Sample records for sample collection storage

  1. NHEXAS PHASE I ARIZONA STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE AND SHIPMENT OF URINE SAMPLES FOR SELECTED METALS AND PESTICIDES (UA-F-20.1)

    EPA Science Inventory

    The purpose of this SOP is to guide the collection, storage, and shipment of urine samples collected for the NHEXAS Arizona project. This SOP provides a brief description of sample, collection, preservation, storage, shipping, and custody procedures. This procedure was followed ...

  2. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF URINE SAMPLES FOR METALS AND PESTICIDES ANALYSIS (UA-F-20.1)

    EPA Science Inventory

    The purpose of this SOP is to guide the collection, storage, and shipment of urine samples collected. This SOP provides a brief description of sample, collection, preservation, storage, shipping, and custody procedures. This procedure was followed to ensure consistent data retri...

  3. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF BLOOD SAMPLES FOR SELECTED METALS, PESTICIDES, AND VOCS (UA-F-19.1)

    EPA Science Inventory

    The purpose of this SOP is to guide the collection, storage, and shipment of blood samples collected. This SOP provides a brief description of sample collection, preservation, storage, and custody procedures. This procedure was followed to ensure consistent data retrieval during...

  4. NHEXAS PHASE I ARIZONA STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE AND SHIPMENT OF BLOOD SAMPLES FOR SELECTED METALS AND VOCS (UA-F-19.1)

    EPA Science Inventory

    The purpose of this SOP is to guide the collection, storage, and shipment of blood samples collected for the NHEXAS Arizona project. This SOP provides a brief description of sample collection, preservation, storage, and custody procedures. This procedure was followed to ensure c...

  5. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR VOCS (EPA METHOD 524.2) ANALYSIS (UA-F-18.1)

    EPA Science Inventory

    The purpose of this SOP is to describe the collection, storage, and shipment of tap and drinking water samples for analysis by EPA method 524.2 (revision 4.0). This SOP provides a brief description of the sample containers, collection, preservation, storage, shipping, and custod...

  6. NHEXAS PHASE I REGION 5 STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF HAIR SAMPLES FOR TRACE METALS AND ARSENIC (RTI/ACS-209-046)

    EPA Science Inventory

    This protocol describes the procedures for the collection, storage, and shipping of human scalp hair samples for trace metals and arsenic or potential adduct analysis. Scalp hair samples were collected from each participant that agreed to provide the sample. Thinning shears were ...

  7. 40 CFR 761.283 - Determination of the number of samples to collect and sample collection locations.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... sites at this example location: a loading dock, a transformer storage lot, and a disposal pit. The... (three samples). The non-liquid PCB remediation wastes present at the transformer storage lot are oily...

  8. 40 CFR 761.283 - Determination of the number of samples to collect and sample collection locations.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... sites at this example location: a loading dock, a transformer storage lot, and a disposal pit. The... (three samples). The non-liquid PCB remediation wastes present at the transformer storage lot are oily...

  9. Precipitation chemistry affected by differences in location of collection sites and storage methods

    NASA Astrophysics Data System (ADS)

    Mahendrappa, M. K.

    An investigation was carried out to evaluate rigorously the possible differences in measured concentrations of hydrogen, nitrate and sulfate ions in 'bulk' precipitation samples that may be caused by variations in location of rain collectors, and duration and temperature of storage. Storage of precipitation samples, up to 1 month, both in the coldroom and in the field resulted in a significant reduction in the concentration of hydrogen ions. Only field storage caused a statistically significant reduction in the concentration of nitrate in the precipitation samples. Levels of sulfate ions were not found to be significantly affected by storage either in the field or in coldrooms. Samples collected from a rain gage located on a building roof were more acidic than those collected in open spaces in forests. Though all samples showed similar seasonal patterns in the concentrations of sulfate and nitrate, the individual values consistently differed from each other.

  10. NHEXAS PHASE I MARYLAND STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF URINE SAMPLES FOR METAL, PESTICIDE, AND CREATININE ANALYSIS (F10)

    EPA Science Inventory

    The purpose of this SOP is to describe the procedures for collection, storage, and shipment of urine samples for metal, pesticides, and creatinine analysis. Samples were collected on Days 2 and 8 of each Cycle. The Day 2 sample was analyzed for metals and creatinine. The Day 8...

  11. NHEXAS PHASE I ARIZONA STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR VOCS (EPA METHOD 524.2) (UA-F-18.1)

    EPA Science Inventory

    The purpose of this SOP is to describe the collection, storage, and shipment of tap and drinking water samples for analysis by EPA method 524.2 (revision 4.0) for the NHEXAS Arizona project. This SOP provides a brief description of the sample containers, collection, preservation...

  12. Evaluation of sampling and storage procedures on preserving the community structure of stool microbiota: A simple at-home toilet-paper collection method.

    PubMed

    Al, Kait F; Bisanz, Jordan E; Gloor, Gregory B; Reid, Gregor; Burton, Jeremy P

    2018-01-01

    The increasing interest on the impact of the gut microbiota on health and disease has resulted in multiple human microbiome-related studies emerging. However, multiple sampling methods are being used, making cross-comparison of results difficult. To avoid additional clinic visits and increase patient recruitment to these studies, there is the potential to utilize at-home stool sampling. The aim of this pilot study was to compare simple self-sampling collection and storage methods. To simulate storage conditions, stool samples from three volunteers were freshly collected, placed on toilet tissue, and stored at four temperatures (-80, 7, 22 and 37°C), either dry or in the presence of a stabilization agent (RNAlater®) for 3 or 7days. Using 16S rRNA gene sequencing by Illumina, the effect of storage variations for each sample was compared to a reference community from fresh, unstored counterparts. Fastq files may be accessed in the NCBI Sequence Read Archive: Bioproject ID PRJNA418287. Microbial diversity and composition were not significantly altered by any storage method. Samples were always separable based on participant, regardless of storage method suggesting there was no need for sample preservation by a stabilization agent. In summary, if immediate sample processing is not feasible, short term storage of unpreserved stool samples on toilet paper offers a reliable way to assess the microbiota composition by 16S rRNA gene sequencing. Copyright © 2017 Elsevier B.V. All rights reserved.

  13. NHEXAS PHASE I ARIZONA STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR TRACE METALS (EPA METHOD 200.8) (UA-F-16.1)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship tap and drinking water samples for analysis by EPA Method 200.8 (revision 4.4) for the NHEXAS Arizona project. This SOP provides a brief description of the sample containers, collection, preservation, storage...

  14. Collecting, archiving and processing DNA from wildlife samples using FTA® databasing paper

    PubMed Central

    Smith, LM; Burgoyne, LA

    2004-01-01

    Background Methods involving the analysis of nucleic acids have become widespread in the fields of traditional biology and ecology, however the storage and transport of samples collected in the field to the laboratory in such a manner to allow purification of intact nucleic acids can prove problematical. Results FTA® databasing paper is widely used in human forensic analysis for the storage of biological samples and for purification of nucleic acids. The possible uses of FTA® databasing paper in the purification of DNA from samples of wildlife origin were examined, with particular reference to problems expected due to the nature of samples of wildlife origin. The processing of blood and tissue samples, the possibility of excess DNA in blood samples due to nucleated erythrocytes, and the analysis of degraded samples were all examined, as was the question of long term storage of blood samples on FTA® paper. Examples of the end use of the purified DNA are given for all protocols and the rationale behind the processing procedures is also explained to allow the end user to adjust the protocols as required. Conclusions FTA® paper is eminently suitable for collection of, and purification of nucleic acids from, biological samples from a wide range of wildlife species. This technology makes the collection and storage of such samples much simpler. PMID:15072582

  15. Archival policies and collections database for the Woods Hole Science Center's marine sediment samples

    USGS Publications Warehouse

    Buczkowski, Brian J.; Kelsey, Sarah A.

    2007-01-01

    The Woods Hole Science Center of the U.S. Geological Survey (USGS) has been an active member of the Woods Hole research community, Woods Hole, Massachusetts, for over 40 years. In that time there have been many projects that involved the collection of sediment samples conducted by USGS scientists and technicians for the research and study of seabed environments and processes. These samples were collected at sea or near shore and then brought back to the Woods Hole Science Center (WHSC) for analysis. While at the center, samples are stored in ambient temperature, refrigerated and freezing conditions ranging from +2º Celsius to -18º Celsius, depending on the best mode of preparation for the study being conducted or the duration of storage planned for the samples. Recently, storage methods and available storage space have become a major concern at the WHSC. The core and sediment archive program described herein has been initiated to set standards for the management, methods, and duration of sample storage. A need has arisen to maintain organizational consistency and define storage protocol. This handbook serves as a reference and guide to all parties interested in using and accessing the WHSC's sample archive and also defines all the steps necessary to construct and maintain an organized collection of geological samples. It answers many questions as to the way in which the archive functions.

  16. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR TRACE METALS (EPA METHOD 200.8) ANALYSIS (UA-F-16.1)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship tap and drinking water samples for analysis by EPA Method 200.8 (revision 4.4) for the NHEXAS Arizona project. This SOP provides a brief description of the sample containers, collection, preservation, storage...

  17. 40 CFR 80.1653 - Recordkeeping.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... this subpart O: (i) The location, date, time, and storage tank or truck identification for each sample... analytical testing: (i) The location, date, time, and storage tank or truck identification for each sample..., time, and storage tank or truck identification for each sample collected. (B) The name and title of the...

  18. NHEXAS PHASE I MARYLAND STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF BLOOD SAMPLES FOR METAL, PESTICIDE, PAH, VOC, AND LIPID ANALYSIS (F11)

    EPA Science Inventory

    The purpose of this SOP is to describe collection, storage, and shipment requirements of blood samples for metal, pesticide, polynuclear aromatic hydrocarbons (PAHs), volatile organic compound (VOC), and lipid analysis. Seven samples were taken from a single puncture: two 3-mL t...

  19. NHEXAS PHASE I MARYLAND STUDY--STANDARD OPERATING PROCEDURE FOR STORAGE AND SHIPPING OF SAMPLES (G05)

    EPA Science Inventory

    The purpose of this SOP is to describe the procedures for storage and shipping of samples. For the NHEXAS project, Harvard School of Public Health/Emory University will collect many different kinds of samples, some of which will require particularly careful storage and shipping ...

  20. Beverage Cans Used for Sediment Collection.

    ERIC Educational Resources Information Center

    Studlick, Joseph R. J.; Trautman, Timothy A.

    1979-01-01

    Beverage cans are well suited for sediment collection and storage containers. Advantages include being free, readily available, and the correct size for many samples. Instruction for selection, preparation, and use of cans in sediment collection and storage is provided. (RE)

  1. Analysis of Samples Collected from the Surface of Interim Storage Canisters at Calvert Cliffs in June 2017: Revision 01.

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

    Bryan, Charles R.; Schindelholz, Eric John

    In June 2017, dust and salt samples were collected from the surface of Spent Nuclear Fuel (SNF) dry storage canisters at the Calvert Cliffs Nuclear Power Plant. The samples were delivered to Sandia National laboratories for analysis. Two types of samples were collected: filter-backed Scotch-Brite TM pads were used to collect dry dust samples for characterization of salt and dust morphologies and distributions; and Saltsmart TM test strips were used to collect soluble salts for determining salt surface loadings per unit area. After collection, the samples were sealed into plastic sleeves for shipping. Condensation within the sleeves containing the Scotch-Britemore » TM samples remobilized the salts, rendering them ineffective for the intended purpose, and also led to mold growth, further compromising the samples; for these reasons, the samples were not analyzed. The SaltSmart TM samples were unaffected and were analyzed by ion chromatography for major anions and cations. The results of those analyses are presented here.« less

  2. Gas chromatographic column for the storage of sample profiles

    NASA Technical Reports Server (NTRS)

    Dimandja, J. M.; Valentin, J. R.; Phillips, J. B.

    1994-01-01

    The concept of a sample retention column that preserves the true time profile of an analyte of interest is studied. This storage system allows for the detection to be done at convenient times, as opposed to the nearly continuous monitoring that is required by other systems to preserve a sample time profile. The sample storage column is essentially a gas chromatography column, although its use is not the separation of sample components. The functions of the storage column are the selective isolation of the component of interest from the rest of the components present in the sample and the storage of this component as a function of time. Using octane as a test substance, the sample storage system was optimized with respect to such parameters as storage and readout temperature, flow rate through the storage column, column efficiency and storage time. A 3-h sample profile was collected and stored at 30 degrees C for 20 h. The profile was then retrieved, essentially intact, in 5 min at 130 degrees C.

  3. Comparative study of modified bitumen binder properties collected from mixing plant and quarry.

    NASA Astrophysics Data System (ADS)

    Mustafa Kamal, M.; Abu Bakar, R.; Hadithon, K. A.

    2017-11-01

    Quality control and assurance are essential in pavement construction. In general, the properties of bitumen change as it ages in bulk storage, transport, and storage on site. The minimization of bituminous hardening during storing, transportation and mixing depends on careful control of binder temperature. Hence therefore, bitumen should always be stored and handled at the lowest temperature possible, consistent with efficient use. The objective of the work is to monitor the quality of bitumen samples collected from mixing plant and quarry. Results showed that, samples modified bitumen which collected from quarry showed some adverse effects on rheological properties and physical properties after subjecting to high temperature storage within a period of time. The dynamic stiffness, elastic properties and other common binder properties were deteriorated too. The chemical changes that occurred during storage were analysed using Fourier transform infra-red spectroscopy (FTIR). Thus studies developed an understanding of bitumen ageing in storage.

  4. Effects of Specimen Collection Methodologies and Storage Conditions on the Short-Term Stability of Oral Microbiome Taxonomy.

    PubMed

    Luo, Ting; Srinivasan, Usha; Ramadugu, Kirtana; Shedden, Kerby A; Neiswanger, Katherine; Trumble, Erika; Li, Jiean J; McNeil, Daniel W; Crout, Richard J; Weyant, Robert J; Marazita, Mary L; Foxman, Betsy

    2016-09-15

    Community profiling of the oral microbiome requires the recovery of quality sequences in order to accurately describe microbial community structure and composition. Our objective was to assess the effects of specimen collection method, storage medium, and storage conditions on the relative abundance of taxa in saliva and plaque identified using 16S rRNA genes. We also assessed short-term changes in taxon composition and relative abundance and compared the salivary and dental plaque communities in children and adults. Over a 2-week period, four successive saliva and dental plaque specimens were collected from four adults with no dental decay (108 samples), and two successive specimens were collected from six children with four or more erupted teeth (48 samples). There were minimal differences in community composition at the phylum and operational taxonomic unit levels between dental plaque collection using a scaler and collection using a CytoSoft brush. Plaque samples stored in OMNIgene medium showed higher within-sample Shannon diversity, were compositionally different, and were more similar to each other than plaque stored in liquid dental transport medium. Saliva samples stored in OMNIgene recovered similar communities for at least a week following storage at room temperature. However, the microbial communities recovered from plaque and saliva stored in OMNIgene were significantly different in composition from their counterparts stored in liquid dental transport medium. Dental plaque communities collected from the same tooth type over four successive visits from the same adult did not significantly differ in structure or composition. Large-scale epidemiologic studies require collection over time and space, often with multiple teams collecting, storing, and processing data. Therefore, it is essential to understand how sensitive study results are to modest changes in collection and storage protocols that may occur with variation in personnel, resources available at a study site, and shipping requirements. The research presented in this paper measures the effects of multiple storage parameters and collection methodologies on the measured ecology of the oral microbiome from healthy adults and children. These results will potentially enable investigators to conduct oral microbiome studies at maximal efficiency by guiding informed administrative decisions pertaining to the necessary field or clinical work. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  5. Effects of Specimen Collection Methodologies and Storage Conditions on the Short-Term Stability of Oral Microbiome Taxonomy

    PubMed Central

    Luo, Ting; Srinivasan, Usha; Ramadugu, Kirtana; Shedden, Kerby A.; Neiswanger, Katherine; Trumble, Erika; Li, Jiean J.; McNeil, Daniel W.; Crout, Richard J.; Weyant, Robert J.; Marazita, Mary L.

