...

  20. 46 CFR 160.058-5 - Labeling and marking.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 6 2011-10-01 2011-10-01 false Labeling and marking. 160.058-5 Section 160.058-5 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-5...

  1. Teaching about the Child and World Environment: Elementary Teacher's Kit.

    ERIC Educational Resources Information Center

    Morris, Donald M.

    Three environmental education units relating environmental concerns to elementary school students' own experiences are described in this materials packet. Emphasis is on helping students become more sensitive to human and personal dimensions of environmental upset. Designed to teach children about themselves and their air, water, and land…

  2. 46 CFR 160.058-5 - Labeling and marking.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 6 2010-10-01 2010-10-01 false Labeling and marking. 160.058-5 Section 160.058-5 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) EQUIPMENT, CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Desalter Kits, Sea Water, for Merchant Vessels § 160.058-5...

  3. A decrease in ubiquitination and resulting prolonged life-span of KIT underlies the KIT overexpression-mediated imatinib resistance of KIT mutation-driven canine mast cell tumor cells.

    PubMed

    Kobayashi, Masato; Kuroki, Shiori; Kurita, Sena; Miyamoto, Ryo; Tani, Hiroyuki; Tamura, Kyoichi; Bonkobara, Makoto

    2017-10-01

    Overexpression of KIT is one of the mechanisms that contributes to imatinib resistance in KIT mutation-driven tumors. Here, the mechanism underlying this overexpression of KIT was investigated using an imatinib-sensitive canine mast cell tumor (MCT) line CoMS, which has an activating mutation in KIT exon 11. A KIT-overexpressing imatinib-resistant subline, rCoMS1, was generated from CoMS cells by their continuous exposure to increasing concentrations of imatinib. Neither a secondary mutation nor upregulated transcription of KIT was detected in rCoMS1 cells. A decrease in KIT ubiquitination, a prolonged KIT life-span, and KIT overexpression were found in rCoMS1 cells. These events were suppressed by withdrawal of imatinib and were re-induced by re‑treatment with imatinib. These findings suggest that imatinib elicited overexpression of KIT via suppression of its ubiquitination. These results also indicated that imatinib-induced overexpression of KIT in rCoMS1 cells was not a permanently acquired feature but was a reversible response of the cells. Moreover, the pan deubiquitinating enzyme inhibitor PR619 prevented imatinib induction of KIT overexpression, suggesting that the imatinib-induced decrease in KIT ubiquitination could be mediated by upregulation and/or activation of deubiquitinating enzyme(s). It may be possible that a similar mechanism of KIT overexpression underlies the acquisition of imatinib resistance in some human tumors that are driven by KIT mutation.

  4. Detecting an elusive invasive species: a diagnostic PCR to detect Burmese python in Florida waters and an assessment of persistence of environmental DNA.

    PubMed

    Piaggio, Antoinette J; Engeman, Richard M; Hopken, Matthew W; Humphrey, John S; Keacher, Kandy L; Bruce, William E; Avery, Michael L

    2014-03-01

    Recent studies have demonstrated that detection of environmental DNA (eDNA) from aquatic vertebrates in water bodies is possible. The Burmese python, Python bivittatus, is a semi-aquatic, invasive species in Florida where its elusive nature and cryptic coloration make its detection difficult. Our goal was to develop a diagnostic PCR to detect P. bivittatus from water-borne eDNA, which could assist managers in monitoring this invasive species. First, we used captive P. bivittatus to determine whether reptilian DNA could be isolated and amplified from water samples. We also evaluated the efficacy of two DNA isolation methods and two DNA extraction kits commonly used in eDNA preparation. A fragment of the mitochondrial cytochrome b gene from P. bivittatus was detected in all water samples isolated with the sodium acetate precipitate and the QIAamp DNA Micro Kit. Next, we designed P. bivittatus-specific primers and assessed the degradation rate of eDNA in water. Our primers did not amplify DNA from closely related species, and we found that P. bivittatus DNA was consistently detectable up to 96 h. Finally, we sampled water from six field sites in south Florida. Samples from five sites, where P. bivittatus has been observed, tested positive for eDNA. The final site was negative and had no prior documented evidence of P. bivittatus. This study shows P. bivittatus eDNA can be isolated from water samples; thus, this method is a new and promising technique for the management of invasive reptiles. Published 2013. This article is a U.S. Government work and is in the public domain in the USA.

  5. Identification of bacteria in drinking and purified water during the monitoring of a typical water purification system

    PubMed Central

    Penna, Vessoni Thereza Christina; Martins, Silva Alzira Maria; Mazzola, Priscila Gava

    2002-01-01

    Background A typical purification system that provides purified water which meets ionic and organic chemical standards, must be protected from microbial proliferation to minimize cross-contamination for use in cleaning and preparations in pharmaceutical industries and in health environments. Methodology Samples of water were taken directly from the public distribution water tank at twelve different stages of a typical purification system were analyzed for the identification of isolated bacteria. Two miniature kits were used: (i) identification system (api 20 NE, Bio-Mérieux) for non-enteric and non-fermenting gram-negative rods; and (ii) identification system (BBL crystal, Becton and Dickson) for enteric and non-fermenting gram-negative rods. The efficiency of the chemical sanitizers used in the stages of the system, over the isolated and identified bacteria in the sampling water, was evaluated by the minimum inhibitory concentration (MIC) method. Results The 78 isolated colonies were identified as the following bacteria genera: Pseudomonas, Flavobacterium and Acinetobacter. According to the miniature kits used in the identification, there was a prevalence of isolation of P. aeruginosa 32.05%, P. picketti (Ralstonia picketti) 23.08%, P. vesiculares 12.82%,P. diminuta 11.54%, F. aureum 6.42%, P. fluorescens 5.13%, A. lwoffi 2.56%, P. putida 2.56%, P. alcaligenes 1.28%, P. paucimobilis 1.28%, and F. multivorum 1.28%. Conclusions We found that research was required for the identification of gram-negative non-fermenting bacteria, which were isolated from drinking water and water purification systems, since Pseudomonas genera represents opportunistic pathogens which disperse and adhere easily to surfaces, forming a biofilm which interferes with the cleaning and disinfection procedures in hospital and industrial environments. PMID:12182763

  6. Oncogenic signaling by Kit tyrosine kinase occurs selectively on the Golgi apparatus in gastrointestinal stromal tumors

    PubMed Central

    Obata, Y; Horikawa, K; Takahashi, T; Akieda, Y; Tsujimoto, M; Fletcher, J A; Esumi, H; Nishida, T; Abe, R

    2017-01-01

    Gastrointestinal stromal tumors (GISTs) are caused by gain-of-function mutations in the Kit receptor tyrosine kinase. Most primary GIST patients respond to the Kit inhibitor imatinib, but this drug often becomes ineffective because of secondary mutations in the Kit kinase domain. The characteristic intracellular accumulation of imatinib-sensitive and -resistant Kit protein is well documented, but its relationship to oncogenic signaling remains unknown. Here, we show that in cancer tissue from primary GIST patients as well as in cell lines, mutant Kit accumulates on the Golgi apparatus, whereas normal Kit localizes to the plasma membrane (PM). In imatinib-resistant GIST with a secondary Kit mutation, Kit localizes predominantly on the Golgi apparatus. Both imatinib-sensitive and imatinib-resistant Kit (Kit(mut)) become fully auto-phosphorylated only on the Golgi and only if in a complex-glycosylated form. Kit(mut) accumulates on the Golgi during the early secretory pathway, but not after endocytosis. The aberrant kinase activity of Kit(mut) prevents its export from the Golgi to the PM. Furthermore, Kit(mut) on the Golgi signals and activates the phosphatidylinositol 3-kinase–Akt (PI3K–Akt) pathway, signal transducer and activator of transcription 5 (STAT5), and the Mek–Erk pathway. Blocking the biosynthetic transport of Kit(mut) to the Golgi from the endoplasmic reticulum inhibits oncogenic signaling. PM localization of Kit(mut) is not required for its signaling. Activation of Src-family tyrosine kinases on the Golgi is essential for oncogenic Kit signaling. These results suggest that the Golgi apparatus serves as a platform for oncogenic Kit signaling. Our study demonstrates that Kit(mut)’s pathogenicity is related to its mis-localization, and may offer a new strategy for treating imatinib-resistant GISTs. PMID:28192400

  7. Protein kinase C-δ-mediated recycling of active KIT in colon cancer.

    PubMed

    Park, Misun; Kim, Won Kyu; Song, Meiying; Park, Minhee; Kim, Hyunki; Nam, Hye Jin; Baek, Sung Hee; Kim, Hoguen

    2013-09-15

    Abnormal signaling through receptor tyrosine kinase (RTK) moieties is important in tumorigenesis and drug targeting of colorectal cancers. Wild-type KIT (WT-KIT), a RTK that is activated upon binding with stem cell factor (SCF), is highly expressed in some colon cancers; however, little is known about the functional role of SCF-dependent KIT activation in colon cancer pathogenesis. We aimed to elucidate the conditions and roles of WT-KIT activation in colon cancer tumorigenesis. Colorectal cancers with KIT expression were characterized by immunoblotting and immunohistochemistry. The biologic alterations after KIT-SCF binding were analyzed with or without protein kinase C (PKC) activation. We found that WT-KIT was expressed in a subset of colon cancer cell lines and was activated by SCF, leading to activation of downstream AKT and extracellular signal-regulated kinase (ERK) signaling pathways. We also showed that KIT expression gradually decreased, after prolonged SCF stimulation, due to lysosomal degradation. Degradation of WT-KIT after SCF binding was significantly rescued when PKC was activated. We also showed the involvement of activated PKC-δ in the recycling of WT-KIT. We further showed that a subset of colorectal cancers exhibit expressions of both WT-KIT and activated PKC-δ and that expression of KIT is correlated with poor patient survival (P = 0.004). Continuous downstream signal activation after KIT-SCF binding is accomplished through PKC-δ-mediated recycling of KIT. This sustained KIT activation may contribute to tumor progression in a subset of colon cancers with KIT expression and might provide the rationale for a therapeutic approach targeting KIT. ©2013 AACR.

  8. Technology Assessment and Strategy for Development of a Rapid Field Water Microbiology Test Kit

    DTIC Science & Technology

    1991-09-01

    fresh and marine waters (Santiago- Mercado and Hazen 1987; Valdes-Collazo et al. 1987; Perez-Rosas and Hazen 1988; Bermudez and Hazen 1988; Rivera et al...Santiago- Mercado and Hazen 1987; Valdes-Collazo et al. 1987). Investigators are constantly improving the coltform-MF technique as it is the... Mercado and Hazen 1987; Valdes-Collazo et al. 1987; Perez-Rosas and Hazen 1988; Bermudez and Hazen 1988; Rivera et al. 1988). The tropical conditions

  9. Symposium on Toxic Substance Control: Decontamination, April 22 - 24, 1980, Columbus, Ohio.

    DTIC Science & Technology

    1981-06-01

    standard decontaminants is used. TABLE 1. Standard Chemical Decontaminants Decontaminant Agents Used On STB Blister and nerve agents DS-2 All chemical... agents M258 Kit Sodium Hydroxide, Ethanol, G-Series nerve agents Phenol, Water Chloramine B, ZnCI2, Blister ana V-Series Ethanol, Water nerve agents A...is a point source alarm that actively samples ambient air and reacts to low concentrations of nerve agents . The M-8 alarm detector also detects several

  10. Documentation for MeshKit - Reactor Geometry (&mesh) Generator

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

    Jain, Rajeev; Mahadevan, Vijay

    2015-09-30

    This report gives documentation for using MeshKit’s Reactor Geometry (and mesh) Generator (RGG) GUI and also briefly documents other algorithms and tools available in MeshKit. RGG is a program designed to aid in modeling and meshing of complex/large hexagonal and rectilinear reactor cores. RGG uses Argonne’s SIGMA interfaces, Qt and VTK to produce an intuitive user interface. By integrating a 3D view of the reactor with the meshing tools and combining them into one user interface, RGG streamlines the task of preparing a simulation mesh and enables real-time feedback that reduces accidental scripting mistakes that could waste hours of meshing.more » RGG interfaces with MeshKit tools to consolidate the meshing process, meaning that going from model to mesh is as easy as a button click. This report is designed to explain RGG v 2.0 interface and provide users with the knowledge and skills to pilot RGG successfully. Brief documentation of MeshKit source code, tools and other algorithms available are also presented for developers to extend and add new algorithms to MeshKit. RGG tools work in serial and parallel and have been used to model complex reactor core models consisting of conical pins, load pads, several thousands of axially varying material properties of instrumentation pins and other interstices meshes.« less

  11. The Qiagen Investigator® Quantiplex HYres as an alternative kit for DNA quantification.

    PubMed

    Frégeau, Chantal J; Laurin, Nancy

    2015-05-01

    The Investigator® Quantiplex HYres kit was evaluated as a potential replacement for dual DNA quantification of casework samples. This kit was determined to be highly sensitive with a limit of quantification and limit of detection of 0.0049ng/μL and 0.0003ng/μL, respectively, for both human and male DNA, using full or half reaction volumes. It was also accurate in assessing the amount of male DNA present in 96 mock and actual casework male:female mixtures (various ratios) processed in this exercise. The close correlation between the male/human DNA ratios expressed in percentages derived from the Investigator® Quantiplex HYres quantification results and the male DNA proportion calculated in mixed AmpFlSTR® Profiler® Plus or AmpFlSTR® Identifiler® Plus profiles, using the Amelogenin Y peak and STR loci, allowed guidelines to be developed to facilitate decisions regarding when to submit samples to Y-STR rather than autosomal STR profiling. The internal control (IC) target was shown to be more sensitive to inhibitors compared to the human and male DNA targets included in the Investigator® Quantiplex HYres kit serving as a good quality assessor of DNA extracts. The new kit met our criteria of enhanced sensitivity, accuracy, consistency, reliability and robustness for casework DNA quantification. Crown Copyright © 2015. Published by Elsevier Ireland Ltd. All rights reserved.

  12. Cardiac c-Kit Biology Revealed by Inducible Transgenesis.

    PubMed

    Gude, Natalie A; Firouzi, Fareheh; Broughton, Kathleen M; Ilves, Kelli; Nguyen, Kristine P; Payne, Christina R; Sacchi, Veronica; Monsanto, Megan M; Casillas, Alexandria R; Khalafalla, Farid G; Wang, Bingyan J; Ebeid, David E; Alvarez, Roberto; Dembitsky, Walter P; Bailey, Barbara A; van Berlo, Jop; Sussman, Mark A

    2018-06-22

    Biological significance of c-Kit as a cardiac stem cell marker and role(s) of c-Kit+ cells in myocardial development or response to pathological injury remain unresolved because of varied and discrepant findings. Alternative experimental models are required to contextualize and reconcile discordant published observations of cardiac c-Kit myocardial biology and provide meaningful insights regarding clinical relevance of c-Kit signaling for translational cell therapy. The main objectives of this study are as follows: demonstrating c-Kit myocardial biology through combined studies of both human and murine cardiac cells; advancing understanding of c-Kit myocardial biology through creation and characterization of a novel, inducible transgenic c-Kit reporter mouse model that overcomes limitations inherent to knock-in reporter models; and providing perspective to reconcile disparate viewpoints on c-Kit biology in the myocardium. In vitro studies confirm a critical role for c-Kit signaling in both cardiomyocytes and cardiac stem cells. Activation of c-Kit receptor promotes cell survival and proliferation in stem cells and cardiomyocytes of either human or murine origin. For creation of the mouse model, the cloned mouse c-Kit promoter drives Histone2B-EGFP (enhanced green fluorescent protein; H2BEGFP) expression in a doxycycline-inducible transgenic reporter line. The combination of c-Kit transgenesis coupled to H2BEGFP readout provides sensitive, specific, inducible, and persistent tracking of c-Kit promoter activation. Tagging efficiency for EGFP+/c-Kit+ cells is similar between our transgenic versus a c-Kit knock-in mouse line, but frequency of c-Kit+ cells in cardiac tissue from the knock-in model is 55% lower than that from our transgenic line. The c-Kit transgenic reporter model reveals intimate association of c-Kit expression with adult myocardial biology. Both cardiac stem cells and a subpopulation of cardiomyocytes express c-Kit in uninjured adult heart, upregulating c-Kit expression in response to pathological stress. c-Kit myocardial biology is more complex and varied than previously appreciated or documented, demonstrating validity in multiple points of coexisting yet heretofore seemingly irreconcilable published findings. © 2018 American Heart Association, Inc.

  13. Mutant KIT as imatinib-sensitive target in metastatic sinonasal carcinoma.

    PubMed

    Dieter, S M; Heining, C; Agaimy, A; Huebschmann, D; Bonekamp, D; Hutter, B; Ehrenberg, K R; Fröhlich, M; Schlesner, M; Scholl, C; Schlemmer, H-P; Wolf, S; Mavratzas, A; Jung, C S; Gröschel, S; von Kalle, C; Eils, R; Brors, B; Penzel, R; Kriegsmann, M; Reuss, D E; Schirmacher, P; Stenzinger, A; Federspil, P A; Weichert, W; Glimm, H; Fröhling, S

    2017-01-01

    Sinonasal carcinomas (SNCs) comprise various rare tumor types that are characterized by marked histologic diversity and largely unknown molecular profiles, yet share an overall poor prognosis owing to an aggressive clinical course and frequent late-stage diagnosis. The lack of effective systemic therapies for locally advanced or metastatic SNC poses a major challenge to therapeutic decision making for individual patients. We here aimed to identify actionable genetic alterations in a patient with metastatic SNC whose tumor, despite all diagnostic efforts, could not be assigned to any known SNC category and was refractory to multimodal therapy. We used whole-exome and transcriptome sequencing to identify a KIT exon 11 mutation (c.1733_1735del, p.D579del) as potentially druggable target in this patient and carried out cancer hotspot panel sequencing to detect secondary resistance-conferring mutations in KIT. Furthermore, as a step towards clinical exploitation of the recently described signatures of mutational processes in cancer genomes, we established and applied a novel bioinformatics algorithm that enables supervised analysis of the mutational catalogs of individual tumors. Molecularly guided treatment with imatinib in analogy to the management of gastrointestinal stromal tumor (GIST) resulted in a dramatic and durable response with remission of nearly all tumor manifestations, indicating a dominant driver function of mutant KIT in this tumor. KIT dependency was further validated by a secondary KIT exon 17 mutation (c.2459_2462delATTCinsG, p.D820_S821delinsG) that was detected upon tumor progression after 10 months of imatinib treatment and provided a rationale for salvage therapy with regorafenib, which has activity against KIT exon 11/17 mutant GIST. These observations highlight the potential of unbiased genomic profiling for uncovering the vulnerabilities of individual malignancies, particularly in rare and unclassifiable tumors, and underscore that KIT exon 11 mutations represent tractable therapeutic targets across different histologies. © The Author 2016. Published by Oxford University Press on behalf of the European Society for Medical Oncology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  14. Basic Science Process Skills. An Inservice Workshop Kit: Outlines and Activities.

    ERIC Educational Resources Information Center

    Rowland, Paul; And Others

    A science process skill project was developed to help elementary teachers meet competency standards in New Mexico for teaching the process approach in their science classes. An outline of the process skills along with recommended activities are presented in this document. Performance objectives are identified and a sample activity form is…

  15. Advantage of using a home-made ELISA kit for detection of Helicobacter pylori infection over commercially imported kits.

    PubMed

    Mohammadi, M; Talebkhan, Y; Khalili, G; Mahboudi, F; Massarrat, S; Zamaninia, L; Oghalaei, A

    2008-01-01

    To evaluate a home-made ELISA kit for detection of Helicobacter pylori (Hp) infection and comparison of its immunologic criteria with those of foreign commercial kits. A home-made IgG ELISA kit was developed using soluble antigenic fractions of Hp proteins. Confirmed sera were tested and serological criteria were evaluated through assessment of 199 serum samples. The accuracy, sensitivity and specificity values of home-made kit were 92, 92 and 90.4%, respectively. These immunologic criteria for Trinity kit were 95.2, 95.2 and 95% in comparison with IBL kit (91.3, 92.2 and 88.5%), BIOHIT kit (72.4, 41.6 and 94.1%) and HelicoBlot2.1 (94.2, 93.4 and 100%). Kappa agreement assessment demonstrated that two of the imported ELISA kits had fair to moderate agreement with the home-made kit while the other one had a poor agreement value. Apart from comparable values between the home-made kit and the most efficient imported kit (Trinity) there was significant cost benefit. Therefore, we recommend the home-made kit as a suitable substitution for detection of Hp infection in the Iranian population.

  16. Evaluation of the kinase domain of c-KIT in canine cutaneous mast cell tumors

    PubMed Central

    Webster, Joshua D; Kiupel, Matti; Yuzbasiyan-Gurkan, Vilma

    2006-01-01

    Background Mutations in the c-KIT proto-oncogene have been implicated in the progression of several neoplastic diseases, including gastrointestinal stromal tumors and mastocytosis in humans, and cutaneous mast cell tumors (MCTs) in canines. Mutations in human mastocytosis patients primarily occur in c-KIT exon 17, which encodes a portion of its kinase domain. In contrast, deletions and internal tandem duplication (ITD) mutations are found in the juxtamembrane domain of c-KIT in approximately 15% of canine MCTs. In addition, ITD c-KIT mutations are significantly associated with aberrant KIT protein localization in canine MCTs. However, some canine MCTs have aberrant KIT localization but lack ITD c-KIT mutations, suggesting that other mutations or other factors may be responsible for aberrant KIT localization in these tumors. Methods In order to characterize the prevalence of mutations in the phospho-transferase portion of c-KIT's kinase domain in canine MCTs exons 16–20 of 33 canine MCTs from 33 dogs were amplified and sequenced. Additionally, in order to determine if mutations in c-KIT exon 17 are responsible for aberrant KIT localization in MCTs that lack juxtamembrane domain c-KIT mutations, c-KIT exon 17 was amplified and sequenced from 18 canine MCTs that showed an aberrant KIT localization pattern but did not have ITD c-KIT mutations. Results No mutations or polymorphisms were identified in exons 16–20 of any of the MCTs examined. Conclusion In conclusion, mutations in the phospho-transferase portion of c-KIT's kinase domain do not play an important role in the progression of canine cutaneous MCTs, or in the aberrant localization of KIT in canine MCTs. PMID:16579858

  17. Adult cardiac stem cells are multipotent and robustly myogenic: c-kit expression is necessary but not sufficient for their identification.

    PubMed

    Vicinanza, Carla; Aquila, Iolanda; Scalise, Mariangela; Cristiano, Francesca; Marino, Fabiola; Cianflone, Eleonora; Mancuso, Teresa; Marotta, Pina; Sacco, Walter; Lewis, Fiona C; Couch, Liam; Shone, Victoria; Gritti, Giulia; Torella, Annalaura; Smith, Andrew J; Terracciano, Cesare Mn; Britti, Domenico; Veltri, Pierangelo; Indolfi, Ciro; Nadal-Ginard, Bernardo; Ellison-Hughes, Georgina M; Torella, Daniele

    2017-12-01

    Multipotent adult resident cardiac stem cells (CSCs) were first identified by the expression of c-kit, the stem cell factor receptor. However, in the adult myocardium c-kit alone cannot distinguish CSCs from other c-kit-expressing (c-kit pos ) cells. The adult heart indeed contains a heterogeneous mixture of c-kit pos cells, mainly composed of mast and endothelial/progenitor cells. This heterogeneity of cardiac c-kit pos cells has generated confusion and controversy about the existence and role of CSCs in the adult heart. Here, to unravel CSC identity within the heterogeneous c-kit-expressing cardiac cell population, c-kit pos cardiac cells were separated through CD45-positive or -negative sorting followed by c-kit pos sorting. The blood/endothelial lineage-committed (Lineage pos ) CD45 pos c-kit pos cardiac cells were compared to CD45 neg (Lineage neg /Lin neg ) c-kit pos cardiac cells for stemness and myogenic properties in vitro and in vivo. The majority (~90%) of the resident c-kit pos cardiac cells are blood/endothelial lineage-committed CD45 pos CD31 pos c-kit pos cells. In contrast, the Lin neg CD45 neg c-kit pos cardiac cell cohort, which represents ⩽10% of the total c-kit pos cells, contain all the cardiac cells with the properties of adult multipotent CSCs. These characteristics are absent from the c-kit neg and the blood/endothelial lineage-committed c-kit pos cardiac cells. Single Lin neg c-kit pos cell-derived clones, which represent only 1-2% of total c-kit pos myocardial cells, when stimulated with TGF-β/Wnt molecules, acquire full transcriptome and protein expression, sarcomere organisation, spontaneous contraction and electrophysiological properties of differentiated cardiomyocytes (CMs). Genetically tagged cloned progeny of one Lin neg c-kit pos cell when injected into the infarcted myocardium, results in significant regeneration of new CMs, arterioles and capillaries, derived from the injected cells. The CSC's myogenic regenerative capacity is dependent on commitment to the CM lineage through activation of the SMAD2 pathway. Such regeneration was not apparent when blood/endothelial lineage-committed c-kit pos cardiac cells were injected. Thus, among the cardiac c-kit pos cell cohort only a very small fraction has the phenotype and the differentiation/regenerative potential characteristics of true multipotent CSCs.

  18. Colorimetric Solid Phase Extraction (CSPE): Using Color to Monitor Spacecraft Water Quality

    NASA Technical Reports Server (NTRS)

    Gazda, Daniel B.; Nolan, Daniel J.; Rutz, Jeffrey A.; Schultz, John R.; Siperko, Lorraine M.; Porter, Marc D.; Lipert, Robert J.; Flint, Stephanie M.; McCoy, J. Torin

    2010-01-01

    In August 2009, an experimental water quality monitoring kit based on Colorimetric Solid Phase Extraction (CSPE) technology was delivered to the International Space Station (ISS). The kit, called the Colorimetric Water Quality Monitoring Kit (CWQMK), was launched as a Station Development Test Objective (SDTO) experiment to evaluate the suitability of CSPE technology for routine use monitoring water quality on the ISS. CSPE is a sorption-spectrophotometric technique that combines colorimetric reagents, solid-phase extraction, and diffuse reflectance spectroscopy to quantify trace analytes in water samples. In CSPE, a known volume of sample is metered through a membrane disk that has been impregnated with an analyte-specific colorimetric reagent and any additives required to optimize the formation of the analyte-reagent complex. As the sample flows through the membrane disk, the target analyte is selectively extracted, concentrated, and complexed. Formation of the analyte-reagent complex causes a detectable change in the color of the membrane disk that is proportional to the amount of analyte present in the sample. The analyte is then quantified by measuring the color of the membrane disk surface using a hand-held diffuse reflectance spectrophotometer (DRS). The CWQMK provides the capability to measure the ionic silver (Ag +) and molecular iodine (I2) in water samples on-orbit. These analytes were selected for the evaluation of CSPE technology because they are the biocides used in the potable water storage and distribution systems on the ISS. Biocides are added to the potable water systems on spacecraft to inhibit microbial growth. On the United States (US) segment of the ISS molecular iodine serves as the biocide, while the Russian space agency utilizes silver as a biocide in their systems. In both cases, the biocides must be maintained at a level sufficient to control bacterial growth, but low enough to avoid any negative effects on crew health. For example, the presence of high levels of iodine in water can cause taste and odor issues that result in decreased water consumption by the crew. There are also concerns about potential impacts on thyroid function following exposure to high levels of iodine. With silver, there is a risk of developing argyria, an irreversible blue-gray discoloration of the skin, associated with long term consumption of water containing high concentrations of silver. The need to ensure that safe, effective levels of biocide are maintained in the potable water systems on the ISS provides a perfect platform for evaluating the suitability of CSPE technology for in-flight water quality monitoring. This paper provides an overview of CSPE technology and details on the silver and iodine methods used in the CWQMK. It also reports results obtained during in-flight analyses performed with the CWQMK and briefly discusses other potential applications for CSPE technology in both the spacecraft and terrestrial environments.

  19. Pathologic Stimulus Determines Lineage Commitment of Cardiac C-kit+ Cells.

    PubMed

    Chen, Zhongming; Zhu, Wuqiang; Bender, Ingrid; Gong, Wuming; Kwak, Il-Youp; Yellamilli, Amritha; Hodges, Thomas J; Nemoto, Natsumi; Zhang, Jianyi; Garry, Daniel J; van Berlo, Jop H

    2017-12-12

    Although cardiac c-kit + cells are being tested in clinical trials, the circumstances that determine lineage differentiation of c-kit + cells in vivo are unknown. Recent findings suggest that endogenous cardiac c-kit + cells rarely contribute cardiomyocytes to the adult heart. We assessed whether various pathological stimuli differentially affect the eventual cell fates of c-kit + cells. We used single-cell sequencing and genetic lineage tracing of c-kit + cells to determine whether various pathological stimuli would result in different fates of c-kit + cells. Single-cell sequencing of cardiac CD45 - c-kit + cells showed innate heterogeneity, indicative of the existence of vascular and mesenchymal c-kit + cells in normal hearts. Cardiac pressure overload resulted in a modest increase in c-kit-derived cardiomyocytes, with significant increases in the numbers of endothelial cells and fibroblasts. Doxorubicin-induced acute cardiotoxicity did not increase c-kit-derived endothelial cell fates but instead induced cardiomyocyte differentiation. Mechanistically, doxorubicin-induced DNA damage in c-kit + cells resulted in expression of p53. Inhibition of p53 blocked cardiomyocyte differentiation in response to doxorubicin, whereas stabilization of p53 was sufficient to increase c-kit-derived cardiomyocyte differentiation. These results demonstrate that different pathological stimuli induce different cell fates of c-kit + cells in vivo. Although the overall rate of cardiomyocyte formation from c-kit + cells is still below clinically relevant levels, we show that p53 is central to the ability of c-kit + cells to adopt cardiomyocyte fates, which could lead to the development of strategies to preferentially generate cardiomyocytes from c-kit + cells. © 2017 American Heart Association, Inc.

  20. Receptor tyrosine kinase (c-Kit) inhibitors: a potential therapeutic target in cancer cells

    PubMed Central

    Abbaspour Babaei, Maryam; Kamalidehghan, Behnam; Saleem, Mohammad; Huri, Hasniza Zaman; Ahmadipour, Fatemeh

    2016-01-01

    c-Kit, a receptor tyrosine kinase, is involved in intracellular signaling, and the mutated form of c-Kit plays a crucial role in occurrence of some cancers. The function of c-Kit has led to the concept that inhibiting c-Kit kinase activity can be a target for cancer therapy. The promising results of inhibition of c-Kit for treatment of cancers have been observed in some cancers such as gastrointestinal stromal tumor, acute myeloid leukemia, melanoma, and other tumors, and these results have encouraged attempts toward improvement of using c-Kit as a capable target for cancer therapy. This paper presents the findings of previous studies regarding c-Kit as a receptor tyrosine kinase and an oncogene, as well as its gene targets and signaling pathways in normal and cancer cells. The c-Kit gene location, protein structure, and the role of c-Kit in normal cell have been discussed. Comprehending the molecular mechanism underlying c-Kit-mediated tumorogenesis is consequently essential and may lead to the identification of future novel drug targets. The potential mechanisms by which c-Kit induces cellular transformation have been described. This study aims to elucidate the function of c-Kit for future cancer therapy. In addition, it has c-Kit inhibitor drug properties and their functions have been listed in tables and demonstrated in schematic pictures. This review also has collected previous studies that targeted c-Kit as a novel strategy for cancer therapy. This paper further emphasizes the advantages of this approach, as well as the limitations that must be addressed in the future. Finally, although c-Kit is an attractive target for cancer therapy, based on the outcomes of treatment of patients with c-Kit inhibitors, it is unlikely that Kit inhibitors alone can lead to cure. It seems that c-Kit mutations alone are not sufficient for tumorogenesis, but do play a crucial role in cancer occurrence. PMID:27536065

  1. Comparison of TT-F-1098 Solvent-Thinned Block Fillers with Water-Thinnable Block Fillers.

    DTIC Science & Technology

    1985-03-01

    saved money , because the latex is less roller were visible. The appearance of the surface expensive than the epoxy it replaced. In both cases...a previous coating. A kit manu- The appearance of all the fillers was satisfactory. factured b, Paul N. Gardner Company, Inc., Lauder - Voids were

  2. A Comparison of Soil Test Kits for Use in the Secondary Classroom

    ERIC Educational Resources Information Center

    Yusten, Jason; Gerber, D. Timothy; Beck, Judy

    2003-01-01

    Because soils provide water, minerals, and a medium to anchor the roots of plants, measuring the basic physical/chemical components of soils is important to maintaining healthy garden plants and agricultural crops. Historically, soil analysis has been practiced to determine fertilizer and lime applications, soil fertility, and soil improvement…

  3. How great a thirst? Assembling a river restoration toolkit

    Treesearch

    Steve Harris

    1999-01-01

    The Rio Grande River's biologically troubled status is clearly linked to present and historic water management. To restore the river to pre-settlement conditions will take a "tool kit" that holds authorities, knowledge, and skills needed to correct historical neglect and abuse. Tools include awareness, planning, partnerships, engineering solutions, and a...

  4. SHP-1 Binds and Negatively Modulates the c-Kit Receptor by Interaction with Tyrosine 569 in the c-Kit Juxtamembrane Domain

    PubMed Central

    Kozlowski, Maya; Larose, Louise; Lee, Fai; Le, Duc Mingh; Rottapel, Robert; Siminovitch, Katherine A.

    1998-01-01

    The SH2 domain-containing SHP-1 tyrosine phosphatase has been shown to negatively regulate a broad spectrum of growth factor- and cytokine-driven mitogenic signaling pathways. Included among these is the cascade of intracellular events evoked by stem cell factor binding to c-Kit, a tyrosine kinase receptor which associates with and is dephosphorylated by SHP-1. Using a series of glutathione S-transferase (GST) fusion proteins containing either tyrosine-phosphorylated segments of the c-Kit cytosolic region or the SH2 domains of SHP-1, we have shown that SHP-1 interacts with c-Kit by binding selectively to the phosphorylated c-Kit juxtamembrane region and that the association of c-Kit with the larger of the two SHP-1 isoforms may be mediated through either the N-terminal or C-terminal SHP-1 SH2 domain. The results of binding assays with mutagenized GST-Kit juxtamembrane fusion proteins and competitive inhibition assays with phosphopeptides encompassing each c-Kit juxtamembrane region identified the tyrosine residue at position 569 as the major site for binding of SHP-1 to c-Kit and suggested that tyrosine 567 contributes to, but is not required for, this interaction. By analysis of Ba/F3 cells retrovirally transduced to express c-Kit receptors, phenylalanine substitution of c-Kit tyrosine residue 569 was shown to be associated with disruption of c-Kit–SHP-1 binding and induction of hyperproliferative responses to stem cell factor. Although phenylalanine substitution of c-Kit tyrosine residue 567 in the Ba/F3–c-Kit cells did not alter SHP-1 binding to c-Kit, the capacity of a second c-Kit-binding tyrosine phosphatase, SHP-2, to associate with c-Kit was markedly reduced, and the cells again showed hyperproliferative responses to stem cell factor. These data therefore identify SHP-1 binding to tyrosine 569 on c-Kit as an interaction pivotal to SHP-1 inhibitory effects on c-Kit signaling, but they indicate as well that cytosolic protein tyrosine phosphatases other than SHP-1 may also negatively regulate the coupling of c-Kit engagement to proliferation. PMID:9528781

  5. Unmanned Evaluation of Select Commercially Available Open Circuit Scuba Regulators for Cold Water Diving

    DTIC Science & Technology

    2010-04-01

    Water Kit (dry system) installed as standard Abyss second stage with integrated 30-inch braided intermediate pressure hose as standard No user...diaphragm system) installed as standard Abyss second stage with integrated 30-inch braided intermediate pressure hose as standard No user adjustments...1st Stage Regulator with Abyss 2nd Stage and Integrated Intermediate Pressure Hose ..………………………….. A-2 A3 Modified Mares Proton Ice Extreme V32

  6. [The effect of mineral water on serotonin and insulin production (an experimental study)].

    PubMed

    Polushina, N D

    1998-01-01

    Radioimmunoassay (DRG kits) and orthotoluidine test were conducted to measure blood serotonin, insulin and glucose in 70 intact Wistar rat males before and after a course of drinking mineral water Essentuki 17 (MW). After the MW drinking course, a single dose of mineral water increases basal levels of serotonin and insulin, sensitivity of endocrine cells to MW. Serotonin and insulin rose maximally on minute 5 after the drink while in contrast to minute 15 and 30 before initiation of the MW drinking course. A direct correlation was found between blood concentrations of serotonin and insulin.

  7. Comparison of three DNA extraction kits to establish maximum yield and quality of coral-associated microbial DNA

    USGS Publications Warehouse

    Baker, Erin J.; Kellogg, Christina A.

    2014-01-01

    Coral microbiology is an expanding field, yet there is no standard DNA extraction protocol. Although many researchers depend on commercial extraction kits, no specific kit has been optimized for use with coral samples. Both soil and plant DNA extraction kits from MO BIO Laboratories, Inc., have been used by many research groups for this purpose. MO BIO recently replaced their PowerPlant® kit with an improved PowerPlantPro kit, but it was unclear how these changes would affect the kit’s use with coral samples. In order to determine which kit produced the best results, we conducted a comparison between the original PowerPlant kit, the new PowerPlantPro kit, and an alternative kit, PowerSoil, using samples from several different coral genera. The PowerPlantPro kit had the highest DNA yields, but the lack of 16S rRNA gene amplification in many samples suggests that much of the yield may be coral DNA rather than microbial DNA. The most consistent positive amplifications came from the PowerSoil kit.

  8. Nanostructured catalysts applied to degrade atrazine in aqueous phase by heterogeneous photo-Fenton process.

    PubMed

    Benzaquén, Tamara B; Barrera, Deicy A; Carraro, Paola M; Sapag, Karim; Alfano, Orlando M; Eimer, Griselda A

    2018-06-02

    SBA-15 and KIT-6 materials have been synthesized and modified with iron salts by the wet impregnation method with different metal loadings. The different mesostructures obtained were characterized by N 2 adsorption-desorption at 77 K, X-ray diffraction, temperature-programmed reduction, and ultraviolet-visible spectroscopy. These iron-containing mesostructured materials have been successfully tested for the heterogeneous photo-Fenton degradation of aqueous solutions of dangerous herbicides, such as atrazine, using UV-visible light irradiation, at room temperature and close to neutral pH. The results showed that the Fe/SBA-15 (10%) and Fe/KIT-6 (5%) catalysts exhibited the highest activities. However, the Fe/KIT-6 (5%) catalyst with minor Fe loading than Fe/SBA-15 (10%) presented a higher degradation of atrazine (above 98% in a reaction time of 240 min). Therefore, the interconnectivity of the cage-like mesopores had an important influence on the catalytic activity, favoring probably mass-transfer effects. Thus, the high performance of these materials indicates that the heterogeneous via of photo-Fenton process can also be efficiently employed to treat wastewaters containing pollutants such as herbicides, in order to reduce them to simplest and less toxic molecules.

  9. [Possibility of the species identification using blood stains located on the material evidences and bone fragments with the method of solid phase enzyme immunoassay with "IgG general-EIA-BEST" kit and human immunoglobulin G].

    PubMed

    Sidorov, V L; Shvetsova, I V; Isakova, I V

    2007-01-01

    The authors give the comparative analysis of Russian and foreign forensic medical methods of species character identification of the blood from the stains on the material evidences and bone fragments. It is shown that for this purpose it is feasible to apply human immunoglobulin G (IgG) and solid phase enzyme immunoassay (EIA) with the kit "IgG general-EIA-BEST". In comparison with the methods used in Russia this method is more sensitive, convenient for objective registration and computer processing. The results of experiments shown that it is possible to use the kit "IgG general-EIA-BEST" in forensic medicine for the species character identification of the blood from the stains on the material evidences and bone fragments.

  10. The Development and Use of Internal Amplification Controls (IACs) with DNA Profiling Kits for Forensic DNA Analysis.

    PubMed

    Zahra, Nathalie; Goodwin, William

    2016-01-01

    Biological samples recovered for forensic investigations are often degraded and/or have low amounts of DNA; in addition, in some instances the samples may be contaminated with chemicals that can act as PCR inhibitors. As a consequence this can make interpretation of the results challenging with the possibility of having partial profiles and false negative results. Because of the impact of DNA analysis on forensic investigations, it is important to monitor the process of DNA profiling, in particular the amplification reaction. In this chapter we describe a method for the in-house generation and use of internal amplification controls (IACs) with DNA profiling kits to monitor the success of the PCR proces. In the example we show the use of the SGM Plus® kit. These controls can also be used to aid the interpretation of the DNA profile.

  11. A Measurement of Vocational Education’s Student Satisfaction in Learning Electronic Appliance Repair and Maintenance Course with CCTV Trainer Kit

    NASA Astrophysics Data System (ADS)

    Rakhmawati, Lusia; Irwansyah Febriyanto, Fariz

    2018-04-01

    This study aimed to investigate satisfaction levels, among of the eleventh-grade students majoring in audio-video technique, towards the instructional media CCTV trainer kit: The CCTV Prototype and job sheet on to and analyzed perspectives of the practical approach to learning using CCTV prototype to improve installation skills in the electronic appliance repair and maintenance class. Survey questionnaires and instructional media were applied to data collection. The students’ satisfaction towards the instructional media in teaching and learning process was presented in positive responses, 91.94 % satisfied. The finding reveals that the instructional media using trainer could better to overcome the need a practical approach to learning. Furthermore, using trainer kit media creates an environment where students can support each other and receive feedback from their peers. They performed practice activities that help them apply the new information from the steps on the job sheet.

  12. Possible effects of drilling operations in Section 6D, Naval Petroleum Reserve No. 2, Kern County, California on the endangered San Joaquin kit fox, blunt-nosed leopard lizard, and other sensitive species

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

    O'Farrell, T.P.; Kato, T.

    1982-12-01

    Getty Oil Company requested permission from the US Department of Energy to drill 10 petroleum wells and one water disposal well in Section 6D, Naval Petroleum Reserve No. 2, Kern County, California, which is thought to provide habitat for the endangered San Joaquin kit fox and blunt-nosed leopard lizard, and two sensitive species: the giant kangaroo rat and San Joaquin antelope ground squirrel. The objective of this study was to assess the possible impacts of development drilling on these species and their essential habitats. Most of the proposed wells will be located on or near existing well pads; therefore, onlymore » 2 hectares of potential habitat will be disturbed. Although 21 kit fox dens were found, none were closer than 10 m from proposed well pads, and most were more than 40 m away. No evidence of either the blunt-nosed leopard lizard or giant kangaroo rat was gathered. Ten San Joaquin antelope ground squirrels were observed. Although 2 hectares of habitat will be disturbed, there is no evidence to indicate that any of the species has burrows that will be lost during landclearing. Loss of vegetation may have some small, unknown impacts on food supplies for species preyed upon by kit fox and blunt-nosed leopard lizards, but the disturbed vegetation represents a small fraction of the food supplies available in the surrounding habitat. Because the project poses few threats to individuals of the endangered or sensitive species surveyed, it was concluded that completion of the drilling is unlikely to jeopardize the continued existence of any of the species or their essential habitats if: (1) present kit fox dens are protected during construction activities, (2) topsoil removed during land-levelling is used to reclaim past disturbances of the habitat, and (3) artificial kit fox dens are installed to compensate for the possible loss of denning sites.« less

  13. Identification of mutations in the coding sequence of the proto-oncogene c-kit in a human mast cell leukemia cell line causing ligand-independent activation of c-kit product.

    PubMed Central

    Furitsu, T; Tsujimura, T; Tono, T; Ikeda, H; Kitayama, H; Koshimizu, U; Sugahara, H; Butterfield, J H; Ashman, L K; Kanayama, Y

    1993-01-01

    The c-kit proto-oncogene encodes a receptor tyrosine kinase. Binding of c-kit ligand, stem cell factor (SCF) to c-kit receptor (c-kitR) is known to activate c-kitR tyrosine kinase, thereby leading to autophosphorylation of c-kitR on tyrosine and to association of c-kitR with substrates such as phosphatidylinositol 3-kinase (PI3K). In a human mast cell leukemia cell line HMC-1, c-kitR was found to be constitutively phosphorylated on tyrosine, activated, and associated with PI3K without the addition of SCF. The expression of SCF mRNA transcript in HMC-1 cells was not detectable by means of PCR after reverse transcription (RT-PCR) analysis, suggesting that the constitutive activation of c-kitR was ligand independent. Sequencing of whole coding region of c-kit cDNA revealed that c-kit genes of HMC-1 cells were composed of a normal, wild-type allele and a mutant allele with two point mutations resulting in intracellular amino acid substitutions of Gly-560 for Val and Val-816 for Asp. Amino acid sequences in the regions of the two mutations are completely conserved in all of mouse, rat, and human c-kit. In order to determine the causal role of these mutations in the constitutive activation, murine c-kit mutants encoding Gly-559 and/or Val-814, corresponding to human Gly-560 and/or Val-816, were constructed by site-directed mutagenesis and expressed in a human embryonic kidney cell line, 293T cells. In the transfected cells, both c-kitR (Gly-559, Val-814) and c-kitR (Val-814) were abundantly phosphorylated on tyrosine and activated in immune complex kinase reaction in the absence of SCF, whereas tyrosine phosphorylation and activation of c-kitR (Gly-559) or wild-type c-kitR was modest or little, respectively. These results suggest that conversion of Asp-816 to Val in human c-kitR may be an activating mutation and responsible for the constitutive activation of c-kitR in HMC-1 cells. Images PMID:7691885

  14. Distinct cellular properties of oncogenic KIT receptor tyrosine kinase mutants enable alternative courses of cancer cell inhibition

    PubMed Central

    Shi, Xiarong; Sousa, Leiliane P.; Mandel-Bausch, Elizabeth M.; Tome, Francisco; Reshetnyak, Andrey V.; Hadari, Yaron; Schlessinger, Joseph; Lax, Irit

    2016-01-01

    Large genomic sequencing analysis as part of precision medicine efforts revealed numerous activating mutations in receptor tyrosine kinases, including KIT. Unfortunately, a single approach is not effective for inhibiting cancer cells or treating cancers driven by all known oncogenic KIT mutants. Here, we show that each of the six major KIT oncogenic mutants exhibits different enzymatic, cellular, and dynamic properties and responds distinctly to different KIT inhibitors. One class of KIT mutants responded well to anti-KIT antibody treatment alone or in combination with a low dose of tyrosine kinase inhibitors (TKIs). A second class of KIT mutants, including a mutant resistant to imatinib treatment, responded well to a combination of TKI with anti-KIT antibodies or to anti-KIT toxin conjugates, respectively. We conclude that the preferred choice of precision medicine treatments for cancers driven by activated KIT and other RTKs may rely on clear understanding of the dynamic properties of oncogenic mutants. PMID:27482095

  15. Megakaryocytes compensate for Kit insufficiency in murine arthritis.

    PubMed

    Cunin, Pierre; Penke, Loka R; Thon, Jonathan N; Monach, Paul A; Jones, Tatiana; Chang, Margaret H; Chen, Mary M; Melki, Imene; Lacroix, Steve; Iwakura, Yoichiro; Ware, Jerry; Gurish, Michael F; Italiano, Joseph E; Boilard, Eric; Nigrovic, Peter A

    2017-05-01

    The growth factor receptor Kit is involved in hematopoietic and nonhematopoietic development. Mice bearing Kit defects lack mast cells; however, strains bearing different Kit alleles exhibit diverse phenotypes. Herein, we investigated factors underlying differential sensitivity to IgG-mediated arthritis in 2 mast cell-deficient murine lines: KitWsh/Wsh, which develops robust arthritis, and KitW/Wv, which does not. Reciprocal bone marrow transplantation between KitW/Wv and KitWsh/Wsh mice revealed that arthritis resistance reflects a hematopoietic defect in addition to mast cell deficiency. In KitW/Wv mice, restoration of susceptibility to IgG-mediated arthritis was neutrophil independent but required IL-1 and the platelet/megakaryocyte markers NF-E2 and glycoprotein VI. In KitW/Wv mice, platelets were present in numbers similar to those in WT animals and functionally intact, and transfer of WT platelets did not restore arthritis susceptibility. These data implicated a platelet-independent role for the megakaryocyte, a Kit-dependent lineage that is selectively deficient in KitW/Wv mice. Megakaryocytes secreted IL-1 directly and as a component of circulating microparticles, which activated synovial fibroblasts in an IL-1-dependent manner. Transfer of WT but not IL-1-deficient megakaryocytes restored arthritis susceptibility to KitW/Wv mice. These findings identify functional redundancy among Kit-dependent hematopoietic lineages and establish an unanticipated capacity of megakaryocytes to mediate IL-1-driven systemic inflammatory disease.

  16. Making Earth Science Relevant in the K-8 Classroom. The Development of an Instructional Soils Module for Pre-Service Elementary Teachers Using the Next Generation Science Standards

    NASA Astrophysics Data System (ADS)

    Baldwin, K. A.; Hauge, R.; Dechaine, J. M.; Varrella, G.; Egger, A. E.

    2013-12-01

    The development and adoption of the Next Generation Science Standards (NGSS) raises a challenge in teacher preparation: few current teacher preparation programs prepare students to teach science the way it is presented in the NGSS, which emphasize systems thinking, interdisciplinary science, and deep engagement in the scientific process. In addition, the NGSS include more geoscience concepts and methods than previous standards, yet this is a topic area in which most college students are traditionally underprepared. Although nationwide, programmatic reform is needed, there are a few targets where relatively small, course-level changes can have a large effect. One of these targets is the 'science methods' course for pre-service elementary teachers, a requirement in virtually all teacher preparation programs. Since many elementary schools, both locally and across the country, have adopted a kit based science curriculum, examining kits is often a part of a science methods course. Unfortunately, solely relying on a kit based curriculum may leave gaps in science content curriculum as one prepares teachers to meet the NGSS. Moreover, kits developed at the national level often fall short in connecting geoscientific content to the locally relevant societal issues that engage students. This highlights the need to train pre-service elementary teachers to supplement kit curriculum with inquiry based geoscience investigations that consider relevant societal issues, promote systems thinking and incorporate connections between earth, life, and physical systems. We are developing a module that teaches geoscience concepts in the context of locally relevant societal issues while modeling effective pedagogy for pre-service elementary teachers. Specifically, we focus on soils, an interdisciplinary topic relevant to multiple geoscience-related societal grand challenges (e.g., water, food) that is difficult to engage students in. Module development is funded through InTeGrate, NSF's STEP Center in the geosciences. The module goals are: 1) Pre-service teachers will apply classification methods, testing procedures and interdisciplinary systems thinking to analyze and evaluate a relevant societal issue in the context of soils, 2) Pre-service teachers will design, develop, and facilitate a standards-based K-8 soils unit, incorporating a relevant broader societal issue that applies authentic geoscientific data, and incorporates geoscientific habits of mind. In addition, pre-service teachers will look toward the NGSS and align activities with content standards, systems thinking, and science and engineering practices. This poster will provide an overview of module development to date as well as a summary of pre-semester survey results indicating pre-service elementary teachers' ideas (beliefs, attitudes, preconceptions, and content knowledge) about teaching soils, and making science relevant in a K-8 classroom.

  17. Signal transduction by normal isoforms and W mutant variants of the Kit receptor tyrosine kinase.

    PubMed

    Reith, A D; Ellis, C; Lyman, S D; Anderson, D M; Williams, D E; Bernstein, A; Pawson, T

    1991-09-01

    Germline mutations at the Dominant White Spotting (W) and Steel (Sl) loci have provided conclusive genetic evidence that c-kit mediated signal transduction pathways are essential for normal mouse development. We have analysed the interactions of normal and mutant W/c-kit gene products with cytoplasmic signalling proteins, using transient c-kit expression assays in COS cells. In addition to the previously identified c-kit gene product (Kit+), a second normal Kit isoform (KitA+) containing an in-frame insertion, Gly-Asn-Asn-Lys, within the extracellular domain, was detected in murine mast cell cultures and mid-gestation placenta. Both Kit+ and KitA+ isoforms showed increased autophosphorylation and enhanced association with phosphatidylinositol (PI) 3' kinase and PLC gamma 1, when stimulated with recombinant soluble Steel factor. No association or increase in phosphorylation of GAP and two GAP-associated proteins, p62 and p190, was observed. The two isoforms had distinct activities in the absence of exogenous soluble Steel factor; Kit+, but not KitA+, showed constitutive tyrosine phosphorylation that was accompanied by a low constitutive level of association with PI-3' kinase and PLC gamma 1. Introduction of the point substitutions associated with W37 (Glu582----Lys) or W41 (Val831----Met) mutant alleles into c-kit expression constructs abolished (W37) or reduced (W41) the Steel factor-induced association of the Kit receptor with signalling proteins in a manner proportional to the overall severity of the corresponding W mutant phenotype. These data suggest a diversity of normal Kit signalling pathways and indicate that W mutant phenotypes result from primary defects in the Kit receptor that affect its interaction with cytoplasmic signalling proteins.

  18. The identification of specialized pacemaking cells in the anal sphincters.

    PubMed

    Shafik, Ahmed; El Sibai, Olfat; Ahmed, Ismail

    2006-07-01

    Interstitial cells of Cajal (ICC) are claimed to generate the electrical activity in the colon and stomach. As the external (EAS) and internal (IAS) anal sphincters exhibit resting electrical activity, we hypothesized the presence of ICC in these sphincters. This hypothesis was investigated in the current study. Specimens from the EAS and IAS were taken from normal areas of the anorectum which had been surgically excised by abdominoperineal operation for rectal cancer of 28 patients (16 men, 12 women, mean age 42.2+/-4.8 years). The specimens were subjected to c-kit immunohistochemistry. Controls for the specificity of the antisera consisted of tissue incubation with normal rabbit serum substituted for the primary antiserum. Fusiform, c-kit positive, ICC-like cells were detected in the anal sphincters; they had dendritic processes. They were clearly distinguishable from the non-branching, c-kit negative smooth and striated muscle cells of the anal sphincters. The specimens contained also c-kit positive mast cells, but they had a rounded body with no dendritic processes. Immunoreactivity was absent in negative controls in which the primary antibody was omitted. We have identified, for the first time, cells in EAS and IAS with morphological and immunological phenotypes similar to ICCs of the gut. These cells appear to be responsible for initiating the slow waves recorded from the anal sphincters and for controlling their activity. A deficiency or absence of these cells may affect the anal motile activity. Studies are needed to explore the role of these cells in anal motility disorders.

  19. STOCK: Structure mapper and online coarse-graining kit for molecular simulations

    DOE PAGES

    Bevc, Staš; Junghans, Christoph; Praprotnik, Matej

    2015-03-15

    We present a web toolkit STructure mapper and Online Coarse-graining Kit for setting up coarse-grained molecular simulations. The kit consists of two tools: structure mapping and Boltzmann inversion tools. The aim of the first tool is to define a molecular mapping from high, e.g. all-atom, to low, i.e. coarse-grained, resolution. Using a graphical user interface it generates input files, which are compatible with standard coarse-graining packages, e.g. VOTCA and DL_CGMAP. Our second tool generates effective potentials for coarse-grained simulations preserving the structural properties, e.g. radial distribution functions, of the underlying higher resolution model. The required distribution functions can be providedmore » by any simulation package. Simulations are performed on a local machine and only the distributions are uploaded to the server. The applicability of the toolkit is validated by mapping atomistic pentane and polyalanine molecules to a coarse-grained representation. Effective potentials are derived for systems of TIP3P (transferable intermolecular potential 3 point) water molecules and salt solution. The presented coarse-graining web toolkit is available at http://stock.cmm.ki.si.« less

  20. Germline mutations of KIT in gastrointestinal stromal tumor (GIST) and mastocytosis.

    PubMed

    Ke, Hengning; Kazi, Julhash U; Zhao, Hui; Sun, Jianmin

    2016-01-01

    Somatic mutations of KIT are frequently found in mastocytosis and gastrointestinal stromal tumor (GIST), while germline mutations of KIT are rare, and only found in few cases of familial GIST and mastocytosis. Although ligand-independent activation is the common feature of KIT mutations, the phenotypes mediated by various germline KIT mutations are different. Germline KIT mutations affect different tissues such as interstitial cells of Cajal (ICC), mast cells or melanocytes, and thereby lead to GIST, mastocytosis, or abnormal pigmentation. In this review, we summarize germline KIT mutations in familial mastocytosis and GIST and discuss the possible cellular context dependent transforming activity of KIT mutations.

  1. No GIST-type c-kit gain of function mutations in neuroblastic tumours

    PubMed Central

    Korja, M; Finne, J; Salmi, T T; Haapasalo, H; Tanner, M; Isola, J

    2005-01-01

    Aims: Neuroblastic tumours (NTs) have been shown to respond to imatinib treatment in vivo and in vitro, possibly via inactivating the c-kit receptor. The purpose of this study was to identify gastrointestinal stromal tumour (GIST)-type c-kit gene associated mutations in exons 9, 11, 13, and 17 in NTs to recognise a subset of tumours that would probably respond to imatinib treatment. Methods: Expression of the c-kit protein was detected immunohistochemically in a total of 37 archival paraffin wax embedded NTs using polyclonal rabbit antihuman c-kit antibody. After immunohistochemistry, c-kit gene associated chromosomal mutations in all cases of NT were detected with denaturing high performance liquid chromatography (HPLC). Results: Denaturing HLPC analysis did not reveal GIST-type mutations in four immunohistochemically detected c-kit positive or in 33 c-kit negative NTs. Conclusions: c-kit receptor expression and GIST-type c-kit gene mutations are rare events in NTs. Oncogenic activation of c-kit in NTs presumably differs from that of GISTs, which may influence their responsiveness to imatinib treatment. Whether c-kit has an essential role in the pathogenesis of NTs remains to be investigated. PMID:15976348

  2. Pim1 kinase regulates c-Kit gene translation.

    PubMed

    An, Ningfei; Cen, Bo; Cai, Houjian; Song, Jin H; Kraft, Andrew; Kang, Yubin

    2016-01-01

    Receptor tyrosine kinase, c-Kit (CD117) plays a pivotal role in the maintenance and expansion of hematopoietic stem/progenitor cells (HSPCs). Additionally, over-expression and/or mutational activation of c-Kit have been implicated in numerous malignant diseases including acute myeloid leukemia. However, the translational regulation of c-Kit expression remains largely unknown. We demonstrated that loss of Pim1 led to specific down-regulation of c-Kit expression in HSPCs of Pim1 -/- mice and Pim1 -/- 2 -/- 3 -/- triple knockout (TKO) mice, and resulted in attenuated ERK and STAT3 signaling in response to stimulation with stem cell factor. Transduction of c-Kit restored the defects in colony forming capacity seen in HSPCs from Pim1 -/- and TKO mice. Pharmacologic inhibition and genetic modification studies using human megakaryoblastic leukemia cells confirmed the regulation of c-Kit expression by Pim1 kinase: i.e., Pim1-specific shRNA knockdown down-regulated the expression of c-Kit whereas overexpression of Pim1 up-regulated the expression of c-Kit. Mechanistically, inhibition or knockout of Pim1 kinase did not affect the transcription of c-Kit gene. Pim1 kinase enhanced c-Kit 35 S methionine labeling and increased the incorporation of c-Kit mRNAs into the polysomes and monosomes, demonstrating that Pim1 kinase regulates c-Kit expression at the translational level. Our study provides the first evidence that Pim1 regulates c-Kit gene translation and has important implications in hematopoietic stem cell transplantation and cancer treatment.

  3. A rapid and efficient assay for extracting DNA from fungi

    USGS Publications Warehouse

    Griffin, Dale W.; Kellogg, C.A.; Peak, K.K.; Shinn, E.A.

    2002-01-01

    Aims: A method for the rapid extraction of fungal DNA from small quantities of tissue in a batch-processing format was investigated. Methods and Results: Tissue (< 3.0 mg) was scraped from freshly-grown fungal isolates. The tissue was suspended in buffer AP1 and subjected to seven rounds of freeze/thaw using a crushed dry ice/ethanol bath and a boiling water bath. After a 30 min boiling step, the tissue was quickly ground against the wall of the microfuge tube using a sterile pipette tip. The Qiagen DNeasy Plant Tissue Kit protocol was then used to purify the DNA for PCR/ sequencing applications. Conclusions: The method allowed batch DNA extraction from multiple fungal isolates using a simple yet rapid and reliable assay. Significance and Impact of the Study: Use of this assay will allow researchers to obtain DNA from fungi quickly for use in molecular assays that previously required specialized instrumentation, was time-consuming or was not conducive to batch processing.

  4. Implementing inquiry-based kits within a professional development school model

    NASA Astrophysics Data System (ADS)

    Jones, Mark Thomas

    2005-07-01

    Implementation of guided inquiry teaching for the first time carries inherent problems for science teachers. Reform efforts on inquiry-based science teaching are often unsustainable and are not sensitive to teachers' needs and abilities as professionals. Professional development schools are meant to provide a research-based partnership between a public school and a university. These collaborations can provide support for the professional development of teachers. This dissertation reports a study focused on the implementation of inquiry-based science kits within the support of one of these collaborations. The researcher describes the difficulties and successful adaptations experienced by science teachers and how a coteaching model provided support. These types of data are needed in order to develop a bottom-up, sustainable process that will allow teachers to implement inquiry-based science. A qualitative methodology with "researcher as participant" was used in this study of two science teachers during 2002--2003. These two teachers were supported by a coteaching model, which included preservice teachers for each teacher as well as a supervising professor. Data were collected from the researcher's direct observations of coteachers' practice. Data were also collected from interviews and reflective pieces from the coteachers. Triangulation of the data on each teacher's case supported the validity of the findings. Case reports were prepared from these data for each classroom teacher. These case reports were used and cross-case analysis was conducted to search for major themes and findings in the study. Major findings described the hurdles teachers encounter, examples of adaptations observed in the teachers' cases and the supportive interactions with their coteachers while implementing the inquiry-based kits. In addition, the data were used to make recommendations for future training and use of the kits and the coteaching model. Results from this study showed that the kit's guided structure of inquiry and the collaboration both affected the inservice teachers in the following ways: The coteaching model supported behavioral and material management issues caused by the implementation of the kits; collaboration with preservice teachers created a "smaller-class-size" effect, which allowed teachers to attend to a smaller number of students for cooperative learning and assessment, and the elementary inservice teachers learned pedagogical strategies and science content from collaborating with secondary preservice teachers in kit use and from the kits' curriculum. Results were used as a self-study for future training and support for implementation of inquiry-based kits.

  5. Early kit mortality and growth in farmed mink are affected by litter size rather than nest climate.

    PubMed

    Schou, T M; Malmkvist, J

    2017-09-01

    We investigated the effects of nest box climate on early mink kit mortality and growth. We hypothesised that litters in warm nest boxes experience less hypothermia-induced mortality and higher growth rates during the 1st week of life. This study included data from 749, 1-year-old breeding dams with access to nesting materials. Kits were weighed on days 1 and 7, dead kits were collected daily from birth until day 7 after birth, and nest climate was measured continuously from days 1 to 6. We tested the influences of the following daily temperature (T) and humidity (H) parameters on the number of live-born kit deaths and kit growth: T mean, T min, T max, T var (fluctuation) and H mean. The nest microclimate experienced by the kits was buffered against the ambient climate, with higher temperatures and reduced climate fluctuation. Most (77.0%) live-born kit deaths in the 1st week occurred on days 0 and 1. Seven of 15 climate parameters on days 1 to 3 had significant effects on live-born kit mortality. However, conflicting effects among days, marginal effects and late effects indicated that climate was not the primary cause of kit mortality. Five of 30 climate parameters had significant effects on kit growth. Few and conflicting effects indicated that the climate effect on growth was negligible. One exception was that large nest temperature fluctuations on day 1 were associated with reduced deaths of live-born kit (P<0.001) and increased kit growth (P=0.003). Litter size affected kit vitality; larger total litter size at birth was associated with greater risks of kit death (P<0.001) and reduced growth (P<0.001). The number of living kits in litters had the opposite effect, as kits in large liveborn litters had a reduced risk of death (P<0.001) and those with large mean litter size on days 1 to 7 had increased growth (P=0.026). Nest box temperature had little effect on early kit survival and growth, which could be due to dams' additional maternal behaviour. Therefore, we cannot confirm that temperature is the primary reason for kit mortality, under the conditions of plenty straw access for maternal nest building. Instead, prenatal and/or parturient litter size is the primary factor influencing early kit vitality. The results indicate that the focus should be on litter size and dam welfare around the times of gestation and birth to increase early kit survival in farmed mink.

  6. Identification of Giardia lamblia and the human infectious-species of Cryptosporidium in drinking water resources in Western Saudi Arabia by nested-PCR assays.

    PubMed

    Hawash, Y; Ghonaim, M; Hussein, Y; Alhazmi, A; Alturkistani, A

    2015-06-01

    The presence of Cryptosporidium and/or Giardia in drinking water represents a major public health problem. This study was the first report concerned with the occurrence of these protozoa in drinking water in Saudi Arabia. The study was undertaken in Al-Taif, a high altitude region, Western Saudi Arabia. Eight underground wells water, six desalinated water and five domestic brands of bottled water samples, 10 liter each, were monthly collected between May 2013 and April 2014. All samples (n = 228), were processed using an automated wash/elution station (IDEXX Laboratories, Inc.). Genomic DNA was directly isolated and purified from samples concentrates with QIAamp® Stool Mini Kit (Qiagen). The target protozoan DNA sequences were amplified using two previously published nested-PCR protocols. Of all the analyzed water, 31 samples (≈14%) were found contaminated with the target protozoa. Giardia lamblia was detected in ≈10% (7/72) of desalinated water and in ≈9% (9/96) of wells water. On the other hand, Cryptosporidium was identified in ≈8% (8/72) of desalinated water and in ≈7% (7/96) of wells water. All bottled water samples (n = 60) were (oo)cysts-free. Protozoan (oo)cysts were more frequently identified in water samples collected in the spring than in other seasons. The methodology established in our study proved sensitive, cost-effective and is amenable for future automation or semi-automation. For better understanding of the current situation that represent an important health threat to the local inhabitants, further studies concerned with (oo)cyst viability, infectivity, concentration and genotype identification are recommended.

  7. The clinical significance of c-Kit mutations in metastatic oral mucosal melanoma in China.

    PubMed

    Ma, Xuhui; Wu, Yunteng; Zhang, Tian; Song, Hao; Jv, Houyu; Guo, Wei; Ren, Guoxin

    2017-10-10

    c-Kit mutations are frequently detected in mucosal melanomas, but their clinical significance in metastatic oral mucosal melanomas (OMM) remains unclear. The main purpose of this study was to investigate the clinical and pathological features of metastatic OMMs with c-Kit mutations and the efficiency of the tyrosine kinase inhibitor imatinib in treating metastatic OMMs. We found thatresidual primary lesion and neck lymph nodes could act as independent prognostic factors in metastatic OMM patients. c-Kit mutations were detected in 22 out of 139 (15.8%) metastatic OMM patients. Under chemotherapy, the overall survival (OS) of c-Kit mutant patients was significantly shorter than that of wild-type patients. The Ki67 expression was significantly higher in c-Kit mutant patients than in wild-type patients. In distant metastatic OMM patients with c-Kit mutations, the treatment with c-Kit inhibitor resulted in a better OS. In conclusion, residual primary lesion, cervical lymph nodes and c-Kit mutations act as adverse prognostic factors of metastatic OMMs. The Kit inhibitor imatinib could benefit metastatic OMM patients with c-Kit mutations.