    2016-01-01

    ABSTRACT Community profiling of the oral microbiome requires the recovery of quality sequences in order to accurately describe microbial community structure and composition. Our objective was to assess the effects of specimen collection method, storage medium, and storage conditions on the relative abundance of taxa in saliva and plaque identified using 16S rRNA genes. We also assessed short-term changes in taxon composition and relative abundance and compared the salivary and dental plaque communities in children and adults. Over a 2-week period, four successive saliva and dental plaque specimens were collected from four adults with no dental decay (108 samples), and two successive specimens were collected from six children with four or more erupted teeth (48 samples). There were minimal differences in community composition at the phylum and operational taxonomic unit levels between dental plaque collection using a scaler and collection using a CytoSoft brush. Plaque samples stored in OMNIgene medium showed higher within-sample Shannon diversity, were compositionally different, and were more similar to each other than plaque stored in liquid dental transport medium. Saliva samples stored in OMNIgene recovered similar communities for at least a week following storage at room temperature. However, the microbial communities recovered from plaque and saliva stored in OMNIgene were significantly different in composition from their counterparts stored in liquid dental transport medium. Dental plaque communities collected from the same tooth type over four successive visits from the same adult did not significantly differ in structure or composition. IMPORTANCE Large-scale epidemiologic studies require collection over time and space, often with multiple teams collecting, storing, and processing data. Therefore, it is essential to understand how sensitive study results are to modest changes in collection and storage protocols that may occur with variation in personnel, resources available at a study site, and shipping requirements. The research presented in this paper measures the effects of multiple storage parameters and collection methodologies on the measured ecology of the oral microbiome from healthy adults and children. These results will potentially enable investigators to conduct oral microbiome studies at maximal efficiency by guiding informed administrative decisions pertaining to the necessary field or clinical work. PMID:27371581

  6. Sample handling for mass spectrometric proteomic investigations of human urine.

    PubMed

    Petri, Anette Lykke; Høgdall, Claus; Christensen, Ib Jarle; Simonsen, Anja Hviid; T'jampens, Davy; Hellmann, Marja-Leena; Kjaer, Susanne Krüger; Fung, Eric T; Høgdall, Estrid

    2008-09-01

    Because of its non-invasive sample collection method, human urine is an attractive biological material both for discovering biomarkers and for use in future screening trials for different diseases. Before urine can be used for these applications, standardized protocols for sample handling that optimize protein stability are required. In this explorative study, we examine the influence of different urine collection methods, storage temperatures, storage times, and repetitive freeze-thaw procedures on the protein profiles obtained by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Prospectively collected urine samples from 11 women were collected as either morning or midday specimens. The effects of storage temperature, time to freezing, and freeze-thaw cycles were assessed by calculating the number, intensity, and reproducibility of peaks visualized by SELDI-TOF-MS. On the CM10 array, 122 peaks were detected and 28 peaks were found to be significantly different between urine types, storage temperature and time to freezing. On the IMAC-Cu array, 65 peaks were detected and 1 peak was found to be significantly different according to time to freezing. No significant differences were demonstrated for freeze-thaw cycles. Optimal handling and storage conditions are necessary in clinical urine proteomic investigations. Collection of urine with a single and consistently performed protocol is needed to reduce analytical bias. Collecting only one urine type, which is stored for a limited period at 4°C until freezing at -80°C prior to analysis will provide the most stable profiles. Copyright © 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. Data on subsurface storage of liquid waste near Pensacola, Florida, 1963-1980

    USGS Publications Warehouse

    Hull, R.W.; Martin, J.B.

    1982-01-01

    Since 1963, when industrial waste was first injected into the subsurface in northwest Florida, considerable data have been collected relating to the geochemistry of subsurface waste storage. This report presents hydrogeologic data on two subsurface waste storage. This report presents hydrogeologic data on two subsurface storage systems near Pensacola, Fla., which inject liquid industrial waste through deep wells into a saline aquifer. Injection sites are described giving a history of well construction, injection, and testing; geologic data from cores and grab samples; hydrographs of injection rates, volume, pressure, and water levels; and chemical and physical data from water-quality samples collected from injection and monitor wells. (USGS)

  8. Impact of urine preservation methods and duration of storage on measured levels of environmental contaminants.

    PubMed

    Hoppin, Jane A; Ulmer, Ross; Calafat, Antonia M; Barr, Dana B; Baker, Susan V; Meltzer, Helle M; Rønningen, Kjersti S

    2006-01-01

    Collection of urine samples in human studies involves choices regarding shipping, sample preservation, and storage that may ultimately influence future analysis. As more studies collect and archive urine samples to evaluate environmental exposures in the future, we were interested in assessing the impact of urine preservative, storage temperature, and time since collection on nonpersistent contaminants in urine samples. In spiked urine samples stored in three types of urine vacutainers (no preservative, boric acid, and chlorhexidine), we measured five groups of contaminants to assess the levels of these analytes at five time points (0, 24, 48, and 72 h, and 1 week) and at two temperatures (room temperature and 4 degrees C). The target chemicals were bisphenol A (BPA), metabolites of organophosphate (OP), carbamate, and pyrethroid insecticides, chlorinated phenols, and phthalate monoesters, and were measured using five different mass spectrometry-based methods. Three samples were analyzed at each time point, with the exception of BPA. Repeated measures analysis of variance was used to evaluate effects of storage time, temperature, and preservative. Stability was summarized with percent change in mean concentration from time 0. In general, most analytes were stable under all conditions with changes in mean concentration over time, temperature, and preservative being generally less than 20%, with the exception of the OP metabolites in the presence of boric acid. The effect of storage temperature was less important than time since collection. The precision of the laboratory measurements was high allowing us to observe small differences, which may not be important when categorizing individuals into broader exposure groups.

  9. Evaluation of storage and filtration protocols for alpine/subalpine lake water quality samples

    Treesearch

    John L. Korfmacher; Robert C. Musselman

    2007-01-01

    Many government agencies and other organizations sample natural alpine and subalpine surface waters using varying protocols for sample storage and filtration. Simplification of protocols would be beneficial if it could be shown that sample quality is unaffected. In this study, samples collected from low ionic strength waters in alpine and subalpine lake inlets...

  10. The UK Biobank sample handling and storage protocol for the collection, processing and archiving of human blood and urine.

    PubMed

    Elliott, Paul; Peakman, Tim C

    2008-04-01

    UK Biobank is a large prospective study in the UK to investigate the role of genetic factors, environmental exposures and lifestyle in the causes of major diseases of late and middle age. Extensive data and biological samples are being collected from 500,000 participants aged between 40 and 69 years. The biological samples that are collected and how they are processed and stored will have a major impact on the future scientific usefulness of the UK Biobank resource. The aim of the UK Biobank sample handling and storage protocol is to specify methods for the collection and storage of participant samples that give maximum scientific return within the available budget. Processing or storage methods that, as far as can be predicted, will preclude current or future assays have been avoided. The protocol was developed through a review of the literature on sample handling and processing, wide consultation within the academic community and peer review. Protocol development addressed which samples should be collected, how and when they should be processed and how the processed samples should be stored to ensure their long-term integrity. The recommended protocol was extensively tested in a series of validation studies. UK Biobank collects about 45 ml blood and 9 ml of urine with minimal local processing from each participant using the vacutainer system. A variety of preservatives, anti-coagulants and clot accelerators is used appropriate to the expected end use of the samples. Collection of other material (hair, nails, saliva and faeces) was also considered but rejected for the full cohort. Blood and urine samples from participants are transported overnight by commercial courier to a central laboratory where they are processed and aliquots of urine, plasma, serum, white cells and red cells stored in ultra-low temperature archives. Aliquots of whole blood are also stored for potential future production of immortalized cell lines. A standard panel of haematology assays is completed on whole blood from all participants, since such assays need to be conducted on fresh samples (whereas other assays can be done on stored samples). By the end of the recruitment phase, 15 million sample aliquots will be stored in two geographically separate archives: 9.5 million in a -80 degrees C automated archive and 5.5 million in a manual liquid nitrogen archive at -180 degrees C. Because of the size of the study and the numbers of samples obtained from participants, the protocol stipulates a highly automated approach for the processing and storage of samples. Implementation of the processes, technology, systems and facilities has followed best practices used in manufacturing industry to reduce project risk and to build in quality and robustness. The data produced from sample collection, processing and storage are highly complex and are managed by a commercially available LIMS system fully integrated with the entire process. The sample handling and storage protocol adopted by UK Biobank provides quality assured and validated methods that are feasible within the available funding and reflect the size and aims of the project. Experience from recruiting and processing the first 40,000 participants to the study demonstrates that the adopted methods and technologies are fit-for-purpose and robust.

  11. The effects of water sample treatment, preparation, and storage prior to cyanotoxin analysis for cylindrospermopsin, microcystin and saxitoxin.

    PubMed

    Kamp, Lisa; Church, Jennifer L; Carpino, Justin; Faltin-Mara, Erin; Rubio, Fernando

    2016-02-25

    Cyanobacterial harmful algal blooms occur in freshwater lakes, ponds, rivers, and reservoirs, and in brackish waters throughout the world. The wide variety of cyanotoxins and their congeners can lead to frequent exposure of humans through consumption of meat, fish, seafood, blue-green algal products and water, accidental ingestion of contaminated water and cyanobacterial scum during recreational activities, and inhalation of cyanobacterial aerosols. Cyanotoxins can also occur in the drinking water supply. In order to monitor human exposure, sensitive analytical methods such as enzyme linked immunosorbent assay and liquid chromatography-mass spectrometry are often used. Regardless of the analytical method of choice, some problems regularly occur during sample collection, treatment, storage, and preparation which cause toxin loss and therefore underestimation of the true concentration. To evaluate the potential influence of sample treatment, storage and preparation materials on surface and drinking water samples, the effects of different types of materials on toxin recovery were compared. Collection and storage materials included glass and various types of plastics. It was found that microcystin congeners LA and LF adsorbed to polystyrene, polypropylene, high density polyethylene and polycarbonate storage containers, leading to low recoveries (<70%), cylindrospermopsin and saxitoxin did not adsorb to the containers tested. Therefore, this study shows that glass or polyethylene terephthalate glycol containers are the materials of choice for collection and storage of samples containing the cyanotoxins cylindrospermopsin, microcystins, and saxitoxin. This study also demonstrated that after 15 min chlorine decreased the concentration of microcystin LR to <40%, microcystin LA and saxitoxin to <15%, therefore quenching of drinking water samples immediately upon sample collection is critical for accurate analysis. In addition, the effect of various drinking water treatment chemicals on toxin recovery and the behavior of those chemicals in the enzyme linked immunosorbent assays were also studied and are summarized. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  12. Effects of storage time and temperature on pH, specific gravity, and crystal formation in urine samples from dogs and cats.

    PubMed

    Albasan, Hasan; Lulich, Jody P; Osborne, Carl A; Lekcharoensuk, Chalermpol; Ulrich, Lisa K; Carpenter, Kathleen A

    2003-01-15

    To determine effects of storage temperature and time on pH and specific gravity of and number and size of crystals in urine samples from dogs and cats. Randomized complete block design. 31 dogs and 8 cats. Aliquots of each urine sample were analyzed within 60 minutes of collection or after storage at room or refrigeration temperatures (20 vs 6 degrees C [68 vs 43 degrees F]) for 6 or 24 hours. Crystals formed in samples from 11 of 39 (28%) animals. Calcium oxalate (CaOx) crystals formed in vitro in samples from 1 cat and 8 dogs. Magnesium ammonium phosphate (MAP) crystals formed in vitro in samples from 2 dogs. Compared with aliquots stored at room temperature, refrigeration increased the number and size of crystals that formed in vitro; however, the increase in number and size of MAP crystals in stored urine samples was not significant. Increased storage time and decreased storage temperature were associated with a significant increase in number of CaOx crystals formed. Greater numbers of crystals formed in urine aliquots stored for 24 hours than in aliquots stored for 6 hours. Storage time and temperature did not have a significant effect on pH or specific gravity. Urine samples should be analyzed within 60 minutes of collection to minimize temperature- and time-dependent effects on in vitro crystal formation. Presence of crystals observed in stored samples should be validated by reevaluation of fresh urine.

  13. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF HUMAN HAIR (UA-F-26.0)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship human hair for analysis of mercury. This SOP provides a brief description of the sample containers, collection, storage, shipping and custody procedures. This procedure was followed to ensure consistent data ...

  14. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF UNKNOWN SUBSTANCES (UA-F-25.0)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship unknown substances for chemical analysis and identification. This SOP provides a brief description of the sample containers, collection, storage, shipping and custody procedures. This procedure was followed ...

  15. Analysis of Dust Samples Collected from an In-Service Interim Storage System at the Maine Yankee Nuclear Site.

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

    Bryan, Charles R.; Enos, David

    In July, 2016, the Electric Power Research Institute and industry partners performed a field test at the Maine Yankee Nuclear Site, located near Wiscasset, Maine. The primary goal of the field test was to evaluate the use of robots in surveying the surface of an in-service interim storage canister within an overpack; however, as part of the demonstration, dust and soluble salt samples were collected from horizontal surfaces within the interim storage system. The storage system is a vertical system made by NAC International, consisting of a steel-lined concrete overpack containing a 304 stainless steel (SS) welded storage canister. Themore » canister did not contain spent fuel but rather greater-than-class-C waste, which did not generate significant heat, limiting airflow through the storage system. The surfaces that were sampled for deposits included the top of the shield plug, the side of the canister, and a shelf at the bottom of the overpack, just below the level of the pillar on which the canister sits. The samples were sent to Sandia National Laboratories for analysis. This report summarizes the results of those analyses. Because the primary goal of the field test was to evaluate the use of robots in surveying the surface of the canister within the overpack, collection of dust samples was carried out in a qualitative fashion, using paper filters and sponges as the sampling media. The sampling focused mostly on determining the composition of soluble salts present in the dust. It was anticipated that a wet substrate would more effectively extract soluble salts from the surface that was sampled, so both the sponges and the filter paper were wetted prior to being applied to the surface of the metal. Sampling was accomplished by simply pressing the damp substrate against the metal surface for two minutes, and then removing it. It is unlikely that the sampling method quantitatively collected dust or salts from the metal surface; however, both substrates did extract a significant amount of material. The paper filters collected both particles, trapped within the cellulose fibers of the filter, and salts, while the sponges collected only the soluble salts, with very few particles. Upon delivery to Sandia, both collection media were analyzed using the same methods. The soluble salts were leached from the substrates and analyzed via ion chromatography, and insoluble minerals were analyzed by scanning electron microscopy and energy dispersive X-ray spectroscopy. The insoluble minerals were found to consist largely of terrestrially-derived mineral fragments, dominantly quartz and biotite. Large (mm-sized) aggregates of calcium carbonate, calcium silicate, and calcium aluminum silicate were also present. The aggregates had one flat, smooth surface and one well crystallized surface, and were interpreted to be efflorescence on the inside of the overpack and in the vent, formed by seepage of cement pore fluids through joints in the steel liner of the overpack. Such efflorescence was commonly observed during the boroscope inspection of the storage system at the site. The material may have flaked off and fallen to the point where the dust was collected, or may have brushed off onto the sponges when the robot was retrieved through the inlet vent. Chemical analysis showed that the soluble salts on the shield plug were Ca- and Na-rich, with lesser K and minor Mg; the anionic component was dominated by SO 4 and Cl, with minor amounts of NO 3 . The cation composition of the soluble salts from the overpack shelf and the canister surface was similar to the filter samples, but the anions differed significantly, being dominantly NO 3 with lesser Cl and only trace SO 4 . The salts appear to represent a mixture of sea-salts (probably partially converted to nitrates and sulfates by particle-gas conversion reactions) and continental salt aerosols. Ammonium, a common component in continental aerosols, was not observed and may have been lost by degassing from the canister surface or after collection during sample storage and transportation. The demonstration at Maine Yankee has shown that the robot and sampling method used for the test can successfully be used to collect soluble salts from metal surfaces within an interim storage system overpack. The results were consistent from sample to sample, suggesting that a representative sample of the soluble salts was being collected. However, it is unlikely that the salt samples collected here represent quantitative sampling of the salts on the surfaces evaluated; for that reason, chloride densities per unit area are not presented here. It should also be noted that the relevance to storage systems at the site that contain SNF may be limited, because a heat- generating canister will result in greater airflow through the overpack, affecting dust deposition rates and possibly salt compositions.« less

  16. Natural biopolymer for preservation of microorganisms during sampling and storage.

    PubMed

    Sorokulova, Iryna; Watt, James; Olsen, Eric; Globa, Ludmila; Moore, Timothy; Barbaree, James; Vodyanoy, Vitaly

    2012-01-01

    Stability of microbial cultures during sampling and storage is a vital issue in various fields of medicine, biotechnology, food science, and forensics. We have developed a unique bacterial preservation process involving a non-toxic, water-soluble acacia gum polymer that eliminates the need for refrigerated storage of samples. The main goal of this study is to characterize the efficacy of acacia gum polymer for preservation of pathogenic bacteria (Bacillus anthracis and methicillin-resistant Staphylococcus aureus-MRSA) on different materials, used for swabbing and filtration: cotton, wool, polyester, rayon, charcoal cloth, and Whatman paper. Acacia gum polymer used for preservation of two pathogens has been shown to significantly protect bacteria during dehydration and storage in all tested samples at the range of temperatures (5-45°C for MRSA and 40-90°C for B. anthracis). Our results showed higher recovery as well as higher viability during the storage of both bacteria in all materials with acacia gum. Addition of acacia gum polymer to swabbing materials or filters will increase efficacy of sample collection and identification of pathogenic bacteria from locations such as hospitals or the environment. Proposed approach can also be used for long-term storage of culture collections, since acacia gum contributes to viability and stability of bacterial cultures. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Effect of sample storage time on detection of hybridization signals in Checkerboard DNA-DNA hybridization.

    PubMed

    do Nascimento, Cássio; Muller, Katia; Sato, Sandra; Albuquerque Junior, Rubens Ferreira

    2012-04-01

    Long-term sample storage can affect the intensity of the hybridization signals provided by molecular diagnostic methods that use chemiluminescent detection. The aim of this study was to evaluate the effect of different storage times on the hybridization signals of 13 bacterial species detected by the Checkerboard DNA-DNA hybridization method using whole-genomic DNA probes. Ninety-six subgingival biofilm samples were collected from 36 healthy subjects, and the intensity of hybridization signals was evaluated at 4 different time periods: (1) immediately after collecting (n = 24) and (2) after storage at -20 °C for 6 months (n = 24), (3) for 12 months (n = 24), and (4) for 24 months (n = 24). The intensity of hybridization signals obtained from groups 1 and 2 were significantly higher than in the other groups (p < 0.001). No differences were found between groups 1 and 2 (p > 0.05). The Checkerboard DNA-DNA hybridization method was suitable to detect hybridization signals from all groups evaluated, and the intensity of signals decreased significantly after long periods of sample storage.

  18. Ex vivo spontaneous generation of 19-norandrostenedione and nandrolone detected in equine plasma and urine.

    PubMed

    Guan, Fuyu; Uboh, Cornelius E; Soma, Lawrence R; You, Youwen; Li, Xiaoqing; McDonnell, Sue

    2012-01-01

    19-Norandrostenedione (NAED) and nandrolone are anabolic-androgenic steroids (AASs). Nandrolone was regarded solely as a synthetic AAS until the 1980s when trace concentrations of apparently endogenous nandrolone were detected in urine samples obtained from intact male horses (stallions). Since then, its endogenous origin has been reported in boars and bulls; endogenous NAED and nandrolone have been identified in plasma and urine samples collected from stallions. More recently, however, it was suggested that NAED and nandrolone detected in urine samples from stallions are primarily artifacts due to the analytical procedure. The present study was undertaken to determine whether NAED and nandrolone detected in plasma and urine samples collected from stallions are truly endogenous or artifacts from sample processing. To answer this question, fresh plasma and urine samples from ≥8 stallions were analyzed for the two AASs, soon after collection, by liquid chromatography hyphenated to tandem mass spectrometry (LC-MS/MS). NAED and nandrolone were not detected in fresh plasma samples but detected in the same samples post storage. Concentrations of both AASs increased with storage time, and the increases were greater at a higher storage temperature (37°C versus 4°C, and ambient temperature versus 4°C). Although NAED was detected in some fresh stallion urine samples, its concentration (<407 pg/mL) was far lower (<0.4%) than that in the same samples post storage (at ambient temperature for 15 days). Nandrolone was not detected in most of fresh urine samples but detected in the same samples post storage. Based on these results, it is concluded that all NAED and nandrolone detected in stored plasma samples of stallions and most of them in the stored urine samples are not from endogenous origins but spontaneously generated during sample storage, most likely from spontaneous decarboxylation of androstenedione-19-oic acid and testosterone-19-oic acid. To our knowledge, it is the first time that all NAED and nandrolone detected in plasma of stallions and most of them detected in the urine have been shown to be spontaneously generated in vitro during sample storage. This finding would have significant implications with regard to the regulation of the two steroids in horse racing. Copyright © 2011 Elsevier Ltd. All rights reserved.

  19. Evaluation of Different Blood Collection Tubes and Blood Storage Conditions for the Preservation and Stability of Cell-Free Circulating DNA for the Analysis of the Methylated mSEPT9 Colorectal Cancer Screening Marker.

    PubMed

    Distler, Jurgen; Tetzner, Reimo; Weiss, Gunter; König, Thomas; Schlegel, Anne; Bagrowski, Michal

    2016-01-01

    For the subsequent analysis of the methylated mSEPT9 colorectal cancer screening marker in plasma, different blood collection tubes and blood storage conditions were investigated. The study demonstrated that methylated Septin 9 ( m SEPT9) can be consistently detected in plasma samples derived from whole blood samples collected with S-Monovette® K3E and BD Vacutainer ® K2EDTA tubes stored at 2-8 °C for a maximum of 24 h and for samples collected in S-Monovette CPDA tubes stored at 18-25 °C for up to 48 h.

  20. [Nonesterified fatty acids and the titrable acidity of breast milk. Consequences for collection conditions in milk bans].

    PubMed

    Luzeau, R; Barrois, V; Odièvre, M

    1983-01-01

    The study of breast-milk samples, fresh or after storage, shows that the titrable acidity (expressed in degrees Dornic) is directly correlated with their nonesterified fatty acid concentration. Those fresh samples which contain a high activity of lipoprotein lipase can develop in situ lipolysis. The resulting elevated titrable acidity may lead to consider these samples as unsuitable for infant nutrition. These results suggest that collection and storage of breast-milk have to be reassessed in order to avoid in situ lipolysis.

  1. Preparation of serum and plasma samples for determination of tricyclic antidepressants: effects of blood collection tubes and storage.

    PubMed

    Nyberg, G; Mårtensson, E

    1986-01-01

    The effects were tested of eight common types of blood collection tubes and two types of "plasma separators" on the stability of the tricyclic antidepressants amitriptyline, imipramine, clomipramine, and their monodemethylated metabolites in venous blood samples. Although EDTA-containing Venoject lavender and Vacutainer lavender tubes seemed to give the most stable plasma samples, and Venoject red the most stable serum samples, the differences were too small to have practical consequences. Vacutainer royal blue collection tubes gave significant losses of greater than 20% of some of the substances. The tubes with serum separator gel or filter proved unsuitable, since they were responsible for losses of greater than 40%. The losses were not caused by redistribution between blood cells and plasma but occurred mainly as a result of contact between the contents and the caps of the tubes. Experiments with freezing, thawing, and storage of samples showed that freshly sampled blood could be stored at room temperature for 24 h in Venoject green tubes without significant losses. Serum samples could be stored at refrigerator temperature for 4 weeks without important losses. Freezing, thawing, and storage at -20 degrees C did not influence the serum or plasma concentrations.

  2. Sample storage conditions significantly influence faecal microbiome profiles

    PubMed Central

    Choo, Jocelyn M; Leong, Lex EX; Rogers, Geraint B

    2015-01-01

    Sequencing-based studies of the human faecal microbiota are increasingly common. Appropriate storage of sample material is essential to avoid the introduction of post-collection bias in microbial community composition. Rapid freezing to −80 °C is commonly considered to be best-practice. However, this is not feasible in many studies, particularly those involving sample collection in participants’ homes. We determined the extent to which a range of stabilisation and storage strategies maintained the composition of faecal microbial community structure relative to freezing to −80 °C. Refrigeration at 4 °C, storage at ambient temperature, and the use of several common preservative buffers (RNAlater, OMNIgene.GUT, Tris-EDTA) were assessed relative to freezing. Following 72 hours of storage, faecal microbial composition was assessed by 16 S rRNA amplicon sequencing. Refrigeration was associated with no significant alteration in faecal microbiota diversity or composition. However, samples stored using other conditions showed substantial divergence compared to −80 °C control samples. Aside from refrigeration, the use of OMNIgene.GUT resulted in the least alteration, while the greatest change was seen in samples stored in Tris-EDTA buffer. The commercially available OMNIgene.GUT kit may provide an important alternative where refrigeration and cold chain transportation is not available. PMID:26572876

  3. Sample storage conditions significantly influence faecal microbiome profiles.

    PubMed

    Choo, Jocelyn M; Leong, Lex E X; Rogers, Geraint B

    2015-11-17

    Sequencing-based studies of the human faecal microbiota are increasingly common. Appropriate storage of sample material is essential to avoid the introduction of post-collection bias in microbial community composition. Rapid freezing to -80 °C is commonly considered to be best-practice. However, this is not feasible in many studies, particularly those involving sample collection in participants' homes. We determined the extent to which a range of stabilisation and storage strategies maintained the composition of faecal microbial community structure relative to freezing to -80 °C. Refrigeration at 4 °C, storage at ambient temperature, and the use of several common preservative buffers (RNAlater, OMNIgene.GUT, Tris-EDTA) were assessed relative to freezing. Following 72 hours of storage, faecal microbial composition was assessed by 16 S rRNA amplicon sequencing. Refrigeration was associated with no significant alteration in faecal microbiota diversity or composition. However, samples stored using other conditions showed substantial divergence compared to -80 °C control samples. Aside from refrigeration, the use of OMNIgene.GUT resulted in the least alteration, while the greatest change was seen in samples stored in Tris-EDTA buffer. The commercially available OMNIgene.GUT kit may provide an important alternative where refrigeration and cold chain transportation is not available.

  4. Liquid chromatography and ultra-performance liquid chromatography-mass spectrometry fingerprinting of human urine: sample stability under different handling and storage conditions for metabonomics studies.

    PubMed

    Gika, Helen G; Theodoridis, Georgios A; Wilson, Ian D

    2008-05-02

    Typically following collection biological samples are kept in a freezer for periods ranging from a few days to several months before analysis. Experience has shown that in LC-MS-based metabonomics research the best analytical practice is to store samples as these are collected, complete the sample set and analyse it in a single run. However, this approach is prudent only if the samples stored in the refrigerator or in the freezer are stable. Another important issue is the stability of the samples following the freeze-thaw process. To investigate these matters urine samples were collected from 6 male volunteers and analysed by LC-MS and ultra-performance liquid chromatography (UPLC)-MS [in both positive and negative electrospray ionization (ESI)] on the day of collection or at intervals of up to 6 months storage at -20 degrees C and -80 degrees C. Other sets of these samples underwent a series of up to nine freeze-thaw cycles. The stability of samples kept at 4 degrees C in an autosampler for up to 6 days was also assessed, with clear differences appearing after 48h. Data was analysed using multivariate statistical analysis (principal component analysis). The results show that sample storage at both -20 and -80 degrees C appeared to ensure sample stability. Similarly up to nine freeze thaw cycles were without any apparent effect on the profile.

  5. Common methods for fecal sample storage in field studies yield consistent signatures of individual identity in microbiome sequencing data.

    PubMed

    Blekhman, Ran; Tang, Karen; Archie, Elizabeth A; Barreiro, Luis B; Johnson, Zachary P; Wilson, Mark E; Kohn, Jordan; Yuan, Michael L; Gesquiere, Laurence; Grieneisen, Laura E; Tung, Jenny

    2016-08-16

    Field studies of wild vertebrates are frequently associated with extensive collections of banked fecal samples-unique resources for understanding ecological, behavioral, and phylogenetic effects on the gut microbiome. However, we do not understand whether sample storage methods confound the ability to investigate interindividual variation in gut microbiome profiles. Here, we extend previous work on storage methods for gut microbiome samples by comparing immediate freezing, the gold standard of preservation, to three methods commonly used in vertebrate field studies: lyophilization, storage in ethanol, and storage in RNAlater. We found that the signature of individual identity consistently outweighed storage effects: alpha diversity and beta diversity measures were significantly correlated across methods, and while samples often clustered by donor, they never clustered by storage method. Provided that all analyzed samples are stored the same way, banked fecal samples therefore appear highly suitable for investigating variation in gut microbiota. Our results open the door to a much-expanded perspective on variation in the gut microbiome across species and ecological contexts.

  6. Mass Spectrometric Rapid Diagnosis of Infectious Diseases.

    DTIC Science & Technology

    1980-01-01

    the analytical procedures to warrant reporting anew the whole analytical procedure. A. Sample Collection and Storage Procedure Urine samples were...positives or false-negatives. Next we have carried out a longitudinal study on urine samples obtained from groups of volunteer subjects vaccinated with...sterilization and storage procedures. 2. Developed new, simpler sample preparation techniques including one to handle tissue culture media. 3. Improved on the

  7. The ex situ conservation strategy for endangered plant species: small samples, storage and lessons from seed collected from US national parks

    USDA-ARS?s Scientific Manuscript database

    Ex situ collections of seeds sampled from wild populations provide germplasm for restoration and for scientific study about biological diversity. Seed collections of endangered species are urgent because they might forestall ever-dwindling population size and genetic diversity. However, collecting ...

  8. Sample collection and preparation of biofluids and extracts for gas chromatography-mass spectrometry.

    PubMed

    Emwas, Abdul-Hamid M; Al-Talla, Zeyad A; Kharbatia, Najeh M

    2015-01-01

    To maximize the utility of gas chromatography-mass spectrometry (GC-MS) in metabonomics research, all stages of the experimental design should be standardized, including sample collection, storage, preparation, and sample separation. Moreover, the prerequisite for any GC-MS analysis is that a compound must be volatile and thermally stable if it is to be analyzed using this technique. Since many metabolites are nonvolatile and polar in nature, they are not readily amenable to analysis by GC-MS and require initial chemical derivatization of the polar functional groups in order to reduce the polarity and to increase the thermal stability and volatility of the analytes. In this chapter, an overview is presented of the optimum approach to sample collection, storage, and preparation for gas chromatography-mass spectrometry-based metabonomics with particular focus on urine samples as example of biofluids.

  9. Impact of Oral Fluid Collection Device on Cannabinoid Stability Following Smoked Cannabis

    PubMed Central

    Anizan, Sébastien; Bergamaschi, Mateus M.; Barnes, Allan J.; Milman, Garry; Desrosiers, Nathalie; Lee, Dayong; Gorelick, David A.; Huestis, Marilyn A.