  8. Intratumoral CD3+ T-Lymphocytes Immunoexpression and Its Association with c-Kit, Angiogenesis, and Overall Survival in Malignant Canine Mammary Tumors

    PubMed Central

    Carvalho, Maria Isabel; Pires, Isabel; Dias, Marlene; Prada, Justina; Gregório, Hugo; Lobo, Luis; Queiroga, Felisbina

    2015-01-01

    In this study 80 malignant CMT were submitted to immunohistochemical detection of CD3, c-kit, VEGF, and CD31, together with clinicopathological parameters of tumor aggressiveness. CD3+ T-cells and c-kit overexpression revealed a positive correlation with VEGF (r = 0.503, P < 0.0001; r = 0.284, P = 0.023 for CD3 and c-kit, resp.) and CD31 (r = 0.654, P < 0.0001; r = 0.365, P = 0.003 for CD3 and c-kit, resp.). A significant association (P = 0.039) and a positive correlation (r = 0.263, P = 0.039) between CD3 and c-kit were also observed. High CD3/VEGF, c-kit/VEGF, and CD3/c-kit tumors were associated with elevated grade of malignancy (P < 0.0001 for all groups), presence of intravascular emboli (P < 0.0001 for CD3/VEGF and CD3/c-kit; P = 0.002 for c-kit/VEGF), and presence of lymph node metastasis (P < 0.0001 for all groups). Tumors with high CD3/VEGF (P = 0.006), c-kit/VEGF (P < 0.0001), and CD3/c-kit (P = 0.002) were associated with poor prognosis. Interestingly high c-kit/VEGF tumors retained their significance by multivariate analysis arising as independent prognostic factor. PMID:26346272

  9. Enzyme-linked immunosorbent assay for ultratrace determination of antibiotics in aqueous samples.

    PubMed

    Kumar, Kuldip; Thompson, Anita; Singh, Ashok K; Chander, Yogesh; Gupta, Satish C

    2004-01-01

    Two commercially available enzyme-linked immunosorbent assay (ELISA) kits that are commonly used for tylosin or tetracycline residues in meat and milk were adapted for ultratrace analysis of these antibiotics in surface and ground waters. These two antibiotics are commonly fed to swine, turkeys, and cattle at subtherapeutic doses for growth promotion purposes. Both ELISA techniques were found to be highly sensitive and selective for the respective antibiotics with detection limits of 0.10 and 0.05 microg L(-1) for tylosin and tetracycline, respectively. The recovery of both tylosin and tetracycline from spiked samples of lake waters, runoff samples, soil saturation extracts, and nanopure water was close to 100%. Tetracycline ELISA was highly specific for tetracycline and chlortetracycline but not for other forms of tetracycline (oxytetracycline, demeclocycline, and doxycycline). Analysis of a few liquid swine manure samples by liquid chromatography-mass spectrometry (LC-MS) showed lower concentrations for chlortetracycline as compared with concentrations obtained using ELISA. However, the concentrations of tylosin from ELISA were comparable with that of LC-MS. The lower concentrations of chlortetracycline obtained by LC-MS in manure samples indicate the presence of other similar or transformed compounds that were detected by ELISA but not determined by LC-MS. These results indicate that both ELISA kits can be useful tools for low-cost screening of tylosin, tetracycline, and chlortetracycline in environmental waters. Furthermore, both ELISA procedures are rapid, portable, and easily adaptable for testing of multiple samples simultaneously.

  10. A Federal Employee's Family Preparedness Guide.

    ERIC Educational Resources Information Center

    Office of Personnel Management, Washington, DC.

    This document outlines what families can do before, during, and after an emergency. The first section covers preparations that can be made before an emergency occurs and includes the creation of a family emergency plan and an "emergency go kit," which can save time if one has to evacuate or go without heat, electricity, or water for a…

  11. Commander Young removes CAP from FDF stowage locker on middeck

    NASA Technical Reports Server (NTRS)

    1981-01-01

    Commander Young removes Crew Activity Plans (CAP) from Flight Data File (FD/FDF) modular stowage locker single tray assembly located in forward middeck locker MF28E. Window shade and filter kit on port side bulkhead and potable water tank on middeck floor appear in view. Photo was taken by Pilot Crippen with a 35mm camera.

  12. Using Fluid Power in the Middle School Classroom

    ERIC Educational Resources Information Center

    Rogers, Stephen

    2013-01-01

    In a middle school classroom, the science of fluidics was introduced using the rig. Afterwards the students were able to make informed choices about using air or water as the medium inside their syringes. The three kits available through the NFPA were explored, reverse-engineered, discussed, and analyzed as a lead-in to the in-house culminating…

  13. Automatic Sprout Grower

    NASA Technical Reports Server (NTRS)

    Sauer, Richard L.; Scheld, H. W.; Magnuson, J. W.

    1989-01-01

    Self-contained seed-sprouting system provides environment for sprouting seeds quickly and easily. Sprouting container standard 6-oz package for dehydrated food and drink mixes in Space Shuttle. About 4 g of dry alfalfa or radish seeds vacuum-sealed in each cup, like freeze-dried foods. Sixteen cups suspended in tray. Air-and-water inlet tube links each cup to system of tubes and solenoid valves alternately furnish air and water and remove stale air. Peristaltic pump supplies water from vinyl medical-fluid bag. Small diaphragm pump supplies and exhausts air. Small circuit board times movements of air and water. Kit offers advantages to home gardeners. Apartment dwellers use it for steady production of homegrown sprouts even though they have no garden space.

  14. Water-level records for the northern High Plains of Colorado, 1973-77

    USGS Publications Warehouse

    Major, Thomas J.; Borman, Ronald G.; Vaught, Kenneth D.

    1977-01-01

    Water-level measurements were made in more than 600 wells during January 1977 in the northern High Plains of Colorado which includes about 9,500 square miles. Most of these wells were irrigation wells, but several stock and government-owned wells were also measured, especially in areas where there is little irrigation. Changes in water levels from January 1976 to January 1977 ranged from a rise of about 12 feet (3.7 meters) to a decline of about 10 feet (3 meters), both of which occurred in Kit Carson County. Measurements for the four preceding winters also are included to serve as references illustrating declining or rising water levels. (Woodard-USGS)

  15. From preclinical development to clinical application: Kit formulation for radiolabelling the minigastrin analogue CP04 with In-111 for a first-in-human clinical trial.

    PubMed

    Pawlak, Dariusz; Rangger, Christine; Kolenc Peitl, Petra; Garnuszek, Piotr; Maurin, Michał; Ihli, Laura; Kroselj, Marko; Maina, Theodosia; Maecke, Helmut; Erba, Paola; Kremser, Leopold; Hubalewska-Dydejczyk, Alicja; Mikołajczak, Renata; Decristoforo, Clemens

    2016-03-31

    A variety of radiolabelled minigastrin analogues targeting the cholecystokinin 2 (CCK2) receptor were developed and compared in a concerted preclinical testing to select the most promising radiotracer for diagnosis and treatment of medullary thyroid carcinoma (MTC). DOTA-DGlu-DGlu-DGlu-DGlu-DGlu-DGlu-Ala-Tyr-Gly-Trp-Met-Asp-Phe-NH2 (CP04) after labelling with (111)In displayed excellent characteristics, such as high stability, receptor affinity, specific and persistent tumour uptake and low kidney retention in animal models. Therefore, it was selected for further clinical evaluation within the ERA-NET project GRAN-T-MTC. Here we report on the development of a pharmaceutical freeze-dried formulation of the precursor CP04 for a first multi-centre clinical trial with (111)In-CP04 in MTC patients. The kit formulation was optimised by adjustment of buffer, additives and radiolabelling conditions. Three clinical grade batches of a final kit formulation with two different amounts of peptide (10 or 50 μg) were prepared and radiolabelled with (111)In. Quality control and stability assays of both the kits and the resulting radiolabelled compound were performed by HPLC analysis. Use of ascorbic acid buffer (pH4.5) allowed freeze-drying of the kit formulation with satisfactory pellet-formation. Addition of methionine and gentisic acid as well as careful selection of radiolabelling temperature was required to avoid extensive oxidation of the Met(11)-residue. Trace metal contamination, in particular Zn, was found to be a major challenge during the pharmaceutical filling process in particular for the 10 μg formulation. The final formulations contained 10 or 50 μg CP04, 25mg ascorbic acid, 0.5mg gentisic acid and 5mg L-methionine. The radiolabelling performed by incubation of 200-250 MBq (111)InCl3 at 90 °C for 15 min resulted in reproducible radiochemical purity (RCP) >94%. Kit-stability was proven for >6 months at +5 °C and at +25 °C. The radiolabelled product was stable for >4h at +25 °C. A kit formulation to prepare (111)In-CP04 for clinical application was developed, showing high stability of the kit as well as high RCP of the final product. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  16. Evaluation of the PrioCHECK™ Trichinella AAD Kit for the digestion and recovery of larvae in pork, horse meat and wild meat.

    PubMed

    Konecsni, Kelly; Scheller, Cheryl; Scandrett, Brad; Buholzer, Patrik; Gajadhar, Alvin

    2017-08-30

    The artificial digestion magnetic stirrer method using pepsin protease and hydrochloric acid is the standard assay for the detection of Trichinella larvae in muscle of infected animals. Recently, an alternative enzyme, serine protease, was employed in the development of a commercially available digestion kit (PrioCHECK™ Trichinella AAD Kit). This assay requires a higher digestion temperature of 60°C which kills the larvae during the digestion process, mitigating the risk of environmental contamination from the parasite. The present study was conducted to determine the performance of the PrioCHECK™ Trichinella AAD Kit compared to the conventional pepsin/HCl digestion. Replicate paired 115g samples of Trichinella-negative pork diaphragm and masseter, and of horse tongue and masseter, were used to compare the two methods for tissue digestibility. Similarly, paired 100g samples of pork diaphragm and horse tongue were spiked with proficiency samples containing known numbers of Trichinella spiralis first stage larvae to compare larval recoveries for the two methods. Masseter samples from wild bears and wolves naturally infected with Trichinella nativa or T6 were also used to compare the performance of the methods. The results of the study showed that the PrioCHECK™ Trichinella AAD Kit, when used according to the manufacturer's instructions, was effective in detecting Trichinella infection in all samples that contained 0.05 or more larvae per gram of tissue. Although there was no significant difference between the Kit method and the standard pepsin/HCl digestion procedure in the average number of larvae recovered from spiked pork diaphragm, 38% fewer larvae were recovered from similarly spiked samples of horse tongue by digestion using serine protease (one way ANOVA, P value <0.001). Additional clarification was also more often required for both horse meat and pork when using the Kit compared to the pepsin/HCl method. The results of testing wildlife samples were similar for the two methods. Overall, the performance of the Kit method was suitable for the digestion of muscle samples and recovery of Trichinella larvae, according to international standards. It also provides advantages of faster digestion, safer reagents and recovered parasites that are non-hazardous for analysts and the environment. Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.

  17. Computer-Based Training Starter Kit.

    ERIC Educational Resources Information Center

    Federal Interagency Group for Computer-Based Training, Washington, DC.

    Intended for use by training professionals with little or no background in the application of automated data processing (ADP) systems, processes, or procurement requirements, this reference manual provides guidelines for establishing a computer based training (CBT) program within a federal agency of the United States government. The manual covers:…

  18. Telescience Resource Kit

    NASA Technical Reports Server (NTRS)

    Schneider, Michelle; Lippincott, Jeff; Chubb, Steve; Whitaker, Jimmy; Rice, Jim; Gillis, Robert; Sims, Chris; Sellers, Donna; Bailey, Darrell (Technical Monitor)

    2002-01-01

    The Telescience Resource Kit (TReK) is a PC based ground control system. It can be used by a single individual or in a group environment to monitor and control spacecraft systems and payloads. Capabilities include data receipt, data processing, data storage, data management, and data transmission. Commercial-Off-The-Shelf (COTS) hardware and software have been employed to reduce development costs, operations and maintenance costs, and to effectively take advantage of new commercial products as they become available. The TReK system is currently being used to monitor and control payloads aboard the International Space Station. It is located at sites around the world.

  19. The value of molecular expression of KIT and KIT ligand analysed using real-time polymerase chain reaction and immunohistochemistry as a prognostic indicator for canine cutaneous mast cell tumours.

    PubMed

    Costa Casagrande, T A; de Oliveira Barros, L M; Fukumasu, H; Cogliati, B; Chaible, L M; Dagli, M L Z; Matera, J M

    2015-03-01

    This study investigated the correlation between KIT gene expression determined by immunohistochemistry and real-time polymerase chain reaction (RT-PCR) and the rate of tumour recurrence and tumour-related deaths in dogs affected with mast cell tumour (MCT). Kaplan-Meier curves were constructed to compare tumour recurrence and tumour-related death between patients. The log-rank test was used to check for significant differences between curves. KIT-I, KIT-II and KIT-III staining patterns were observed in 9 (11.11%), 50 (61.73%) and 22 (27.16%) tumours, respectively. Tumour recurrence rates and tumour-related deaths were not associated with KIT staining patterns (P = 0278, P > 0.05), KIT (P = 0.289, P > 0.05) or KIT ligand (P = 0.106, P > 0.05) gene expression. Despite the lack of association between KIT staining pattern and patient survival time, the results suggest a correlation between aberrant KIT localization and increased proliferative activity of MCTs. RT-PCR seems to be a sensible method for quantitative detection of KIT gene expression in canine MCT, although expressions levels are not correlated with prognosis. © 2013 Blackwell Publishing Ltd.

  20. A meta-analysis of prognostic value of KIT mutation status in gastrointestinal stromal tumors

    PubMed Central

    Jiang, Zhiqiang; Zhang, Jian; Li, Zhi; Liu, Yingjun; Wang, Daohai; Han, Guangsen

    2016-01-01

    Numerous types of KIT mutations have been reported in gastrointestinal stromal tumors (GISTs); however, controversy still exists regarding their clinicopathological significance. In this study, we reviewed the publicly available literature to assess the data by a meta-analysis to characterize KIT mutations and different types of KIT mutations in prognostic prediction in patients with GISTs. Twenty-eight studies that included 4,449 patients were identified and analyzed. We found that KIT mutation status was closely correlated with size of tumors and different mitosis indexes, but not with tumor location. KIT mutation was also observed to be significantly correlated with tumor recurrence, metastasis, as well as the overall survival of patients. Interestingly, there was higher risk of progression in KIT exon 9-mutated patients than in exon 11-mutated patients. Five-year relapse-free survival (RFS) rate was significantly higher in KIT exon 11-deleted patients than in those with other types of KIT exon 11 mutations. In addition, RFS for 5 years was significantly worse in patients bearing KIT codon 557–558 deletions than in those bearing other KIT exon 11 deletions. Our results strongly support the hypothesis that KIT mutation status is another evaluable factor for prognosis prediction in GISTs. PMID:27350754

  1. Targeting c-KIT (CD117) by dasatinib and radotinib promotes acute myeloid leukemia cell death.

    PubMed

    Heo, Sook-Kyoung; Noh, Eui-Kyu; Kim, Jeong Yi; Jeong, Yoo Kyung; Jo, Jae-Cheol; Choi, Yunsuk; Koh, SuJin; Baek, Jin Ho; Min, Young Joo; Kim, Hawk

    2017-11-10

    Dasatinib and radotinib are oral BCR-ABL tyrosine kinase inhibitors that were developed as drugs for the treatment of chronic myeloid leukemia. We report here that the c-KIT (CD117) targeting with dasatinib and radotinib promotes acute myeloid leukemia (AML) cell death, and c-KIT endocytosis is essential for triggering c-KIT-positive AML cell death by dasatinib and radotinib during the early stages. In addition, dasatinib and radotinib reduce heat shock protein 90β (HSP90β) expression and release Apaf-1 in c-KIT-positive AML cells. Finally, this activates a caspase-dependent apoptotic pathway in c-KIT-positive AML cells. Moreover, the inhibition of c-KIT endocytosis by dynamin inhibitor (DY) reversed cell viability and c-KIT expression by dasatinib and radotinib. HSP90β expression was recovered by DY in c-KIT-positive AML cells as well. Furthermore, the effect of radotinib on c-KIT and HSP90β showed the same pattern in a xenograft animal model using HEL92.1.7 cells. Therefore, dasatinib and radotinib promote AML cell death by targeting c-KIT. Taken together, these results indicate that dasatinib and radotinib treatment have a potential role in anti-leukemic therapy on c-KIT-positive AML cells.

  2. RanBPM (RanBP9) regulates mouse c-Kit receptor level and is essential for normal development of bone marrow progenitor cells

    PubMed Central

    Singh, Satyendra; Klarmann, Kimberly D.; Coppola, Vincenzo; Keller, Jonathan R.; Tessarollo, Lino

    2016-01-01

    c-Kit is a tyrosine kinase receptor important for gametogenesis, hematopoiesis, melanogenesis and mast cell biology. Dysregulation of c-Kit function is oncogenic and its expression in the stem cell niche of a number of tissues has underlined its relevance for regenerative medicine and hematopoietic stem cell biology. Yet, very little is known about the mechanisms that control c-Kit protein levels. Here we show that the RanBPM/RanBP9 scaffold protein binds to c-Kit and is necessary for normal c-Kit protein expression in the mouse testis and subset lineages of the hematopoietic system. RanBPM deletion causes a reduction in c-Kit protein but not its mRNA suggesting a posttranslational mechanism. This regulation is specific to the c-Kit receptor since RanBPM reduction does not affect other membrane proteins examined. Importantly, in both mouse hematopoietic system and testis, RanBPM deficiency causes defects consistent with c-Kit loss of expression suggesting that RanBPM is an important regulator of c-Kit function. The finding that this regulatory mechanism is also present in human cells expressing endogenous RanBPM and c-Kit suggests a potential new strategy to target oncogenic c-Kit in malignancies. PMID:27835883

  3. RanBPM (RanBP9) regulates mouse c-Kit receptor level and is essential for normal development of bone marrow progenitor cells.

    PubMed

    Puverel, Sandrine; Kiris, Erkan; Singh, Satyendra; Klarmann, Kimberly D; Coppola, Vincenzo; Keller, Jonathan R; Tessarollo, Lino

    2016-12-20

    c-Kit is a tyrosine kinase receptor important for gametogenesis, hematopoiesis, melanogenesis and mast cell biology. Dysregulation of c-Kit function is oncogenic and its expression in the stem cell niche of a number of tissues has underlined its relevance for regenerative medicine and hematopoietic stem cell biology. Yet, very little is known about the mechanisms that control c-Kit protein levels. Here we show that the RanBPM/RanBP9 scaffold protein binds to c-Kit and is necessary for normal c-Kit protein expression in the mouse testis and subset lineages of the hematopoietic system. RanBPM deletion causes a reduction in c-Kit protein but not its mRNA suggesting a posttranslational mechanism. This regulation is specific to the c-Kit receptor since RanBPM reduction does not affect other membrane proteins examined. Importantly, in both mouse hematopoietic system and testis, RanBPM deficiency causes defects consistent with c-Kit loss of expression suggesting that RanBPM is an important regulator of c-Kit function. The finding that this regulatory mechanism is also present in human cells expressing endogenous RanBPM and c-Kit suggests a potential new strategy to target oncogenic c-Kit in malignancies.

  4. Changes in c-Kit expression levels during the course of radiation therapy for nasopharyngeal carcinoma.

    PubMed

    Jiang, Feng; Hu, Wei; Zhang, Bicheng; Xu, Jing; Shui, Yongjie; Zhou, Xiaofeng; Ren, Xiaoqiu; Chen, Xiaozhong; Shen, Li; Wei, Qichun

    2016-10-01

    In the era of intensity-modulated radiotherapy, distant metastasis is currently the main cause of treatment failure for nasopharyngeal carcinoma (NPC). Additional therapeutic strategies are required to control the metastasis and improve survival. One strategy is targeted therapy, for example against c-Kit. In the current study, the frequency of c-Kit expression was determined immunohistochemically in 106 NPC patients. c-Kit expression changes during the course of radiation therapy were detected in 41 cases via weekly biopsy. Twelve cases (11.3%) had c-Kit expression scores of 3 + and 16 (15.1%) had scores of 2 + . Thus, c-Kit overexpression (2 + or 3 + ) was observed in 28 (26.4%) patients. There were 35 (33.0%) and 43 (40.6%) patients with c-Kit expression scores of 1 + and 0, respectively. Furthermore, a trend of decreased c-Kit expression was observed after commencing radiotherapy according to the 41 NPC patients who were biopsied weekly. Therefore, c-Kit overexpression was identified to be common in NPC, and evaluating c-Kit as a therapeutic target for metastatic NPC via c-Kit overexpression subsequent to first line treatment may be of interest. To the best of our knowledge, the present study is the first to demonstrate a trend of decreased c-Kit expression during the course of radiotherapy.

  5. [Development of surgical antibioprophylaxis kits: evaluation of the impact on prescribing habits].

    PubMed

    Aouizerate, P; Guizard, M

    2002-01-01

    In our hospital, surgical antibioprophylaxis (ATBP) was too often administered too late, thus raising the infectious risk. Antibiotic stocks of the anaesthesia department were also systematically used, instead of nominal prescriptions of these drugs. The pharmacy could neither charge antibiotics to each surgical department nor quantify and differentiate ATBP from curative antibiotic therapy. The pharmacy and anaesthesia departments therefore set out to standardize surgical ATBP, in order to adapt this treatment to each surgical indication, and particularly in the case of allergy to beta-lactamase antibiotics (second line treatment kits). Consequently, prescription forms were developed and supplied to each surgery department, as well as ATBP kits. The kits were prepared and distributed by the pharmacy, and comprised boxes containing antibiotics in sufficient quantities to respect the protocols approved by the French Society of Anaesthesia and Resuscitation (SFAR). A protocol describing prescriptions, dispensation and administration has been presented to physicians and nurses. Fifteen surgical departments were included in our study and 30 different kits were prepared. From 1998 to 2001, 5586 surgical operations required administration of a kit (second line treatment kits in 5% of cases): 1848 (33%) in visceral surgery; 764 (13.8%) in urology; 802 (14%) in orthopaedics; 13 (0.2%) in vascular and thoracic surgery; 1236 (22%) in ear-nose-throat (ENT), periodontics and ophtalmology, and 923 (17%) in gynaecology and obstetrics. 93% of filled prescriptions forms were spontaneously returned to the pharmacy, the others were obtained during the renewal of kit stocks. The cost (over 4 years) of ATBP was quantified: 157,871 F for the 15 departments included, 26,123 F in visceral surgery, 13,520 F in urology, 73,741 F in orthopaedics, 569 F in vascular surgery, 39,720 F in ENT/ophthalmology/periodontics and 4,198 F in gynaecology and obstetrics. According to the Altemeier classification, 2226 class I, 3151 class II, and 209 class III surgical operations were performed. Since the kits have been brought into use, the committee for the protection against nosocomial infections (CLIN) has observed a reduction in the incidence of post-operative infections, according to the Altemeier classification: from 1.6% to 0.5% in class I, from 6.5% to 4.3% in class II, and from 11% to 8.5% in class III. The difference was statistically significant only for classes I (p < 0.01) and II (p < 0.001), and unchanged for class III (p = 0.3). No analysis was carried out for class IV (curative treatments). Both nurses and physicians have greatly appreciated the implementation of this organization. The advantage in terms of post-operative infections, administration exhaustiveness and stock management is obvious. The prescribed kits were systematically appropriate for the surgical interventions. In orthopaedics, cefamandole was used over 24 h (188 kits) in ligament plasty and osteotomy, or for 48 h (499 kits) in prosthetic surgery; 24 amoxicillin/clavulanic acid (first line) and 9 clindamycin/gentamicin (second line) single dose kits have been prescribed in traumatic indications. In ophthalmology, kits were only prescribed in endophtalmitis (24 ofloxacin/fosfomycin single amount kits), implant replacement or cornea graft (1076 ofloxacin 24 h kits) and cataract surgery in diabetic patients (12 ofloxacin single amount kits). In ENT and periodontics, 124 surgical operations required cefazolin single dose kits. In vascular surgery, 5 pefloxacin/gentamicin 48 h kits and 1 amoxicillin/clavulanic acid 48 h kit were used in contaminated limb amputation, 1 cefamandole 48 h kit in class I surgery and 1 vancomycin 24 h kit (betalactamase antibiotic allergy); in thoracic surgery, 1 cefamandole 24 h kit was used for a thoracic wound. In visceral surgery, 9 different kits have been used, depending on the opening (class II) or not (class I) of the digestive tract. 797 cefazolin (first line) and 68 clindamycin/gentamicin (second line) single dose kits were used in class I surgery, and 689 amoxicillin/clavulanic acid single dose (SD) kits in class II surgery. Specific protocols consisted of 18 ceftriaxone/metronidazole and 48 metronidazole/gentamicin SD kits in oesophagus surgery, 11 ceftriaxone and 17 gentamicin SD kits in biliary endoscopy, 137 metronidazole SD kits in proctology and 34 amoxicillin/gentamicin 6 h kits for prevention of endocarditis. In urology, 133 cefotaxime and 20 pefloxacin/gentamicin SD kits were precribed in renal lithiasis, 102 amoxicillin/clavulanic acid SD kits in cystectomy, 27 amoxicillin/gentamicin 6 h kits in endocarditis prevention and 58 cefamandole SD kits in all other indications. In gynaecology and obstetrics, 534 cefazoline and 19 clindamycin/gentamicin (second line) SD kits were used, and 370 doxycyclin SD kits were prescribed in pregnancy termination. Some departments (orthopaedics and visceral surgery) adapted the protocols to their needs, specifically with regard to treatment duration. However, these situations were quickly corrected. A constant follow-up and update of this system, associated with routine audits, should allow the maintenance and possibly the improvement of these results, hence shortening treatment duration.

  6. 49 CFR 173.165 - Polyester resin kits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 2 2012-10-01 2012-10-01 false Polyester resin kits. 173.165 Section 173.165... Polyester resin kits. (a) Except for transportation by aircraft, polyester resin kits consisting of a base... resin kits consisting of a base material component (Class 3, Packing Group II or III) and an activator...

  7. The Use of Kits in the Analysis of Tissue Lipids Requires Validation.

    PubMed

    Rider, T; LeBoeuf, R C; Tso, Patrick; Jandacek, R J

    2016-04-01

    The ready availability and ease of use of kits for the measurement of serum lipids has greatly facilitated these measurements. In many cases it would be convenient to use these kits in the determination of lipid concentrations in tissues. The successful application of serum kits in tissue analysis requires that two important issues be considered. First, the solvent system for the extraction of the lipids and the solvent used for analysis by the kit must be compatible with the reactions in the kit. Second, the concentration range in the analyzed solution must be within the range for which the kit is used. We report here that lipids in liver and adipose tissues may be significantly underestimated by the use of some kits. We recommend that the use of kits for tissue analysis of lipids be validated for the specific analysis.

  8. Oncogenic activation of v-kit involves deletion of a putative tyrosine-substrate interaction site.

    PubMed

    Herbst, R; Munemitsu, S; Ullrich, A

    1995-01-19

    The transforming gene of the Hardy-Zuckerman-4 strain of feline sarcoma virus, v-kit, arose by transduction of the cellular c-kit gene, which encodes the receptor tyrosine kinase (RTK) p145c-kit. To gain insight into the molecular basis of the v-kit transforming potential, we characterized the feline c-kit by cDNA cloning. Comparison of the feline v-kit and c-kit sequences revealed, in addition to deletions of the extracellular and transmembrane domains, three additional mutations in the v-kit oncogene product: deletion of tyrosine-569 and valine-570, the exchange of aspartate at position 761 to glycine, and replacement of the C-terminal 50 amino acids by five unrelated residues. Examinations of individual v-kit mutations in the context of chimeric receptors yielded inhibitory effects for some mutants on both autophosphorylation and substrate phosphorylation functions. In contrast, deletion of tyrosine-569 and valine-570 significantly enhanced transforming and mitogenic activities of p145c-kit, while the other mutations had no significant effects. Conservation in subclass III RTKs and the identification of the corresponding residue in beta PDGF-R, Y579, as a binding site for src family tyrosine kinases suggests an important role for Y568 in kit signal regulation and the definition of its oncogenic potential. Repositioning of Y571 by an inframe two codon deletion may be the crucial alteration resulting in enhancement of v-kit oncogenic activity.

  9. M-COPA suppresses endolysosomal Kit-Akt oncogenic signalling through inhibiting the secretory pathway in neoplastic mast cells.

    PubMed

    Hara, Yasushi; Obata, Yuuki; Horikawa, Keita; Tasaki, Yasutaka; Suzuki, Kyohei; Murata, Takatsugu; Shiina, Isamu; Abe, Ryo

    2017-01-01

    Gain-of-function mutations in Kit receptor tyrosine kinase result in the development of a variety of cancers, such as mast cell tumours, gastrointestinal stromal tumours (GISTs), acute myeloid leukemia, and melanomas. The drug imatinib, a selective inhibitor of Kit, is used for treatment of mutant Kit-positive cancers. However, mutations in the Kit kinase domain, which are frequently found in neoplastic mast cells, confer an imatinib resistance, and cancers expressing the mutants can proliferate in the presence of imatinib. Recently, we showed that in neoplastic mast cells that endogenously express an imatinib-resistant Kit mutant, Kit causes oncogenic activation of the phosphatidylinositol 3-kinase-Akt (PI3K-Akt) pathway and the signal transducer and activator of transcription 5 (STAT5) but only on endolysosomes and on the endoplasmic reticulum (ER), respectively. Here, we show a strategy for inhibition of the Kit-PI3K-Akt pathway in neoplastic mast cells by M-COPA (2-methylcoprophilinamide), an inhibitor of this secretory pathway. In M-COPA-treated cells, Kit localization in the ER is significantly increased, whereas endolysosomal Kit disappears, indicating that M-COPA blocks the biosynthetic transport of Kit from the ER. The drug greatly inhibits oncogenic Akt activation without affecting the association of Kit with PI3K, indicating that ER-localized Kit-PI3K complex is unable to activate Akt. Importantly, M-COPA but not imatinib suppresses neoplastic mast cell proliferation through inhibiting anti-apoptotic Akt activation. Results of our M-COPA treatment assay show that Kit can activate Erk not only on the ER but also on other compartments. Furthermore, Tyr568/570, Tyr703, Tyr721, and Tyr936 in Kit are phosphorylated on the ER, indicating that these five tyrosine residues are all phosphorylated before mutant Kit reaches the plasma membrane (PM). Our study provides evidence that Kit is tyrosine-phosphorylated soon after synthesis on the ER but is unable to activate Akt and also demonstrates that M-COPA is efficacious for growth suppression of neoplastic mast cells.

  10. M-COPA suppresses endolysosomal Kit-Akt oncogenic signalling through inhibiting the secretory pathway in neoplastic mast cells

    PubMed Central

    Hara, Yasushi; Obata, Yuuki; Horikawa, Keita; Tasaki, Yasutaka; Suzuki, Kyohei; Murata, Takatsugu; Shiina, Isamu; Abe, Ryo

    2017-01-01

    Gain-of-function mutations in Kit receptor tyrosine kinase result in the development of a variety of cancers, such as mast cell tumours, gastrointestinal stromal tumours (GISTs), acute myeloid leukemia, and melanomas. The drug imatinib, a selective inhibitor of Kit, is used for treatment of mutant Kit-positive cancers. However, mutations in the Kit kinase domain, which are frequently found in neoplastic mast cells, confer an imatinib resistance, and cancers expressing the mutants can proliferate in the presence of imatinib. Recently, we showed that in neoplastic mast cells that endogenously express an imatinib-resistant Kit mutant, Kit causes oncogenic activation of the phosphatidylinositol 3-kinase-Akt (PI3K-Akt) pathway and the signal transducer and activator of transcription 5 (STAT5) but only on endolysosomes and on the endoplasmic reticulum (ER), respectively. Here, we show a strategy for inhibition of the Kit-PI3K-Akt pathway in neoplastic mast cells by M-COPA (2-methylcoprophilinamide), an inhibitor of this secretory pathway. In M-COPA-treated cells, Kit localization in the ER is significantly increased, whereas endolysosomal Kit disappears, indicating that M-COPA blocks the biosynthetic transport of Kit from the ER. The drug greatly inhibits oncogenic Akt activation without affecting the association of Kit with PI3K, indicating that ER-localized Kit-PI3K complex is unable to activate Akt. Importantly, M-COPA but not imatinib suppresses neoplastic mast cell proliferation through inhibiting anti-apoptotic Akt activation. Results of our M-COPA treatment assay show that Kit can activate Erk not only on the ER but also on other compartments. Furthermore, Tyr568/570, Tyr703, Tyr721, and Tyr936 in Kit are phosphorylated on the ER, indicating that these five tyrosine residues are all phosphorylated before mutant Kit reaches the plasma membrane (PM). Our study provides evidence that Kit is tyrosine-phosphorylated soon after synthesis on the ER but is unable to activate Akt and also demonstrates that M-COPA is efficacious for growth suppression of neoplastic mast cells. PMID:28403213

  11. A morphological study of the pacemaker cells of the aganglionic intestine in Hirschsprung's disease utilizing ls/ls model mice.

    PubMed

    Taniguchi, Kan; Matsuura, Kimio; Matsuoka, Takanori; Nakatani, Hajime; Nakano, Takumi; Furuya, Yasuo; Sugimoto, Takeki; Kobayashi, Michiya; Araki, Keijiro

    2005-06-01

    Hirschsprung's disease is a congenital aganglionic neural disorder of the segmental distal intestine characterized by unsettled pathogenesis. The relationship between Hirschsprung's disease and pacemaker cells (PMC), which almost corresponds to that of the interstitial cells of Cajal (ICC), was morphologically observed at the level of the intermuscular layer corresponding to Auerbach's plexus using ls/ls mice. These mice are an ideal model because of their large intestinal aganglionosis and gene abnormalities, which are similar to the human form of the disease. Immunostaining using anti-c-kit receptor antibody (ACK2), a marker of PMC, applied to whole-mount muscle-layer specimens, revealed the presence of c-kit immunopositive multipolar cells with many cytoplasmic processes in normal mice. For ls/ls mice, however, there were significantly fewer processes. The average number of processes per positive cell of 2.5 for the aganglionic large intestine was fewer than 3.5 for the large and small intestine of normal mice, indicating the inability to form connections between nerves and PMC in the aganglionic intestine. For normal mice with an Auerbach's plexus, the process attachment of ICC to the Auerbach's plexus was observed by scanning electron microscopy. However, for ls/ls mice no attachment to the intermuscular nerve without Auerbach's plexus was found, although transmission electron microscopy showed no difference in the cell structure and organelles of the c-kit immunopositive cells between the normal and ls/ls mice. These findings suggest that in the aganglionic intestine of Hirschsprung's disease, aplasia of enteric ganglia induces secondary disturbances during the normal development of intestinal PMC.