    2014-01-01

    Evaluation of cannabinoid stability in authentic oral fluid (OF) is critical, as most OF stability studies employed fortified or synthetic OF. Participants (n=16) smoked a 6.8% delta-9-tetrahydrocannabinol (THC) cigarette, and baseline concentrations of THC, 11-nor-9-carboxy-THC (THCCOOH), cannabidiol (CBD), and cannabinol (CBN) were determined within 24h in 16 separate pooled samples (collected 1h before to 10.5 or 13h after smoking). OF was collected with the StatSure Saliva Sampler™ and Oral-Eze® devices. Oral-Eze samples were re-analyzed after room temperature (RT) storage for 1 week, and for both devices after 4°C for 1 and 4 weeks, and –20°C for 4 and 24 weeks. Concentrations ±20% from initial concentrations were considered stable. With the StatSure device, all cannabinoids were within 80-120% median %baseline for all storage conditions. Individual THC, CBD, CBN and THCCOOH pool concentrations were stable in 100%, 100%, 80-94% and >85%, respectively, across storage conditions. With the Oral-Eze device, at RT or refrigerated storage (for 1 and 4 weeks), THC, CBD and THCCOOH were stable in 94-100%, 78-89% and 93-100% of samples, respectively, while CBN concentrations were 53–79% stable. However, after 24 weeks at -20°C, stability decreased, especially for CBD, with a median of 56% stability. Overall, the collection devices’ elution/stabilizing buffers provided good stability for OF cannabinoids, with the exception of the more labile CBN. To ensure OF cannabinoid concentration accuracy, these data suggest analysis within 4 weeks at 4°C storage for Oral-Eze collection and within 4 weeks at 4°C or 24 weeks at -20°C for StatSure collection. PMID:24995604

  10. Uncertainties in stormwater runoff data collection from a small urban catchment, Southeast China.

    PubMed

    Huang, Jinliang; Tu, Zhenshun; Du, Pengfei; Lin, Jie; Li, Qingsheng

    2010-01-01

    Monitoring data are often used to identify stormwater runoff characteristics and in stormwater runoff modelling without consideration of their inherent uncertainties. Integrated with discrete sample analysis and error propagation analysis, this study attempted to quantify the uncertainties of discrete chemical oxygen demand (COD), total suspended solids (TSS) concentration, stormwater flowrate, stormwater event volumes, COD event mean concentration (EMC), and COD event loads in terms of flow measurement, sample collection, storage and laboratory analysis. The results showed that the uncertainties due to sample collection, storage and laboratory analysis of COD from stormwater runoff are 13.99%, 19.48% and 12.28%. Meanwhile, flow measurement uncertainty was 12.82%, and the sample collection uncertainty of TSS from stormwater runoff was 31.63%. Based on the law of propagation of uncertainties, the uncertainties regarding event flow volume, COD EMC and COD event loads were quantified as 7.03%, 10.26% and 18.47%.

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

    DOE Data Explorer

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

    2003-04-01

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

  12. The Effect of Sampling and Storage on the Fecal Microbiota Composition in Healthy and Diseased Subjects

    PubMed Central

    Tedjo, Danyta I.; Jonkers, Daisy M. A. E.; Savelkoul, Paul H.; Masclee, Ad A.; van Best, Niels; Pierik, Marieke J.; Penders, John

    2015-01-01

    Large-scale cohort studies are currently being designed to investigate the human microbiome in health and disease. Adequate sampling strategies are required to limit bias due to shifts in microbial communities during sampling and storage. Therefore, we examined the impact of different sampling and storage conditions on the stability of fecal microbial communities in healthy and diseased subjects. Fecal samples from 10 healthy controls, 10 irritable bowel syndrome and 8 inflammatory bowel disease patients were collected on site, aliquoted immediately after defecation and stored at -80°C, -20°C for 1 week, at +4°C or room temperature for 24 hours. Fecal transport swabs (FecalSwab, Copan) were collected and stored for 48-72 hours at room temperature. We used pyrosequencing of the 16S gene to investigate the stability of microbial communities. Alpha diversity did not differ between all storage methods and -80°C, except for the fecal swabs. UPGMA clustering and principal coordinate analysis showed significant clustering by test subject (p<0.001) but not by storage method. Bray-Curtis dissimilarity and (un)weighted UniFrac showed a significant higher distance between fecal swabs and -80°C versus the other methods and -80°C samples (p<0.009). The relative abundance of Ruminococcus and Enterobacteriaceae did not differ between the storage methods versus -80°C, but was higher in fecal swabs (p<0.05). Storage up to 24 hours (at +4°C or room temperature) or freezing at -20°C did not significantly alter the fecal microbial community structure compared to direct freezing of samples from healthy subjects and patients with gastrointestinal disorders. PMID:26024217

  13. [Sampling, storage and transport of biological materials collected from living and deceased subjects for determination of concentration levels of ethyl alcohol and similarly acting substances. A proposal of updating the blood and urine sampling protocol].

    PubMed

    Wiergowski, Marek; Reguła, Krystyna; Pieśniak, Dorota; Galer-Tatarowicz, Katarzyna; Szpiech, Beata; Jankowski, Zbigniew

    2007-01-01

    The present paper emphasizes the most common mistakes committed at the beginning of an analytical procedure. To shorten the time and decrease the cost of determinations of substances with similar to alcohol activity, it is postulated to introduce mass-scale screening analysis of saliva collected from a living subject at the site of the event, with all positive results confirmed in blood or urine samples. If no saliva sample is collected for toxicology, a urine sample, allowing for a stat fast screening analysis, and a blood sample, to confirm the result, should be ensured. Inappropriate storage of a blood sample in the tube without a preservative can cause sample spilling and its irretrievable loss. The authors propose updating the "Blood/urine sampling protocol", with the updated version to be introduced into practice following consultations and revisions.

  14. Critical study of common conditions of storage of glucocorticoids and catecholamines in 24-h urine collected during resting and exercising conditions.

    PubMed

    Gouarne, C; Foury, A; Duclos, M

    2004-10-01

    Except immediate freezing of the samples, no practical method has been validated for preservation of glucocorticoids and catecholamines in 24-h urine collection. Furthermore, the influence of urine storage at bladder temperature during periods of different lengths and the effect of prior exercise on preservation of these hormones in the bladder have not been investigated until now. Ten healthy volunteers collected their urine both after a resting and after an exercise session. Urine was aliquoted into tubes which were stored during 24 h in the presence or in the absence of preservatives and at different temperatures. Two samples were stored either 3 or 9 h at 37 degrees C (bladder temperature) without additive. When collecting 24-h urine samples for glucocorticoids determination, sample can be stored at room temperature during the 24-h collection period without compromising glucocorticoids preservation. When collecting 24-h urine samples for catecholamines determination, samples have to be chilled without preservative during the whole of the collection period. If the samples have to be stored at room temperature, HCl should be used. Moreover, we report for the first time that catecholamines can be degraded in the bladder and therefore that subjects should urinate every 3 h during either a resting or an exercising day.

  15. Impact of collection conditions on the metabolite content of human urine samples as analyzed by liquid chromatography coupled to mass spectrometry and nuclear magnetic resonance spectroscopy.

    PubMed

    Roux, Aurélie; Thévenot, Etienne A; Seguin, François; Olivier, Marie-Françoise; Junot, Christophe

    There is a lack of comprehensive studies documenting the impact of sample collection conditions on metabolic composition of human urine. To address this issue, two experiments were performed at a 3-month interval, in which midstream urine samples from healthy individuals were collected, pooled, divided into several aliquots and kept under specific conditions (room temperature, 4 °C, with or without preservative) up to 72 h before storage at -80 °C. Samples were analyzed by high-performance liquid chromatography coupled to high-resolution mass spectrometry and bacterial contamination was monitored by turbidimetry. Multivariate analyses showed that urinary metabolic fingerprints were affected by the presence of preservatives and also by storage at room temperature from 24 to 72 h, whereas no change was observed for urine samples stored at 4 °C over a 72-h period. Investigations were then focused on 280 metabolites previously identified in urine: 19 of them were impacted by the kind of sample collection protocol in both experiments, including 12 metabolites affected by bacterial contamination and 7 exhibiting poor chemical stability. Finally, our results emphasize that the use of preservative prevents bacterial overgrowth, but does not avoid metabolite instability in solution, whereas storage at 4 °C inhibits bacterial overgrowth at least over a 72-h period and slows the chemical degradation process. Consequently, and for further LC/MS analyses, human urine samples should be kept at 4 °C if their collection is performed over 24 h.

  16. Reproducibility of NMR Analysis of Urine Samples: Impact of Sample Preparation, Storage Conditions, and Animal Health Status

    PubMed Central

    Schreier, Christina; Kremer, Werner; Huber, Fritz; Neumann, Sindy; Pagel, Philipp; Lienemann, Kai; Pestel, Sabine

    2013-01-01

    Introduction. Spectroscopic analysis of urine samples from laboratory animals can be used to predict the efficacy and side effects of drugs. This employs methods combining 1H NMR spectroscopy with quantification of biomarkers or with multivariate data analysis. The most critical steps in data evaluation are analytical reproducibility of NMR data (collection, storage, and processing) and the health status of the animals, which may influence urine pH and osmolarity. Methods. We treated rats with a solvent, a diuretic, or a nephrotoxicant and collected urine samples. Samples were titrated to pH 3 to 9, or salt concentrations increased up to 20-fold. The effects of storage conditions and freeze-thaw cycles were monitored. Selected metabolites and multivariate data analysis were evaluated after 1H NMR spectroscopy. Results. We showed that variation of pH from 3 to 9 and increases in osmolarity up to 6-fold had no effect on the quantification of the metabolites or on multivariate data analysis. Storage led to changes after 14 days at 4°C or after 12 months at −20°C, independent of sample composition. Multiple freeze-thaw cycles did not affect data analysis. Conclusion. Reproducibility of NMR measurements is not dependent on sample composition under physiological or pathological conditions. PMID:23865070

  17. Reproducibility of NMR analysis of urine samples: impact of sample preparation, storage conditions, and animal health status.

    PubMed

    Schreier, Christina; Kremer, Werner; Huber, Fritz; Neumann, Sindy; Pagel, Philipp; Lienemann, Kai; Pestel, Sabine

    2013-01-01

    Spectroscopic analysis of urine samples from laboratory animals can be used to predict the efficacy and side effects of drugs. This employs methods combining (1)H NMR spectroscopy with quantification of biomarkers or with multivariate data analysis. The most critical steps in data evaluation are analytical reproducibility of NMR data (collection, storage, and processing) and the health status of the animals, which may influence urine pH and osmolarity. We treated rats with a solvent, a diuretic, or a nephrotoxicant and collected urine samples. Samples were titrated to pH 3 to 9, or salt concentrations increased up to 20-fold. The effects of storage conditions and freeze-thaw cycles were monitored. Selected metabolites and multivariate data analysis were evaluated after (1)H NMR spectroscopy. We showed that variation of pH from 3 to 9 and increases in osmolarity up to 6-fold had no effect on the quantification of the metabolites or on multivariate data analysis. Storage led to changes after 14 days at 4°C or after 12 months at -20°C, independent of sample composition. Multiple freeze-thaw cycles did not affect data analysis. Reproducibility of NMR measurements is not dependent on sample composition under physiological or pathological conditions.

  18. Evaluation of sample stability for a quantitative faecal immunochemical test and comparison of two sample collection approaches.

    PubMed

    Mellen, Samantha; de Ferrars, Maria; Chapman, Claire; Bevan, Sarah; Turvill, James; Turnock, Daniel

    2018-01-01

    Background Faecal immunochemical testing is increasingly being used to triage symptomatic patients for suspected colorectal cancer. However, there are limited data on the effect of preanalytical factors on faecal haemoglobin when measured by faecal immunochemical testing. The aim of this work was to evaluate the stability of faecal haemoglobin in faeces and to compare two methods of faecal haemoglobin sampling for faecal immunochemical testing. Methods Six patients provided faeces for faecal haemoglobin measurement which were transferred into specialized collection devices at baseline and at 1, 2, 3 and 7 days after storage at either room temperature or 4°C. A total of 137 patients returned both faeces transferred into the specialized collection device and faeces in a standard collection pot. A quantitative immunoturbidometric method was used to measure faecal haemoglobin and results were compared categorically. Discrepant results were assessed against diagnosis. Results Faecal haemoglobin concentration declined rapidly within a day of storage at room temperature but results remained ≥10 μg Hb/g faeces in 5/6 patients after two days. A faecal haemoglobin result ≥10 μg Hb/g faeces was obtained in 4/6 patients after storage for seven days at 4°C. Results obtained when patients used specialized collection devices were significantly different from results obtained when faeces was transferred into the specialized collection device in the laboratory. Conclusion There is considerable heterogeneity in the sample stability of faecal haemoglobin; therefore, samples should be transferred rapidly into specialized collection devices to prevent false-negative results. Use of collection devices by patients can lead to false-positive results compared with their use in a laboratory.

  19. The effectiveness of large household water storage tanks for protecting the quality of drinking water.

    PubMed

    Graham, Jay P; VanDerslice, James

    2007-06-01

    Many communities along the US-Mexico border remain without infrastructure for water and sewage. Residents in these communities often collect and store their water in open 55-gallon drums. This study evaluated changes in drinking water quality resulting from an intervention that provided large closed water storage tanks (2,500-gallons) to individual homes lacking a piped water supply. After the intervention, many of the households did not change the source of their drinking water to the large storage tanks. Therefore, water quality results were first compared based on the source of the household's drinking water: store or vending machine, large tank, or collected from a public supply and transported by the household. Of the households that used the large storage tank as their drinking water supply, drinking water quality was generally of poorer quality. Fifty-four percent of samples collected prior to intervention had detectable levels of total coliforms, while 82% of samples were positive nine months after the intervention (p < 0.05). Exploratory analyses were also carried out to measure water quality at different points between collection by water delivery trucks and delivery to the household's large storage tank. Thirty percent of the samples taken immediately after water was delivered to the home had high total coliforms (> 10 CFU/100 ml). Mean free chlorine levels dropped from 0.43 mg/l, where the trucks filled their tanks, to 0.20 mg/l inside the household's tank immediately after delivery. Results of this study have implications for interventions that focus on safe water treatment and storage in the home, and for guidelines regarding the level of free chlorine required in water delivered by water delivery trucks.

  20. Experimental evaluation of sand fly collection and storage methods for the isolation and molecular detection of Phlebotomus-borne viruses.

    PubMed

    Remoli, Maria Elena; Bongiorno, Gioia; Fortuna, Claudia; Marchi, Antonella; Bianchi, Riccardo; Khoury, Cristina; Ciufolini, Maria Grazia; Gramiccia, Marina

    2015-11-09

    Several viruses have been recently isolated from Mediterranean phlebotomine sand flies; some are known to cause human disease while some are new to science. To monitor the Phlebotomus-borne viruses spreading, field studies are in progress using different sand fly collection and storage methods. Two main sampling techniques consist of CDC light traps, an attraction method allowing collection of live insects in which the virus is presumed to be fairly preserved, and sticky traps, an interception method suitable to collect dead specimens in high numbers, with a risk for virus viability or integrity. Sand flies storage requires a "deep cold chain" or specimen preservation in ethanol. In the present study the influence of sand fly collection and storage methods on viral isolation and RNA detection performances was evaluated experimentally. Specimens of laboratory-reared Phlebotomus perniciosus were artificially fed with blood containing Toscana virus (family Bunyaviridae, genus Phlebovirus). Various collection and storage conditions of blood-fed females were evaluated to mimic field procedures using single and pool samples. Isolation on VERO cell cultures, quantitative Real time-Retro-transcriptase (RT)-PCR and Nested-RT-PCR were performed according to techniques commonly used in surveillance studies. Live engorged sand flies stored immediately at -80 °C were the most suitable sample for phlebovirus identification by both virus isolation and RNA detection. The viral isolation rate remained very high (26/28) for single dead engorged females frozen after 1 day, while it was moderate (10/30) for specimens collected by sticky traps maintained up to 3 days at room temperature and then stored frozen without ethanol. Opposed to viral isolation, molecular RNA detection kept very high on dead sand flies collected by sticky traps when left at room temperature up to 6 days post blood meal and then stored frozen in presence (88/95) or absence (87/88) of ethanol. Data were confirmed using sand fly pools. While the collection and storage methods investigated had not much impact on the ability to detect viral RNA by molecular methods, they affected the capacity to recover viable viruses. Consequently, sand fly collection and handling procedures should be established in advance depending on the goal of the surveillance studies.

  1. A robust ambient temperature collection and stabilization strategy: Enabling worldwide functional studies of the human microbiome

    PubMed Central

    Anderson, Ericka L.; Li, Weizhong; Klitgord, Niels; Highlander, Sarah K.; Dayrit, Mark; Seguritan, Victor; Yooseph, Shibu; Biggs, William; Venter, J. Craig; Nelson, Karen E.; Jones, Marcus B.

    2016-01-01

    As reports on possible associations between microbes and the host increase in number, more meaningful interpretations of this information require an ability to compare data sets across studies. This is dependent upon standardization of workflows to ensure comparability both within and between studies. Here we propose the standard use of an alternate collection and stabilization method that would facilitate such comparisons. The DNA Genotek OMNIgene∙Gut Stool Microbiome Kit was compared to the currently accepted community standard of freezing to store human stool samples prior to whole genome sequencing (WGS) for microbiome studies. This stabilization and collection device allows for ambient temperature storage, automation, and ease of shipping/transfer of samples. The device permitted the same data reproducibility as with frozen samples, and yielded higher recovery of nucleic acids. Collection and stabilization of stool microbiome samples with the DNA Genotek collection device, combined with our extraction and WGS, provides a robust, reproducible workflow that enables standardized global collection, storage, and analysis of stool for microbiome studies. PMID:27558918

  2. A quantitative evaluation of two methods for preserving hair samples

    USGS Publications Warehouse

    Roon, David A.; Waits, L.P.; Kendall, K.C.

    2003-01-01

    Hair samples are an increasingly important DNA source for wildlife studies, yet optimal storage methods and DNA degradation rates have not been rigorously evaluated. We tested amplification success rates over a one-year storage period for DNA extracted from brown bear (Ursus arctos) hair samples preserved using silica desiccation and -20C freezing. For three nuclear DNA microsatellites, success rates decreased significantly after a six-month time point, regardless of storage method. For a 1000 bp mitochondrial fragment, a similar decrease occurred after a two-week time point. Minimizing delays between collection and DNA extraction will maximize success rates for hair-based noninvasive genetic sampling projects.

  3. Evaluation of the impact of RNA preservation methods of spiders for de novo transcriptome assembly.

    PubMed

    Kono, Nobuaki; Nakamura, Hiroyuki; Ito, Yusuke; Tomita, Masaru; Arakawa, Kazuharu

    2016-05-01

    With advances in high-throughput sequencing technologies, de novo transcriptome sequencing and assembly has become a cost-effective method to obtain comprehensive genetic information of a species of interest, especially in nonmodel species with large genomes such as spiders. However, high-quality RNA is essential for successful sequencing, and sample preservation conditions require careful consideration for the effective storage of field-collected samples. To this end, we report a streamlined feasibility study of various storage conditions and their effects on de novo transcriptome assembly results. The storage parameters considered include temperatures ranging from room temperature to -80°C; preservatives, including ethanol, RNAlater, TRIzol and RNAlater-ICE; and sample submersion states. As a result, intact RNA was extracted and assembly was successful when samples were preserved at low temperatures regardless of the type of preservative used. The assemblies as well as the gene expression profiles were shown to be robust to RNA degradation, when 30 million 150-bp paired-end reads are obtained. The parameters for sample storage, RNA extraction, library preparation, sequencing and in silico assembly considered in this work provide a guideline for the study of field-collected samples of spiders. © 2015 John Wiley & Sons Ltd.

  4. An evaluation of long-term preservation methods for brown bear (Ursus arctos) faecal DNA samples

    USGS Publications Warehouse

    Murphy, M.A.; Waits, L.P.; Kendall, K.C.; Wasser, S.K.; Higbee, J.A.; Bogden, R.

    2002-01-01

    Relatively few large-scale faecal DNA studies have been initiated due to difficulties in amplifying low quality and quantity DNA template. To improve brown bear faecal DNA PCR amplification success rates and to determine post collection sample longevity, five preservation methods were evaluated: 90% ethanol, DETs buffer, silica-dried, oven-dried stored at room temperature, and oven-dried stored at -20??C. Preservation effectiveness was evaluated for 50 faecal samples by PCR amplification of a mitochondrial DNA (mtDNA) locus (???146 bp) and a nuclear DNA (nDNA) locus (???200 bp) at time points of one week, one month, three months and six months. Preservation method and storage time significantly impacted mtDNA and nDNA amplification success rates. For mtDNA, all preservation methods had ??? 75% success at one week, but storage time had a significant impact on the effectiveness of the silica preservation method. Ethanol preserved samples had the highest success rates for both mtDNA (86.5%) and nDNA (84%). Nuclear DNA amplification success rates ranged from 26-88%, and storage time had a significant impact on all methods but ethanol. Preservation method and storage time should be important considerations for researchers planning projects utilizing faecal DNA. We recommend preservation of faecal samples in 90% ethanol when feasible, although when collecting in remote field conditions or for both DNA and hormone assays a dry collection method may be advantageous.

  5. Evaluation of Streck BCT and PAXgene Stabilised Blood Collection Tubes for Cell-Free Circulating DNA Studies in Plasma.

    PubMed

    Warton, Kristina; Yuwono, Nicole L; Cowley, Mark J; McCabe, Mark J; So, Alwin; Ford, Caroline E

    2017-10-01

    Blood samples for studies of circulating DNA in disease are often collected in clinical settings where prompt processing of samples is not possible. In order to avoid problems associated with leukocyte lysis after prolonged blood storage, stabilised blood tubes have been developed containing preservatives that prevent cell lysis. We evaluated Streck BCT tubes and PAXgene ccfDNA tubes, as well as standard EDTA blood collection tubes, in terms of DNA yield and fragment size. Blood was collected in EDTA, Streck BCT or PAXgene ccfDNA tubes and stored for 1 h at 4 °C, or 4 days at room temperature. DNA was extracted using the QIAamp Circulating Nucleic Acids kit, and visualised on an agarose gel or quantitated by qPCR. Ratios of a 247-base and a 115-base amplicon of the Alu repetitive element were used to infer size distribution. While plasma DNA in EDTA tube blood samples increased by ~10- to 20-fold after 4 days of storage at room temperature, both Streck BCT tubes and PAXgene ccfDNA tubes maintained stable plasma DNA concentrations. A slight decrease in DNA yield following 1 h of blood storage at 4 °C was observed in Streck BCT and PAXgene ccfDNA tubes relative to EDTA tubes. This decrease was reversed by increasing the proteinase digest step of the DNA extraction protocol to 60 min, as recommended by Streck tube product literature. Visualisation of the extracted DNA on an agarose gel showed that after 4 days of room temperature storage, samples collected in EDTA tubes contained abundant high-molecular weight DNA, which was partially fragmented in a ladder pattern. A slight increase in high-molecular weight DNA in samples stored for 4 days at room temperature in Streck BCT tubes was also observed, but this was not reflected in a change in large and small Alu fragment ratios as measured by qPCR. Tubes containing preservative to prevent cell lysis can extend the scope for blood collection in clinical settings; however, slight differences between samples collected in different tube types underscore the requirement for standardised protocols, as well as attention to sample handling.

  6. Recalcitrant Behavior of Temperate Forest Tree Seeds: Storage, Biochemistry, and Physiology

    Treesearch

    Kristina F. Connor; Sharon Sowa

    2002-01-01

    The recalcitrant behavior of seeds of live oak (Quercus virginiana Mill.), and Durand oak (Quercus durandii Buckl.) was examined after hydrated storage at two temperatures, +4°C and -2°C for up to 1 year. Samples were collected and analyses performed at monthly intervals. At each sampling time, seeds were tested for viability and...

  7. Quality assessment of wild Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) semen under conditions of short-term storage

    USDA-ARS?s Scientific Manuscript database

    Short-term storage trials were conducted with Atlantic sturgeon semen collected from a total of nine wild males during the 2008 and 2009 spawning seasons on the Hudson River. Semen samples were kept refrigerated (4 plus or minus 1 degree C) and stored in different gaseous atmospheres and storage ext...

  8. World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonisation Project: IV. Tissue collection, processing, and storage in endometriosis research

    PubMed Central

    Fassbender, Amelie; Rahmioglu, Nilufer; Vitonis, Allison F.; Viganò, Paola; Giudice, Linda C.; D’Hooghe, Thomas M.; Hummelshoj, Lone; Adamson, G. David; Becker, Christian M.; Missmer, Stacey A.; Zondervan, Krina T.; Adamson, G.D.; Allaire, C.; Anchan, R.; Becker, C.M.; Bedaiwy, M.A.; Buck Louis, G.M.; Calhaz-Jorge, C.; Chwalisz, K.; D'Hooghe, T.M.; Fassbender, A.; Faustmann, T.; Fazleabas, A.T.; Flores, I.; Forman, A.; Fraser, I.; Giudice, L.C.; Gotte, M.; Gregersen, P.; Guo, S.-W.; Harada, T.; Hartwell, D.; Horne, A.W.; Hull, M.L.; Hummelshoj, L.; Ibrahim, M.G.; Kiesel, L.; Laufer, M.R.; Machens, K.; Mechsner, S.; Missmer, S.A.; Montgomery, G.W.; Nap, A.; Nyegaard, M.; Osteen, K.G.; Petta, C.A.; Rahmioglu, N.; Renner, S.P.; Riedlinger, J.; Roehrich, S.; Rogers, P.A.; Rombauts, L.; Salumets, A.; Saridogan, E.; Seckin, T.; Stratton, P.; Sharpe-Timms, K.L.; Tworoger, S.; Vigano, P.; Vincent, K.; Vitonis, A.F.; Wienhues-Thelen, U.-H.; Yeung, P.P.; Yong, P.; Zondervan, K.T.

    2014-01-01

    Objective To harmonize standard operating procedures (SOPs) and standardize the recording of associated data for collection, processing, and storage of human tissues relevant to endometriosis. Design An international collaboration involving 34 clinical/academic centers and three industry collaborators from 16 countries on five continents. Setting In 2013, two workshops were conducted followed by global consultation, bringing together 54 leaders in endometriosis research and sample processing from around the world. Patient(s) None. Intervention(s) Consensus SOPs were based on: 1) systematic comparison of SOPs from 24 global centers collecting tissue samples from women with and without endometriosis on a medium or large scale (publication on >100 cases); 2) literature evidence where available, or consultation with laboratory experts otherwise; and 3) several global consultation rounds. Main Outcome Measure(s) Standard recommended and minimum required SOPs for tissue collection, processing, and storage in endometriosis research. Result(s) We developed “recommended standard” and “minimum required” SOPs for the collection, processing, and storage of ectopic and eutopic endometrium, peritoneum, and myometrium, and a biospecimen data collection form necessary for interpretation of sample-derived results. Conclusion(s) The EPHect SOPs allow endometriosis research centers to decrease variability in tissue-based results, facilitating between-center comparisons and collaborations. The procedures are also relevant to research into other gynecologic conditions involving endometrium, myometrium, and peritoneum. The consensus SOPs are based on the best available evidence; areas with limited evidence are identified as requiring further pilot studies. The SOPs will be reviewed based on investigator feedback and through systematic triannual follow-up. Updated versions will be made available at: http://endometriosisfoundation.org/ephect. PMID:25256928

  9. Sweat Sodium, Potassium, and Chloride Concentrations Analyzed Same Day as Collection Versus After 7 Days Storage in a Range of Temperatures.

    PubMed

    Baker, Lindsay B; Barnes, Kelly A; Sopeña, Bridget C; Nuccio, Ryan P; Reimel, Adam J; Ungaro, Corey T

    2018-05-22

    The purpose of this study was to determine the effect of storage temperature on sodium ([Na + ]), potassium ([K + ]), and chloride ([Cl - ]) concentrations of sweat samples analyzed 7 days after collection. Using the absorbent patch technique, 845 sweat samples were collected from 39 subjects (32 ± 7 years, 72.9 ± 10.5 kg) during exercise. On the same day as collection (PRESTORAGE), 609 samples were analyzed for [Na + ], [Cl - ], and [K + ] by ion chromatography (IC) and 236 samples were analyzed for [Na + ] using a compact ion-selective electrode (ISE). Samples were stored at one of the four conditions: -20 °C (IC, n = 138; ISE, n = 60), 8 °C (IC, n = 144; ISE, n = 59), 23 °C (IC, n = 159; ISE, n = 59), or alternating between 8 °C and 23 °C (IC, n = 168; ISE, n = 58). After 7 days in storage (POSTSTORAGE), samples were reanalyzed using the same technique as PRESTORAGE. PRESTORAGE sweat electrolyte concentrations were highly related to that of POSTSTORAGE (intraclass correlation coefficient: .945-.989, p < .001). Mean differences (95% confidence intervals) between PRESTORAGE and POSTSTORAGE were statistically, but not practically, significant for most comparisons: IC [Na + ]: -0.5(0.9) to -2.1(0.9) mmol/L; IC [K + ]: -0.1(0.1) to -0.2(0.1) mmol/L; IC [Cl - ]: -0.4(1.4) to -1.3(1.3) mmol/L; ISE [Na + ]: -2.0(1.1) to 1.3(1.1) mmol/L. Based on typical error of measurement results, 95% of the time PRESTORAGE and POSTSTORAGE sweat [Na + ], [K + ], and [Cl - ] by IC analysis fell within ±7-9, ±0.6-0.7, and ±9-13 mmol/L, respectively, while sweat [Na + ] by ISE was ±6 mmol/L. All conditions produced high reliability and acceptable levels of agreement in electrolyte concentrations of sweat samples analyzed on the day of collection versus after 7 days in storage.

  10. Practical Considerations of Moisture in Baled Biomass Feedstocks

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

    William A. Smith; Ian J. Bonner; Kevin L. Kenney

    2013-01-01

    Agricultural residues make up a large portion of the immediately available biomass feedstock for renewable energy markets. Current collection and storage methods rely on existing feed and forage practices designed to preserve nutrients and properties of digestibility. Low-cost collection and storage practices that preserve carbohydrates across a range of inbound moisture contents are needed to assure the economic and technical success of the emerging biomass industry. This study examines the movement of moisture in storage and identifies patterns of migration resulting from several on-farm storage systems and their impacts on moisture measurement and dry matter recovery. Baled corn stover andmore » energy sorghum were stored outdoors in uncovered, tarp-covered, or wrapped stacks and sampled periodically to measure moisture and dry matter losses. Interpolation between discrete sampling locations in the stack improved bulk moisture content estimates and showed clear patterns of accumulation and re-deposition. Atmospheric exposure, orientation, and contact with barriers (i.e., soil, tarp, and wrap surfaces) were found to cause the greatest amount of moisture heterogeneity within stacks. Although the bulk moisture content of many stacks remained in the range suitable for aerobic stability, regions of high moisture were sufficient to support microbial activity, thus support dry matter loss. Stack configuration, orientation, and coverage methods are discussed relative to impact on moisture management and dry matter preservation. Additionally, sample collection and data analysis are discussed relative to assessment at the biorefinery as it pertains to stability in storage, queuing, and moisture carried into processing.« less

  11. Stability of Proteins in Dried Blood Spot Biobanks*

    PubMed Central

    Björkesten, Johan; Enroth, Stefan; Shen, Qiujin; Wik, Lotta; Hougaard, David M.; Cohen, Arieh S.; Sörensen, Lene; Giedraitis, Vilmantas; Ingelsson, Martin; Larsson, Anders; Kamali-Moghaddam, Masood; Landegren, Ulf

    2017-01-01

    An important motivation for the construction of biobanks is to discover biomarkers that identify diseases at early, potentially curable stages. This will require biobanks from large numbers of individuals, preferably sampled repeatedly, where the samples are collected and stored under conditions that preserve potential biomarkers. Dried blood samples are attractive for biobanking because of the ease and low cost of collection and storage. Here we have investigated their suitability for protein measurements. Ninety-two proteins with relevance for oncology were analyzed using multiplex proximity extension assays (PEA) in dried blood spots collected on paper and stored for up to 30 years at either +4 °C or −24 °C. Our main findings were that (1) the act of drying only slightly influenced detection of blood proteins (average correlation of 0.970), and in a reproducible manner (correlation of 0.999), (2) detection of some proteins was not significantly affected by storage over the full range of three decades (34 and 76% of the analyzed proteins at +4 °C and −24 °C, respectively), whereas levels of others decreased slowly during storage with half-lives in the range of 10 to 50 years, and (3) detectability of proteins was less affected in dried samples stored at −24 °C compared with at +4 °C, as the median protein abundance had decreased to 80 and 93% of starting levels after 10 years of storage at +4 °C or −24 °C, respectively. The results of our study are encouraging as they suggest an inexpensive means to collect large numbers of blood samples, even by the donors themselves, and to transport, and store biobanked samples as spots of whole blood dried on paper. Combined with emerging means to measure hundreds or thousands of protein, such biobanks could prove of great medical value by greatly enhancing discovery as well as routine analysis of blood biomarkers. PMID:28501802

  12. Making It Last: Storage Time and Temperature Have Differential Impacts on Metabolite Profiles of Airway Samples from Cystic Fibrosis Patients.