  12. Effects of gestational maternal undernutrition on growth, carcass composition and meat quality of rabbit offspring.

    PubMed

    Symeon, George K; Goliomytis, Michael; Bizelis, Iosif; Papadomichelakis, George; Pagonopoulou, Olga; Abas, Zafeiris; Deligeorgis, Stelios G; Chadio, Stella E

    2015-01-01

    An experiment was conducted in order to evaluate the effects of gestational undernutrition of rabbit does on growth, carcass composition and meat quality of the offsprings. Thirty primiparous non lactating rabbit does were artificially inseminated and randomly divided in three treatment groups: Control (C; fed to 100% of maintenance requirements throughout gestation, n = 10), early undernourished (EU; fed to 50% of maintenance requirements during days 7-19 of gestation, n = 10) and late undernourished (LU; fed to 50% of maintenance requirements during days 20-27 of gestation, n = 10). During the 4th week of the gestation period, LU does significantly lost weight compared to C and EU groups (P<0.05). At kindling, C does produced litters with higher proportions of stillborn kits (P<0.05) while the total litter size (alive and stillborn kits) was not different among groups (10.7, 12.8 and 12.7 kits in C, EU and LU groups, respectively). Kit birth weight tended to be lower in the LU group. During fattening, body weight and feed intake were not different among offsprings of the three experimental groups. Moreover, the maternal undernutrition did not have any impact on carcass composition of the offsprings in terms of carcass parts and internal organs weights as well as meat quality of L. lumborum muscle (pH24, colour, water holding capacity and shear values) at slaughter (70 days of age). Therefore, it can be concluded that the gestational undernutrition of the mother does not have detrimental effects on the productive and quality traits of the offsprings.

  13. Field soil aggregate stability kit for soil quality and rangeland health evaluations

    USGS Publications Warehouse

    Herrick, J.E.; Whitford, W.G.; de Soyza, A. G.; Van Zee, J. W.; Havstad, K.M.; Seybold, C.A.; Walton, M.

    2001-01-01

    Soil aggregate stability is widely recognized as a key indicator of soil quality and rangeland health. However, few standard methods exist for quantifying soil stability in the field. A stability kit is described which can be inexpensively and easily assembled with minimal tools. It permits up to 18 samples to be evaluated in less than 10 min and eliminates the need for transportation, minimizing damage to soil structure. The kit consists of two 21??10.5??3.5 cm plastic boxes divided into eighteen 3.5??3.5 cm sections, eighteen 2.5-cm diameter sieves with 1.5-mm distance openings and a small spatula used for soil sampling. Soil samples are rated on a scale from one to six based on a combination of ocular observations of slaking during the first 5 min following immersion in distilled water, and the percent remaining on a 1.5-mm sieve after five dipping cycles at the end of the 5-min period. A laboratory comparison yielded a correlation between the stability class and percent aggregate stability based on oven dry weight remaining after treatment using a mechanical sieve. We have applied the method in a wide variety of agricultural and natural ecosystems throughout western North America, including northern Mexico, and have found that it is highly sensitive to differences in management and plant community composition. Although the field kit cannot replace the careful laboratory-based measurements of soil aggregate stability, it can clearly provide valuable information when these more intensive procedures are not possible.

  14. Effects of Gestational Maternal Undernutrition on Growth, Carcass Composition and Meat Quality of Rabbit Offspring

    PubMed Central

    Symeon, George K.; Goliomytis, Michael; Bizelis, Iosif; Papadomichelakis, George; Pagonopoulou, Olga; Abas, Zafeiris; Deligeorgis, Stelios G.; Chadio, Stella E.

    2015-01-01

    An experiment was conducted in order to evaluate the effects of gestational undernutrition of rabbit does on growth, carcass composition and meat quality of the offsprings. Thirty primiparous non lactating rabbit does were artificially inseminated and randomly divided in three treatment groups: Control (C; fed to 100% of maintenance requirements throughout gestation, n = 10), early undernourished (EU; fed to 50% of maintenance requirements during days 7–19 of gestation, n = 10) and late undernourished (LU; fed to 50% of maintenance requirements during days 20-27 of gestation, n = 10). During the 4th week of the gestation period, LU does significantly lost weight compared to C and EU groups (P<0.05). At kindling, C does produced litters with higher proportions of stillborn kits (P<0.05) while the total litter size (alive and stillborn kits) was not different among groups (10.7, 12.8 and 12.7 kits in C, EU and LU groups, respectively). Kit birth weight tended to be lower in the LU group. During fattening, body weight and feed intake were not different among offsprings of the three experimental groups. Moreover, the maternal undernutrition did not have any impact on carcass composition of the offsprings in terms of carcass parts and internal organs weights as well as meat quality of L. lumborum muscle (pH24, colour, water holding capacity and shear values) at slaughter (70 days of age). Therefore, it can be concluded that the gestational undernutrition of the mother does not have detrimental effects on the productive and quality traits of the offsprings. PMID:25671602

  15. Targeting Low-arsenic Groundwater with Mobile-phone Technology in Araihazar, Bangladesh

    PubMed Central

    Trevisani, M.; Immel, J.; Jakariya, Md.; Osman, N.; Cheng, Z.; Gelman, A.; Ahmed, K.M.

    2006-01-01

    The Bangladesh Arsenic Mitigation and Water Supply Program (BAMWSP) has compiled field-kit measurements of the arsenic content of groundwater for nearly five million wells. By comparing the spatial distribution of arsenic inferred from these field-kit measurements with geo-referenced laboratory data in a portion of Araihazar upazila, it is shown here that the BAMWSP data could be used for targeting safe aquifers for the installation of community wells in many villages of Bangladesh. Recent experiences with mobile-phone technology to access and update the BAMWSP data in the field are also described. It is shown that the technology, without guaranteeing success, could optimize interventions by guiding the choice of the drilling method that is likely to reach a safe aquifer and identifying those villages where exploratory drilling is needed. PMID:17366770

  16. A national survey of home-based care kits for palliative HIV/AIDS care in South Africa.

    PubMed

    Mabude, Z A; Beksinska, M E; Ramkissoon, A; Wood, S; Folsom, M

    2008-09-01

    The objective of this study was to assess home-based care (HBC) kits and programs in South Africa to evaluate the feasibility of scaling up kit production and distribution. South African HBC organizations received structured questionnaires; key informant interviews and a literature review were completed to assess systems for production, distribution and supply of HBC kits. Meetings with stakeholders were held in two Provinces to share and analyze the study findings. The study team distributed questionnaires to 466 organizations and conducted interviews with representatives from 45 organizations, the Provincial Department of Health (DoH) and manufacturers of kits. All identifiable HBC organizations in South Africa were included in the survey. As a result 215 HBC organizations returned questionnaires; including non-governmental organizations (56%), community-based organizations (32%) and organizations affiliated with government health departments. Two types of kits were available: a home kit and a professional kit. The demand for HBC kits exceeded availability, kit contents and availability varied considerably and the supply chain was irregular. Kit production and distribution systems were fragmented. Replenishment of kit items was problematic. End-users are mostly caregivers who have not received adequate training on their use. The study shows that substantial work has been done by HBC organizations in South Africa to respond to the need for palliative care supplies within resource constraints. The growing demand for kits exceeds the supply. There is a need to improve the supply chain management of HBC kits, strengthen referral systems and links between community-based organizations and government departments, expand training opportunities for care givers, and develop monitoring and evaluation systems.

  17. Single-cell analysis of the fate of c-kit-positive bone marrow cells

    NASA Astrophysics Data System (ADS)

    Czarna, Anna; Sanada, Fumihiro; Matsuda, Alex; Kim, Junghyun; Signore, Sergio; Pereira, João D.; Sorrentino, Andrea; Kannappan, Ramaswamy; Cannatà, Antonio; Hosoda, Toru; Rota, Marcello; Crea, Filippo; Anversa, Piero; Leri, Annarosa

    2017-10-01

    The plasticity of c-kit-positive bone marrow cells (c-kit-BMCs) in tissues different from their organ of origin remains unclear. We tested the hypothesis that c-kit-BMCs are functionally heterogeneous and only a subgroup of these cells possesses cardiomyogenic potential. Population-based assays fall short of identifying the properties of individual stem cells, imposing on us the introduction of single cell-based approaches to track the fate of c-kit-BMCs in the injured heart; they included viral gene-tagging, multicolor clonal-marking and transcriptional profiling. Based on these strategies, we report that single mouse c-kit-BMCs expand clonally within the infarcted myocardium and differentiate into specialized cardiac cells. Newly-formed cardiomyocytes, endothelial cells, fibroblasts and c-kit-BMCs showed in their genome common sites of viral integration, providing strong evidence in favor of the plasticity of a subset of BMCs expressing the c-kit receptor. Similarly, individual c-kit-BMCs, which were infected with multicolor reporters and injected in infarcted hearts, formed cardiomyocytes and vascular cells organized in clusters of similarly colored cells. The uniform distribution of fluorescent proteins in groups of specialized cells documented the polyclonal nature of myocardial regeneration. The transcriptional profile of myogenic c-kit-BMCs and whole c-kit-BMCs was defined by RNA sequencing. Genes relevant for engraftment, survival, migration, and differentiation were enriched in myogenic c-kit-BMCs, a cell subtype which could not be assigned to a specific hematopoietic lineage. Collectively, our findings demonstrate that the bone marrow comprises a category of cardiomyogenic, vasculogenic and/or fibrogenic c-kit-positive cells and a category of c-kit-positive cells that retains an undifferentiated state within the damaged heart.

  18. Single-cell analysis of the fate of c-kit-positive bone marrow cells.

    PubMed

    Czarna, Anna; Sanada, Fumihiro; Matsuda, Alex; Kim, Junghyun; Signore, Sergio; Pereira, João D; Sorrentino, Andrea; Kannappan, Ramaswamy; Cannatà, Antonio; Hosoda, Toru; Rota, Marcello; Crea, Filippo; Anversa, Piero; Leri, Annarosa

    2017-01-01

    The plasticity of c-kit-positive bone marrow cells (c-kit-BMCs) in tissues different from their organ of origin remains unclear. We tested the hypothesis that c-kit-BMCs are functionally heterogeneous and only a subgroup of these cells possesses cardiomyogenic potential. Population-based assays fall short of identifying the properties of individual stem cells, imposing on us the introduction of single cell-based approaches to track the fate of c-kit-BMCs in the injured heart; they included viral gene-tagging, multicolor clonal-marking and transcriptional profiling. Based on these strategies, we report that single mouse c-kit-BMCs expand clonally within the infarcted myocardium and differentiate into specialized cardiac cells. Newly-formed cardiomyocytes, endothelial cells, fibroblasts and c-kit-BMCs showed in their genome common sites of viral integration, providing strong evidence in favor of the plasticity of a subset of BMCs expressing the c-kit receptor. Similarly, individual c-kit-BMCs, which were infected with multicolor reporters and injected in infarcted hearts, formed cardiomyocytes and vascular cells organized in clusters of similarly colored cells. The uniform distribution of fluorescent proteins in groups of specialized cells documented the polyclonal nature of myocardial regeneration. The transcriptional profile of myogenic c-kit-BMCs and whole c-kit-BMCs was defined by RNA sequencing. Genes relevant for engraftment, survival, migration, and differentiation were enriched in myogenic c-kit-BMCs, a cell subtype which could not be assigned to a specific hematopoietic lineage. Collectively, our findings demonstrate that the bone marrow comprises a category of cardiomyogenic, vasculogenic and/or fibrogenic c-kit-positive cells and a category of c-kit-positive cells that retains an undifferentiated state within the damaged heart.

  19. In-frame Val216-Ser217 deletion of KIT in mild piebaldism causes aberrant secretion and SCF response.

    PubMed

    Hattori, Mai; Ishikawa, Osamu; Oikawa, Daisuke; Amano, Hiroo; Yasuda, Masahito; Kaira, Kyoichi; Ishida-Yamamoto, Akemi; Nakano, Hajime; Sawamura, Daisuke; Terawaki, Shin-Ichi; Wakamatsu, Kaori; Tokunaga, Fuminori; Shimizu, Akira

    2018-03-21

    Piebaldism is a pigmentary disorder characterized by a white forelock and depigmented patches. Although the loss-of-function mutations in the KIT gene underlie the disease, the intracellular dynamics of the mutant KIT are largely unknown. We herein report a Japanese family with piebaldism in which the affected members showed a mild phenotype. The objective of this study is to investigate the functions and intracellular dynamics of the mutant KIT protein. We performed genetic analyses of the KIT gene using peripheral blood cells. We analyzed the intracellular localization of the mutant KIT protein in HEK293T cells transfected with wild-type (Wt) and/or mutant KIT genes. Immunoprecipitation analyses, immunoblotting and immunofluorescence studies were performed using antibodies against KIT and downstream signaling proteins. Glycosidase digestion analysis was performed to clarify the intracellular localization of KIT protein. A genetic analysis revealed a novel heterozygous mutation c.645_650delTGTGTC which results in the in-frame deletion of Val 216 and Ser 217 in the extracellular domain of KIT. Immunoprecipitation analyses confirmed that the wild and mutant KIT formed a heterodimer after treatment with stem cell factor (SCF); however, the phosphorylation of the downstream signaling factors was decreased. In an immunofluorescence study, the mutant KIT accumulated predominantly in the endoplasmic reticulum (ER) and was sparsely expressed on the cell surface. A glycosidase digestion study revealed that the mutant KIT is predominantly localized in the ER. These data reveal an aberrant function and intracellular localization of mutant KIT protein in piebaldism. Copyright © 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.

  20. Membrane estrogen receptor alpha is an important modulator of bone marrow C-Kit+ cells mediated cardiac repair after myocardial infarction

    PubMed Central

    Su, Feng; Zhang, Wentian; Liu, Jianfang

    2015-01-01

    It has been validated that c-kit positive (c-kit+) cells in infarcted myocardium are from bone marrow (BM). Given the recent study that in the heart, estrogen receptor alpha (ERα) is involved in adaptive mechanisms by supporting cardiomyocytes survival via post-infarct cardiac c-kit+ cells, we tested a novel hypothesis that membrane ERα (mERа) supports survival of BM c-kit+ cells and enhance protective paracrine function for cardiac repair. Our data showed that myocardial infarction (MI) leads to an increase in c-kit+ first in bone marrow and then specifically within the infarcted myocardium. Also up-regulated mERа in post-infarct BM c-kit+ cells was found in day 3 post MI. In vitro co-culture system, mERа+ enhances the beneficial effects of BM c-kit+ cells by increasing their viability and reducing apoptosis. Post-infarct c-kit+ mERа+ cells population expresses predominant ERα and holds self-renewal as well as cardiac differentiation potentials after MI. In vivo, BM c-kit+ cells reduced infarct size, fibrosis and improved cardiac function. In conclusion, BM c-kit+ mERа+ exerted significantly cardiac protection after MI. A potential important implication of this study is that the manipulation of BM c-kit+ stem cells with ERа-dependent fashion may be helpful in recovering functional performance after cardiac tissue injury. PMID:26191121

  1. Direct engagement of the PI3K pathway by mutant KIT dominates oncogenic signaling in gastrointestinal stromal tumor.

    PubMed

    Bosbach, Benedikt; Rossi, Ferdinand; Yozgat, Yasemin; Loo, Jennifer; Zhang, Jennifer Q; Berrozpe, Georgina; Warpinski, Katherine; Ehlers, Imke; Veach, Darren; Kwok, Andrew; Manova, Katia; Antonescu, Cristina R; DeMatteo, Ronald P; Besmer, Peter

    2017-10-03

    Gastrointestinal stromal tumors (GISTs) predominantly harbor activating mutations in the receptor tyrosine kinase KIT. To genetically dissect in vivo the requirement of different signal transduction pathways emanating from KIT for tumorigenesis, the oncogenic Kit V558Δ mutation was combined with point mutations abrogating specific phosphorylation sites on KIT. Compared with single-mutant Kit V558Δ/+ mice, double-mutant Kit V558Δ;Y567F/Y567F knock-in mice lacking the SRC family kinase-binding site on KIT (pY567) exhibited attenuated MAPK signaling and tumor growth. Surprisingly, abrogation of the PI3K-binding site (pY719) in Kit V558Δ;Y719F/Y719F mice prevented GIST development, although the interstitial cells of Cajal (ICC), the cells of origin of GIST, were normal. Pharmacologic inhibition of the PI3K pathway in tumor-bearing Kit V558Δ/+ mice with the dual PI3K/mTOR inhibitor voxtalisib, the pan-PI3K inhibitor pilaralisib, and the PI3K-alpha-restricted inhibitor alpelisib each diminished tumor proliferation. The addition of the MEK inhibitor PD-325901 or binimetinib further decreased downstream KIT signaling. Moreover, combining PI3K and MEK inhibition was effective against imatinib-resistant Kit V558Δ;T669I/+ tumors.

  2. Direct engagement of the PI3K pathway by mutant KIT dominates oncogenic signaling in gastrointestinal stromal tumor

    PubMed Central

    Bosbach, Benedikt; Rossi, Ferdinand; Yozgat, Yasemin; Loo, Jennifer; Zhang, Jennifer Q.; Berrozpe, Georgina; Warpinski, Katherine; Ehlers, Imke; Kwok, Andrew; Manova, Katia; Antonescu, Cristina R.; DeMatteo, Ronald P.; Besmer, Peter

    2017-01-01

    Gastrointestinal stromal tumors (GISTs) predominantly harbor activating mutations in the receptor tyrosine kinase KIT. To genetically dissect in vivo the requirement of different signal transduction pathways emanating from KIT for tumorigenesis, the oncogenic KitV558Δ mutation was combined with point mutations abrogating specific phosphorylation sites on KIT. Compared with single-mutant KitV558Δ/+ mice, double-mutant KitV558Δ;Y567F/Y567F knock-in mice lacking the SRC family kinase-binding site on KIT (pY567) exhibited attenuated MAPK signaling and tumor growth. Surprisingly, abrogation of the PI3K-binding site (pY719) in KitV558Δ;Y719F/Y719F mice prevented GIST development, although the interstitial cells of Cajal (ICC), the cells of origin of GIST, were normal. Pharmacologic inhibition of the PI3K pathway in tumor-bearing KitV558Δ/+ mice with the dual PI3K/mTOR inhibitor voxtalisib, the pan-PI3K inhibitor pilaralisib, and the PI3K-alpha–restricted inhibitor alpelisib each diminished tumor proliferation. The addition of the MEK inhibitor PD-325901 or binimetinib further decreased downstream KIT signaling. Moreover, combining PI3K and MEK inhibition was effective against imatinib-resistant KitV558Δ;T669I/+ tumors. PMID:28923937

  3. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples...

  4. 40 CFR 745.88 - Recognized test kits.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 40 Protection of Environment 32 2012-07-01 2012-07-01 false Recognized test kits. 745.88 Section... Renovation § 745.88 Recognized test kits. (a) Effective June 23, 2008, EPA recognizes the test kits that have... publicizes its recognition of the first test kit that meets both the negative response and positive response...

  5. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples...

  6. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples...

  7. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples...

  8. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples...

  9. 46 CFR 121.710 - First-aid kits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents and instructions. For equivalent...

  10. 19 CFR 122.132 - Sealing of aircraft liquor kits.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 19 Customs Duties 1 2010-04-01 2010-04-01 false Sealing of aircraft liquor kits. 122.132 Section... OF THE TREASURY AIR COMMERCE REGULATIONS Aircraft Liquor Kits § 122.132 Sealing of aircraft liquor kits. (a) Sealing required. Aircraft liquor kits shall be sealed on board the aircraft by crewmembers...

  11. 46 CFR 184.710 - First-aid kits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 7 2012-10-01 2012-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...

  12. 21 CFR 870.1350 - Catheter balloon repair kit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Catheter balloon repair kit. 870.1350 Section 870... repair kit. (a) Identification. A catheter balloon repair kit is a device used to repair or replace the balloon of a balloon catheter. The kit contains the materials, such as glue and balloons, necessary to...

  13. 21 CFR 870.1350 - Catheter balloon repair kit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Catheter balloon repair kit. 870.1350 Section 870... repair kit. (a) Identification. A catheter balloon repair kit is a device used to repair or replace the balloon of a balloon catheter. The kit contains the materials, such as glue and balloons, necessary to...

  14. 46 CFR 184.710 - First-aid kits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 7 2013-10-01 2013-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...

  15. 46 CFR 121.710 - First-aid kits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents and instructions. For equivalent...

  16. 19 CFR 122.132 - Sealing of aircraft liquor kits.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 19 Customs Duties 1 2011-04-01 2011-04-01 false Sealing of aircraft liquor kits. 122.132 Section... OF THE TREASURY AIR COMMERCE REGULATIONS Aircraft Liquor Kits § 122.132 Sealing of aircraft liquor kits. (a) Sealing required. Aircraft liquor kits shall be sealed on board the aircraft by crewmembers...

  17. 21 CFR 870.1350 - Catheter balloon repair kit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Catheter balloon repair kit. 870.1350 Section 870... repair kit. (a) Identification. A catheter balloon repair kit is a device used to repair or replace the balloon of a balloon catheter. The kit contains the materials, such as glue and balloons, necessary to...

  18. 46 CFR 121.710 - First-aid kits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents and instructions. For equivalent...

  19. 46 CFR 184.710 - First-aid kits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 7 2014-10-01 2014-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...

  20. 46 CFR 121.710 - First-aid kits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents and instructions. For equivalent...

  1. 19 CFR 122.132 - Sealing of aircraft liquor kits.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 19 Customs Duties 1 2013-04-01 2013-04-01 false Sealing of aircraft liquor kits. 122.132 Section... OF THE TREASURY AIR COMMERCE REGULATIONS Aircraft Liquor Kits § 122.132 Sealing of aircraft liquor kits. (a) Sealing required. Aircraft liquor kits shall be sealed on board the aircraft by crewmembers...

  2. 19 CFR 122.132 - Sealing of aircraft liquor kits.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 1 2012-04-01 2012-04-01 false Sealing of aircraft liquor kits. 122.132 Section... OF THE TREASURY AIR COMMERCE REGULATIONS Aircraft Liquor Kits § 122.132 Sealing of aircraft liquor kits. (a) Sealing required. Aircraft liquor kits shall be sealed on board the aircraft by crewmembers...

  3. 46 CFR 184.710 - First-aid kits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 7 2011-10-01 2011-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...

  4. 19 CFR 122.132 - Sealing of aircraft liquor kits.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 19 Customs Duties 1 2014-04-01 2014-04-01 false Sealing of aircraft liquor kits. 122.132 Section... OF THE TREASURY AIR COMMERCE REGULATIONS Aircraft Liquor Kits § 122.132 Sealing of aircraft liquor kits. (a) Sealing required. Aircraft liquor kits shall be sealed on board the aircraft by crewmembers...

  5. DEMONSTRATION BULLETIN: CLOR-N-SOIL PCB TEST KIT L2000 PCB/CHLORIDE ANALYZER - DEXSIL CORP.

    EPA Science Inventory

    DEXSIL CORP(Environmental Test Kits)The Dexsil Corporation (Dexsil) produces two test kits that detect polychlorinated biphenyls (PCB) in soil: the Dexsil Clor-N-Soil PCB Screening Kit, and the Dexsil L2000 PCB/Chloride Analyzer. The Dexsil Clor-N-Soil PCB Screening Kit extr...

  6. 46 CFR 184.710 - First-aid kits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false First-aid kits. 184.710 Section 184.710 Shipping COAST... CONTROL AND MISCELLANEOUS SYSTEMS AND EQUIPMENT Miscellaneous § 184.710 First-aid kits. A vessel must carry either a first-aid kit approved under approval series 160.041 or a kit with equivalent contents...

  7. A Magnetic Nanoparticle Based Nucleic Acid Isolation and Purification Instrument for DNA Extraction of Escherichia Coli O157: H7.

    PubMed

    Chen, Yahui; Lin, Jianhan; Jiang, Qin; Chen, Qi; Zhang, Shengjun; Li, Li

    2016-03-01

    The objective of this study was to evaluate the performance of a nucleic acid isolation and purification instrument using Escherichia coli O157:H7 as the model. The instrument was developed with magnetic nanoparticles for efficiently capturing nucleic acids and an intelligent mechanical unit for automatically performing the whole nucleic acid extraction process. A commercial DNA extraction kit from Huier Nano Company was used as reference. Nucleic acids in 1 ml of E. coli O157: H7 at a concentration of 5 x 10(8) CFU/mL were extracted by using this instrument and the kit in parallel and then detected by an ultraviolet spectrophotometer to obtain A260 values and A260/A280 values for the determination of the extracted DNA's quantity and purity, respectively. The A260 values for the instrument and the kit were 0.78 and 0.61, respectively, and the A260/A280 values were 1.98 and 1.93. The coefficient of variations of these parallel tests ranged from 10.5% to 16.7%. The results indicated that this nucleic acid isolation and purification instrument could extract a comparable level of nucleic acid within 50 min compared to the commercial DNA extraction kit.

  8. Immunohistochemistry of a choroidal melanoma: nestin, CD34 and CD117÷c-kit labeling.

    PubMed

    Vrapciu, Alexandra Diana; Rusu, Mugurel Constantin; Voinea, Liliana Mary

    2014-01-01

    In a case of choroidal melanoma (CM) in a 70-year-old male patient, was firstly aimed at studying the processes of angiogenesis by use of nestin and CD34 antibodies. Anti-CD117÷c-kit antibodies were further considered for their progenitor cells specificity. Choroidal melanoma was histopathologically confirmed. Nestin-positive endothelia were found in the CM and the adjacent retina, but not in endothelia elsewhere in that eye. Nestin-positive non-pigmentary cells were found within the CM. Filopodia-projecting endothelial tip cells (ETCs), nestin- and CD34-positive were found in the CM. CD34-positive ETCs were also found in the iridial stroma. There were found two different immune patterns of the retinal Müller cells (MCs). They were nestin-positive in the retina adjacent to the tumor, but negative in any other part of retina. On the other hand, CD117÷c-kit antibodies labeled MCs as follows: (a) discontinuously, or continuously, in the retina adjacent to the CM; (b) only the inner segments of the MCs were labeled in the retina unrelated to the CM. While nestin could be a reliable marker for retinal damage, the CD117÷c-kit phenotype of MCs still needs further investigations. Antiangiogenic therapy appears as a good choice for tumor therapy.

  9. Insulin-like growth factor-1-mediated regulation of miR-193a expression promotes the migration and proliferation of c-kit-positive mouse cardiac stem cells.

    PubMed

    Sun, Yuning; Xu, Rongfeng; Huang, Jia; Yao, Yuyu; Pan, Xiaodong; Chen, Zhongpu; Ma, Genshan

    2018-02-21

    C-kit-positive cardiac stem cells (CSCs) have been shown to be a promising candidate treatment for myocardial infarction and heart failure. Insulin-like growth factor (IGF)-1 is an anabolic growth hormone that regulates cellular proliferation, differentiation, senescence, and death in various tissues. Although IGF-1 promotes the migration and proliferation of c-kit-positive mouse CSCs, the underlying mechanism remains unclear. Cells were isolated from adult mouse hearts, and c-kit-positive CSCs were separated using magnetic beads. The cells were cultured with or without IGF-1, and c-kit expression was measured by Western blotting. IGF-1 induced CSC proliferation and migration, as measured through Cell Counting Kit-8 (CCK-8) and Transwell assays, respectively. The miR-193a expression was measured by quantitative real-time PCR (qPCR) assays. IGF-1 enhanced c-kit expression in c-kit-positive CSCs. The activities of the phosphoinositol 3-kinase (PI3K)/AKT signaling pathway and DNA methyltransferases (DNMTs) were enhanced, and their respective inhibitors LY294002 and 5-azacytidine (5-AZA) blunted c-kit expression. Based on the results of quantitative real-time PCR (qPCR) assays, the expression of miR-193a, which is embedded in a CpG island, was down-regulated in the IGF-1-stimulated group and negatively correlated with c-kit expression, whereas c-kit-positive CSCs infected with lentivirus carrying micro-RNA193a displayed reduced c-kit expression, migration and proliferation. IGF-1 upregulated c-kit expression in c-kit-positive CSCs resulting in enhanced CSC proliferation and migration by activating the PI3K/AKT/DNMT signaling pathway to epigenetically silence miR-193a, which negatively modifies the c-kit expression level.

  10. Investigation of c-KIT and Ki67 expression in normal, preneoplastic and neoplastic canine prostate.

    PubMed

    Fonseca-Alves, Carlos Eduardo; Kobayashi, Priscilla Emiko; Palmieri, Chiara; Laufer-Amorim, Renée

    2017-12-06

    c-KIT expression has been related to bone metastasis in human prostate cancer, but whether c-KIT expression can be similarly classified in canine prostatic tissue is unknown. This study assessed c-KIT and Ki67 expression in canine prostate cancer (PC). c-KIT gene and protein expression and Ki67 expression were evaluated in forty-four canine prostatic tissues by immunohistochemistry, RT-qPCR and western blot. Additionally, we have investigated c-KIT protein expression by immunoblotting in two primary canine prostate cancer cell lines. Eleven normal prostates, 12 proliferative inflammatory atrophy (PIA) prostates, 18 PC, 3 metastatic lesions and two prostate cancer cell cultures (PC1 and PC2) were analysed. The prostatic tissue exhibited varying degrees of membranous, cytoplasmic or membranous/cytoplasmic c-KIT staining. Four normal prostates, 4 PIA and 5 prostatic carcinomas showed positive c-KIT expression. No c-KIT immunoexpression was observed in metastases. Canine prostate cancer and PIA samples contained a higher number of Ki67-positive cells compared to normal samples. The median relative quantification (RQ) for c-KIT expression in normal, PIA and prostate cancer and metastatic samples were 0.6 (0.1-2.5), 0.7 (0.09-2.1), 0.7 (0.09-5.1) and 0.1 (0.07-0.6), respectively. A positive correlation between the number of Ki67-positive cells and c-KIT transcript levels was observed in prostate cancer samples. In the cell line, PC1 was negative for c-KIT protein expression, while PC2 was weakly positive. The present study identified a strong correlation between c-KIT expression and proliferative index, suggesting that c-KIT may influence cell proliferation. Therefore, c-KIT heterogeneous protein expression among the samples (five positive and thirteen negative prostate cancer samples) indicates a personalized approach for canine prostate cancer.

  11. Validation of a commercial kit aimed to the detection of pathogenic anisakid nematodes in fish products.

    PubMed

    Cavallero, Serena; Bruno, Alessandro; Arletti, Enrico; Caffara, Monica; Fioravanti, Maria Letizia; Costa, Antonella; Cammilleri, Gaetano; Graci, Stefania; Ferrantelli, Vincenzo; D'Amelio, Stefano

    2017-09-18

    Anisakids are parasitic nematodes responsible for a zoonosis that occurs following the ingestion of fish and fish products infected with larvae belonging to the genera Anisakis and Pseudoterranova. Rarely Contracaecum is found in association with gastric/intestinal illness, while Hysterothylacium is commonly considered not pathogenic. Although Real Time PCR assays have been recently used with the aim to detect and quantify these parasites in food products, methods applied did not undergo through extensive validation process, a feature highly desirable or mandatory in the case of testing laboratories accredited for the ISO EN 17025:2005. Here, a comprehensive study has been performed to validate a commercial kit based on multiplex real time PCR for the qualitative detection of Anisakis and Pseudoterranova. Inclusivity/exclusivity trials were carried out on DNA from species of the genera Anisakis, Pseudoterranova, Contracaecum, Hysterothylacium and Ascaris, on fish intentionally contaminated with Anisakis spp. and Pseudoterranova spp. and on marine organisms as fish, crustacean and squid to test the commercial kit on a large sample. The assay gave positive amplification for several Anisakis and Pseudoterranova species, while providing no signal for the members of the remaining genera. Each sample was correctly assigned either to Anisakis or Pseudoterranova, thus indicating that no cross-reaction occurred. The LOD was determined using two independent standard curves. Robustness was assayed by using two different thermocyclers in three distinct laboratories with different operators. The establishment of a validation dossier will permit the use of the commercial kit for the detection of Anisakis and Pseudoterranova DNA in fish and fish products intended for human consumption by public or private laboratories, following the requirements regarding the quality assurance processes described in the ISO EN 17025:2005. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Disruption of c-Kit Signaling in Kit(W-sh/W-sh) Growing Mice Increases Bone Turnover.

    PubMed

    Lotinun, Sutada; Krishnamra, Nateetip

    2016-08-16

    c-Kit tyrosine kinase receptor has been identified as a regulator of bone homeostasis. The c-Kit loss-of-function mutations in WBB6F1/J-Kit(W/W-v) mice result in low bone mass. However, these mice are sterile and it is unclear whether the observed skeletal phenotype is secondary to a sex hormone deficiency. In contrast, C57BL/6J-Kit(W-sh)/(W-sh) (W(sh)/W(sh)) mice, which carry an inversion mutation affecting the transcriptional regulatory elements of the c-Kit gene, are fertile. Here, we showed that W(sh)/W(sh) mice exhibited osteopenia with elevated bone resorption and bone formation at 6- and 9-week-old. The c-Kit W(sh) mutation increased osteoclast differentiation, the number of committed osteoprogenitors, alkaline phosphatase activity and mineralization. c-Kit was expressed in both osteoclasts and osteoblasts, and c-Kit expression was decreased in W(sh)/W(sh)osteoclasts, but not osteoblasts, suggesting an indirect effect of c-Kit on bone formation. Furthermore, the osteoclast-derived coupling factor Wnt10b mRNA was increased in W(sh)/W(sh) osteoclasts. Conditioned medium from W(sh)/W(sh) osteoclasts had elevated Wnt10b protein levels and induced increased alkaline phosphatase activity and mineralization in osteoblast cultures. Antagonizing Wnt10b signaling with DKK1 or Wnt10b antibody inhibited these effects. Our data suggest that c-Kit negatively regulates bone turnover, and disrupted c-Kit signaling couples increased bone resorption with bone formation through osteoclast-derived Wnt 10 b.