    PubMed

    Wandro, Stephen; Carmody, Lisa; Gallagher, Tara; LiPuma, John J; Whiteson, Katrine

    2017-01-01

    Metabolites of human or microbial origin have the potential to be important biomarkers of the disease state in cystic fibrosis (CF). Clinical sample collection and storage conditions may impact metabolite abundances with clinical relevance. We measured the change in metabolite composition based on untargeted gas chromatography-mass spectrometry (GC-MS) when CF sputum samples were stored at 4°C, -20°C, or -80°C with one or two freeze-thaw cycles. Daily measurements were taken for 1 week and then weekly for 4 weeks (4°C) and 8 weeks (-20°C). The metabolites in samples stored at -20°C maintained abundances similar to those found at-80°C over the course of 8 weeks (average change in Bray-Curtis distance, 0.06 ± 0.04) and were also stable after one or two freeze-thaw cycles. However, the metabolite profiles of samples stored at 4°C shifted after 1 day and continued to change over the course of 4 weeks (average change in Bray-Curtis distance, 0.31 ± 0.12). The abundances of several amino acids and other metabolites increased with time of storage at 4°C but remained constant at -20°C. Storage temperature was a significant factor driving the metabolite composition (permutational multivariate analysis of variance: r 2 = 0.32 to 0.49, P < 0.001). CF sputum samples stored at -20°C at the time of sampling maintain a relatively stable untargeted GC-MS profile. Samples should be frozen on the day of collection, as more than 1 day at 4°C impacts the global composition of the metabolites in the sample. IMPORTANCE Metabolomics has great potential for uncovering biomarkers of the disease state in CF and many other contexts. However, sample storage timing and temperature may alter the abundance of clinically relevant metabolites. To assess whether existing samples are stable and to direct future study design, we conducted untargeted GC-MS metabolomic analysis of CF sputum samples after one or two freeze-thaw cycles and storage at 4°C and -20°C for 4 to 8 weeks. Overall, storage at -20°C and freeze-thaw cycles had little impact on metabolite profiles; however, storage at 4°C shifted metabolite abundances significantly. GC-MS profiling will aid in our understanding of the CF lung, but care should be taken in studies using sputum samples to ensure that samples are properly stored.

  13. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE, AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR PESTICIDES (EPA 225.2) AND CARBARYL (EPA 531.1) ANALYSIS (UA-F-17.1)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship tap and drinking water samples for analysis by EPA Method 525.2 (revision 1.0) and EPA method 531.1 (revision 3). This SOP provides a brief description of the sample containers, collection, preservation, stor...

  14. Storage Time and Urine Biomarker Levels in the ASSESS-AKI Study

    PubMed Central

    Liu, Kathleen D.; Siew, Edward D.; Reeves, W. Brian; Himmelfarb, Jonathan; Go, Alan S.; Hsu, Chi-yuan; Bennett, Michael R.; Devarajan, Prasad; Ikizler, T. Alp; Kaufman, James S.; Kimmel, Paul L.; Chinchilli, Vernon M.; Parikh, Chirag R.

    2016-01-01

    Background Although stored urine samples are often used in biomarker studies focused on acute and chronic kidney disease, how storage time impacts biomarker levels is not well understood. Methods 866 subjects enrolled in the NIDDK-sponsored ASsessment, Serial Evaluation, and Subsequent Sequelae in Acute Kidney Injury (ASSESS-AKI) Study were included. Samples were processed under standard conditions and stored at -70°C until analyzed. Kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), and liver fatty acid binding protein (L-FABP) were measured in urine samples collected during the index hospitalization or an outpatient visit 3 months later. Mixed effects models were used to determine the effect of storage time on biomarker levels and stratified by visit. Results Median storage was 17.8 months (25–75% IQR 10.6–23.7) for samples from the index hospitalization and 14.6 months (IQR 7.3–20.4) for outpatient samples. In the mixed effects models, the only significant association between storage time and biomarker concentration was for KIM-1 in outpatient samples, where each month of storage was associated with a 1.7% decrease (95% CI -3% to -0.3%). There was no relationship between storage time and KIM-1 levels in samples from the index hospitalization. Conclusion There was no significant impact of storage time over a median of 18 months on urine KIM-1, NGAL, IL-18 or L-FABP in hospitalized samples; a statistically significant effect towards a decrease over time was noted for KIM-1 in outpatient samples. Additional studies are needed to determine whether longer periods of storage at -70°C systematically impact levels of these analytes. PMID:27788160

  15. Storage Time and Urine Biomarker Levels in the ASSESS-AKI Study.

    PubMed

    Liu, Kathleen D; Siew, Edward D; Reeves, W Brian; Himmelfarb, Jonathan; Go, Alan S; Hsu, Chi-Yuan; Bennett, Michael R; Devarajan, Prasad; Ikizler, T Alp; Kaufman, James S; Kimmel, Paul L; Chinchilli, Vernon M; Parikh, Chirag R

    2016-01-01

    Although stored urine samples are often used in biomarker studies focused on acute and chronic kidney disease, how storage time impacts biomarker levels is not well understood. 866 subjects enrolled in the NIDDK-sponsored ASsessment, Serial Evaluation, and Subsequent Sequelae in Acute Kidney Injury (ASSESS-AKI) Study were included. Samples were processed under standard conditions and stored at -70°C until analyzed. Kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin (NGAL), interleukin-18 (IL-18), and liver fatty acid binding protein (L-FABP) were measured in urine samples collected during the index hospitalization or an outpatient visit 3 months later. Mixed effects models were used to determine the effect of storage time on biomarker levels and stratified by visit. Median storage was 17.8 months (25-75% IQR 10.6-23.7) for samples from the index hospitalization and 14.6 months (IQR 7.3-20.4) for outpatient samples. In the mixed effects models, the only significant association between storage time and biomarker concentration was for KIM-1 in outpatient samples, where each month of storage was associated with a 1.7% decrease (95% CI -3% to -0.3%). There was no relationship between storage time and KIM-1 levels in samples from the index hospitalization. There was no significant impact of storage time over a median of 18 months on urine KIM-1, NGAL, IL-18 or L-FABP in hospitalized samples; a statistically significant effect towards a decrease over time was noted for KIM-1 in outpatient samples. Additional studies are needed to determine whether longer periods of storage at -70°C systematically impact levels of these analytes.

  16. Hematology and pathology devices; reclassification; restricted devices; OTC test sample collection systems for drugs of abuse testing. Food and Drug Administration, HHS. Final rule.

    PubMed

    2000-04-07

    The Food and Drug Administration (FDA) is reclassifying over-the-counter (OTC) test sample collection systems for drugs of abuse testing from class III (premarket approval) into class I (general controls) and exempting them from premarket notification (510(k)) and current good manufacturing practice (CGMP) requirements. FDA is also designating OTC test sample collection systems for drugs of abuse testing as restricted devices under the Federal Food, Drug, and Cosmetic Act (the act) and establishing restrictions intended to assure consumers that: The underlying laboratory test(s) are accurate and reliable; the laboratory performing the test(s) has adequate expertise and competency; and the product has adequate labeling and methods of communicating test results to consumers. Finally, FDA is adding a conforming amendment to the existing classification regulation for specimen transport and storage containers to clarify that it does not apply to specimen transport and storage containers that are part of an OTC test sample collection system for the purpose of testing for the presence of drugs of abuse or their metabolites in a laboratory.

  17. Stability Study of Cervical Specimens Collected by Swab and Stored Dry Followed by Human Papillomavirus DNA Detection Using the cobas 4800 Test.

    PubMed

    Lin, Chun-Qing; Zeng, Xi; Cui, Jian-Feng; Liao, Guang-Dong; Wu, Ze-Ni; Gao, Qian-Qian; Zhang, Xun; Yu, Xiu-Zhang; Chen, Wen; Xi, Ming-Rong; Qiao, You-Lin

    2017-02-01

    Safer, more convenient methods for cervical sample collection and storage are necessary to facilitate human papillomavirus (HPV) DNA testing in low-resource settings. Our study aimed to evaluate the stability of cervical specimens collected with dry swabs and stored dry, compared to liquid-based cytology (LBC) samples, as detected by HPV DNA testing. Women with abnormal cytological findings or HPV-positive results at colposcopy were recruited from the West China Second University Hospital, Sichuan University, between October 2013 and March 2014. From each woman, physicians collected cervical specimens with a swab placed into a Sarstedt tube and a CytoBrush placed into LBC medium. Samples were randomly assigned to be stored at uncontrolled ambient temperature for 2, 7, 14, or 28 days and then were tested for 14 high-risk HPV (HR-HPV) types using the cobas HPV test. The rates of agreement between dry swab and LBC samples for any HR-HPV type, HPV16, HPV18, and the 12 pooled HR-HPV types were 93.8%, 97.8%, 99.4%, and 93.2%, respectively, with kappa values of 0.87 (95% confidence interval [CI], 0.83 to 0.91), 0.94 (95% CI, 0.91 to 0.97), 0.94 (95% CI, 0.87 to 1.00), and 0.86 (95% CI, 0.82 to 0.90). The performance of swab samples for detection of cervical precancerous lesions by means of cobas HPV testing was equal to that of LBC samples, even with stratification by storage time. Dry storage of swab-collected cervical samples can last for 1 month without loss of test performance by cobas HPV testing, compared to LBC samples, which may offer a simple inexpensive approach for cervical cancer screening in low-resource settings. Copyright © 2017 American Society for Microbiology.

  18. Stability of Drugs of Abuse in Urine Samples at Room Temperature by Use of a Salts Mixture.

    PubMed

    Pellegrini, Manuela; Graziano, Silvia; Mastrobattista, Luisa; Minutillo, Adele; Busardo, Francesco Paolo; Scarsella, Gianfranco

    2017-01-01

    It has long been recognized that ensuring analyte stability is of crucial importance in the use of any quantitative bioanalytical method. As analyses are usually not performed directly after collection of the biological samples, but after these have been processed and stored, it is essential that analyte stability can be maintained at storage conditions to ensure that the obtained concentration results adequately reflect those directly after sampling. The conservation of urine samples in refrigerated/ frozen conditions is strongly recommended; but not always feasible. The aim of this study was to assess the stability of some well-known drugs of abuse methamphetamine (MA), 11-nor-9-carboxy-Δ9- tetrahydrocannabinol (THC-COOH), benzoylecgonine (BE), and morphine (MOR) in urine samples kept at room temperature by adding a salt mixture (sodium citrate, sodium ascorbate, borax). Two different urine samples were prepared with and without salt mixture, stored at room temperature and then analyzed by gas chromatography-mass spectrometry at 0, 1, 7, 15, and 30 days after collection/preparation to look for eventual analyte degradation. Methamphetamine showed no significant changes with respect to the time of collection/ preparation (T0) up to 7 days later (T7), with or without salt mixture addiction. Then a significant degradation occurred in both salted and non salted urine. BE decrease was observed starting from day 1 after sample collection in salted and not salted samples, respectively. Salt addition seemed to reduce at least the initial BE degradation, with a significant difference (p<0.001) at 7 and 15 days of storage. However, the degradation was not more prevented in salted samples at 30 days of storage. A 20% decrease of MOR concentration was observed starting from day 1 after collection/preparation, both in salted and not salted samples with no subsequent decrease. With regard to THCCOOH, a significant decrease was observed starting from 7 days after collection/preparation, with of without adding the salt mixture. However, when comparing salted versus non salted samples at each time point, a statistically significant difference was observed at 7 and 30 days of storage. The results obtained indicate that the degradation of MA, THC-COOH and BE in urine samples kept at room temperature can be slowed by the addition of the salt mixture, whereas it seems to be ineffective in samples containing MOR. This evidence has to be taken into account, in the eventuality of using salted urine to prevent in a certain extent abuse of above-reported drugs of abuse. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

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

    Alan W. Scaroni; David C. Glick

    This project generated and provided coal samples and accompanying analytical data for research by DOE contractors and others. The five-year contract and a six-month no-cost extension have been completed. The Final Technical Progress Report is being prepared. All activities specified under the five-year contract and its six-month no-cost extension have been completed. Eleven DECS samples were collected, processed to a variety of particle sizes, heat-sealed in foil laminate bags under argon, and placed in refrigerated storage. All were analyzed for basic chemical composition, inorganic major and trace element composition including hazardous air pollutant elements, petrographic composition and characteristics, thermoplastic behaviormore » (if applicable), and other properties relevant to commercial utilization. Most were also analyzed by NMR, py/gc/ms, and a standardized liquefaction test; trends and relationships observed were evaluated and summarized. Twenty-two DECS samples collected under the previous contract received further processing, and most of these were subjected to organic geochemical and standardized liquefaction tests as well. Selected DECS samples were monitored annually to evaluate the effectiveness of foil laminate bags for long-term sample storage. Twenty-three PSOC samples collected under previous contracts and purged with argon before storage were also maintained and distributed, for a total of 56 samples covered by the contract. During the contract, 804 samples in 1586 containers, 2109 data printouts, and individual data items from 34208 samples were distributed. In the subject quarter, 25 samples, 18 data printouts, and individual data items from 1374 samples were distributed. All DECS samples are now available for immediate distribution at minus 6 mm (-1/4 inch), minus 0.85 mm (-20 mesh U.S.), and minus 0.25 mm (-60 mesh U.S.).« less

  20. Collection of Aerosolized Human Cytokines Using Teflon® Filters

    PubMed Central

    McKenzie, Jennifer H.; McDevitt, James J.; Fabian, M. Patricia; Hwang, Grace M.; Milton, Donald K.

    2012-01-01

    Background Collection of exhaled breath samples for the analysis of inflammatory biomarkers is an important area of research aimed at improving our ability to diagnose, treat and understand the mechanisms of chronic pulmonary disease. Current collection methods based on condensation of water vapor from exhaled breath yield biomarker levels at or near the detection limits of immunoassays contributing to problems with reproducibility and validity of biomarker measurements. In this study, we compare the collection efficiency of two aerosol-to-liquid sampling devices to a filter-based collection method for recovery of dilute laboratory generated aerosols of human cytokines so as to identify potential alternatives to exhaled breath condensate collection. Methodology/Principal Findings Two aerosol-to-liquid sampling devices, the SKC® Biosampler and Omni 3000™, as well as Teflon® filters were used to collect aerosols of human cytokines generated using a HEART nebulizer and single-pass aerosol chamber setup in order to compare the collection efficiencies of these sampling methods. Additionally, methods for the use of Teflon® filters to collect and measure cytokines recovered from aerosols were developed and evaluated through use of a high-sensitivity multiplex immunoassay. Our results show successful collection of cytokines from pg/m3 aerosol concentrations using Teflon® filters and measurement of cytokine levels in the sub-picogram/mL concentration range using a multiplex immunoassay with sampling times less than 30 minutes. Significant degradation of cytokines was observed due to storage of cytokines in concentrated filter extract solutions as compared to storage of dry filters. Conclusions Use of filter collection methods resulted in significantly higher efficiency of collection than the two aerosol-to-liquid samplers evaluated in our study. The results of this study provide the foundation for a potential new technique to evaluate biomarkers of inflammation in exhaled breath samples. PMID:22574123

  1. Assessment of changes in plasma hemoglobin and potassium levels in red cell units during processing and storage.

    PubMed

    Saini, Nishant; Basu, Sabita; Kaur, Ravneet; Kaur, Jasbinder

    2015-06-01

    Red cell units undergo changes during storage and processing. The study was planned to assess plasma potassium, plasma hemoglobin, percentage hemolysis during storage and to determine the effects of outdoor blood collection and processing on those parameters. Blood collection in three types of blood storage bags was done - single CPDA bag (40 outdoor and 40 in-house collection), triple CPD + SAGM bag (40 in-house collection) and quadruple CPD + SAGM bag with integral leukoreduction filter (40 in-house collection). All bags were sampled on day 0 (day of collection), day 1 (after processing), day 7, day 14 and day 28 for measurement of percentage hemolysis and potassium levels in the plasma of bag contents. There was significant increase in percentage hemolysis, plasma hemoglobin and plasma potassium level in all the groups during storage (p < 0.001). No significant difference was found between any parameter analyzed for outdoor and in-house collected single CPDA red cell units. There was significant lower percentage hemolysis (p < 0.001) and potassium (day 7 to day 14 - p < 0.05 and day 14 to day 28 - p < 0.001) in red cell units from day 7 onward until day 28 of storage in the leukoreduced quadruple bag as compared to the triple bag. The in-house single CPDA red cell units showed significantly more hemolysis (p < 0.001) as compared to the triple bags with SAGM additive solution after 28 days of storage. There is gradual increase in plasma hemoglobin and plasma potassium levels during the storage of red blood cells. Blood collection can be safely undertaken in outdoor blood donation camps even in hot summer months in monitored blood transport boxes. SAGM additive solution decreases the red cell hemolysis and allows extended storage of red cells. Prestorage leukoreduction decreases the red cell hemolysis and improves the quality of blood. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Temperature fluctuations during deep temperature cryopreservation reduce PBMC recovery, viability and T-cell function.