  13. Exacerbated experimental autoimmune encephalomyelitis in mast-cell-deficient Kit W-sh/W-sh mice.

    PubMed

    Piconese, Silvia; Costanza, Massimo; Musio, Silvia; Tripodo, Claudio; Poliani, Pietro L; Gri, Giorgia; Burocchi, Alessia; Pittoni, Paola; Gorzanelli, Andrea; Colombo, Mario P; Pedotti, Rosetta

    2011-04-01

    Mast cell (MC)-deficient c-Kit mutant Kit(W/W-v) mice are protected against experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, suggesting a detrimental role for MCs in this disease. To further investigate the role of MCs in EAE, we took advantage of a recently characterized model of MC deficiency, Kit(W-sh/W-sh). Surprisingly, we observed that myelin oligodendrocyte glycoprotein (MOG)(35-55)-induced chronic EAE was exacerbated in Kit(W-sh/W-sh) compared with Kit(+/+) mice. Kit(W-sh/W-sh) mice showed more inflammatory foci in the central nervous system (CNS) and increased T-cell response against myelin. To understand whether the discrepant results obtained in Kit(W-sh/W-sh) and in Kit(W/W-v) mice were because of the different immunization protocols, we induced EAE in these two strains with varying doses of MOG(35-55) and adjuvants. Although Kit(W-sh/W-sh) mice exhibited exacerbated EAE under all immunization protocols, Kit(W/W-v) mice were protected from EAE only when immunized with high, but not low, doses of antigen and adjuvants. Kit(W-sh/W-sh) mice reconstituted systemically, but not in the CNS, with bone marrow-derived MCs still developed exacerbated EAE, indicating that protection from disease could be exerted by MCs mainly in the CNS, and/or by other cells possibly dysregulated in Kit(W-sh/W-sh) mice. In summary, these data suggest to reconsider MC contribution to EAE, taking into account the variables of using different experimental models and immunization protocols.

  14. [Outline and effectiveness of support system in the surgical center by supply, processing and distribution center (SPD)].

    PubMed

    Ito, Nobuko; Chinzei, Mieko; Fujiwara, Haruko; Usui, Hisako; Hanaoka, Kazuo; Saitoh, Eisho

    2006-04-01

    Supply, Processing and Distribution system had been introduced to surgical center (the University of Tokyo Hospital) since October of 2002. This system had reduced stock for medicine and materials and decreased medical cost dramatically. We designed some kits for therapeutic drugs related to anesthesia. They were prepared for general anesthesia, epidural and spinal anesthesia, and cardiovascular anesthesia, respectively. One kit had been used for one patient, and new kits were prepared in the anesthesia preparation room by pharmaceutical department staffs. Equipment, for general anesthesia as well as epidural and spinal anesthesia, and central catheter set were also designed and provided for each patient by SPD system. According to the questionnaire of anesthesia residents before and after introduction of SPD system, the time spent for anesthesia preparation had been reduced and 92.3% residents had answered that preparation for anesthesia on the previous day was getting easier. Most of the anesthesia residents had been less stressed after introduction of SPD system. Beside the dramatic economical effect, coordination with SPD system and pharmaceutical department reduced anesthesia preparation time and stress of the staff. Introduction of Support system of SPD to surgical center is important for safe and effective management of operating rooms.

  15. Discovery of amido-benzisoxazoles as potent c-Kit inhibitors

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

    Kunz, Roxanne K.; Rumfelt, Shannon; Chen, Ning

    2010-01-12

    Deregulation of the receptor tyrosine kinase c-Kit is associated with an increasing number of human diseases, including certain cancers and mast cell diseases. Interference of c-Kit signaling with multi-kinase inhibitors has been shown clinically to successfully treat gastrointestinal stromal tumors and mastocytosis. Targeted therapy of c-Kit activity may provide therapeutic advantages against off-target effects for non-oncology applications. A new structural class of c-Kit inhibitors is described, including in vitro c-Kit potency, kinase selectivity, and the observed binding mode.

  16. Comparison of DNA extraction kits and modification of DNA elution procedure for the quantitation of subdominant bacteria from piggery effluents with real-time PCR

    PubMed Central

    Desneux, Jérémy; Pourcher, Anne-Marie

    2014-01-01

    Four commercial DNA extraction kits and a minor modification in the DNA elution procedure were evaluated for the quantitation of bacteria in pig manure samples. The PowerSoil®, PowerFecal®, NucleoSpin® Soil kits and QIAamp® DNA Stool Mini kit were tested on raw manure samples and on lagoon effluents for their ability to quantify total bacteria and a subdominant bacteria specific of pig manure contamination: Lactobacillus amylovorus. The NucleoSpin® Soil kit (NS kit), and to a lesser extent the PowerFecal® kit were the most efficient methods. Regardless of the kit utilized, the modified elution procedure increased DNA yield in the lagoon effluent by a factor of 1.4 to 1.8. When tested on 10 piggery effluent samples, compared to the QIAamp kit, the NS kit combined with the modified elution step, increased by a factor up to 1.7 log10 the values of the concentration of L. amylovorus. Regardless of the type of manure, the best DNA quality and the highest concentrations of bacteria were obtained using the NS kit combined with the modification of the elution procedure. The method recommended here significantly improved quantitation of subdominant bacteria in manure. PMID:24838631

  17. Automated forensic DNA purification optimized for FTA card punches and identifiler STR-based PCR analysis.

    PubMed

    Tack, Lois C; Thomas, Michelle; Reich, Karl

    2007-03-01

    Forensic labs globally face the same problem-a growing need to process a greater number and wider variety of samples for DNA analysis. The same forensic lab can be tasked all at once with processing mixed casework samples from crime scenes, convicted offender samples for database entry, and tissue from tsunami victims for identification. Besides flexibility in the robotic system chosen for forensic automation, there is a need, for each sample type, to develop new methodology that is not only faster but also more reliable than past procedures. FTA is a chemical treatment of paper, unique to Whatman Bioscience, and is used for the stabilization and storage of biological samples. Here, the authors describe optimization of the Whatman FTA Purification Kit protocol for use with the AmpFlSTR Identifiler PCR Amplification Kit.

  18. [Denaturation of egg antigens by cooking].

    PubMed

    Watanabe, Hiroko; Akaboshi, Chie; Sekido, Haruko; Tanaka, Kouki; Tanaka, Kazuko; Shimojo, Naoki

    2012-01-01

    Changes in egg protein contents by cooking were measured with an ELISA kit using Tris-HCl buffer in model foods including cake, meatballs, pasta and pudding made with whole egg, egg-white and egg-yolk. The egg protein contents were lowest in the deep-fried model foods of cakes and meatballs. Ovalbumin (OVA) was undetectable (<1 µg/g) and ovomucoid (OVM) was lowest in pouched meatballs, suggesting that processing temperature and uniform heat-treatment affect the detection of egg protein. Furthermore, egg protein contents were below 6 µg/g in the pouched meatballs and pasta made with egg-yolk, and OVA and OVM were not detected by Western blotting analysis with human IgE from patients' serum. On the other hand, processed egg proteins were detected with an ELISA kit using a surfactant and reductant in the extract buffer.

  19. Characterization of various types of mast cells derived from model mice of familial gastrointestinal stromal tumors with KIT-Asp818Tyr mutation

    PubMed Central

    Kajimoto, Noriko; Nakai, Norihiro; Ohkouchi, Mizuka; Hashikura, Yuka; Liu-Kimura, Ning-Ning; Isozaki, Koji; Hirota, Seiichi

    2015-01-01

    Sporadic mast cell neoplasms and gastrointestinal stromal tumors (GISTs) often have various types of somatic gain-of-function mutations of the c-kit gene which encodes a receptor tyrosine kinase, KIT. Several types of germline gain-of-function mutations of the c-kit gene have been detected in families with multiple GISTs. All three types of model mice for the familial GISTs with germline c-kit gene mutations at exon 11, 13 or 17 show development of GIST, while they are different from each other in skin mast cell number. Skin mast cell number in the model mice with exon 17 mutation was unchanged compared to the corresponding wild-type mice. In the present study, we characterized various types of mast cells derived from the model mice with exon 17 mutation (KIT-Asp818Tyr) corresponding to human familial GIST case with human KIT-Asp820Tyr to clarify the role of the c-kit gene mutation in mast cells. Bone marrow-derived cultured mast cells (BMMCs) derived from wild-type mice, heterozygotes and homozygotes were used for the experiments. Immortalized BMMCs, designated as IMC-G4 cells, derived from BMMCs of a homozygote during long-term culture were also used. Ultrastructure, histamine contents, proliferation profiles and phosphorylation of various signaling molecules in those cells were examined. In IMC-G4 cells, presence of additional mutation(s) of the c-kit gene and effect of KIT inhibitors on both KIT autophosphorylation and cell proliferation were also analyzed. We demonstrated that KIT-Asp818Tyr did not affect ultrastructure and proliferation profiles but did histamine contents in BMMCs. IMC-G4 cells had an additional novel c-kit gene mutation of KIT-Tyr421Cys which is considered to induce neoplastic transformation of mouse mast cells and the mutation appeared to be resistant to a KIT inhibitor of imatinib but sensitive to another KIT inhibitor of nilotinib. IMC-G4 cells might be a useful mast cell line to investigate mast cell biology. PMID:26722383

  20. 46 CFR 121.710 - First-aid kits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false First-aid kits. 121.710 Section 121.710 Shipping COAST... SYSTEMS AND EQUIPMENT Miscellaneous § 121.710 First-aid kits. A vessel must carry either a first-aid kit... kits, the contents must be stowed in a suitable, watertight container that is marked “First-Aid Kit”. A...

  1. Supporting and Resourcing Secondary Science Teachers in Rural and Regional Schools

    ERIC Educational Resources Information Center

    Kenny, John; Seen, Andrew; Purser, John

    2008-01-01

    This paper reports on the outcomes of a pilot project to support secondary teachers of science in rural and regional Tasmania. The pilot project involved eight regional schools and was based on the provision of a kit of materials and an associated learning program that used brine shrimp or "sea-monkeys" to test for water quality. The…

  2. Genetic lineage tracing identifies in situ Kit-expressing cardiomyocytes

    PubMed Central

    Liu, Qiaozhen; Yang, Rui; Huang, Xiuzhen; Zhang, Hui; He, Lingjuan; Zhang, Libo; Tian, Xueying; Nie, Yu; Hu, Shengshou; Yan, Yan; Zhang, Li; Qiao, Zengyong; Wang, Qing-Dong; Lui, Kathy O; Zhou, Bin

    2016-01-01

    Cardiac cells marked by c-Kit or Kit, dubbed cardiac stem cells (CSCs), are in clinical trials to investigate their ability to stimulate cardiac regeneration and repair. These studies were initially motivated by the purported cardiogenic activity of these cells. Recent lineage tracing studies using Kit promoter to drive expression of the inducible Cre recombinase showed that these CSCs had highly limited cardiogenic activity, inadequate to support efficient cardiac repair. Here we reassess the lineage tracing data by investigating the identity of cells immediately after Cre labeling. Our instant lineage tracing approach identifies Kit-expressing cardiomyocytes, which are labeled immediately after tamoxifen induction. In combination with long-term lineage tracing experiments, these data reveal that the large majority of long-term labeled cardiomyocytes are pre-existing Kit-expressing cardiomyocytes rather than cardiomyocytes formed de novo from CSCs. This study presents a new interpretation for the contribution of Kit+ cells to cardiomyocytes and shows that Kit genetic lineage tracing over-estimates the cardiogenic activity of Kit+ CSCs. PMID:26634606

  3. Oncogenic Kit signals on endolysosomes and endoplasmic reticulum are essential for neoplastic mast cell proliferation

    PubMed Central

    Obata, Yuuki; Toyoshima, Shota; Wakamatsu, Ei; Suzuki, Shunichi; Ogawa, Shuhei; Esumi, Hiroyasu; Abe, Ryo

    2014-01-01

    Kit is a receptor-type tyrosine kinase found on the plasma membrane. It can transform mast cells through activating mutations. Here, we show that a mutant Kit from neoplastic mast cells from mice, Kit(D814Y), is permanently active and allows cells to proliferate autonomously. It does so by activating two signalling pathways from different intracellular compartments. Mutant Kit from the cell surface accumulates on endolysosomes through clathrin-mediated endocytosis, which requires Kit’s kinase activity. Kit(D814Y) is constitutively associated with phosphatidylinositol 3-kinase, but the complex activates Akt only on the cytoplasmic surface of endolysosomes. It resists destruction because it is under-ubiquitinated. Kit(D814Y) also appears in the endoplasmic reticulum soon after biosynthesis, and there, can activate STAT5 aberrantly. These mechanisms of oncogenic signalling are also seen in rat and human mast cell leukemia cells. Thus, oncogenic Kit signalling occurs from different intracellular compartments, and the mutation acts by altering Kit trafficking as well as activation. PMID:25493654

  4. Structural Basis for Activation of the Receptor Tyrosine Kinase KIT by Stem Cell Factor

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

    Yuzawa,S.; Opatowsky, Y.; Zhang, Z.

    2007-01-01

    Stem Cell Factor (SCF) initiates its multiple cellular responses by binding to the ectodomain of KIT, resulting in tyrosine kinase activation. We describe the crystal structure of the entire ectodomain of KIT before and after SCF stimulation. The structures show that KIT dimerization is driven by SCF binding whose sole role is to bring two KIT molecules together. Receptor dimerization is followed by conformational changes that enable lateral interactions between membrane proximal Ig-like domains D4 and D5 of two KIT molecules. Experiments with cultured cells show that KIT activation is compromised by point mutations in amino acids critical for D4-D4more » interaction. Moreover, a variety of oncogenic mutations are mapped to the D5-D5 interface. Since key hallmarks of KIT structures, ligand-induced receptor dimerization, and the critical residues in the D4-D4 interface, are conserved in other receptors, the mechanism of KIT stimulation unveiled in this report may apply for other receptor activation.« less

  5. Evaluation of DNA extraction methods for PCR-based detection of Listeria monocytogenes from vegetables.

    PubMed

    Vojkovska, H; Kubikova, I; Kralik, P

    2015-03-01

    Epidemiological data indicate that raw vegetables are associated with outbreaks of Listeria monocytogenes. Therefore, there is a demand for the availability of rapid and sensitive methods, such as PCR assays, for the detection and accurate discrimination of L. monocytogenes. However, the efficiency of PCR methods can be negatively affected by inhibitory compounds commonly found in vegetable matrices that may cause false-negative results. Therefore, the sample processing and DNA isolation steps must be carefully evaluated prior to the introduction of such methods into routine practice. In this study, we compared the ability of three column-based and four magnetic bead-based commercial DNA isolation kits to extract DNA of the model micro-organism L. monocytogenes from raw vegetables. The DNA isolation efficiency of all isolation kits was determined using a triplex real-time qPCR assay designed to specifically detect L. monocytogenes. The kit with best performance, the PowerSoil(™) Microbial DNA Isolation Kit, is suitable for the extraction of amplifiable DNA from L. monocytogenes cells in vegetable with efficiencies ranging between 29.6 and 70.3%. Coupled with the triplex real-time qPCR assay, this DNA isolation kit is applicable to the samples with bacterial loads of 10(3) bacterial cells per gram of L. monocytogenes. Several recent outbreaks of Listeria monocytogenes have been associated with the consumption of fruits and vegetables. Real-time PCR assays allow fast detection and accurate quantification of microbes. However, the success of real-time PCR is dependent on the success with which template DNA can be extracted. The results of this study suggest that the PowerSoil(™) Microbial DNA Isolation Kit can be used for the extraction of amplifiable DNA from L. monocytogenes cells in vegetable with efficiencies ranging between 29.6 and 70.3%. This method is applicable to samples with bacterial loads of 10(3) bacterial cells per gram of L. monocytogenes. © 2014 The Society for Applied Microbiology.

  6. Modified CTAB and TRIzol protocols improve RNA extraction from chemically complex Embryophyta1

    PubMed Central

    Jordon-Thaden, Ingrid E.; Chanderbali, Andre S.; Gitzendanner, Matthew A.; Soltis, Douglas E.

    2015-01-01

    Premise of the study: Here we present a series of protocols for RNA extraction across a diverse array of plants; we focus on woody, aromatic, aquatic, and other chemically complex taxa. Methods and Results: Ninety-one taxa were subjected to RNA extraction with three methods presented here: (1) TRIzol/TURBO DNA-free kits using the manufacturer’s protocol with the addition of sarkosyl; (2) a combination method using cetyltrimethylammonium bromide (CTAB) and TRIzol/sarkosyl/TURBO DNA-free; and (3) a combination of CTAB and QIAGEN RNeasy Plant Mini Kit. Bench-ready protocols are given. Conclusions: After an iterative process of working with chemically complex taxa, we conclude that the use of TRIzol supplemented with sarkosyl and the TURBO DNA-free kit is an effective, efficient, and robust method for obtaining RNA from 100 mg of leaf tissue of land plant species (Embryophyta) examined. Our protocols can be used to provide RNA of suitable stability, quantity, and quality for transcriptome sequencing. PMID:25995975

  7. Effect of planecta and ROSE™ on the frequency characteristics of blood pressure-transducer kits.

    PubMed

    Fujiwara, Shigeki; Kawakubo, Yoshifumi; Mori, Satoshi; Tachihara, Keiichi; Toyoguchi, Izumi; Yokoyama, Takeshi

    2015-12-01

    Pressure-transducer kits have frequency characteristics such as natural frequency and damping coefficient, which affect the monitoring accuracy. The aim of the present study was to investigate the effect of planecta ports and a damping device (ROSE™, Argon Medical Devices, TX, USA) on the frequency characteristics of pressure-transducer kits. The FloTrac sensor kit (Edwards Lifesciences, CA, USA) and the DTXplus transducer kit (Argon Medical Devices) were prepared with planecta ports, and their frequency characteristics were tested with or without ROSE™. The natural frequency and damping coefficient of each kit were obtained using frequency characteristics analysis software and evaluated by plotting them on the Gardner's chart. By inserting a planecta port, the natural frequency markedly decreased in both the FloTrac sensor kit (from 40 to 22 Hz) and the DTXplus transducer kit (from 35 to 22 Hz). In both kits with one planecta port, the damping coefficient markedly increased by insertion of ROSE™ from 0.2 to 0.5, optimising frequency characteristics. In both kits with two planecta ports, however, the natural frequency decreased from 22 to 12 Hz. The damping coefficient increased from 0.2 to 0.8 by insertion of ROSE™; however, optimisation was not achieved even by ROSE™ insertion. Planecta ports decrease the natural frequency of the kit. ROSE™ is useful to optimise the frequency characteristics in the kits without or with one planecta port. However, optimisation is difficult with two or more planecta ports, even with the ROSE™ device.

  8. KIT Mutations Are Common in Testicular Seminomas

    PubMed Central

    Kemmer, Kathleen; Corless, Christopher L.; Fletcher, Jonathan A.; McGreevey, Laura; Haley, Andrea; Griffith, Diana; Cummings, Oscar W.; Wait, Cecily; Town, Ajia; Heinrich, Michael C.

    2004-01-01

    Expression of KIT tyrosine kinase is critical for normal germ cell development and is observed in the majority of seminomas. Activating mutations in KIT are common in gastrointestinal stromal tumors and mastocytosis. In this study we examined the frequency and spectrum of KIT mutations in 54 testicular seminomas, 1 ovarian dysgerminoma and 37 non-seminomatous germ cell tumors (NSGCT). Fourteen seminomas (25.9%) contained exon 17 point mutations including D816V (6 cases), D816H (3 cases), Y823D (2 cases), and single examples of Y823C, N822K, and T801I. No KIT mutations were found in the ovarian dysgerminoma or the NSGCTs. In transient transfection assays, mutant isoforms D816V, D816H, Y823D, and N822K were constitutively phosphorylated in the absence of the natural ligand for KIT, stem cell factor (SCF). In contrast, activation of T801I and wild-type KIT required SCF. Mutants N822K and Y823D were inhibited by imatinib mesylate (Gleevec, previously STI571) whereas D816V and D816H were both resistant to imatinib mesylate. Biochemical evidence of KIT activation, as assessed by KIT phosphorylation and KIT association with phosphatidylinositol (PI) 3-kinase in tumor cell lysates, was largely confined to seminomas with a genomic KIT mutation. These findings suggest that activating KIT mutations may contribute to tumorigenesis in a subset of seminomas, but are not involved in NSGCT. PMID:14695343

  9. FES kinase participates in KIT-ligand induced chemotaxis

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

    Voisset, Edwige, E-mail: Edwige.Voisset@inserm.fr; Institut Paoli-Calmettes, Marseille; Universite de la Mediterranee, Aix-Marseille II

    2010-02-26

    FES is a cytoplasmic tyrosine kinase activated by several membrane receptors, originally identified as a viral oncogene product. We have recently identified FES as a crucial effector of oncogenic KIT mutant receptor. However, FES implication in wild-type KIT receptor function was not addressed. We report here that FES interacts with KIT and is phosphorylated following activation by its ligand SCF. Unlike in the context of oncogenic KIT mutant, FES is not involved in wild-type KIT proliferation signal, or in cell adhesion. Instead, FES is required for SCF-induced chemotaxis. In conclusion, FES kinase is a mediator of wild-type KIT signalling implicatedmore » in cell migration.« less

  10. Telescience Resource Kit

    NASA Technical Reports Server (NTRS)

    Schneider, Michelle

    2003-01-01

    This viewgraph representation provides an overview of the Telescience Resource Kit. The Telescience Resource Kit is a pc-based telemetry and command system that will be used by scientists and engineers to monitor and control experiments located on-board the International Space Station (ISS). Topics covered include: ISS Payload Telemetry and Command Flow, kit computer applications, kit telemetry capabilities, command capabilities, and training/testing capabilities.

  11. Performances of Four Helicobacter pylori Serological Detection Kits Using Stool Antigen Test as Gold Standard.

    PubMed

    Biranjia-Hurdoyal, Susheela D; Seetulsingh-Goorah, Sharmila P

    2016-01-01

    The aim was to determine the performances of four Helicobacter pylori serological detection kits in different target groups, using Amplified IDEIA™ Hp StAR™ as gold standard. Kits studied were Rapid Immunochromatoghraphic Hexagon, Helicoblot 2.1, an EIA IgG kit and EIA IgA kit. Stool and blood samples were collected from 162 apparently healthy participants (control) and 60 Type 2 diabetes mellitus (T2DM) patients. The performances of the four serological detection kits were found to be affected by gender, age, health status and ethnicity of the participants. In the control group, the Helicoblot 2.1 kit had the best performance (AUC = 0.85; p<0.05, accuracy = 86.4%), followed by EIA IgG (AUC = 0.75; p<0.05, accuracy = 75.2%). The Rapid Hexagon and EIA IgA kits had relatively poor performances. In the T2DM subgroup, the kits H2.1 and EIA IgG had best performances, with accuracies of 96.5% and 93.1% respectively. The performance of EIA IgG improved with adjustment of its cut-off value. The performances of the detection kits were affected by various factors which should be taken into consideration.

  12. Hotspot Mutations in KIT Receptor Differentially Modulate Its Allosterically Coupled Conformational Dynamics: Impact on Activation and Drug Sensitivity

    PubMed Central

    Chauvot de Beauchêne, Isaure; Allain, Ariane; Panel, Nicolas; Laine, Elodie; Trouvé, Alain; Dubreuil, Patrice; Tchertanov, Luba

    2014-01-01

    Receptor tyrosine kinase KIT controls many signal transduction pathways and represents a typical allosterically regulated protein. The mutation-induced deregulation of KIT activity impairs cellular physiological functions and causes serious human diseases. The impact of hotspots mutations (D816H/Y/N/V and V560G/D) localized in crucial regulatory segments, the juxtamembrane region (JMR) and the activation (A-) loop, on KIT internal dynamics was systematically studied by molecular dynamics simulations. The mutational outcomes predicted in silico were correlated with in vitro and in vivo activation rates and drug sensitivities of KIT mutants. The allosteric regulation of KIT in the native and mutated forms is described in terms of communication between the two remote segments, JMR and A-loop. A strong correlation between the communication profile and the structural and dynamical features of KIT in the native and mutated forms was established. Our results provide new insight on the determinants of receptor KIT constitutive activation by mutations and resistance of KIT mutants to inhibitors. Depiction of an intra-molecular component of the communication network constitutes a first step towards an integrated description of vast communication pathways established by KIT in physiopathological contexts. PMID:25079768

  13. KIT pathway alterations in mucosal melanomas of the vulva and other sites.

    PubMed

    Omholt, Katarina; Grafström, Eva; Kanter-Lewensohn, Lena; Hansson, Johan; Ragnarsson-Olding, Boel K

    2011-06-15

    A significant proportion of mucosal melanomas contain alterations in KIT. The aim of this study was to characterize the pattern of KIT, NRAS, and BRAF mutations in mucosal melanomas at specific sites and to assess activation of the KIT downstream RAF/MEK/extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K)/AKT pathways in mucosal melanoma specimens. Seventy-one primary mucosal melanomas from various sites were studied. Mutation analysis was done by DNA sequencing. Expression of KIT, phosphorylated (p)-ERK, and p-AKT was evaluated by immunohistochemistry. KIT mutations were detected in 35% (8 of 23) of vulvar, 9% (2 of 22) of anorectal, 7% (1 of 14) of nasal cavity, and 20% (1 of 5) of penile melanomas. No KIT mutations were found in 7 vaginal melanomas. The difference in KIT mutation frequency between vulvar and nonvulvar cases was statistically significant (P = 0.014). The overall frequencies of NRAS and BRAF mutations were 10% and 6%, respectively. Notably, vaginal melanomas showed a NRAS mutation rate of 43%. KIT gene amplification (≥4 copies), as assessed by quantitative real-time PCR, was observed in 19% of cases. KIT expression was associated with KIT mutation status (P < 0.001) and was more common in vulvar than nonvulvar tumors (P = 0.016). Expression of p-ERK and p-AKT was observed in 42% and 59% of tumors, respectively, and occurred irrespective of KIT/NRAS/BRAF mutation status. NRAS mutation was associated with worse overall survival in univariate analysis. Results show that KIT mutations are more common in vulvar melanomas than other types of mucosal melanomas and that both the RAF/MEK/ERK and PI3K/AKT pathways are activated in mucosal melanoma specimens. ©2011 AACR.

  14. Imatinib for melanomas harboring mutationally activated or amplified KIT arising on mucosal, acral, and chronically sun-damaged skin.

    PubMed

    Hodi, F Stephen; Corless, Christopher L; Giobbie-Hurder, Anita; Fletcher, Jonathan A; Zhu, Meijun; Marino-Enriquez, Adrian; Friedlander, Philip; Gonzalez, Rene; Weber, Jeffrey S; Gajewski, Thomas F; O'Day, Steven J; Kim, Kevin B; Lawrence, Donald; Flaherty, Keith T; Luke, Jason J; Collichio, Frances A; Ernstoff, Marc S; Heinrich, Michael C; Beadling, Carol; Zukotynski, Katherine A; Yap, Jeffrey T; Van den Abbeele, Annick D; Demetri, George D; Fisher, David E

    2013-09-10

    Amplifications and mutations in the KIT proto-oncogene in subsets of melanomas provide therapeutic opportunities. We conducted a multicenter phase II trial of imatinib in metastatic mucosal, acral, or chronically sun-damaged (CSD) melanoma with KIT amplifications and/or mutations. Patients received imatinib 400 mg once per day or 400 mg twice per day if there was no initial response. Dose reductions were permitted for treatment-related toxicities. Additional oncogene mutation screening was performed by mass spectroscopy. Twenty-five patients were enrolled (24 evaluable). Eight patients (33%) had tumors with KIT mutations, 11 (46%) with KIT amplifications, and five (21%) with both. Median follow-up was 10.6 months (range, 3.7 to 27.1 months). Best overall response rate (BORR) was 29% (21% excluding nonconfirmed responses) with a two-stage 95% CI of 13% to 51%. BORR was significantly greater than the hypothesized null of 5% and statistically significantly different by mutation status (7 of 13 or 54% KIT mutated v 0% KIT amplified only). There were no statistical differences in rates of progression or survival by mutation status or by melanoma site. The overall disease control rate was 50% but varied significantly by KIT mutation status (77% mutated v 18% amplified). Four patients harbored pretreatment NRAS mutations, and one patient acquired increased KIT amplification after treatment. Melanomas that arise on mucosal, acral, or CSD skin should be assessed for KIT mutations. Imatinib can be effective when tumors harbor KIT mutations, but not if KIT is amplified only. NRAS mutations and KIT copy number gain may be mechanisms of therapeutic resistance to imatinib.

  15. Facilitators and Barriers to Naloxone Kit Use Among Opioid-Dependent Patients Enrolled in Medication Assisted Therapy Clinics in North Carolina.

    PubMed

    Khatiwoda, Prasana; Proeschold-Bell, Rae Jean; Meade, Christina S; Park, Lawrence P; Proescholdbell, Scott

    2018-01-01

    BACKGROUND Naloxone-an opioid antagonist that reverses the effects of opioids-is increasingly being distributed in non-medical settings. We sought to identify the facilitators of, and barriers to, opioid users using naloxone kits in North Carolina. METHODS In 2015, we administered a 15-item survey to a convenience sample of 100 treatment seekers at 4 methadone/buprenorphine Medication Assisted Therapy (MAT) clinics in North Carolina. RESULTS Seventy-four percent of participants reported having ever gotten a naloxone kit; this percentage was higher for females (81%) than males (63%) ( P = .06). The primary reason given for not having a kit was not knowing where to get one. Only 6% had heard of kits from the media and only 5% received one from a medical provider. Among kit recipients, 56% of both females and males reported mostly or sometimes carrying the kit, with additional participants reporting always. Reasons for not carrying a kit were no longer being around drugs, forgetting it, and the kit being too large. Men discussed the difficulties of carrying the naloxone kits, which are currently too large to fit in a pocket. Ninety-four percent of naloxone users reported intending to call emergency services in case of an overdose emergency. LIMITATIONS Study limitations included a small sample, participants limited to MAT clinics, and a predominantly white sample. CONCLUSIONS MAT treatment seekers reported a willingness to carry and use naloxone kits. Education, outreach, media, and medical providers need to promote naloxone kits. A smaller kit may increase the likelihood of men carrying one. ©2018 by the North Carolina Institute of Medicine and The Duke Endowment. All rights reserved.

  16. Identification of a population of cells with hematopoietic stem cell properties in mouse aorta-gonad-mesonephros cultures

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

    Nobuhisa, Ikuo; Ohtsu, Naoki; Okada, Seiji

    The aorta-gonad-mesonephros (AGM) region is a primary source of definitive hematopoietic cells in the midgestation mouse embryo. In cultures of dispersed AGM regions, adherent cells containing endothelial cells are observed first, and then non-adherent hematopoietic cells are produced. Here we report on the characterization of hematopoietic cells that emerge in the AGM culture. Based on the expression profiles of CD45 and c-Kit, we defined three cell populations: CD45{sup low} c-Kit{sup +} cells that had the ability to form hematopoietic cell colonies in methylcellulose media and in co-cultures with stromal cells; CD45{sup low} c-Kit{sup -} cells that showed a granulocyte morphology;more » CD45{sup high} c-Kit{sup low/-} that exhibited a macrophage morphology. In co-cultures of OP9 stromal cells and freshly prepared AGM cultures, CD45{sup low} c-Kit{sup +} cells from the AGM culture had the abilities to reproduce CD45{sup low} c-Kit{sup +} cells and differentiate into CD45{sup low} c-Kit{sup -} and CD45{sup high} c-Kit{sup low/-} cells, whereas CD45{sup low} c-Kit{sup -} and CD45{sup high} c-Kit{sup low/-} did not produce CD45{sup low} c-Kit{sup +} cells. Furthermore, CD45{sup low} c-Kit{sup +} cells displayed a long-term repopulating activity in adult hematopoietic tissue when transplanted into the liver of irradiated newborn mice. These results indicate that CD45{sup low} c-Kit{sup +} cells from the AGM culture have the potential to reconstitute multi-lineage hematopoietic cells.« less

  17. Evaluation of four commercial quantitative real-time PCR kits with inhibited and degraded samples.

    PubMed

    Holmes, Amy S; Houston, Rachel; Elwick, Kyleen; Gangitano, David; Hughes-Stamm, Sheree

    2018-05-01

    DNA quantification is a vital step in forensic DNA analysis to determine the optimal input amount for DNA typing. A quantitative real-time polymerase chain reaction (qPCR) assay that can predict DNA degradation or inhibitors present in the sample prior to DNA amplification could aid forensic laboratories in creating a more streamlined and efficient workflow. This study compares the results from four commercial qPCR kits: (1) Investigator® Quantiplex® Pro Kit, (2) Quantifiler® Trio DNA Quantification Kit, (3) PowerQuant® System, and (4) InnoQuant® HY with high molecular weight DNA, low template samples, degraded samples, and DNA spiked with various inhibitors.The results of this study indicate that all kits were comparable in accurately predicting quantities of high quality DNA down to the sub-picogram level. However, the InnoQuant(R) HY kit showed the highest precision across the DNA concentration range tested in this study. In addition, all kits performed similarly with low concentrations of forensically relevant PCR inhibitors. However, in general, the Investigator® Quantiplex® Pro Kit was the most tolerant kit to inhibitors and provided the most accurate quantification results with higher concentrations of inhibitors (except with salt). PowerQuant® and InnoQuant® HY were the most sensitive to inhibitors, but they did indicate significant levels of PCR inhibition. When quantifying degraded samples, each kit provided different degradation indices (DI), with Investigator® Quantiplex® Pro indicating the largest DI and Quantifiler® Trio indicating the smallest DI. When the qPCR kits were paired with their respective STR kit to genotype highly degraded samples, the Investigator® 24plex QS and GlobalFiler® kits generated more complete profiles when the small target concentrations were used for calculating input amount.

  18. Aberrant expressions of c-KIT and DOG-1 in mucinous and nonmucinous colorectal carcinomas and relation to clinicopathologic features and prognosis.

    PubMed

    Foda, Abd Al-Rahman Mohammad; Mohamed, Mie Ali

    2015-10-01

    c-KIT and DOG-1 are 2 highly expressed proteins in gastrointestinal stromal tumors. Few studies had investigated c-KIT, but not DOG-1, expression in colorectal carcinoma (CRC). This study aims to investigate expressions of c-KIT and DOG-1 in colorectal mucinous carcinoma and nonmucinous carcinoma using manual tissue microarray technique. In this work, we studied tumor tissue specimens from 150 patients with colorectal mucinous (MA) and nonmucinous adenocarcinoma (NMA). High-density manual tissue microarrays were constructed using modified mechanical pencil tip technique, and immunohistochemistry for c-KIT and DOG-1 was done. We found that aberrant c-KIT expression was detected in 12 cases (8%); 6 cases (4%) showed strong expression. Aberrant DOG-1 expression was detected in 15 cases (10%); among them, only 4 cases (2.7%) showed strong expression. Nonmucinous adenocarcinoma showed a significantly high expression of c-KIT, but not DOG-1, than MA. Aberrant c-KIT and DOG-1 expressions were significantly unrelated but were associated with excessive microscopic abscess formation. Neither c-KIT nor DOG-1 expression showed a significant impact on disease-free survival or overall survival. In conclusion, aberrant c-KIT and DOG-1 expressions in CRC are rare events, either in NMA or MA. Nonmucinous adenocarcinoma showed a significantly higher expression of c-KIT, but not DOG-1, than MA. The expressions of both in CRC are significantly unrelated but are associated with microscopic abscess formation. Neither c-KIT nor DOG-1 expression showed a significant impact on disease-free survival or overall survival. So, c-KIT and DOG-1 immunostaining is not a cost-effective method of identifying patients with CRC who may benefit from treatment with tyrosine kinase inhibitors. Copyright © 2015 Elsevier Inc. All rights reserved.

  19. Adapted hydropower-driven water supply system: assessment of an underground application in an Indonesian karst area

    NASA Astrophysics Data System (ADS)

    Oberle, P.; Ikhwan, M.; Stoffel, D.; Nestmann, F.

    2016-09-01

    Populated karst landscapes can be found all over the world, although their natural boundary conditions mostly lead to distinct challenges regarding a sustainable water supply. Especially in developing and emerging countries, this situation aggravates since appropriate technologies and water management concepts are rarely available. Against this background, the interdisciplinary, German-Indonesian joint project " Integrated Water Resources Management (IWRM) Indonesia", funded by the German Federal Ministry of Education and Research (BMBF), focused on the development and exemplary implementation of adapted techniques to remedy the partly severe water scarcity in the region Gunung Sewu. This karst area, widely known as " Java's poorhouse", is located on the southern coast of Java Island and distinctly suffers from the mentioned constraints. Under the aegis of the Karlsruhe Institute of Technology (KIT), the conceptual and technical achievements of the "IWRM Indonesia" joint research project are characterized by a high potential for multiplication not only for karst areas but also for non-karst regions. One of the project's major accomplishments is the erection of an innovative hydropower-driven water supply facility located in a karst cave 100 m below ground and continuously supplying tens of thousands of people with fresh water. Referring to the plant's innovative character and the demanding conditions on-site, the implementation was a highly iterative process leading to today's autonomous operation by an Indonesian public authority. Based on the experiences gained during design, construction, operation and monitoring phase, this paper introduces an implementation approach for adapted technologies as well as a comprising technical and economical assessment of the plant's operation.

  20. A phase 2 trial of dasatinib in patients with locally advanced or stage IV mucosal, acral, or vulvovaginal melanoma: A trial of the ECOG-ACRIN Cancer Research Group (E2607).

    PubMed

    Kalinsky, Kevin; Lee, Sandra; Rubin, Krista M; Lawrence, Donald P; Iafrarte, Anthony J; Borger, Darell R; Margolin, Kim A; Leitao, Mario M; Tarhini, Ahmad A; Koon, Henry B; Pecora, Andrew L; Jaslowski, Anthony J; Cohen, Gary I; Kuzel, Timothy M; Lao, Christopher D; Kirkwood, John M

    2017-07-15

    KIT-directed tyrosine kinase inhibitors such as imatinib have demonstrated benefits in KIT-mutant (KIT+) mucosal, acral, vulvovaginal, and chronically sun-damaged (CSD) melanoma. Dasatinib has superior preclinical activity in comparison with other tyrosine kinase inhibitors against cells with the most common KIT mutation, exon 11 L576P . The ECOG-ACRIN E2607 trial assessed dasatinib in patients with these melanoma subtypes. Patients received 70 mg of oral dasatinib twice daily. The primary objective for this 2-stage phase 2 trial was response rate. Stage I was open to KIT+ and wild-type KIT (KIT-) mucosal, acral, and CSD melanoma (n = 57). Stage II accrued only KIT+ tumors (n = 30). To enrich the trial for KIT+ tumors, vulvovaginal melanoma was added, and CSD melanoma was removed from eligibility. Secondary objectives included progression-free survival (PFS), overall survival (OS), and safety. From May 2009 to December 2010, the first stage enrolled 57 patients. Among the evaluable patients, 3 of 51 (5.9%) achieved a partial response: all were KIT-. Stage II closed early because of slow accrual (November 2011 to December 2015). In stage II, 4 of 22 evaluable patients (18.2%) had a partial response; the median duration was 4.2 months. The median PFS was 2.1 months (n = 73; 95% confidence interval [CI], 1.5-2.9 months). The median OS was 7.5 months (95% CI, 6.0-11.9 months). In exploratory analyses, no differences were seen in PFS or OS with the KIT status or subtype. Dasatinib was discontinued because of adverse events in 9 of 75 patients (12%). The dasatinib response rate among KIT+ melanoma patients was low. In view of its clinical activity, it is recommended that imatinib remain the KIT tyrosine kinase inhibitor of choice for unresectable KIT+ melanoma. Cancer 2017;123:2688-97. © 2017 American Cancer Society. © 2017 American Cancer Society.

  1. CD45{sup low}c-Kit{sup high} cells have hematopoietic properties in the mouse aorta-gonad-mesonephros region

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

    Nobuhisa, Ikuo, E-mail: nobuhisa.scr@mri.tmd.ac.jp; Department of Cell Fate Modulation, Institute of Molecular Embryology and Genetics/Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, 860-0811; Yamasaki, Shoutarou

    Long-term reconstituting hematopoietic stem cells first arise from the aorta of the aorta-gonad-mesonephros (AGM) region in a mouse embryo. We have previously reported that in cultures of the dispersed AGM region, CD45{sup low}c-Kit{sup +} cells possess the ability to reconstitute multilineage hematopoietic cells, but investigations are needed to show that this is not a cultured artifact and to clarify when and how this population is present. Based on the expression profile of CD45 and c-Kit in freshly dissociated AGM cells from embryonic day 9.5 (E9.5) to E12.5 and aorta cells in the AGM from E13.5 to E15.5, we defined sixmore » cell populations (CD45{sup -}c-Kit{sup -}, CD45{sup -}c-Kit{sup low}, CD45{sup -}c-Kit{sup high}, CD45{sup low}c-Kit{sup high}, CD45{sup high}c-Kit{sup high}, and CD45{sup high}c-Kit{sup very} {sup low}). Among these six populations, CD45{sup low}c-Kit{sup high} cells were most able to form hematopoietic cell colonies, but their ability decreased after E11.5 and was undetectable at E13.5 and later. The CD45{sup low}c-Kit{sup high} cells showed multipotency in vitro. We demonstrated further enrichment of hematopoietic activity in the Hoechst dye-effluxing side population among the CD45{sup low}c-Kit{sup high} cells. Here, we determined that CD45{sup low}c-Kit{sup high} cells arise from the lateral plate mesoderm using embryonic stem cell-derived differentiation system. In conclusion, CD45{sup low}c-Kit{sup high} cells are the major hematopoietic cells of mouse AGM.« less

  2. Development and evaluation of a "working together" framework and a tool kit to enhance inter-organizational relationships in healthcare.

    PubMed

    Edwards, Lynn; Gibson, Rick; Carson, Shannon Ryan; Sampalli, Tara

    2013-01-01

    Healthcare is in a constant state of change and evolution driven by a multitude of complex factors and interactions. Consequently, organizations, teams and individuals in healthcare have to habitually realign their working relationships. Furthermore, research has shown that "working together" relationships fail in the absence of a defined framework. In this research, a novel framework and a tool kit for working together have been developed and evaluated. The framework has a formal process to articulate the intended purpose/outcome, clearly align the type of working relationship with the purpose and identify the barriers and facilitators to working relationships in healthcare.

  3. Bone-induced c-kit expression in prostate cancer: a driver of intraosseous tumor growth

    PubMed Central

    Mainetti, Leandro E.; Zhe, Xiaoning; Diedrich, Jonathan; Saliganan, Allen D.; Cho, Won Jin; Cher, Michael L.; Heath, Elisabeth; Fridman, Rafael; Kim, Hyeong-Reh Choi; Bonfil, R. Daniel

    2014-01-01

    Loss of BRCA2 function stimulates prostate cancer (PCa) cell invasion and is associated with more aggressive and metastatic tumors in PCa patients. Concurrently, the receptor tyrosine kinase c-kit is highly expressed in skeletal metastases of PCa patients and induced in PCa cells placed into the bone microenvironment in experimental models. However, the precise requirement of c-kit for intraosseous growth of PCa and its relation to BRCA2 expression remain unexplored. Here, we show that c-kit expression promotes migration and invasion of PCa cells. Alongside, we found that c-kit expression in PCa cells parallels BRCA2 downregulation. Gene rescue experiments with human BRCA2 transgene in c-kit-transfected PCa cells resulted in reduction of c-kit protein expression and migration and invasion, suggesting a functional significance of BRCA2 downregulation by c-kit. The inverse association between c-kit and BRCA2 gene expressions in PCa cells was confirmed using laser capture microdissection in experimental intraosseous tumors and bone metastases of PCa patients. Inhibition of bone-induced c-kit expression in PCa cells transduced with lentiviral short hairpin RNA reduced intraosseous tumor incidence and growth. Overall, our results provide evidence of a novel pathway that links bone-induced c-kit expression in PCa cells to BRCA2 downregulation and supports bone metastasis. PMID:24798488

  4. c-Kit modifies the inflammatory status of smooth muscle cells

    PubMed Central

    Song, Lei; Martinez, Laisel; Zigmond, Zachary M.; Hernandez, Diana R.; Lassance-Soares, Roberta M.; Selman, Guillermo

    2017-01-01

    Background c-Kit is a receptor tyrosine kinase present in multiple cell types, including vascular smooth muscle cells (SMC). However, little is known about how c-Kit influences SMC biology and vascular pathogenesis. Methods High-throughput microarray assays and in silico pathway analysis were used to identify differentially expressed genes between primary c-Kit deficient (KitW/W–v) and control (Kit+/+) SMC. Quantitative real-time RT-PCR and functional assays further confirmed the differences in gene expression and pro-inflammatory pathway regulation between both SMC populations. Results The microarray analysis revealed elevated NF-κB gene expression secondary to the loss of c-Kit that affects both the canonical and alternative NF-κB pathways. Upon stimulation with an oxidized phospholipid as pro-inflammatory agent, c-Kit deficient SMC displayed enhanced NF-κB transcriptional activity, higher phosphorylated/total p65 ratio, and increased protein expression of NF-κB regulated pro-inflammatory mediators with respect to cells from control mice. The pro-inflammatory phenotype of mutant cells was ameliorated after restoring c-Kit activity using lentiviral transduction. Functional assays further demonstrated that c-Kit suppresses NF-κB activity in SMC in a TGFβ-activated kinase 1 (TAK1) and Nemo-like kinase (NLK) dependent manner. Discussion Our study suggests a novel mechanism by which c-Kit suppresses NF-κB regulated pathways in SMC to prevent their pro-inflammatory transformation. PMID:28626608

  5. c-Kit modifies the inflammatory status of smooth muscle cells.

    PubMed

    Song, Lei; Martinez, Laisel; Zigmond, Zachary M; Hernandez, Diana R; Lassance-Soares, Roberta M; Selman, Guillermo; Vazquez-Padron, Roberto I

    2017-01-01

    c-Kit is a receptor tyrosine kinase present in multiple cell types, including vascular smooth muscle cells (SMC). However, little is known about how c-Kit influences SMC biology and vascular pathogenesis. High-throughput microarray assays and in silico pathway analysis were used to identify differentially expressed genes between primary c-Kit deficient (Kit W/W-v ) and control (Kit +/+ ) SMC. Quantitative real-time RT-PCR and functional assays further confirmed the differences in gene expression and pro-inflammatory pathway regulation between both SMC populations. The microarray analysis revealed elevated NF-κB gene expression secondary to the loss of c-Kit that affects both the canonical and alternative NF-κB pathways. Upon stimulation with an oxidized phospholipid as pro-inflammatory agent, c-Kit deficient SMC displayed enhanced NF-κB transcriptional activity, higher phosphorylated/total p65 ratio, and increased protein expression of NF-κB regulated pro-inflammatory mediators with respect to cells from control mice. The pro-inflammatory phenotype of mutant cells was ameliorated after restoring c-Kit activity using lentiviral transduction. Functional assays further demonstrated that c-Kit suppresses NF-κB activity in SMC in a TGFβ-activated kinase 1 (TAK1) and Nemo-like kinase (NLK) dependent manner. Our study suggests a novel mechanism by which c-Kit suppresses NF-κB regulated pathways in SMC to prevent their pro-inflammatory transformation.

  6. Oncogenic Kit signalling on the Golgi is suppressed by blocking secretory trafficking with M-COPA in gastrointestinal stromal tumours.

    PubMed

    Obata, Yuuki; Horikawa, Keita; Shiina, Isamu; Takahashi, Tsuyoshi; Murata, Takatsugu; Tasaki, Yasutaka; Suzuki, Kyohei; Yonekura, Keita; Esumi, Hiroyasu; Nishida, Toshirou; Abe, Ryo

    2018-02-28

    Most gastrointestinal stromal tumours (GISTs) are caused by constitutively active mutations in Kit tyrosine kinase. The drug imatinib, a specific Kit inhibitor, improves the prognosis of metastatic GIST patients, but these patients become resistant to the drug by acquiring secondary mutations in the Kit kinase domain. We recently reported that a Kit mutant causes oncogenic signals only on the Golgi apparatus in GISTs. In this study, we show that in GIST, 2-methylcoprophilinamide (M-COPA, also known as "AMF-26"), an inhibitor of biosynthetic protein trafficking from the endoplasmic reticulum (ER) to the Golgi, suppresses Kit autophosphorylation at Y703/Y721/Y730/Y936, resulting in blockade of oncogenic signalling. Results of our M-COPA treatment assay show that Kit Y703/Y730/Y936 in the ER are dephosphorylated by protein tyrosine phosphatases (PTPs), thus the ER-retained Kit is unable to activate downstream molecules. ER-localized Kit Y721 is not phosphorylated, but not due to PTPs. Importantly, M-COPA can inhibit the activation of the Kit kinase domain mutant, resulting in suppression of imatinib-resistant GIST proliferation. Our study demonstrates that Kit autophosphorylation is spatio-temporally regulated and may offer a new strategy for treating imatinib-resistant GISTs. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

  7. Levitation Kits Demonstrate Superconductivity.

    ERIC Educational Resources Information Center

    Worthy, Ward

    1987-01-01

    Describes the "Project 1-2-3" levitation kit used to demonstrate superconductivity. Summarizes the materials included in the kit. Discusses the effect demonstrated and gives details on how to obtain kits. Gives an overview of the documentation that is included. (CW)

  8. Reviews Toy: Air swimmers Book: Their Arrows will Darken the Sun: The Evolution and Science of Ballistics Book: Physics Experiments for your Bag Book: Quantum Physics for Poets Equipment: SEP colour wheel kit Equipment: SEP colour mixing kit Software: USB DrDAQ App: iHandy Level Equipment: Photonics Explorer kit Web Watch

    NASA Astrophysics Data System (ADS)

    2012-01-01

    WE RECOMMEND Air swimmers Helium balloon swims like a fish Their Arrows will Darken the Sun: The Evolution and Science of Ballistics Ballistics book hits the spot Physics Experiments for your Bag Handy experiments for your lessons Quantum Physics for Poets Book shows the economic importance of physics SEP colour wheel kit Wheels investigate colour theory SEP colour mixing kit Cheap colour mixing kit uses red, green and blue LEDs iHandy Level iPhone app superbly measures angles Photonics Explorer kit Free optics kit given to schools WORTH A LOOK DrDAQ DrDAQ software gets an upgrade WEB WATCH Websites show range of physics

  9. FUNCTIONAL DEREGULATION OF KIT: LINK TO MAST CELL PROLIFERATIVE DISEASES AND OTHER NEOPLASMS

    PubMed Central

    Cruse, Glenn; Metcalfe, Dean D.; Olivera, Ana

    2014-01-01

    SYNOPSIS Signaling through the receptor tyrosine kinase KIT mediates differentiation, proliferation and survival of hematopoietic precursor cells and mast cells. Constitutive KIT signaling due to somatic point mutations in c-Kit is an important occurrence in the development of mast cell proliferation disorders and other hematological malignancies. In this review, we discuss the common gain-of-function mutations found in these malignancies, particularly in mast cell proliferation disorders, and summarize the current understanding of the molecular mechanisms by which transforming point mutations in KIT may affect KIT structure and function and lead to altered downstream signaling and cellular transformation. Drugs targeting KIT have shown mixed success in the treatment of these diseases. A brief overview of the most common KIT inhibitors currently used, the reasons for the varied clinical results of such inhibitors and a discussion of potential new strategies are provided. PMID:24745671

  10. Against All Odds: Problem-Solving Strategies and Behavioural Characteristics of Novice Students

    ERIC Educational Resources Information Center

    Chang, Pei-Fen; Lin, Miao-Chen

    2015-01-01

    This study investigates problem-solving difficulties of novices in a classroom setting, using a German instructional tool, the Fischertechnik kit of approximately 400 parts. In order to analyse the students' thinking processes as they solved the problems, verbal protocol analysis (VPA) was used to record the students'' thinking processes and…

  11. Long-term impact of integration of household water treatment and hygiene promotion with antenatal services on maternal water treatment and hygiene practices in Malawi.

    PubMed

    Loharikar, Anagha; Russo, Elizabeth; Sheth, Anandi; Menon, Manoj; Kudzala, Amose; Tauzie, Blessius; Masuku, Humphreys D; Ayers, Tracy; Hoekstra, Robert M; Quick, Robert

    2013-02-01

    A clinic-based program to integrate antenatal services with distribution of hygiene kits including safe water storage containers, water treatment solution (brand name WaterGuard), soap, and hygiene education, was implemented in Malawi in 2007 and evaluated in 2010. We surveyed 389 participants at baseline in 2007, and found and surveyed 232 (60%) participants to assess water treatment, test stored drinking water for residual chlorine (an objective measure of treatment), and observe handwashing technique at follow-up in 2010. Program participants were more likely to know correct water treatment procedures (67% versus 36%; P < 0.0001), treat drinking water with WaterGuard (24% versus 2%; P < 0.0001), purchase and use WaterGuard (21% versus 1%; P < 0.001), and demonstrate correct handwashing technique (50% versus 21%; P < 0.001) at the three-year follow-up survey than at baseline. This antenatal-clinic-based program may have contributed to sustained water treatment and proper handwashing technique among program participants.

  12. Pharmacists' views on and experiences with bowel cancer screening kits in Auckland, New Zealand.

    PubMed

    Martini, Nataly; Basdew, Kamlika; Kammona, Ala; Shen, Amy; Taylor, Caragh; McIntosh, Timothy R; Barnes, Joanne

    2014-08-01

    To explore the views of New Zealand pharmacists on bowel cancer screening, particularly with regards to faecal occult blood testing (FOBT) kits, self-perceived knowledge on FOBT kits and barriers, motivators and experiences with selling and counselling consumers with respect to FOBT kits. Semi-structured interviews were conducted face to face or by telephone with 20 community pharmacists in the Auckland region. Interviews were recorded and transcribed verbatim and data were coded and analysed using NVivo software to identify key themes. Participant pharmacists believed that they were well placed to provide advice on FOBT kits to consumers. Barriers to selling the kits included cost and perceived lack of test sensitivity of the kits, poor consumer demand, pharmacists' lack of training and information, and a belief that selling FOBT kits was outside the pharmacists' scope of practice. Motivators to selling the kits included customer convenience, ease of use, confidence in the kits and embracing new roles for pharmacists. Pharmacists were concerned that use of the kits may increase the burden on the public health system through customer anxiety over test results; however, they agreed that there was a need for bowel cancer screening and awareness and that people concerned about bowel cancer should make visiting their general practitioner a priority. Pharmacists' views were mixed. Pharmacists' training and competence with respect to the provision of bowel cancer kits, and how a bowel cancer screening service can be developed to optimise public health outcomes, need to be addressed. © 2013 Royal Pharmaceutical Society.

  13. Repeatability and validity of a field kit for estimation of cholinesterase in whole blood.

    PubMed Central

    London, L; Thompson, M L; Sacks, S; Fuller, B; Bachmann, O M; Myers, J E

    1995-01-01

    OBJECTIVES--To evaluate a spectrophotometric field kit (Test-Mate-OP) for repeatability and validity in comparison with reference laboratory methods and to model its anticipated sensitivity and specificity based on these findings. METHODS--76 farm workers between the age of 20 and 55, of whom 30 were pesticide applicators exposed to a range of organophosphates in the preceding 10 days, had blood taken for plasma cholinesterase (PCE) and erythrocyte cholinesterase (ECE) measurement by field kit or laboratory methods. Paired blinded duplicate samples were taken from subgroups in the sample to assess repeatability of laboratory and field kit methods. Field kits were also used to test venous blood in one subgroup. The variance obtained for the field kit tests was then applied to two hypothetical scenarios that used published action guidelines to model the kit's sensitivity and specificity. RESULTS--Repeatability for PCE was much poorer and for ECE slightly poorer than that of laboratory measures. A substantial upward bias for field kit ECE relative to laboratory measurements was found. Sensitivity of the kit to a 40% drop in PCE was 67%, whereas that for ECE was 89%. Specificity of the kit with no change in mean of the population was 100% for ECE and 91% for PCE. CONCLUSION--Field kit ECE estimation seems to be sufficiently repeatable for surveillance activities, whereas PCE does not. Repeatability of both tests seems to be too low for use in epidemiological dose-response investigations. Further research is indicated to characterise the upward bias in ECE estimation on the kit. PMID:7697143

  14. Anal mucosal melanoma with KIT-activating mutation and response to imatinib therapy--case report and review of the literature.

    PubMed

    Satzger, Imke; Küttler, Uta; Völker, Bernward; Schenck, Florian; Kapp, Alexander; Gutzmer, Ralf

    2010-01-01

    Previously an increased frequency of KIT aberrations in mucosal melanomas was reported, whereas c-KIT in most types of cutaneous melanomas does not appear to be of pathogenetic importance. Imatinib has become the standard of care in other cancers with KIT mutations such as gastrointestinal stromal tumors. Recently 12 cases of metastatic melanoma and KIT-activating mutations have been published to be successfully treated with c-KIT blockers such as imatinib, sunitinib, dasatinib or sorafenib. We report here on one of our patients with KIT-activating mutation in metastatic anal mucosal melanoma, who showed a response to imatinib therapy and summarize the available literature regarding this new therapeutic option. Copyright 2009 S. Karger AG, Basel.

  15. Evaluation of a Commercial Latex Agglutination Test Kit for Cryptococcal Antigen

    PubMed Central

    Kaufman, Leo; Cowart, Glenda; Blumer, Sharon; Stine, Amy; Wood, Ross

    1974-01-01

    Two dozen Crypto-LA kits for detecting Cryptococcus neoformans capsular polysaccharide antigens were evaluated. Ten kits proved reliable for detecting and titering antigen in clinical materials. Fourteen kits were found to be inadequate. PMID:4596394

  16. KIT Signaling Promotes Growth of Colon Xenograft Tumors in Mice and Is Up-Regulated in a Subset of Human Colon Cancers.

    PubMed

    Chen, Evan C; Karl, Taylor A; Kalisky, Tomer; Gupta, Santosh K; O'Brien, Catherine A; Longacre, Teri A; van de Rijn, Matt; Quake, Stephen R; Clarke, Michael F; Rothenberg, Michael E

    2015-09-01

    Receptor tyrosine kinase (RTK) inhibitors have advanced colon cancer treatment. We investigated the role of the RTK KIT in development of human colon cancer. An array of 137 patient-derived colon tumors and their associated xenografts were analyzed by immunohistochemistry to measure levels of KIT and its ligand KITLG. KIT and/or KITLG was stably knocked down by expression of small hairpin RNAs from lentiviral vectors in DLD1, HT29, LS174T, and COLO320 DM colon cancer cell lines, and in UM-COLON#8 and POP77 xenografts; cells transduced with only vector were used as controls. Cells were analyzed by real-time quantitative reverse transcription polymerase chain reaction, single-cell gene expression analysis, flow cytometry, and immunohistochemical, immunoblot, and functional assays. Xenograft tumors were grown from control and KIT-knockdown DLD1 and UM-COLON#8 cells in immunocompromised mice and compared. Some mice were given the RTK inhibitor imatinib after injection of cancer cells; tumor growth was measured based on bioluminescence. We assessed tumorigenicity using limiting dilution analysis. KIT and KITLG were expressed heterogeneously by a subset of human colon tumors. Knockdown of KIT decreased proliferation of colon cancer cell lines and growth of xenograft tumors in mice compared with control cells. KIT knockdown cells had increased expression of enterocyte markers, decreased expression of cycling genes, and, unexpectedly, increased expression of LGR5 associated genes. No activating mutations in KIT were detected in DLD1, POP77, or UM-COLON#8 cells. However, KITLG-knockdown DLD1 cells formed smaller xenograft tumors than control cells. Gene expression analysis of single CD44(+) cells indicated that KIT can promote growth via KITLG autocrine and/or paracrine signaling. Imatinib inhibited growth of KIT(+) colon cancer organoids in culture and growth of xenograft tumors in mice. Cancer cells with endogenous KIT expression were more tumorigenic in mice. KIT and KITLG are expressed by a subset of human colon tumors. KIT signaling promotes growth of colon cancer cells and organoids in culture and xenograft tumors in mice via its ligand, KITLG, in an autocrine or paracrine manner. Patients with KIT-expressing colon tumors can benefit from KIT RTK inhibitors. Copyright © 2015 AGA Institute. Published by Elsevier Inc. All rights reserved.

  17. KIT Signaling Promotes Growth of Colon Xenograft Tumors in Mice and is Upregulated in a Subset of Human Colon Cancers

    PubMed Central

    Chen, Evan C.; Karl, Taylor A.; Kalisky, Tomer; Gupta, Santosh K.; O’Brien, Catherine A.; Longacre, Teri A.; van de Rijn, Matt; Quake, Stephen R.; Clarke, Michael F.; Rothenberg, Michael E.

    2015-01-01

    Background & Aims Receptor tyrosine kinase (RTK) inhibitors have advanced colon cancer treatment. We investigated the role of the RTK KIT in development of human colon cancer. Methods An array of 137 patient-derived colon tumors and their associated xenografts were analyzed by immunohistochemistry to measure levels of KIT and its ligand KITLG. KIT and/or KITLG was stably knocked down by expression of small hairpin RNAs from lentiviral vectors in DLD1, HT29, LS174T, and COLO320 colon cancer cell lines, and in UM-COLON#8 and POP77 xenografts; cells transduced with only vector were used as controls. Cells were analyzed by real-time quantitative reverse transcription PCR, single-cell gene expression analysis, flow cytometry, and immunohistochemical, immunoblot, and functional assays. Xenograft tumors were grown from control and KIT-knockdown DLD1 and UM-COLON#8 cells in immunocompromised mice and compared. Some mice were given the RTK inhibitor imatinib following injection of cancer cells; tumor growth was measured based on bioluminescence. We assessed tumorigenicity using limiting dilution analysis. Results KIT and KITLG were expressed heterogeneously by a subset of human colon tumors. Knockdown of KIT decreased proliferation of colon cancer cell lines and growth of xenograft tumors in mice, compared with control cells. KIT knockdown cells had increased expression of enterocyte markers, decreased expression of cycling genes, and, unexpectedly, increased expression of LGR5-associated genes. No activating mutations in KIT were detected in DLD1, POP77, or UM-COLON#8 cell lines. However, KITLG-knockdown DLD1 cells formed smaller xenograft tumors than control cells. Gene expression analysis of single CD44+ cells indicated that KIT may promote growth via KITLG autocrine and/or paracrine signaling. Imatinib inhibited growth of KIT+ colon cancer organoids in culture and growth of xenograft tumors in mice. Cancer cells with endogenous KIT expression were more tumorigenic in mice. Conclusions KIT and KITLG are expressed by a subset of human colon tumors. KIT signaling promotes growth of colon cancer cells and organoids in culture and xenograft tumors in mice via its ligand, KITLG, in an autocrine or paracrine manner. Patients with KIT-expressing colon tumors may benefit from KIT RTK inhibitors. PMID:26026391

  18. miR-137 downregulates c-kit expression in acute myeloid leukemia.

    PubMed

    Hu, Yanping; Dong, Xiaolong; Chu, Guoming; Lai, Guangrui; Zhang, Bijun; Wang, Leitong; Zhao, Yanyan

    2017-06-01

    The oncogene c-kit plays a vital role in the pathogenesis of acute myeloid leukemia (AML). However, the mechanism of microRNAs targeting c-kit in AML has not been determined in detail. Moreover, the role miR-137 in tumor cell proliferation remains controversial. The aim of this work was to verify whether miR-137 targets c-kit and to research the biological effects of restoring miR-137 expression in leukemia cells. We found that miR-137 binds specifically to the 3'-UTR of c-kit and suppresses the expression and activities of c-kit. There is a negative correlation between miR-137 and c-kit expression in both patients and cell lines determined by screening large clinical samples. We found that miR-137 can inhibit proliferation, promote apoptosis, and induce differentiation of c-kit+ AML cells. We determined that miR-137 can participate in the leukemogenesis by regulating c-kit, which could be used as a therapeutic target for acute myeloid leukemia. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Combined KIT and FGFR2b Signaling Regulates Epithelial Progenitor Expansion during Organogenesis

    PubMed Central

    Lombaert, Isabelle M.A.; Abrams, Shaun R.; Li, Li; Eswarakumar, Veraragavan P.; Sethi, Aditya J.; Witt, Robert L.; Hoffman, Matthew P.

    2013-01-01

    Summary Organ formation and regeneration require epithelial progenitor expansion to engineer, maintain, and repair the branched tissue architecture. Identifying the mechanisms that control progenitor expansion will inform therapeutic organ (re)generation. Here, we discover that combined KIT and fibroblast growth factor receptor 2b (FGFR2b) signaling specifically increases distal progenitor expansion during salivary gland organogenesis. FGFR2b signaling upregulates the epithelial KIT pathway so that combined KIT/FGFR2b signaling, via separate AKT and mitogen-activated protein kinase (MAPK) pathways, amplifies FGFR2b-dependent transcription. Combined KIT/FGFR2b signaling selectively expands the number of KIT+K14+SOX10+ distal progenitors, and a genetic loss of KIT signaling depletes the distal progenitors but also unexpectedly depletes the K5+ proximal progenitors. This occurs because the distal progenitors produce neurotrophic factors that support gland innervation, which maintains the proximal progenitors. Furthermore, a rare population of KIT+FGFR2b+ cells is present in adult glands, in which KIT signaling also regulates epithelial-neuronal communication during homeostasis. Our findings provide a framework to direct regeneration of branched epithelial organs. PMID:24371813

  20. LogiKit - assisting complex logic specification and implementation for embedded control systems

    NASA Astrophysics Data System (ADS)

    Diglio, A.; Nicolodi, B.

    2002-07-01

    LogiKit provides an overall lifecycle solution. LogiKit is a powerful software engineering case toolkit for requirements specification, simulation and documentation. LogiKit also provides an automatic ADA software design, code and unit test generator.

  1. Psychometric Properties of the Concept Assessment Kit-Conservation.

    ERIC Educational Resources Information Center

    Lehnert, Linda; And Others

    1986-01-01

    This study investigated the psychometric properties of the Educational and Industrial Testing Service Concept Assessment Kit-Conservation (EITS Kit). Presented are an overview of the concept of conservation, a description of the EITS Kit, and results of the study. (MT)

  2. [Comparison and evaluation of the Binax EIA and Biotest EIA urinary antigen kits for detection of Legionella pneumophila antigen in urine samples].

    PubMed

    Rastawicki, Waldemar; Rokosz, Natalia; Jagielski, Marek

    2011-01-01

    The Binax and the Biotest urinary antigen kits for detection of L. pneumophila antigen were compared by testing of selected 67 urine samples obtained from EWGLI as reference samples in External Quality Assessment Scheme. Thirty nine were positive with the Binax kit (100% of sensitivity), and 33 were positive with the Biotest (84.6% of sensitivity). The test specificities were 100% for the both kits. It was concluded that the Binax kit was more suitable for the routine diagnosis of Legionella infections than the Biotest kit.

  3. Data on quantification of signaling pathways activated by KIT and PDGFRA mutants.

    PubMed

    Bahlawane, Christelle; Schmitz, Martine; Letellier, Elisabeth; Arumugam, Karthik; Nicot, Nathalie; Nazarov, Petr V; Haan, Serge

    2016-12-01

    The present data are related to the article entitled "Insights into ligand stimulation effects on gastro-intestinal stromal tumors signaling" (C. Bahlawane, M. Schmitz, E. Letellier, K. Arumugam, N. Nicot, P.V. Nazarov, S. Haan, 2016) [1]. Constitutive and ligand-derived signaling pathways mediated by KIT and PDGFRA mutated proteins found in gastrointestinal stromal tumors (GIST) were compared. Expression of mutant proteins was induced by doxycycline in an isogenic background (Hek293 cells). Kit was identified by FACS at the cell surface and found to be quickly degraded or internalized upon SCF stimulation for both Kit Wild type and Kit mutant counterparts. Investigation of the main activated pathways in GIST unraveled a new feature specific for oncogenic KIT mutants, namely their ability to be further activated by Kit ligand, the stem cell factor (scf). We were also able to identify the MAPK pathway as the most prominent target for a common inhibition of PDGFRA and KIT oncogenic signaling. Western blotting and micro-array analysis were applied to analyze the capacities of the mutant to induce an effective STATs response. Among all Kit mutants, only Kit Ex11 deletion mutant was able to elicit an effective STATs response whereas all PDGFRA were able to do so.