    PubMed

    Germann, Anja; Oh, Young-Joo; Schmidt, Tomm; Schön, Uwe; Zimmermann, Heiko; von Briesen, Hagen

    2013-10-01

    The ability to analyze cryopreserved peripheral blood mononuclear cell (PBMC) from biobanks for antigen-specific immunity is necessary to evaluate response to immune-based therapies. To ensure comparable assay results, collaborative research in multicenter trials needs reliable and reproducible cryopreservation that maintains cell viability and functionality. A standardized cryopreservation procedure is comprised of not only sample collection, preparation and freezing but also low temperature storage in liquid nitrogen without any temperature fluctuations, to avoid cell damage. Therefore, we have developed a storage approach to minimize suboptimal storage conditions in order to maximize cell viability, recovery and T-cell functionality. We compared the influence of repeated temperature fluctuations on cell health from sample storage, sample sorting and removal in comparison to sample storage without temperature rises. We found that cyclical temperature shifts during low temperature storage reduce cell viability, recovery and immune response against specific-antigens. We showed that samples handled under a protective hood system, to avoid or minimize such repeated temperature rises, have comparable cell viability and cell recovery rates to samples stored without any temperature fluctuations. Also T-cell functionality could be considerably increased with the use of the protective hood system compared to sample handling without such a protection system. This data suggests that the impact of temperature fluctuation on cell integrity should be carefully considered in future clinical vaccine trials and consideration should be given to optimal sample storage conditions. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  3. Method and apparatus for data sampling

    DOEpatents

    Odell, Daniel M. C.

    1994-01-01

    A method and apparatus for sampling radiation detector outputs and determining event data from the collected samples. The method uses high speed sampling of the detector output, the conversion of the samples to digital values, and the discrimination of the digital values so that digital values representing detected events are determined. The high speed sampling and digital conversion is performed by an A/D sampler that samples the detector output at a rate high enough to produce numerous digital samples for each detected event. The digital discrimination identifies those digital samples that are not representative of detected events. The sampling and discrimination also provides for temporary or permanent storage, either serially or in parallel, to a digital storage medium.

  4. Ascaris lumbricoides egg die-off in an experimental excreta storage system and public health implication in Vietnam.

    PubMed

    Vu-Van, Tu; Pham-Duc, Phuc; Winkler, Mirko S; Zurbrügg, Christian; Zinsstag, Jakob; Le Thi Thanh, Huong; Bich, Tran Huu; Nguyen-Viet, Hung

    2017-02-01

    We studied the influence of different additive materials (lime, and rice husk) and aeration conditions on Ascaris lumbricoides egg die-off in 24 vaults of an experimental excreta storage unit. Excreta samples were collected once every two weeks over a 181-day period. Temperature, pH, and moisture content were recorded. A. lumbricoides eggs were quantitatively analyzed by the Romanenko method, which identified and counted live and dead eggs. From the first sampling (0 storage day) to the final sampling (181 storage days) the average percentage of viable A. lumbricoides eggs decreased gradually from 76.72 ± 11.23% (mean ± SD) to 8.26 ± 5.20%. The storage time and the high pH value significantly increased the die-off of helminth eggs. Over 181 storage days, all vaults option effectively reduced A. lumbricoides eggs die-off. The best vault option, with aeration and 10% lime per total weight, met the WHO standard for excreta treatment on the 111th storage day.

  5. Analytical precision of the Urolizer for the determination of the BONN-Risk-Index (BRI) for calcium oxalate urolithiasis and evaluation of the influence of 24-h urine storage at moderate temperatures on BRI.

    PubMed

    Berg, Wolfgang; Bechler, Robin; Laube, Norbert

    2009-01-01

    Since its first publication in 2000, the BONN-Risk-Index (BRI) has been successfully used to determine the calcium oxalate (CaOx) crystallization risk from urine samples. To date, a BRI-measuring device, the "Urolizer", has been developed, operating automatically and requiring only a minimum of preparation. Two major objectives were pursued: determination of Urolizer precision, and determination of the influence of 24-h urine storage at moderate temperatures on BRI. 24-h urine samples from 52 CaOx stone-formers were collected. A total of 37 urine samples were used for the investigation of Urolizer precision by performing six independent BRI determinations in series. In total, 30 samples were taken for additional investigation of urine storability. Each sample was measured thrice: directly after collection, after 24-h storage at T=21 degrees C, and after 24-h cooling at T=4 degrees C. Outcomes were statistically tested for identity with regard to the immediately obtained results. Repeat measurements for evaluation of Urolizer precision revealed statistical identity of data (p-0.05). 24-h storage of urine at both tested temperatures did not significantly affect BRI (p-0.05). The pilot-run Urolizer shows high analytical reliability. The innovative analysis device may be especially suited for urologists specializing in urolithiasis treatment. The possibility for urine storage at moderate temperatures without loss of analysis quality further demonstrates the applicability of the BRI method.

  6. Automated biowaste sampling system, solids subsystem operating model, part 2

    NASA Technical Reports Server (NTRS)

    Fogal, G. L.; Mangialardi, J. K.; Stauffer, R. E.

    1973-01-01

    The detail design and fabrication of the Solids Subsystem were implemented. The system's capacity for the collection, storage or sampling of feces and vomitus from six subjects was tested and verified.

  7. Influence of storage vial material on measurement of organophosphate flame retardant metabolites in urine.

    PubMed

    Carignan, Courtney C; Butt, Craig M; Stapleton, Heather M; Meeker, John D; Minguez-Alarcón, Lidia; Williams, Paige L; Hauser, Russ

    2017-08-01

    Use of organophosphate flame retardants (PFRs) has increased over the past decade with the phase out of polybrominated diphenyl ethers. Urinary metabolites of PFRs are used as biomarkers of exposure in epidemiologic research, which typically uses samples collected and stored in polypropylene plastic cryovials. However, a small study suggested that the storage vial material may influence reported concentrations. Therefore, we aimed to examine the influence of the storage vial material on analytical measurement of PFR urinary metabolites. Using urine samples collected from participants in the Environment and Reproductive Health (EARTH) Study, we analyzed the PFR metabolites in duplicate aliquots that were stored in glass and plastic vials (n = 31 pairs). Bis(1,3-dichloro-2-propyl) phosphate (BDCIPP), diphenyl phosphate (DPHP) and isopropyl-phenyl phenyl phosphate (ip-PPP) were detected in 98%, 97% and 87% of duplicates. We observed high correlations between glass-plastic duplicates for BDCIPP (r s  = 0.95), DPHP (r s  = 0.79) and ip-PPP (r s  = 0.82) (p < 0.0001). Urinary ip-PPP was an average of 0.04 ng/ml (p = 0.04) higher among samples stored in glass, with a mean relative difference of 14%. While this difference is statistically significant, it is small in magnitude. No differences were observed for BDCIPP or DPHP, however future research should seek to reduce the potential for type II error (false negatives). We conclude that storing urine samples in polypropylene plastic cryovials may result in slightly reduced concentrations of urinary ip-PPP relative to storage in glass vials and future research should seek to increase the sample size, reduce background variability and consider the material of the urine collection cup. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. The UK Biobank sample handling and storage validation studies.

    PubMed

    Peakman, Tim C; Elliott, Paul

    2008-04-01

    and aims UK Biobank is a large prospective study in the United Kingdom to investigate the role of genetic factors, environmental exposures and lifestyle in the causes of major diseases of late and middle age. It involves the collection of blood and urine from 500 000 individuals aged between 40 and 69 years. How the samples are collected, processed and stored will have a major impact on the future scientific usefulness of the UK Biobank resource. A series of validation studies was recommended to test the robustness of the draft sample handling and storage protocol. Samples of blood and urine were collected from 40 healthy volunteers and either processed immediately according to the protocol or maintained at specified temperatures (4 degrees C for all tubes with the exception of vacutainers containing acid citrate dextrose that were maintained at 18 degrees C) for 12, 24 or 36 h prior to processing. A further sample was maintained for 24 h at 4 degrees C, processed and the aliquots frozen at -80 degrees C for 20 days and then thawed under controlled conditions. The stability of the samples was compared for the different times in a wide variety of assays. The samples maintained at 4 degrees C were stable for at least 24 h after collection for a wide range of assays. Small but significant changes were observed in metabonomic studies in samples maintained at 4 degrees C for 36 h. There was no degradation of the samples for a range of biochemical assays after short-term freezing and thawing under controlled conditions. Whole blood maintained at 18 degrees C for 24 h in vacutainers containing acid citrate dextrose is suitable for viral immortalization techniques. The validation studies reported in this supplement provide justification for the sample handling and storage procedures adopted in the UK Biobank project.

  9. Influence of tube type, storage time, and temperature on the total and free concentration of valproic acid.

    PubMed

    Tarasidis, C G; Garnett, W R; Kline, B J; Pellock, J M

    1986-01-01

    The influence of storage conditions on the total and free concentration of valproic acid (VPA) was studied in six normal male subjects who ingested 750 mg of VPA (3 X 250 mg Depakene capsules; Abbott Laboratories). Blood samples were collected in various types of Vacutainer tubes (red top, no additives; green top, sodium heparin; blue top, sodium citrate; and purple top, EDTA) 2 h post administration of VPA. Either these samples were centrifuged immediately or stored for various periods of time at room temperature or refrigerated, or the supernate was frozen prior to analysis. Free VPA samples were obtained utilizing the Amicon ultrafiltration system. All VPA samples were analyzed by gas-liquid chromatography. Total VPA concentrations obtained from plasma collected with sodium citrate were lower (p less than 0.05) than either serum or plasma collected with other anticoagulants. There were no differences (p greater than 0.05) in total or free VPA concentrations between samples collected in serum or in plasma collected with heparin or EDTA. Storing samples for 96 h at room temperature did not alter the total VPA concentrations but was found to increase the free fraction of VPA (p less than 0.05). The refrigeration or freezing of the supernate from the blood samples for 7 days did not alter (p greater than 0.05) the total or the free fraction of VPA. The results of this study demonstrate that total and/or free VPA may be collected from either serum or plasma, provided sodium citrate is not used to collect plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

  10. Impact of oral fluid collection device on cannabinoid stability following smoked cannabis.

    PubMed

    Anizan, Sébastien; Bergamaschi, Mateus M; Barnes, Allan J; Milman, Garry; Desrosiers, Nathalie; Lee, Dayong; Gorelick, David A; Huestis, Marilyn A

    2015-02-01

    Evaluation of cannabinoid stability in authentic oral fluid (OF) is critical, as most OF stability studies employed fortified or synthetic OF. Participants (n = 16) smoked a 6.8% delta-9-tetrahydrocannabinol (THC) cigarette, and baseline concentrations of THC, 11-nor-9-carboxy-THC (THCCOOH), cannabidiol (CBD), and cannabinol (CBN) were determined within 24 h in 16 separate pooled samples (collected 1 h before to 10.5 or 13 h after smoking). OF was collected with the StatSure Saliva Sampler™ and Oral-Eze® devices. Oral-Eze samples were re-analyzed after room temperature (RT) storage for 1 week, and for both devices after 4 °C for 1 and 4 weeks, and -20 °C for 4 and 24 weeks. Concentrations ±20% from initial concentrations were considered stable. With the StatSure device, all cannabinoids were within 80-120% median %baseline for all storage conditions. Individual THC, CBD, CBN and THCCOOH pool concentrations were stable in 100%, 100%, 80-94% and >85%, respectively, across storage conditions. With the Oral-Eze device, at RT or refrigerated storage (for 1 and 4 weeks), THC, CBD and THCCOOH were stable in 94-100%, 78-89%, and 93-100% of samples, respectively, while CBN concentrations were 53-79% stable. However, after 24 weeks at -20 °C, stability decreased, especially for CBD, with a median of 56% stability. Overall, the collection devices' elution/stabilizing buffers provided good stability for OF cannabinoids, with the exception of the more labile CBN. To ensure OF cannabinoid concentration accuracy, these data suggest analysis within 4 weeks at 4 °C storage for Oral-Eze collection and within 4 weeks at 4 °C or 24 weeks at -20 °C for StatSure collection. Published 2014. This article is a U.S. Government work and is in the public domain in the USA. Published 2014. This article is a U.S. Government work and is in the public domain in the USA.

  11. Paper-based archiving of biological samples from fish for detecting betanodavirus.

    PubMed

    Navaneeth Krishnan, A; Bhuvaneswari, T; Ezhil Praveena, P; Jithendran, K P

    2016-07-01

    This study was carried out to evaluate the efficiency of the Flinders Technology Associates (FTA(®)) card (Whatman(®)) as a sampling device and storage platform for RNA from betanodavirus-infected biological samples (viz., larvae, broodstock, cell culture supernatants and rearing seawater spiked with infected materials). The study showed that FTA cards can be used to detect betanodaviruses by reverse transcription-polymerase chain reaction (RT-PCR). The diagnostic efficiency of RT-PCR from all sample types on FTA cards decreased after 21 days of storage at 4 °C, although the virus could be detected up to 28 days by nested RT-PCR. The FTA card protocol thus provides a supplementary method for quick and easy collection of samples, preservation of RNA on a dry storage basis, and detection of betanodavirus-infected fish.

  12. IMPROVED METHOD FOR THE STORAGE OF GROUND WATER SAMPLES CONTAINING VOLATILE ORGANIC ANALYTES

    EPA Science Inventory

    The sorption of volatile organic analytes from water samples by the Teflon septum surface used with standard glass 40-ml sample collection vials was investigated. Analytes tested included alkanes, isoalkanes, olefins, cycloalkanes, a cycloalkene, monoaromatics, a polynuclear arom...

  13. In vitro production of GHB in blood and serum samples under various storage conditions.

    PubMed

    Zörntlein, S W; Kopp, A; Becker, J; Kaufmann, T J; Röhrich, J; Urban, R

    2012-01-10

    The in vitro production of GHB was observed in freshly collected, untreated whole blood samples using glass BD-Vacutainers and polypropylene S-monovettes. GHB concentrations were determined daily over a period of one week and after 3, 6 and 9 weeks again. Furthermore, the GHB concentration in 40 untreated random whole blood samples stored at 4°C for a longer period of time (10 samples 12 month, 10 samples 24 month and 20 samples 36 month) was also determined. For comparison, the in vitro production of GHB in freshly collected and prepared serum samples was observed. GHB serum concentrations were determined three times over a period of one week and once again after six weeks. Sample preparation was performed by means of methanolic extraction following the precipitation of whole blood and serum samples. A methanolic standard calibration was done in a low range of 0.005-0.1 μg/mL (LOD: 0.004, LLOQ: 0.013). For quantification a spiked blood bank serum with a determined GHB concentration of 0.09 μg/mL was used. Corrected calibrations in the range of 0.09-5.09 μg/mL were used (LOD: 0.08 μg/mL, LLOQ: 0.30 μg/mL), recovery: 91.3% (high level: 4.09 μg/mL) 50.5% (low level: 0.19 μg/mL). Relevant elevation of GHB was observed in all whole blood samples stored in liquid form (4°C or room temperature). In two of the 40 whole blood samples stored over a longer period of time at 4°C, GHB concentrations in the range of 13 μg/mL were even determined. These findings constitute grounds for caution. Even a GHB cut-off level of 5 μg/mL cannot be considered as "absolutely positive" proof of a case of exogenous administration, at least in untreated liquid blood samples in long time storage. However, no significant elevations of GHB were otherwise observed in any of the serum samples independently of storage temperature nor in the whole blood samples that were frozen for storage. The results suggest that the cut-off for exogenous GHB of 5 μg/mL could be lowered significantly, with the consequence of winning valuable time for the potential victim, but only if serum is collected for GHB determination or if the whole blood sample is frozen immediately after collection and the procedure well documented. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

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

    Ottesen, Elizabeth A.; Marin, Roman; Preston, Christina M.