  4. Protocol: a rapid and economical procedure for purification of plasmid or plant DNA with diverse applications in plant biology

    PubMed Central

    2010-01-01

    Research in plant molecular biology involves DNA purification on a daily basis. Although different commercial kits enable convenient extraction of high-quality DNA from E. coli cells, PCR and agarose gel samples as well as plant tissues, each kit is designed for a particular type of DNA extraction work, and the cost of purchasing these kits over a long run can be considerable. Furthermore, a simple method for the isolation of binary plasmid from Agrobacterium tumefaciens cells with satisfactory yield is lacking. Here we describe an easy protocol using homemade silicon dioxide matrix and seven simple solutions for DNA extraction from E. coli and A. tumefaciens cells, PCR and restriction digests, agarose gel slices, and plant tissues. Compared with the commercial kits, this protocol allows rapid DNA purification from diverse sources with comparable yield and purity at negligible cost. Following this protocol, we have demonstrated: (1) DNA fragments as small as a MYC-epitope tag coding sequence can be successfully recovered from an agarose gel slice; (2) Miniprep DNA from E. coli can be eluted with as little as 5 μl water, leading to high DNA concentrations (>1 μg/μl) for efficient biolistic bombardment of Arabidopsis seedlings, polyethylene glycol (PEG)-mediated Arabidopsis protoplast transfection and maize protoplast electroporation; (3) Binary plasmid DNA prepared from A. tumefaciens is suitable for verification by restriction analysis without the need for large scale propagation; (4) High-quality genomic DNA is readily isolated from several plant species including Arabidopsis, tobacco and maize. Thus, the silicon dioxide matrix-based DNA purification protocol offers an easy, efficient and economical way to extract DNA for various purposes in plant research. PMID:20180960

  5. Choline Alleviates Parenteral Nutrition-Associated Duodenal Motility Disorder in Infant Rats.

    PubMed

    Zhu, Jie; Wu, Yang; Guo, Yonggao; Tang, Qingya; Lu, Ting; Cai, Wei; Huang, Haiyan

    2016-09-01

    Parenteral nutrition (PN) has been found to influence duodenal motility in animals. Choline is an essential nutrient, and its deficiency is related to PN-associated organ diseases. Therefore, this study was aimed to investigate the role of choline supplementation in an infant rat model of PN-associated duodenal motility disorder. Three-week-old Sprague-Dawley male rats were fed chow and water (controls), PN solution (PN), or PN plus intravenous choline (600 mg/kg) (PN + choline). Rats underwent jugular vein cannulation for infusion of PN solution or 0.9% saline (controls) for 7 days. Duodenal oxidative stress status, concentrations of plasma choline, phosphocholine, and betaine and serum tumor necrosis factor (TNF)-α were assayed. The messenger RNA (mRNA) and protein expression of c-Kit proto-oncogene protein (c-Kit) and membrane-bound stem cell factor (mSCF) together with the electrophysiological features of slow waves in the duodenum were also evaluated. Rats on PN showed increased reactive oxygen species; decreased total antioxidant capacity in the duodenum; reduced plasma choline, phosphocholine, and betaine; and enhanced serum TNF-α concentrations, which were reversed by choline intervention. In addition, PN reduced mRNA and protein expression of mSCF and c-Kit, which were inversed under choline administration. Moreover, choline attenuated depolarized resting membrane potential and declined the frequency and amplitude of slow waves in duodenal smooth muscles of infant rats induced by PN, respectively. The addition of choline to PN may alleviate the progression of duodenal motor disorder through protecting smooth muscle cells from injury, promoting mSCF/c-Kit signaling, and attenuating impairment of interstitial cells of Cajal in the duodenum during PN feeding. © 2015 American Society for Parenteral and Enteral Nutrition.

  6. [Forensic Application of HuaxiaTM Platinum Kit].

    PubMed

    Wang, Y L; Sheng, X; Li, M; Chen, Y L; Lin, Y; Chen, L Q

    2017-04-01

    To investigate the genetic polymorphism of 23 autosomal STR loci of Huaxia™ Platinum kit in Chinese Han population, and to evaluate the forensic efficiency of Huaxia™ Platinum kit. A total of 500 unrelated healthy individuals from Han population were genotyped with Huaxia™ Platinum kit. The frequency distribution and the parameter of population genetics of STR loci were analysed statistically. Huaxia™ Platinum kit was compared with other 7 commercial STR kits commonly seen at home and abroad in the number of STR loci, interior label, fluorescent mark, total number of alleles in Ladder and system effectiveness. All the 23 autosomal STR loci were consistent with Hardy-Weinberg equilibrium ( P >0.05). The discrimination power was 0.791 5-0.986 2. The polymorphism information content (PIC) was 0.559 0-0.914 0. The combined discrimination power (CDP) was 1-4.1×10⁻²⁸, while combined probability of paternity exclusion in trio (CPET) and in duo (CPED) were 1-4.1×10⁻¹⁰ and 1-8.4×10⁻⁷, respectively. Compared with other 7 kits, Huaxia™ Platinum kit contained the most number of alleles within the Ladder. All the 23 autosomal STR loci of Huaxia™ Platinum kit with highly polymorphic in Han population can be used for paternity testing and individual identification. Compared with other 7 kits, it appears that Huaxia™ Platinum kit can provide more genetic information. Copyright© by the Editorial Department of Journal of Forensic Medicine

  7. SensorKit: An End-to-End Solution for Environmental Sensor Networking

    NASA Astrophysics Data System (ADS)

    Silva, F.; Graham, E.; Deschon, A.; Lam, Y.; Goldman, J.; Wroclawski, J.; Kaiser, W.; Benzel, T.

    2008-12-01

    Modern day sensor network technology has shown great promise to transform environmental data collection. However, despite the promise, these systems have remained the purview of the engineers and computer scientists who design them rather than a useful tool for the environmental scientists who need them. SensorKit is conceived of as a way to make wireless sensor networks accessible to The People: it is an advanced, powerful tool for sensor data collection that does not require advanced technological know-how. We are aiming to make wireless sensor networks for environmental science as simple as setting up a standard home computer network by providing simple, tested configurations of commercially-available hardware, free and easy-to-use software, and step-by-step tutorials. We designed and built SensorKit using a simplicity-through-sophistication approach, supplying users a powerful sensor to database end-to-end system with a simple and intuitive user interface. Our objective in building SensorKit was to make the prospect of using environmental sensor networks as simple as possible. We built SensorKit from off the shelf hardware components, using the Compact RIO platform from National Instruments for data acquisition due to its modular architecture and flexibility to support a large number of sensor types. In SensorKit, we support various types of analog, digital and networked sensors. Our modular software architecture allows us to abstract sensor details and provide users a common way to acquire data and to command different types of sensors. SensorKit is built on top of the Sensor Processing and Acquisition Network (SPAN), a modular framework for acquiring data in the field, moving it reliably to the scientist institution, and storing it in an easily-accessible database. SPAN allows real-time access to the data in the field by providing various options for long haul communication, such as cellular and satellite links. Our system also features reliable data storage and transmission, using a custody transfer mechanism that ensures data is retained until successful delivery to the scientist can be confirmed. The ability for the scientist to communicate in real-time with the sensor network in the field enables remote sensor reconfiguration and system health and status monitoring. We use a spiral approach of design, test, deploy and revise, and, by going to the field frequently and getting feedback from field scientists, we have been able to include additional functionality that is useful to the scientist while ensuring SensorKit remains intuitive to operate. Users can configure, control, and monitor SensorKit using a number of tools we have developed. An intuitive user interface running on a desktop or laptop allows scientists to setup the system, add and configure sensors, and specify when and how the data will be collected. We also have a mobile version of our interface that runs on a PDA and lets scientists calibrate sensors and "tune" the system while in the field, allowing for data validation before leaving the field and returning to the research lab. SensorKit also features SensorBase, an intuitive user interface built on top of a standard SQL database, which allows scientists to store and share their data with other researchers. SensorKit has been used for diverse scientific applications and deployed throughout the world: from studying mercury cycling in rice paddies in China, to ecological research in the neotropical rainforests of Costa Rica, to monitoring the contamination of salt lakes in Argentina.

  8. Research and Development for Health and Environmental Hazard Assessment -Task Order 2. Problem Definition of R and D Requirements for Field Sanitation and Water Supply.

    DTIC Science & Technology

    1982-09-01

    34Development of a Kit for Hazardous Material Spills into Waterways," Edgewood Arsenal Special Publication ED- SP - 76023, 1976. Department of the Army...hepatitis, entamoeba hystolitica are more resistant and require free available chlorine residuals of 10 mg/l in cold weather and levels of 2, 5, 10

  9. High throughput DNA damage quantification of human tissue with home-based collection device

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

    Costes, Sylvain V.; Tang, Jonathan; Yannone, Steven M.

    Kits, methods and systems for providing a service to provide a subject with information regarding the state of a subject's DNA damage. Collection, processing and analysis of samples are also described.

  10. Magnolol pretreatment prevents sepsis-induced intestinal dysmotility by maintaining functional interstitial cells of Cajal.

    PubMed

    Miao, Bin; Zhang, Shuwen; Wang, Hong; Yang, Tiecheng; Zhou, Deshan; Wang, Bao-en

    2013-08-01

    The purpose of this study was to investigate the mechanism by which magnolol treatment prevents lipopolysaccharide (LPS)-induced septic dysmotility in mice. Sepsis was induced by intravenous tail vein injection of LPS (4 mg/kg body weight). Animals were divided into three groups: the magnolol-treated septic group, the placebo-treated septic group, and the control group. Intestinal transit and circular smooth muscle contraction were measured 12 h after LPS injection, and immunocytochemisty was performed to study the morphology of interstitial cells of Cajal (ICCs). Stem cell factor (SCF) expression and c-kit phosphorylation were determined by Western blot analysis, and the mRNA levels of inducible NO synthase (iNOS) were determined by RT-PCR. Nitric oxide (NO) content, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) concentration were detected using commercial kits. Intestinal transit and muscular contractility were significantly lower in the LPS-treated group than in the control group. Immunocytochemical experiments showed that the total number of ICCs, and the total and average lengths of the ICC processes were significantly decreased in the LPS-treated group compared with those in the control group. In LPS-treated animals, magnolol pretreatment significantly accelerated intestinal transit, increased circular muscle contraction, and prevented ICC morphology changes. Phosphorylation of c-kit and expression of SCF were significantly downregulated in LPS-treated animals compared with control animals. Magnolol pretreatment prevented sepsis-induced decreases in c-kit phosphorylation and SCF expression in LPS-treated animals. Magnolol pretreatment prevented the sepsis-induced increase in NO concentration, iNOS expression, and MDA concentration, and decrease in SOD activity in LPS-treated animals. Our results suggest that magnolol treatment prevents sepsis-induced intestinal dysmotility by regulating SCF/c-kit and NO signaling to maintain functional ICCs.

  11. Innovation Online Teaching Module Plus Digital Engineering Kit with Proteus Software through Hybrid Learning Method to Improve Student Skills

    NASA Astrophysics Data System (ADS)

    Kholis, Nur; Syariffuddien Zuhrie, Muhamad; Rahmadian, Reza

    2018-04-01

    Demands the competence (competence) needs of the industry today is a competent workforce to the field of work. However, during this lecture material Digital Engineering (Especially Digital Electronics Basics and Digital Circuit Basics) is limited to the delivery of verbal form of lectures (classical method) is dominated by the Lecturer (Teacher Centered). Though the subject of Digital Engineering requires learning tools and is required understanding of electronic circuits, digital electronics and high logic circuits so that learners can apply in the world of work. One effort to make it happen is by creating an online teaching module and educational aids (Kit) with the help of Proteus software that can improve the skills of learners. This study aims to innovate online teaching modules plus kits in Proteus-assisted digital engineering courses through hybrid learning approaches to improve the skills of learners. The process of innovation is done by considering the skills and mastery of the technology of students (students) Department of Electrical Engineering - Faculty of Engineering – Universitas Negeri Surabaya to produce quality graduates Use of online module plus Proteus software assisted kit through hybrid learning approach. In general, aims to obtain adequate results with affordable cost of investment, user friendly, attractive and interactive (easily adapted to the development of Information and Communication Technology). With the right design, implementation and operation, both in the form of software both in the form of Online Teaching Module, offline teaching module, Kit (Educational Viewer), and e-learning learning content (both online and off line), the use of the three tools of the expenditure will be able to adjust the standard needs of Information and Communication Technology world, both nationally and internationally.

  12. Comparative sensitivity and inhibitor tolerance of GlobalFiler® PCR Amplification and Investigator® 24plex QS kits for challenging samples.

    PubMed

    Elwick, Kyleen; Mayes, Carrie; Hughes-Stamm, Sheree

    2018-05-01

    In cases such as mass disasters or missing persons, human remains are challenging to identify as they may be fragmented, burnt, been buried, decomposed, and/or contain inhibitory substances. This study compares the performance of a relatively new STR kit in the US market (Investigator® 24plex QS kit; Qiagen) with the GlobalFiler® PCR Amplification kit (Thermo Fisher Scientific) when genotyping highly inhibited and low level DNA samples. In this study, DNA samples ranging from 1 ng to 7.8 pg were amplified to define the sensitivity of two systems. In addition, DNA (1 ng and 0.1 ng input amounts) was spiked with various concentrations of five inhibitors common to human remains (humic acid, melanin, hematin, collagen, calcium). Furthermore, bone (N = 5) and tissue samples from decomposed human remains (N = 6) were used as mock casework samples for comparative analysis with both STR kits. The data suggest that the GlobalFiler® kit may be slightly more sensitive than the Investigator® kit. On average STR profiles appeared to be more balanced and average peak heights were higher when using the GlobalFiler® kit. However, the data also show that the Investigator® kit may be more tolerant to common PCR inhibitors. While both STR kits showed a decrease in alleles as the inhibitor concentration increased, more complete profiles were obtained when the Investigator® kit was used. Of the 11 bone and decomposed tissue samples tested, 8 resulted in more complete and balanced STR profiles when amplified with the GlobalFiler® kit. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Comparison of lesional skin c-KIT mutations with clinical phenotype in patients with mastocytosis.

    PubMed

    Chan, I J; Tharp, M D

    2018-06-01

    Activating c-KIT mutations cause abnormal mast cell growth and appear to play a role in mastocytosis. However, the correlation of c-KIT mutations with disease phenotypes is poorly characterized. To evaluate the correlation of c-KIT mutations with clinical presentations and laboratory findings. Total cellular RNA was isolated from the skin lesions of 43 adults and 7 children with mastocytosis, and PCR amplicons of cDNA were sequenced for c-KIT mutations. The most common activating mutation, KIT-D816V, was identified in 72% of adults and 57% of children. Additional activating mutations, namely, V560G and the internal tandem duplications (ITDs) 502-503dupAY, were detected in 12% of adults and 8% of children. V560G occurred more commonly in our patients than previously reported, and it appeared to be associated with more advanced disease. Otherwise, the presence or absence of activating mutations did not correlate with skin lesion morphology, disease extent or total serum tryptase levels. Four adults had expression only of wild-type KIT, while two others had expression of a truncated KIT lacking tyrosine kinase activity; yet these patients were clinically indistinguishable from those patients with activating c-KIT mutations. Activating c-KIT mutations exist in a significant portion of patients with mastocytosis, but not all patients showed expression of these mutations. Except for V560G, the presence or absence of activating c-KIT mutations did not predict the extent of disease. These observations suggest that although activating c-KIT mutations are associated with mast cell growth, other genes probably play a role in the cause of mastocytosis. © 2018 British Association of Dermatologists.

  14. Impact and Effectiveness of a Stand-Alone NRT Starter Kit in a Statewide Tobacco Cessation Program.

    PubMed

    Kerr, Amy N; Schillo, Barbara A; Keller, Paula A; Lachter, Randi B; Lien, Rebecca K; Zook, Heather G

    2018-01-01

    To examine 2-week nicotine replacement therapy (NRT) starter kit quit outcomes and predictors and the impact of adding this new service on treatment reach. Observational study of a 1-year cohort of QUITPLAN Services enrollees using registration and utilization data and follow-up outcome survey data of a subset of enrollees who received NRT starter kits. ClearWay Minnesota's QUITPLAN Services provides a quit line that is available to uninsured and underinsured Minnesotans and NRT starter kits (a free 2-week supply of patches, gum, or lozenges) that are available to all Minnesota tobacco users. A total of 15 536 adult QUITPLAN Services enrollees and 818 seven-month follow-up survey NRT starter kit respondents. Treatment reach for all services and tobacco quit outcomes and predictors for starter kit recipients. Descriptive analyses, χ 2 analyses, and logistic regression. Treatment reach increased 3-fold after adding the 2-week NRT starter kit service option to QUITPLAN Services compared to the prior year (1.86% vs 0.59%). Among all participants enrolling in QUITPLAN services during a 1-year period, 83.8% (13 026/15 536) registered for a starter kit. Among starter kit respondents, 25.6% reported being quit for 30 days at the 7-month follow-up. After controlling for other factors, using all NRT and selecting more cessation services predicted quitting. An NRT starter kit brought more tobacco users to QUITPLAN services, demonstrating interest in cessation services separate from phone counseling. The starter kit produced high quit rates, comparable to the quit line in the same time period. Cessation service providers may want to consider introducing starter kits to reach more tobacco users and ultimately improve population health.

  15. Histone deacetylase inhibitor SAHA mediates mast cell death and epigenetic silencing of constitutively active D816V KIT in systemic mastocytosis.

    PubMed

    Lyberg, Katarina; Ali, Hani Abdulkadir; Grootens, Jennine; Kjellander, Matilda; Tirfing, Malin; Arock, Michel; Hägglund, Hans; Nilsson, Gunnar; Ungerstedt, Johanna

    2017-02-07

    Systemic mastocytosis (SM) is a clonal bone marrow disorder, where therapeutical options are limited. Over 90% of the patients carry the D816V point mutation in the KIT receptor that renders this receptor constitutively active. We assessed the sensitivity of primary mast cells (MC) and mast cell lines HMC1.2 (D816V mutated), ROSA (KIT WT) and ROSA (KIT D816V) cells to histone deacetylase inhibitor (HDACi) treatment. We found that of four HDACi, suberoyl anilide hydroxamic acid (SAHA) was the most effective in killing mutated MC. SAHA downregulated KIT, followed by major MC apoptosis. Primary SM patient MC cultured ex vivo were even more sensitive to SAHA than HMC1.2 cells, whereas primary MC from healthy subjects were less affected. There was a correlation between cell death and SM disease severity, where cell death was more pronounced in the case of aggressive SM, with almost 100% cell death among MC from the mast cell leukemia patient. Additionally, ROSA (KIT D816V) was more affected by HDACi than ROSA (KIT WT) cells. Using ChIP qPCR, we found that the level of active chromatin mark H3K18ac/H3 decreased significantly in the KIT region. This epigenetic silencing was seen only in the KIT region and not in control genes upstream and downstream of KIT, indicating that the downregulation of KIT is exerted by specific epigenetic silencing. In conclusion, KIT D816V mutation sensitized MC to HDACi mediated killing, and SAHA may be of value as specific treatment for SM, although the specific mechanism of action requires further investigation.

  16. Histone deacetylase inhibitor SAHA mediates mast cell death and epigenetic silencing of constitutively active D816V KIT in systemic mastocytosis

    PubMed Central

    Lyberg, Katarina; Ali, Hani Abdulkadir; Grootens, Jennine; Kjellander, Matilda; Tirfing, Malin; Arock, Michel; Hägglund, Hans

    2017-01-01

    Systemic mastocytosis (SM) is a clonal bone marrow disorder, where therapeutical options are limited. Over 90% of the patients carry the D816V point mutation in the KIT receptor that renders this receptor constitutively active. We assessed the sensitivity of primary mast cells (MC) and mast cell lines HMC1.2 (D816V mutated), ROSA (KIT WT) and ROSA (KIT D816V) cells to histone deacetylase inhibitor (HDACi) treatment. We found that of four HDACi, suberoyl anilide hydroxamic acid (SAHA) was the most effective in killing mutated MC. SAHA downregulated KIT, followed by major MC apoptosis. Primary SM patient MC cultured ex vivo were even more sensitive to SAHA than HMC1.2 cells, whereas primary MC from healthy subjects were less affected. There was a correlation between cell death and SM disease severity, where cell death was more pronounced in the case of aggressive SM, with almost 100% cell death among MC from the mast cell leukemia patient. Additionally, ROSA (KIT D816V) was more affected by HDACi than ROSA (KIT WT) cells. Using ChIP qPCR, we found that the level of active chromatin mark H3K18ac/H3 decreased significantly in the KIT region. This epigenetic silencing was seen only in the KIT region and not in control genes upstream and downstream of KIT, indicating that the downregulation of KIT is exerted by specific epigenetic silencing. In conclusion, KIT D816V mutation sensitized MC to HDACi mediated killing, and SAHA may be of value as specific treatment for SM, although the specific mechanism of action requires further investigation. PMID:28038453

  17. EnergyPlus™

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

    Originally developed in 1999, an updated version 8.8.0 with bug fixes was released on September 30th, 2017. EnergyPlus™ is a whole building energy simulation program that engineers, architects, and researchers use to model both energy consumption—for heating, cooling, ventilation, lighting and plug and process loads—and water use in buildings. EnergyPlus is a console-based program that reads input and writes output to text files. It ships with a number of utilities including IDF-Editor for creating input files using a simple spreadsheet-like interface, EP-Launch for managing input and output files and performing batch simulations, and EP-Compare for graphically comparing the results ofmore » two or more simulations. Several comprehensive graphical interfaces for EnergyPlus are also available. DOE does most of its work with EnergyPlus using the OpenStudio® software development kit and suite of applications. DOE releases major updates to EnergyPlus twice annually.« less

  18. Cationic pentaheteroaryls as selective G-quadruplex ligands by solvent-free microwave-assisted synthesis.

    PubMed

    Petenzi, Michele; Verga, Daniela; Largy, Eric; Hamon, Florian; Doria, Filippo; Teulade-Fichou, Marie-Paule; Guédin, Aurore; Mergny, Jean-Louis; Mella, Mariella; Freccero, Mauro

    2012-11-05

    We report herein a solvent-free and microwaved-assisted synthesis of several water soluble acyclic pentaheteroaryls containing 1,2,4-oxadiazole moieties (1-7). Their binding interactions with DNA quadruplex structures were thoroughly investigated by FRET melting, fluorescent intercalator displacement assay (G4-FID) and CD spectroscopy. Among the G-quadruplexes considered, attention was focused on telomeric repeats together with the proto-oncogenic c-kit sequences and the c-myc oncogene promoter. Compound 1, and to a lesser extent 2 and 5, preferentially stabilise an antiparallel structure of the telomeric DNA motif, and exhibit an opposite binding behaviour to structurally related polyoxazole (TOxaPy), and do not bind duplex DNA. The efficiency and selectivity of the binding process was remarkably controlled by the structure of the solubilising moieties. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. 40 CFR 59.506 - How do I demonstrate compliance if I manufacture multi-component kits?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... multi-component kits as defined in § 59.503, then the Kit PWR must not exceed the Total Reactivity Limit. (b) You must calculate the Kit PWR and the Total Reactivity Limit as follows: (1) KIT PWR = (PWR(1) × W1) + (PWR(2) × W2) +. ...+ (PWR(n) × Wn) (2) Total Reactivity Limit = (RL1 × W1) + (RL2 × W2...

  20. 40 CFR 59.506 - How do I demonstrate compliance if I manufacture multi-component kits?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... multi-component kits as defined in § 59.503, then the Kit PWR must not exceed the Total Reactivity Limit. (b) You must calculate the Kit PWR and the Total Reactivity Limit as follows: (1) KIT PWR = (PWR(1) × W1) + (PWR(2) × W2) +. ...+ (PWR(n) × Wn) (2) Total Reactivity Limit = (RL1 × W1) + (RL2 × W2...

  1. 40 CFR 59.506 - How do I demonstrate compliance if I manufacture multi-component kits?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... multi-component kits as defined in § 59.503, then the Kit PWR must not exceed the Total Reactivity Limit. (b) You must calculate the Kit PWR and the Total Reactivity Limit as follows: (1) KIT PWR = (PWR(1) × W1) + (PWR(2) × W2) +. ...+ (PWR(n) × Wn) (2) Total Reactivity Limit = (RL1 × W1) + (RL2 × W2...

  2. 40 CFR 59.506 - How do I demonstrate compliance if I manufacture multi-component kits?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... multi-component kits as defined in § 59.503, then the Kit PWR must not exceed the Total Reactivity Limit. (b) You must calculate the Kit PWR and the Total Reactivity Limit as follows: (1) KIT PWR = (PWR(1) × W1) + (PWR(2) × W2) +. ...+ (PWR(n) × Wn) (2) Total Reactivity Limit = (RL1 × W1) + (RL2 × W2...

  3. Effects of endoplasmic reticulum stressors on maturation and signaling of hemizygous and heterozygous wild-type and mutant forms of KIT.

    PubMed

    Brahimi-Adouane, Sabrina; Bachet, Jean-Baptiste; Tabone-Eglinger, Séverine; Subra, Frédéric; Capron, Claude; Blay, Jean-Yves; Emile, Jean-François

    2013-06-01

    Gain of function mutations of KIT are frequent in some human tumors, and are sensible to tyrosine kinase inhibitors. In most tumors, oncogenic mutations are heterozygous, however most in vitro data of KIT activation have been obtained with hemizygous mutation. This study aimed to investigate the maturation and activation of wild-type (WT) and mutant (M) forms of KIT in hemizygous and heterozygous conditions. WT and two types of exon 11 deletions M forms of human KIT were expressed in NIH3T3 cell lines. Membrane expression of KIT was quantified by flow cytometry. Quantification of glycosylated forms of KIT and phosphorylated forms of AKT and ERK were performed by western blot. Simultaneous activation of WT KIT and treatment with endoplasmic reticulum (ER) inhibitors, tunicamycin or brefeldin A induced a complete inhibition of membrane expression of the 145 kDa form of KIT. By contrast activation or ER inhibitors alone, only partly inhibited this form. ER inhibitors also inhibited KIT activation-dependent phosphorylation of AKT and ERK1/2. Brefeldin A induced a complete down regulation of the 145 kDa form in hemizygous M, and induced an intra-cellular accumulation of the 125 kDa form in WT but not in hemizygous M. Heterozygous cells had glycosylation and response to ER inhibitors patterns more similar to WT than to hemizygous M. Phosphorylated AKT was reduced in hemizygous cells in comparison to WT KIT cells and heterozygous cells, and in the presence of brefeldin A in all cell lines. Effects of ER inhibitors are significantly different in hemizygous and heterozygous mutants. Differences in intra-cellular trafficking of KIT forms result in differences in downstream signaling pathways, and activation of PI3K/AKT pathway appears to be tied to the presence of the mature 145 kDa form of KIT at the membrane surface. Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.

  4. Clinicopathological characteristics of KIT and protein kinase C-δ expression in adenoid cystic carcinoma: comparison with chromophobe renal cell carcinoma and gastrointestinal stromal tumour.

    PubMed

    Park, Cheol Keun; Kim, Won Kyu; Kim, Hoguen

    2017-10-01

    KIT overexpression is frequently observed in adenoid cystic carcinomas (AdCCs), chromophobe renal cell carcinomas (ChRCCs), and gastrointestinal stromal tumours (GISTs). Persistent KIT activation has been reported to be mediated by protein kinase C (PKC)-δ in a subset of colon cancers with wild-type KIT overexpression, and by PKC-θ in GISTs with mutant KIT overexpression. To elucidate the clinical implications of PKC-δ and PKC-θ expression in KIT-expressing tumours, we investigated the expression of KIT, PKC-δ and PKC-θ in AdCCs and ChRCCs in comparison with GISTs. KIT expression, PKC-δ expression and PKC-θ expression were analysed in whole sections from 41 AdCCs, 40 ChRCCs and 56 GISTs by immunohistochemistry. Membranous expression of KIT was found in 34 AdCCs and all ChRCCs, whereas cytoplasmic expression of KIT was found in 46 GISTs. In AdCCs, PKC-δ expression was associated with histological grade (P = 0.049), lymphovascular invasion (P = 0.004), perineural invasion (P = 0.002), and KIT positivity (P = 0.002). PKC-δ positivity was associated with shorter relapse-free survival (RFS) (P = 0.017) and a tendency for there to be shorter overall survival (OS) (P = 0.090) in patients with AdCCs. No clinicopathological associations were observed between PKC-δ and KIT expression in ChRCCs. In GISTs, PKC-θ expression was associated with higher mitotic count (P = 0.011) and high grade according to the modified National Institutes of Health criteria (P < 0.001). PKC-θ positivity was associated with shorter RFS (P = 0.016) and a tendency for there to be shorter OS (P = 0.051) in patients with GISTs. PKC-δ expression is associated with KIT expression and the prognosis of patients with AdCCs, suggesting that PKC-δ may be a potential therapeutic target for AdCCs. © 2017 The Authors. Histopathology published by John Wiley & Sons Ltd.

  5. Activated c-Kit receptor in the heart promotes cardiac repair and regeneration after injury

    PubMed Central

    Di Siena, S; Gimmelli, R; Nori, S L; Barbagallo, F; Campolo, F; Dolci, S; Rossi, P; Venneri, M A; Giannetta, E; Gianfrilli, D; Feigenbaum, L; Lenzi, A; Naro, F; Cianflone, E; Mancuso, T; Torella, D; Isidori, A M; Pellegrini, M

    2016-01-01

    The role of endogenous c-Kit receptor activation on cardiac cell homeostasis and repair remains largely unexplored. Transgenic mice carrying an activating point mutation (TgD814Y) in the kinase domain of the c-Kit gene were generated. c-KitTgD814Y receptor was expressed in the heart during embryonic development and postnatal life, in a similar timing and expression pattern to that of the endogenous gene, but not in the hematopoietic compartment allowing the study of a cardiac-specific phenotype. c-KitTgD814Y mutation produced a constitutive active c-Kit receptor in cardiac tissue and cells from transgenic mice as demonstrated by the increased phosphorylation of ERK1/2 and AKT, which are the main downstream molecular effectors of c-Kit receptor signaling. In adult transgenic hearts, cardiac morphology, size and total c-Kit+ cardiac cell number was not different compared with wt mice. However, when c-KitTgD814Y mice were subjected to transmural necrotic heart damage by cryoinjury (CI), all transgenic survived, compared with half of wt mice. In the sub-acute phase after CI, transgenic and wt mice showed similar heart damage. However, 9 days after CI, transgenic mice exhibited an increased number of c-Kit+CD31+ endothelial progenitor cells surrounding the necrotic area. At later follow-up, a consistent reduction of fibrotic area, increased capillary density and increased cardiomyocyte replenishment rate (as established by BrdU incorporation) were observed in transgenic compared with wt mice. Consistently, CD45−c-Kit+ cardiac stem cells isolated from transgenic c-KitTgD814Y mice showed an enhanced endothelial and cardiomyocyte differentiation potential compared with cells isolated from the wt. Constitutive activation of c-Kit receptor in mice is associated with an increased cardiac myogenic and vasculogenic reparative potential after injury, with a significant improvement of survival. PMID:27468693

  6. Repression of c-Kit by p53 is mediated by miR-34 and is associated with reduced chemoresistance, migration and stemness

    PubMed Central

    Siemens, Helge; Jackstadt, Rene; Kaller, Markus; Hermeking, Heiko

    2013-01-01

    The c-Kit receptor tyrosine kinase is commonly over-expressed in different types of cancer. p53 activation is known to result in the down-regulation of c-Kit. However, the underlying mechanism has remained unknown. Here, we show that the p53-induced miR-34 microRNA family mediates repression of c-Kit by p53 via a conserved seed-matching sequence in the c-Kit 3'-UTR. Ectopic miR-34a resulted in a decrease in Erk signaling and transformation, which was dependent on the down-regulation of c-Kit expression. Furthermore, ectopic expression of c-Kit conferred resistance of colorectal cancer (CRC) cells to treatment with 5-fluorouracil (5-FU), whereas ectopic miR-34a sensitized the cells to 5-FU. After stimulation with c-Kit ligand/stem cell factor (SCF) Colo320 CRC cells displayed increased migration/invasion, whereas ectopic miR-34a inhibited SCF-induced migration/invasion. Activation of a conditional c-Kit allele induced several stemness markers in DLD-1 CRC cells. In primary CRC samples elevated c-Kit expression also showed a positive correlation with markers of stemness, such as Lgr5, CD44, OLFM4, BMI-1 and β-catenin. On the contrary, activation of a conditional miR-34a allele in DLD-1 cells diminished the expression of c-Kit and several stemness markers (CD44, Lgr5 and BMI-1) and suppressed sphere formation. MiR-34a also suppressed enhanced sphere-formation after exposure to SCF. Taken together, our data establish c-Kit as a new direct target of miR-34 and demonstrate that this regulation interferes with several c-Kit-mediated effects on cancer cells. Therefore, this regulation may be potentially relevant for future diagnostic and therapeutic approaches. PMID:24009080

  7. HNU-HANBY PCP IMMUNOASSAY TEST KIT - INNOVATIVE TECHNOLOGY EVALUATION REPORT

    EPA Science Inventory

    The HNU-Hanby pentachlorophenol (PCP) test kit rapidly analyzes for PCP in soil samples. The test kit can only detect those PCP carriers that contain aromatic compounds. The test kit estimates PCP concentrations in soil samples indirectly by measuring petroleum hydrocarbon carrie...

  8. Evaluation of a new rapid kit, BD MGIT TBc identification test for confirmation of Mycobacterium tuberculosis complex.

    PubMed

    Kandhakumari, Gandhi; Stephen, Selvaraj

    2017-01-01

    At present, three rapid kits are available globally for the confirmation of Mycobacterium tuberculosis complex (MTBC) in cultures by MPT64 antigen (MPT64 Ag) detection. These include Capilia TB, SD Bioline, and BD MGIT TBc Identification (TBcID). The third kit is yet to be validated in India. We have tested this kit and compared with SD Bioline using conventional tests as gold standard. Seventy-one MTBC (70 M. tuberculosis and one Mycobacterium bovis) and four nontuberculous mycobacteria (NTM) were isolated from 649 clinical specimens in MGIT 960 and/or Lowenstein-Jensen slants (LJ). MPT64 Ag was detected by both TBcID and SD Bioline kits in all the 71 clinical isolates and the reference strain M. tuberculosis H37Rv. All NTM species tested were negative by the two different kits. Thus, TBcID kit showed 100% concordance in terms of sensitivity and specificity. Rapid kits confirm MTBC cultures within 15 min in contrast to several weeks' time required by conventional techniques.

  9. [Consistency study of PowerPlex 21 kit and Goldeneye 20A kit and forensic application].

    PubMed

    Ren, He; Liu, Ying; Zhang, Qing-Xia; Jiao, Zhang-Ping

    2014-06-01

    To ensure the consistency of genotype results for PowerPlex 21 kit and Goldeneye 20A kit. The STR loci were amplified in DNA samples from 205 unrelated individuals in Beijing Han population. And consistency of 19 overlap STR loci typing were observed. The genetic polymorphism of D1S1656 locus was obtained. All 19 overlap loci typing showed consistent. The proportion of peak height of heterozygous loci in two kits showed no statistical difference (P > 0.05). The observed heterozygosis of D1S1656 was 0.878. The discrimination power was 0.949. The excluding probability of paternity of triplet was 0.751. The excluding probability of paternity of diploid was 0.506. The polymorphism information content was 0.810. PowerPlex 21 kit and Goldeneye 20A kit present a good consistency. The primer design is reasonable. The polymorphism of D1S1656 is good. The two kits can be used for human genetic analysis, paternity test, and individual identification in forensic practice.

  10. Hematopoietic Kit Deficiency, rather than Lack of Mast Cells, Protects Mice from Obesity and Insulin Resistance.

    PubMed

    Gutierrez, Dario A; Muralidhar, Sathya; Feyerabend, Thorsten B; Herzig, Stephan; Rodewald, Hans-Reimer

    2015-05-05

    Obesity, insulin resistance, and related pathologies are associated with immune-mediated chronic inflammation. Kit mutant mice are protected from diet-induced obesity and associated co-morbidities, and this phenotype has previously been attributed to their lack of mast cells. We performed a comprehensive metabolic analysis of Kit-dependent Kit(W/Wv) and Kit-independent Cpa3(Cre/+) mast-cell-deficient mouse strains, employing diet-induced or genetic (Lep(Ob/Ob) background) models of obesity. Our results show that mast cell deficiency, in the absence of Kit mutations, plays no role in the regulation of weight gain or insulin resistance. Moreover, we provide evidence that the metabolic phenotype observed in Kit mutant mice, while independent of mast cells, is immune regulated. Our data underscore the value of definitive mast cell deficiency models to conclusively test the involvement of this enigmatic cell in immune-mediated pathologies and identify Kit as a key hematopoietic factor in the pathogenesis of metabolic syndrome. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Commercial Milk Enzyme-Linked Immunosorbent Assay (ELISA) Kit Reactivities to Purified Milk Proteins and Milk-Derived Ingredients.

    PubMed

    Ivens, Katherine O; Baumert, Joseph L; Taylor, Steve L

    2016-07-01

    Numerous commercial enzyme-linked immunosorbent assay (ELISA) kits exist to quantitatively detect bovine milk residues in foods. Milk contains many proteins that can serve as ELISA targets including caseins (α-, β-, or κ-casein) and whey proteins (α-lactalbumin or β-lactoglobulin). Nine commercially-available milk ELISA kits were selected to compare the specificity and sensitivity with 5 purified milk proteins and 3 milk-derived ingredients. All of the milk kits were capable of quantifying nonfat dry milk (NFDM), but did not necessarily detect all individual protein fractions. While milk-derived ingredients were detected by the kits, their quantitation may be inaccurate due to the use of different calibrators, reference materials, and antibodies in kit development. The establishment of a standard reference material for the calibration of milk ELISA kits is increasingly important. The appropriate selection and understanding of milk ELISA kits for food analysis is critical to accurate quantification of milk residues and informed risk management decisions. © 2016 Institute of Food Technologists®

  12. MicroSEQ® Salmonella spp. Detection Kit Using the Pathatrix® 10-Pooling Salmonella spp. Kit Linked Protocol Method Modification.

    PubMed

    Wall, Jason; Conrad, Rick; Latham, Kathy; Liu, Eric

    2014-03-01

    Real-time PCR methods for detecting foodborne pathogens offer the advantages of simplicity and quick time to results compared to traditional culture methods. The addition of a recirculating pooled immunomagnetic separation method prior to real-time PCR analysis increases processing output while reducing both cost and labor. This AOAC Research Institute method modification study validates the MicroSEQ® Salmonella spp. Detection Kit [AOAC Performance Tested Method (PTM) 031001] linked with the Pathatrix® 10-Pooling Salmonella spp. Kit (AOAC PTM 090203C) in diced tomatoes, chocolate, and deli ham. The Pathatrix 10-Pooling protocol represents a method modification of the enrichment portion of the MicroSEQ Salmonella spp. The results of the method modification were compared to standard cultural reference methods for diced tomatoes, chocolate, and deli ham. All three matrixes were analyzed in a paired study design. An additional set of chocolate test portions was analyzed using an alternative enrichment medium in an unpaired study design. For all matrixes tested, there were no statistically significant differences in the number of positive test portions detected by the modified candidate method compared to the appropriate reference method. The MicroSEQ Salmonella spp. protocol linked with the Pathatrix individual or 10-Pooling procedure demonstrated reliability as a rapid, simplified, method for the preparation of samples and subsequent detection of Salmonella in diced tomatoes, chocolate, and deli ham.

  13. Transplantation of Epigenetically Modified Adult Cardiac c-Kit+ Cells Retards Remodeling and Improves Cardiac Function in Ischemic Heart Failure Model

    PubMed Central

    Zakharova, Liudmila; Nural-Guvener, Hikmet; Feehery, Lorraine; Popovic-Sljukic, Snjezana

    2015-01-01

    Cardiac c-Kit+ cells have a modest cardiogenic potential that could limit their efficacy in heart disease treatment. The present study was designed to augment the cardiogenic potential of cardiac c-Kit+ cells through class I histone deacetylase (HDAC) inhibition and evaluate their therapeutic potency in the chronic heart failure (CHF) animal model. Myocardial infarction (MI) was created by coronary artery occlusion in rats. c-Kit+ cells were treated with mocetinostat (MOCE), a specific class I HDAC inhibitor. At 3 weeks after MI, CHF animals were retrogradely infused with untreated (control) or MOCE-treated c-Kit+ cells (MOCE/c-Kit+ cells) and evaluated at 3 weeks after cell infusion. We found that class I HDAC inhibition in c-Kit+ cells elevated the level of acetylated histone H3 (AcH3) and increased AcH3 levels in the promoter regions of pluripotent and cardiac-specific genes. Epigenetic changes were accompanied by increased expression of cardiac-specific markers. Transplantation of CHF rats with either control or MOCE/c-Kit+ cells resulted in an improvement in cardiac function, retardation of CHF remodeling made evident by increased vascularization and scar size, and cardiomyocyte hypertrophy reduction. Compared with CHF infused with control cells, infusion of MOCE/c-Kit+ cells resulted in a further reduction in left ventricle end-diastolic pressure and total collagen and an increase in interleukin-6 expression. The low engraftment of infused cells suggests that paracrine effects might account for the beneficial effects of c-Kit+ cells in CHF. In conclusion, selective inhibition of class I HDACs induced expression of cardiac markers in c-Kit+ cells and partially augmented the efficacy of these cells for CHF repair. Significance The study has shown that selective class 1 histone deacetylase inhibition is sufficient to redirect c-Kit+ cells toward a cardiac fate. Epigenetically modified c-Kit+ cells improved contractile function and retarded remodeling of the congestive heart failure heart. This study provides new insights into the efficacy of cardiac c-Kit+ cells in the ischemic heart failure model. PMID:26240433

  14. Adapting to an innovation: Solar cooking in the urban households of Ouagadougou (Burkina Faso)

    NASA Astrophysics Data System (ADS)

    Toonen, Hilde M.

    Most households in Sub-Saharan Africa rely on wood as primary energy source. The availability of wood is decreasing and deforestation is a major ecological problem in Sub-Saharan Africa. The scarcity of wood is demanding for a sustainable solution. The sun seems to provide a good alternative. Solar energy is free, without unhealthy smoke or chances to burns. The idea of using solar energy for cooking is not new: many different techniques have already been tested. Most variants are expensive, and therefore not available for most families in Sub-Saharan Africa. A cheap solar cooking device is the CooKit, a cardboard panel cooker covered with aluminium foil. In the adaptation to the CooKit, as to all innovations, it is important that the users are convinced of the advantages. An important step in the adaptation process is learning how to use the cooking device; the best way to do this is by home practice. Monitoring and evaluating the real use is needed, for it is interesting to know if the CooKit is actually used, and also to find out how women have implemented the new technique in their kitchens. In 2005, the SUPO foundation started a project in Burkina Faso: Programme Energie Solaire Grand-Ouaga (PESGO). The aim of PESGO is to introduce the CooKit in the urban households in Ouagadougou by providing training sessions and home assistance. In this paper, a mid-term review on this small-scale cooking project is presented. The possibilities and challenges of solar cooking are outlined, taking the urban context of Ouagadougou in account. In PESGO, dependence on weather conditions is found to be one of the challenges: if sunrays are blocked by clouds or dust in the air, the cooking will be slowed down. The CooKit cannot replace firewood entirely, and a complementary element has to be found. SUPO is exploring the use of Jatropha oil as a complement to the CooKit. The Jatropha plant is drought tolerant and its fruits contain oil which can be used as fuel substitute. Further research on its use is interesting, because the combination of the CooKit and Jatropha oil seems to have high potential in the kitchens of West-Africa.

  15. KIT gene mutations and patterns of protein expression in mucosal and acral melanoma.

    PubMed

    Abu-Abed, Suzan; Pennell, Nancy; Petrella, Teresa; Wright, Frances; Seth, Arun; Hanna, Wedad

    2012-01-01

    Recently characterized KIT (CD117) gene mutations have revealed new pathways involved in melanoma pathogenesis. In particular, certain subtypes harbor mutations similar to those observed in gastrointestinal stromal tumors, which are sensitive to treatment with tyrosine kinase inhibitors. The purpose of this study was to characterize KIT gene mutations and patterns of protein expression in mucosal and acral melanoma. Formalin-fixed, paraffin-embedded tissues were retrieved from our archives. Histologic assessment included routine hematoxylin-eosin stains and immunohistochemical staining for KIT. Genomic DNA was used for polymerase chain reaction-based amplification of exons 11 and 13. We identified 59 acral and mucosal melanoma cases, of which 78% showed variable levels of KIT expression. Sequencing of exons 11 and 13 was completed on all cases, and 4 (6.8%) mutant cases were isolated. We successfully optimized conditions for the detection of KIT mutations and showed that 8.6% of mucosal and 4.2% of acral melanoma cases at our institution harbor KIT mutations; all mutant cases showed strong, diffuse KIT protein expression. Our case series represents the first Canadian study to characterize KIT gene mutations and patterns of protein expression in acral and mucosal melanoma.

  16. Conditional Deletion of Kit in Melanocytes: White Spotting Phenotype Is Cell Autonomous.

    PubMed

    Aoki, Hitomi; Tomita, Hiroyuki; Hara, Akira; Kunisada, Takahiro

    2015-07-01

    It is well established that cell-intrinsic signaling through the receptor tyrosine kinase KIT is critical for the development of neural crest-derived melanocytes. Nevertheless, it is not entirely clear whether Kit acts exclusively in a melanocyte-autonomous manner or in addition indirectly through other cell types. To address this question in vivo, we generated a targeted allele of Kit that allowed for CRE recombinase-mediated deletion of the transmembrane domain of KIT. Mice carrying one copy of the targeted allele and expressing CRE under the melanoblast/melanocyte-specific tyrosinase promoter exhibited a white spotting phenotype that was even more extensive compared with that found in mice heterozygous for a Kit-null allele. This phenotype is unlikely the result of sequestration of KIT ligand by neighboring cells or by potentially secreted forms of KIT because the spotting phenotype could not be rescued by overexpression of KITL. Likewise, overexpression of endothelin-3 or hepatocyte growth factor was unable to rescue melanocytes in these mice. Although the severity of the observed phenotype remains to be explained, the findings indicate that melanocyte-selective impairment of Kit is sufficient to interfere with normal melanocyte development.

  17. Endothelial NOS is required for SDF-1alpha/CXCR4-mediated peripheral endothelial adhesion of c-kit+ bone marrow stem cells.

    PubMed

    Kaminski, Alexander; Ma, Nan; Donndorf, Peter; Lindenblatt, Nicole; Feldmeier, Gregor; Ong, Lee-Lee; Furlani, Dario; Skrabal, Christian A; Liebold, Andreas; Vollmar, Brigitte; Steinhoff, Gustav

    2008-01-01

    In the era of intravascular approaches for regenerative cell therapy, the underlying mechanisms of stem cell migration to non-marrow tissue have not been clarified. We hypothesized that next to a local inflammatory response implying adhesion molecule expression, endothelial nitric oxide synthase (eNOS)-dependent signaling is required for stromal- cell-derived factor-1 alpha (SDF-1alpha)-induced adhesion of c-kit+ cells to the vascular endothelium. SDF-1alpha/tumor necrosis factor-alpha (TNF-alpha)-induced c-kit+-cell shape change and migration capacity was studied in vitro using immunohistochemistry and Boyden chamber assays. In vivo interaction of c-kit+ cells from bone marrow with the endothelium in response to SDF-1alpha/TNF-alpha stimulation was visualized in the cremaster muscle microcirculation of wild-type (WT) and eNOS (-/-) mice using intravital fluorescence microscopy. In addition, NOS activity was inhibited with N-nitro-L-arginine-methylester-hydrochloride in WT mice. To reveal c-kit+-specific adhesion behavior, endogenous leukocytes (EL) and c-kit+ cells from peripheral blood served as control. Moreover, intercellular adhesion molecule-1 (ICAM-1) and CXCR4 were blocked systemically to determine their role in inflammation-related c-kit+-cell adhesion. In vitro, SDF-1alpha enhanced c-kit+-cell migration. In vivo, SDF-1alpha alone triggered endothelial rolling-not firm adherence-of c-kit+ cells in WT mice. While TNF-alpha alone had little effect on adhesion of c-kit+ cells, it induced maximum endothelial EL adherence. However, after combined treatment with SDF-1alpha+TNF-alpha, endothelial adhesion of c-kit+ cells increased independent of their origin, while EL adhesion was not further incremented. Systemic treatment with anti-ICAM-1 and anti-CXCR4-monoclonal antibody completely abolished endothelial c-kit+-cell adhesion. In N-nitro-L-arginine-methylester-hydrochloride-treated WT mice as well as in eNOS (-/-) mice, firm endothelial adhesion of c-kit+ cells was entirely abrogated, while EL adhesion was significantly increased. The chemokine SDF-1alpha mediates firm adhesion c-kit+ cells only in the presence of TNF-alpha stimulation via an ICAM-1- and CXCR4-dependent mechanism. The presence of eNOS appears to be a crucial and specific factor for firm c-kit+-cell adhesion to the vascular endothelium.

  18. The usefulness of c-Kit in the immunohistochemical assessment of melanocytic lesions

    PubMed Central

    Pilloni, L.; Bianco, P.; Difelice, E.; Cabras, S.; Castellanos, M.E.; Atzori, L.; Ferreli, C.; Mulas, P.; Nemolato, S.; Faa, G.

    2011-01-01

    C-Kit (CD117), the receptor for the stem cell factor, a growth factor for melanocyte migration and proliferation, has shown differential immunostaining in various benign and malignant melanocytic lesions. The purpose of this study is to compare c-Kit immunostaining in benign nevi and in primary and metastatic malignant melanomas, to determine whether c-Kit can aid in the differential diagnosis of these lesions. c-Kit immunostaining was performed in 60 cases of pigmented lesions, including 39 benign nevi (5 blue nevi, 5 intra-dermal nevi, 3 junctional nevi, 15 cases of primary compound nevus, 11 cases of Spitz nevus), 18 cases of primary malignant melanoma and 3 cases of metastatic melanoma. The vast majority of nevi and melanomas examined in this study were positive for c-Kit, with minimal differences between benign and malignant lesions. C-Kit cytoplasmatic immunoreactivity in the intraepidermal proliferating nevus cells, was detected in benign pigmented lesions as well as in malignant melanoma, increasing with the age of patients (P=0.007) in both groups. The patient’s age at presentation appeared to be the variable able to cluster benign and malignant pigmented lesions. The percentage of c-Kit positive intraepidermal nevus cells was better associated with age despite other variables (P=0.014). The intensity and percentage of c-Kit positivity in the proliferating nevus cells in the dermis was significantly increased in malignant melanocytic lesions (P=0.015 and P=0.008) compared to benign lesions (compound melanocytic nevi, Spitz nevi, intradermal nevi, blue nevi). Immunostaning for c-Kit in metastatic melanomas was negative. Interestingly in two cases of melanoma occurring on a pre-existent nevus, the melanoma tumor cells showed strong cytoplasmatic and membranous positivity for c-kit, in contrast with the absence of any immunoreactivity in pre-existent intradermal nevus cells. C-Kit does not appear to be a strong immunohistochemical marker for distinguishing melanoma from melanocytic nevi, if we consider c-Kit expression in intraepidermal proliferating cells. The c-Kit expression in proliferating melanocytes in the dermis could help in the differential diagnosis between a superficial spreading melanoma (with dermis invasion) and a compound nevus or an intradermal nevus. Finally, c-Kit could be a good diagnostic tool for distinguishing benign compound nevi from malignant melanocytic lesions with dermis invasion and to differentiate metastatic melanoma from primary melanoma. PMID:22193299

  19. Emergency preparedness for those who care for infants in developed country contexts

    PubMed Central

    2011-01-01

    Emergency management organisations recognise the vulnerability of infants in emergencies, even in developed countries. However, thus far, those who care for infants have not been provided with detailed information on what emergency preparedness entails. Emergency management authorities should provide those who care for infants with accurate and detailed information on the supplies necessary to care for them in an emergency, distinguishing between the needs of breastfed infants and the needs of formula fed infants. Those who care for formula fed infants should be provided with detailed information on the supplies necessary for an emergency preparedness kit and with information on how to prepare formula feeds in an emergency. An emergency preparedness kit for exclusively breastfed infants should include 100 nappies and 200 nappy wipes. The contents of an emergency preparedness for formula fed infants will vary depending upon whether ready-to-use liquid infant formula or powdered infant formula is used. If ready-to-use liquid infant formula is used, an emergency kit should include: 56 serves of ready-to-use liquid infant formula, 84 L water, storage container, metal knife, small bowl, 56 feeding bottles and teats/cups, 56 zip-lock plastic bags, 220 paper towels, detergent, 120 antiseptic wipes, 100 nappies and 200 nappy wipes. If powdered infant formula is used, an emergency preparedness kit should include: two 900 g tins powdered infant formula, 170 L drinking water, storage container, large cooking pot with lid, kettle, gas stove, box of matches/lighter, 14 kg liquid petroleum gas, measuring container, metal knife, metal tongs, feeding cup, 300 large sheets paper towel, detergent, 100 nappies and 200 nappy wipes. Great care with regards hygiene should be taken in the preparation of formula feeds. Child protection organisations should ensure that foster carers responsible for infants have the resources necessary to formula feed in the event of an emergency. Exclusive and continued breastfeeding should be promoted as an emergency preparedness activity by emergency management organisations as well as health authorities. The greater the proportion of infants exclusively breastfed when an emergency occurs, the more resilient the community, and the easier it will be to provide effective aid to the caregivers of formula fed infants. PMID:22059481

  20. 47 CFR 15.25 - Kits.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 1 2010-10-01 2010-10-01 false Kits. 15.25 Section 15.25 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.25 Kits. A TV interface device, including a cable system terminal device, which is marketed as a kit shall comply with the...

  1. 47 CFR 15.25 - Kits.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 47 Telecommunication 1 2012-10-01 2012-10-01 false Kits. 15.25 Section 15.25 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.25 Kits. A TV interface device, including a cable system terminal device, which is marketed as a kit shall comply with the...

  2. 47 CFR 15.25 - Kits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 1 2011-10-01 2011-10-01 false Kits. 15.25 Section 15.25 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.25 Kits. A TV interface device, including a cable system terminal device, which is marketed as a kit shall comply with the...

  3. 47 CFR 15.25 - Kits.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 1 2013-10-01 2013-10-01 false Kits. 15.25 Section 15.25 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.25 Kits. A TV interface device, including a cable system terminal device, which is marketed as a kit shall comply with the...

  4. 47 CFR 15.25 - Kits.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 1 2014-10-01 2014-10-01 false Kits. 15.25 Section 15.25 Telecommunication FEDERAL COMMUNICATIONS COMMISSION GENERAL RADIO FREQUENCY DEVICES General § 15.25 Kits. A TV interface device, including a cable system terminal device, which is marketed as a kit shall comply with the...

  5. The stem cell factor (SCF)/c-KIT signalling in testis and prostate cancer.

    PubMed

    Cardoso, Henrique J; Figueira, Marília I; Socorro, Sílvia

    2017-12-01

    The stem cell factor (SCF) is a cytokine that specifically binds the tyrosine kinase receptor c-KIT. The SCF/c-KIT interaction leads to receptor dimerization, activation of kinase activity and initiation of several signal transduction pathways that control cell proliferation, apoptosis, differentiation and migration in several tissues. The activity of SCF/c-KIT system is linked with the phosphatidylinositol 3-kinase (PI3-K), the Src, the Janus kinase/signal transducers and activators of transcription (JAK/STAT), the phospholipase-C (PLC-γ) and the mitogen-activated protein kinase (MAPK) pathways. Moreover, it has been reported that cancer cases display an overactivation of c-KIT due to the presence of gain-of-function mutations or receptor overexpression, which renders c-KIT a tempting target for cancer treatment. In the case of male cancers the most documented activated pathways are the PI3-K and Src, both enhancing abnormal cell proliferation. It is also known that the Src activity in prostate cancer cases depends on the presence of tr-KIT, the cytoplasmic truncated variant of c-KIT that is specifically expressed in tumour tissues and, thus, a very interesting target for drug development. The present review provides an overview of the signalling pathways activated by SCF/c-KIT and discusses the potential application of c-KIT inhibitors for treatment of testicular and prostatic cancers.

  6. Major inorganic elements in tap water samples in Peninsular Malaysia.

    PubMed

    Azrina, A; Khoo, H E; Idris, M A; Amin, I; Razman, M R

    2011-08-01

    Quality drinking water should be free from harmful levels of impurities such as heavy metals and other inorganic elements. Samples of tap water collected from 24 locations in Peninsular Malaysia were determined for inorganic element content. Minerals and heavy metals were analysed by spectroscopy methods, while non-metal elements were analysed using test kits. Minerals and heavy metals determined were sodium, magnesium, potassium, calcium, chromium, manganese, iron, nickel, copper, zinc, arsenic, cadmium and lead while the non-metal elements were fluoride, chloride, nitrate and sulphate. Most of the inorganic elements found in the samples were below the maximum permitted levels recommended by inter-national drinking water standard limits, except for iron and manganese. Iron concentration of tap water from one of the locations was higher than the standard limit. In general, tap water from different parts of Peninsular Malaysia had low concentrations of heavy metals and inorganic elements.

  7. KIT Suppresses BRAFV600E-Mutant Melanoma by Attenuating Oncogenic RAS/MAPK Signaling.

    PubMed

    Neiswender, James V; Kortum, Robert L; Bourque, Caitlin; Kasheta, Melissa; Zon, Leonard I; Morrison, Deborah K; Ceol, Craig J

    2017-11-01

    The receptor tyrosine kinase KIT promotes survival and migration of melanocytes during development, and excessive KIT activity hyperactivates the RAS/MAPK pathway and can drive formation of melanomas, most notably of rare melanomas that occur on volar and mucosal surfaces of the skin. The much larger fraction of melanomas that occur on sun-exposed skin is driven primarily by BRAF- or NRAS-activating mutations, but these melanomas exhibit a surprising loss of KIT expression, which raises the question of whether loss of KIT in these tumors facilitates tumorigenesis. To address this question, we introduced a kit(lf) mutation into a strain of Tg(mitfa:BRAF V600E ); p53(lf) melanoma-prone zebrafish. Melanoma onset was accelerated in kit(lf); Tg(mitfa:BRAF V600E ); p53(lf) fish. Tumors from kit(lf) animals were more invasive and had higher RAS/MAPK pathway activation. KIT knockdown also increased RAS/MAPK pathway activation in a BRAF V600E -mutant human melanoma cell line. We found that pathway stimulation upstream of BRAF V600E could paradoxically reduce signaling downstream of BRAF V600E , and wild-type BRAF was necessary for this effect, suggesting that its activation can dampen oncogenic BRAF V600E signaling. In vivo , expression of wild-type BRAF delayed melanoma onset, but only in a kit -dependent manner. Together, these results suggest that KIT can activate signaling through wild-type RAF proteins, thus interfering with oncogenic BRAF V600E -driven melanoma formation. Cancer Res; 77(21); 5820-30. ©2017 AACR . ©2017 American Association for Cancer Research.

  8. c-kit Positive Cardiac Outgrowth Cells Demonstrate Better Ability for Cardiac Recovery Against Ischemic Myopathy.

    PubMed

    Li, Chuan; Matsushita, Satoshi; Li, Zhengqing; Guan, Jianjun; Amano, Atsushi

    2017-10-01

    Resident cardiac stem cells are expected to be a therapeutic option for patients who suffer from severe heart failure. However, uncertainty remains over whether sorting cells for c-kit, a stem cell marker, improves therapeutic outcomes. Cardiac outgrowth cells cultured from explants of rat heart atrium were sorted according to their positivity (+) or negativity (-) for c-kit. These cells were exposed to hypoxia for 3 d, and subsequently harvested for mRNA expression measurement. The cell medium was also collected to assess cytokine secretion. To test for a functional benefit in animals, myocardial infarction (MI) was induced in rats, and c-kit+ or c-kit- cells were injected. The left ventricular ejection fraction (LVEF) was measured for up to 4 weeks, after which the heart was harvested for biological and histological analyses. Expression of the angiogenesis-related genes, VEGF and ANGPTL2, was significantly higher in c-kit+ cells after 3 d of hypoxic culture, although we found no such difference prior to hypoxia. Secretion of VEGF and ANGPTL2 was greater in the c-kit+ group than in the c-kit- group, while hypoxia tended to increase cytokine expression in both groups. In addition, IGF-1 was significantly increased in the c-kit+ group, consistent with the relatively low expression of cleaved-caspase 3 revealed by western blot assay, and the relatively low count of apoptotic cells revealed by histochemical analysis. Administration of c-kit+cells into the MI heart improved the LVEF and increased neovascularization. These results indicate that c-kit+cells may be useful in cardiac stem cell therapy.

  9. Overexpression of c-kit(CD117), relevant with microvessel density, is an independent survival prognostic factor for patients with HBV-related hepatocellular carcinoma.

    PubMed

    Yan, Weiwei; Zhu, Zhenyu; Pan, Fei; Huang, Ang; Dai, Guang-Hai

    2018-01-01

    To explore new biomarkers for indicating the recurrence and prognosis in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) patients after tumor resection, we investigated the expression and prognostic value of c-kit(CD117) in HBV-related HCC. Immunohistochemistry was used to estimate the expression of c-kit(CD117) and CD34 in the liver cancer tissues. The correlations between the expression of these biomarkers and the clinicopathologic characteristics were analyzed. The positive rate of c-kit(CD117) expression in 206 HCC cases was 48.1%, and c-kit expression was significantly related with CD34-positive microvessel density. CD34-microvessel density numbers were much higher in c-kit(+) HCC tissues than in c-kit(-) HCC tissues (44.13±17.01 vs 26.87±13.16, P =0.003). The expression of c-kit was significantly higher in patients with Edmondson grade III-IV ( P <0.001) and TNM stage III ( P <0.001). Moreover, Kaplan-Meier survival analysis showed that c-kit ( P <0.001) expression was correlated with reduced disease-free survival (DFS). Multivariate analysis identified c-kit as an independent poor prognostic factor of DFS in HCC patients ( P <0.001). Increased c-kit expression could be considered as an independent unfavorable prognostic factor for predicting DFS in HBV-related HCC patients after surgery. These results could be used to identify patients at a higher risk of early tumor recurrence and poor prognosis.

  10. Estrogen-induced transcription factor EGR1 regulates c-Kit transcription in the mouse uterus to maintain uterine receptivity for embryo implantation.

    PubMed

    Park, Mira; Kim, Hye-Ryun; Kim, Yeon Sun; Yang, Seung Chel; Yoon, Jung Ah; Lyu, Sang Woo; Lim, Hyunjung Jade; Hong, Seok-Ho; Song, Haengseok

    2018-07-15

    Early growth response 1 (Egr1) is a key transcription factor that mediates the action of estrogen (E 2 ) to establish uterine receptivity for embryo implantation. However, few direct target genes of EGR1 have been identified in the uterus. Here, we demonstrated that E 2 induced EGR1-regulated transcription of c-Kit, which plays a crucial role in cell fate decisions. Spatiotemporal expression of c-Kit followed that of EGR1 in uteri of ovariectomized mice at various time points after E 2 treatment. E 2 activated ERK1/2 and p38 to induce EGR1, which then activated c-Kit expression in the uterus. EGR1 transfection produced rapid and transient induction of c-KIT in a time- and dose-dependent manner. Furthermore, luciferase assays to measure c-Kit promoter activity confirmed that a functional EGR1 binding site(s) (EBS) was located within -1 kb of the c-Kit promoter. Site-directed mutagenesis and chromatin immunoprecipitation-PCR for three putative EBS within -1 kb demonstrated that the EBS at -818/-805 was critical for EGR1-dependent c-Kit transcription. c-Kit expression was significantly increased in the uterus on day 4 and administration of Masitinib, a c-Kit inhibitor, effectively interfered with embryo implantation. Collectively, our results showed that estrogen induces transcription factor EGR1 to regulate c-Kit transcription for uterine receptivity for embryo implantation in the mouse uterus. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Elevated expression of the proto-oncogene c-kit in patients with mastocytosis.

    PubMed

    Nagata, H; Worobec, A S; Semere, T; Metcalfe, D D

    1998-02-01

    The stem cell factor (SCF)c-kit receptor interaction plays a critical role in the development and survival of mast cells. Several studies have also associated c-kit receptor mutations with the human diseases, mastocytosis and piebaldism. Overexpression of c-kit has been reported to be associated with myeloproliferative disorders and myelodysplastic syndromes. Using peripheral blood mononuclear cells (PBMCs) from 11 patients with indolent mastocytosis (category I), mastocytosis with an associated hematologic disorder (category II), or aggressive mastocytosis (category III); a patient with CMML unassociated with mastocytosis, and PBMCs from 13 normal subjects, we examined the level of expression of c-kit mRNA along with other c-kit isoforms to determine if overexpression of the c-kit receptor was associated with mastocytosis. Using quantitative competitive PCR, c-kit mRNA levels on average were found to be statistically elevated in the five patients with mastocytosis with an associated hematologic disorder and in the patient with aggressive mastocytosis as compared with controls, but not elevated in patients with indolent mastocytosis. The relative mRNA expression for the two c-kit isoforms was not significantly different in the mastocytosis patients compared with controls. This demonstration of the overexpression of c-kit mRNA in mastocytosis, and particularly those patients with clinical evidence of myelodysplastic syndrome, adds evidence to support the conclusion that mastocytosis, at least in some patients, is a feature of myelodysplasia; and suggests that determination of c-kit mRNA expression in PBMCs may provide an additional approach to assessing prognosis.

  12. 49 CFR 173.165 - Polyester resin kits.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 2 2011-10-01 2011-10-01 false Polyester resin kits. 173.165 Section 173.165... Polyester resin kits. (a) Except for transportation by aircraft, polyester resin kits consisting of a base... will not interact dangerously in the event of leakage. (b) For transportation by aircraft, polyester...

  13. YourSELF. Middle School Nutrition Education Kit [Multimedia].

    ERIC Educational Resources Information Center

    Department of Agriculture, Washington, DC.

    This multimedia kit provides information and materials for teaching nutrition to middle school students (grades 7 and 8). The kit supports schools' efforts to make school meals healthier and more appealing to students. The materials provide information about the relationships between food, nutrition, growth, and health. The kit speaks directly to…

  14. 21 CFR 868.5140 - Anesthesia conduction kit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Anesthesia conduction kit. 868.5140 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Therapeutic Devices § 868.5140 Anesthesia conduction kit. (a) Identification. An anesthesia conduction kit is a device used to administer to a patient conduction, regional, or...

  15. 21 CFR 868.5140 - Anesthesia conduction kit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Anesthesia conduction kit. 868.5140 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Therapeutic Devices § 868.5140 Anesthesia conduction kit. (a) Identification. An anesthesia conduction kit is a device used to administer to a patient conduction, regional, or...

  16. 40 CFR 745.88 - Recognized test kits.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 30 2010-07-01 2010-07-01 false Recognized test kits. 745.88 Section... Renovation § 745.88 Recognized test kits. (a) Effective June 23, 2008, EPA recognizes the test kits that have been determined by National Institute of Standards and Technology research to meet the negative...

  17. 40 CFR 745.88 - Recognized test kits.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 40 Protection of Environment 31 2011-07-01 2011-07-01 false Recognized test kits. 745.88 Section... Renovation § 745.88 Recognized test kits. (a) Effective June 23, 2008, EPA recognizes the test kits that have been determined by National Institute of Standards and Technology research to meet the negative...

  18. 40 CFR 745.88 - Recognized test kits.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 40 Protection of Environment 32 2013-07-01 2013-07-01 false Recognized test kits. 745.88 Section... Renovation § 745.88 Recognized test kits. (a) Effective June 23, 2008, EPA recognizes the test kits that have been determined by National Institute of Standards and Technology research to meet the negative...

  19. 40 CFR 745.88 - Recognized test kits.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 31 2014-07-01 2014-07-01 false Recognized test kits. 745.88 Section... Renovation § 745.88 Recognized test kits. (a) Effective June 23, 2008, EPA recognizes the test kits that have been determined by National Institute of Standards and Technology research to meet the negative...

  20. 46 CFR 160.041-1 - Applicable specification and publication.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ..., CONSTRUCTION, AND MATERIALS: SPECIFICATIONS AND APPROVAL LIFESAVING EQUIPMENT Kits, First-Aid, for Merchant... specification, of the issue in effect on the date first-aid kits are manufactured, forms a part of this subpart: (1) Federal specification: GG-K-391, Kits (Empty), First Aid, Burn Treatment, and Snake Bite; and Kit...

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