    Planktonic microbial activity and community structure is dynamic, and can change dramatically on time scales of hours to days. Yet for logistical reasons, this temporal scale is typically undersampled in the marine environment. In order to facilitate higher-resolution, long-term observation of microbial diversity and activity, we developed a protocol for automated collection and fixation of marine microbes using the Environmental Sample Processor (ESP) platform. The protocol applies a preservative (RNALater) to cells collected on filters, for long-term storage and preservation of total cellular RNA. Microbial samples preserved using this protocol yielded high-quality RNA after 30 days of storage at roommore » temperature, or onboard the ESP at in situ temperatures. Pyrosequencing of complementary DNA libraries generated from ESP-collected and preserved samples yielded transcript abundance profiles nearly indistinguishable from those derived from conventionally treated replicate samples. To demonstrate the utility of the method, we used a moored ESP to remotely and autonomously collect Monterey Bay seawater for metatranscriptomic analysis. Community RNA was extracted and pyrosequenced from samples collected at four time points over the course of a single day. In all four samples, the oxygenic photoautotrophs were predominantly eukaryotic, while the bacterial community was dominated by Polaribacter-like Flavobacteria and a Rhodobacterales bacterium sharing high similarity with Rhodobacterales sp. HTCC2255. However, each time point was associated with distinct species abundance and gene transcript profiles. These laboratory and field tests confirmed that autonomous collection and preservation is a feasible and useful approach for characterizing the expressed genes and environmental responses of marine microbial communities.« less

  15. Filter Paper-based Nucleic Acid Storage in High-throughput Solid Tumor Genotyping.

    PubMed

    Stachler, Matthew; Jia, Yonghui; Sharaf, Nematullah; Wade, Jacqueline; Longtine, Janina; Garcia, Elizabeth; Sholl, Lynette M

    2015-01-01

    Molecular testing of tumors from formalin-fixed paraffin-embedded (FFPE) tissue blocks is central to clinical practice; however, it requires histology support and increases test turnaround time. Prospective fresh frozen tissue collection requires special handling, additional storage space, and may not be feasible for small specimens. Filter paper-based collection of tumor DNA reduces the need for histology support, requires little storage space, and preserves high-quality nucleic acid. We investigated the performance of tumor smears on filter paper in solid tumor genotyping, as compared with paired FFPE samples. Whatman FTA Micro Card (FTA preps) smears were prepared from 21 fresh tumor samples. A corresponding cytology smear was used to assess tumor cellularity and necrosis. DNA was isolated from FTA preps and FFPE core samples using automated methods and quantified using SYBR green dsDNA detection. Samples were genotyped for 471 mutations on a mass spectrophotometry-based platform (Sequenom). DNA concentrations from FTA preps and FFPE correlated for untreated carcinomas but not for mesenchymal tumors (Spearman σ=0.39 and σ=-0.1, respectively). Average DNA concentrations were lower from FTA preps as compared with FFPE, but DNA quality was higher with less fragmentation. Seventy-six percent of FTA preps and 86% of FFPE samples generated adequate DNA for genotyping. FTA preps tended to perform poorly for collection of DNA from pretreated carcinomas and mesenchymal neoplasms. Of the 16 paired DNA samples that were genotyped, 15 (94%) gave entirely concordant results. Filter paper-based sample preservation is a feasible alternative to FFPE for use in automated, high-throughput genotyping of carcinomas.

  16. Stability of odorants from pig production in sampling bags for olfactometry.

    PubMed

    Hansen, Michael J; Adamsen, Anders P S; Feilberg, Anders; Jonassen, Kristoffer E N

    2011-01-01

    Odor from pig production facilities is typically measured with olfactometry, whereby odor samples are collected in sampling bags and assessed by human panelists within 30 h. In the present study, the storage stability of odorants in two types of sampling bags that are often used for olfactometry was investigated. The bags were made of Tedlar or Nalophan. In a field experiment, humid and dried air samples were collected from a pig production facility with growing-finishing pigs and analyzed with a gas chromatograph with an amperometric sulfur detector at 4, 8, 12, 28, 52, and 76 h after sampling. In a laboratory experiment, the bags were filled with a humid gas mixture containing carboxylic acids, phenols, indoles, and sulfur compounds and analyzed with proton-transfer-reaction mass spectrometry after 0, 4, 8, 12, and 24 h. The results demonstrated that the concentrations of carboxylic acids, phenols, and indoles decreased by 50 to >99% during the 24 h of storage in Tedlar and Nalophan bags. The concentration of hydrogen sulfide decreased by approximately 30% during the 24 h of storage in Nalophan bags, whereas in Tedlar bags the concentration of sulfur compounds decreased by <5%. In conclusion, the concentrations of odorants in air samples from pig production facilities significantly decrease during storage in Tedlar and Nalophan bags, and the composition changes toward a higher relative presence of sulfur compounds. This can result in underestimation of odor emissions from pig production facilities and of the effect of odor reduction technologies. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

  17. NHEXAS PHASE I REGION 5 STUDY--QA ANALYTICAL RESULTS FOR VOCS IN BLANKS

    EPA Science Inventory

    This data set includes analytical results for measurements of VOCs in 88 blank samples. Measurements were made for up to 23 VOCs in blank samples of air, water, and blood. Blank samples were used to assess the potential for sample contamination during collection, storage, shipmen...

  18. Method and apparatus for data sampling

    DOEpatents

    Odell, D.M.C.

    1994-04-19

    A method and apparatus for sampling radiation detector outputs and determining event data from the collected samples is described. The method uses high speed sampling of the detector output, the conversion of the samples to digital values, and the discrimination of the digital values so that digital values representing detected events are determined. The high speed sampling and digital conversion is performed by an A/D sampler that samples the detector output at a rate high enough to produce numerous digital samples for each detected event. The digital discrimination identifies those digital samples that are not representative of detected events. The sampling and discrimination also provides for temporary or permanent storage, either serially or in parallel, to a digital storage medium. 6 figures.

  19. Effects of post-sampling conditions on ambient carbon aerosol filter measurements

    NASA Astrophysics Data System (ADS)

    Dillner, Ann M.; Phuah, Chin H.; Turner, Jay R.

    2009-12-01

    Ambient carbonaceous material collected on quartz filters is prone to measurement artifacts due to material gained or lost during post-sampling field latency, shipping, and storage. In seventeen sampling events over a one year period, ambient PM 2.5 aerosols were collected on quartz filters (without denuders) and subjected to various filter treatments to assess the potential for and extent of artifacts. The filter treatments simulated post-sampling environments that filters may be exposed to and included: storage at 40 °C for up to 96 h, storage at -16 °C for 48 h, and storage at room temperature (˜21 °C) for 48 h. Carbon mass on the filters was measured using a thermal-optical method. The total carbon (TC), total organic carbon (TOC) and total elemental carbon (TEC) as well as carbon thermal fraction masses were obtained. Statistical analyses were performed to identify significant differences in carbon fraction concentrations between filters analyzed immediately after sampling and after being subjected to treatment. TOC and TC concentrations decreased by on average 15 ± 5% and 10 ± 4%, respectively, for filters maintained at 40 °C for 96 h but did not change for filters stored at room temperature or frozen for 48 h. TEC did not change for any of the filter treatments. The mass concentration for the organic carbon thermal fraction that evolves at the lowest temperature step (OC1) decreased with increasing storage time at 40 °C with average losses of 70 ± 7% after 96 h. Therefore, OC1 is not a stable measurement due to post-sampling conditions that may be encountered. This work demonstrates that TOC and TC can have substantial measurement artifacts on filters subjected to field latency and other non-temperature controlled post-sampling handling, compared to the carbon loadings on the filter at the end of the sampling period.

  20. Breast Milk Collection and Storage in the Neonatal Intensive Care Unit: Nurses' Knowledge, Practice, and Perceived Barriers.

    PubMed

    Gharaibeh, Huda Falah; Al-Sheyab, Nihaya; Malkawi, Shefa Yousef

    2016-12-01

    Temporary storage of human milk under appropriate conditions encourages prolonged breast-feeding. This study aimed to assess neonatal nurses' knowledge and practice, as well as barriers, related to breast milk collection and storage and to investigate the association between nurses' knowledge and practice and other variables. A cross-sectional design was used. Consecutive samplings of 75 nurses were recruited. The questions were based on a literature review of guidelines for collecting and storing breast milk from various sources, such as the Centers for Disease Control and Prevention and the Academy of Breastfeeding Medicine. Nurses' knowledge and practice of breast milk collection and storage were adequate in general, but inadequate in relation to issues such as discarding breast milk, the thawing process, and storage temperatures. Poor practices related to absence of required equipment and unit policy. Barriers were increased workload, inappropriate milk room (i.e., a small room with uncomfortable chairs and no privacy), inactive policy, and inadequate time. Nurses should be orientated to the guidelines and a unit policy to enhance evidence-based practice. J Contin Educ Nurs. 2016;47(12):551-557. Copyright 2016, SLACK Incorporated.

  1. Effect of ascorbic acid on storage of Greyhound erythrocytes.

    PubMed

    Fontes, Jorge A; Banerjee, Uddyalok; Iazbik, M Cristina; Marín, Liliana M; Couto, C Guillermo; Palmer, Andre F

    2015-09-01

    To assess changes in biochemical and biophysical properties of canine RBCs during cold (1° to 6°C) storage in a licensed RBC additive solution (the RBC preservation solution designated AS-1) supplemented with ascorbic acid. Blood samples from 7 neutered male Greyhounds; all dogs had negative results when tested for dog erythrocyte antigen 1.1. Blood was collected into citrate-phosphate-dextrose and stored in AS-1. Stored RBCs were supplemented with 7.1mM ascorbic acid or with saline (0.9% NaCl) solution (control samples). Several biochemical and biophysical properties of RBCs were measured, including percentage hemolysis, oxygen-hemoglobin equilibrium, and the kinetic rate constants for O2 dissociation, carbon monoxide association, and nitric oxide dioxygenation. Greyhound RBCs stored in AS-1 supplemented with ascorbic acid did not have significantly decreased hemolysis, compared with results for the control samples, during the storage period. In this study, ascorbic acid did not reduce hemolysis during storage. Several changes in stored canine RBCs were identified as part of the hypothermic storage lesion.

  2. Effect of Processing Delay and Storage Conditions on Urine Albumin-to-Creatinine Ratio.

    PubMed

    Herrington, William; Illingworth, Nicola; Staplin, Natalie; Kumar, Aishwarya; Storey, Ben; Hrusecka, Renata; Judge, Parminder; Mahmood, Maria; Parish, Sarah; Landray, Martin; Haynes, Richard; Baigent, Colin; Hill, Michael; Clark, Sarah

    2016-10-07

    Because there is substantial biologic intraindividual variation in albumin excretion, randomized trials of albuminuria-reducing therapies may need multiple urine samples to estimate daily urinary albumin excretion. Mailing spot urine samples could offer a convenient and cost-effective method to collect multiple samples, but urine albumin-to-creatinine ratio stability in samples stored at ambient temperatures for several days is unknown. Patients with kidney disease provided fresh urine samples in two tubes (with and without boric acid preservative). Reference aliquots from each participant were analyzed immediately, whereas remaining aliquots were subject to different handling/storage conditions before analysis, including delayed processing for up to 7 days at three different storage temperatures (4°C, 18°C, and 30°C), multiple freeze-thaw cycles, and long-term frozen storage at -80°C, -40°C, and -20°C. We calculated the mean percentage change in urine albumin-to-creatinine ratio for each condition, and we considered samples stable if the 95% confidence interval was within a ±5% threshold. Ninety-three patients provided samples with detectable albuminuria in the reference aliquot. Median (interquartile range) urine albumin-to-creatinine ratio was 87 (20-499) mg/g. The inclusion of preservative had minimal effect on fresh urine albumin-to-creatinine ratio measurements but reduced the changes in albumin and creatinine in samples subject to processing delay and storage conditions. The urine albumin-to-creatinine ratio was stable for 7 days in samples containing preservative at 4°C and 18°C and 2 days when stored at 30°C. It was also stable in samples with preservative after three freeze-thaw cycles and in frozen storage for 6 months at -80°C or -40°C but not at -20°C. Mailed urine samples collected with preservative and received within 7 days if ambient temperature is ≤18°C, or within 2 days if the temperature is higher but does not exceed 30°C, are suitable for the measurement of urine albumin-to-creatinine ratio in randomized trials. Preserved samples frozen to -40°C or -80°C for 6 months before analysis also seem suitable. Copyright © 2016 by the American Society of Nephrology.

  3. Effect of Processing Delay and Storage Conditions on Urine Albumin-to-Creatinine Ratio

    PubMed Central

    Illingworth, Nicola; Staplin, Natalie; Kumar, Aishwarya; Storey, Ben; Hrusecka, Renata; Judge, Parminder; Mahmood, Maria; Parish, Sarah; Landray, Martin; Haynes, Richard; Baigent, Colin; Hill, Michael; Clark, Sarah

    2016-01-01

    Background and objectives Because there is substantial biologic intraindividual variation in albumin excretion, randomized trials of albuminuria-reducing therapies may need multiple urine samples to estimate daily urinary albumin excretion. Mailing spot urine samples could offer a convenient and cost-effective method to collect multiple samples, but urine albumin-to-creatinine ratio stability in samples stored at ambient temperatures for several days is unknown. Design, setting, participants, & measurements Patients with kidney disease provided fresh urine samples in two tubes (with and without boric acid preservative). Reference aliquots from each participant were analyzed immediately, whereas remaining aliquots were subject to different handling/storage conditions before analysis, including delayed processing for up to 7 days at three different storage temperatures (4°C, 18°C, and 30°C), multiple freeze-thaw cycles, and long–term frozen storage at −80°C, −40°C, and −20°C. We calculated the mean percentage change in urine albumin-to-creatinine ratio for each condition, and we considered samples stable if the 95% confidence interval was within a ±5% threshold. Results Ninety-three patients provided samples with detectable albuminuria in the reference aliquot. Median (interquartile range) urine albumin-to-creatinine ratio was 87 (20–499) mg/g. The inclusion of preservative had minimal effect on fresh urine albumin-to-creatinine ratio measurements but reduced the changes in albumin and creatinine in samples subject to processing delay and storage conditions. The urine albumin-to-creatinine ratio was stable for 7 days in samples containing preservative at 4°C and 18°C and 2 days when stored at 30°C. It was also stable in samples with preservative after three freeze-thaw cycles and in frozen storage for 6 months at −80°C or −40°C but not at −20°C. Conclusions Mailed urine samples collected with preservative and received within 7 days if ambient temperature is ≤18°C, or within 2 days if the temperature is higher but does not exceed 30°C, are suitable for the measurement of urine albumin-to-creatinine ratio in randomized trials. Preserved samples frozen to −40°C or −80°C for 6 months before analysis also seem suitable. PMID:27654930

  4. Long-term storage of salivary cortisol samples at room temperature

    NASA Technical Reports Server (NTRS)

    Chen, Yu-Ming; Cintron, Nitza M.; Whitson, Peggy A.

    1992-01-01

    Collection of saliva samples for the measurement of cortisol during space flights provides a simple technique for studying changes in adrenal function due microgravity. In the present work, several methods for preserving saliva cortisol at room temperature were investigated using radioimmunoassays for determining cortisol in saliva samples collected on a saliva-collection device called Salivettes. It was found that a pretreatment of Salivettes with citric acid resulted in preserving more than 85 percent of the salivary cortisol for as long as six weeks. The results correlated well with those for a sample stored in a freezer on an untreated Salivette.

  5. Research and development of a field-ready protocol for sampling of phosgene from stationary source emissions: Diethylamine reagent studies. Research report, 11 July 1995--30 September 1996

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

    Steger, J.L.; Bursey, J.T.; Merrill, R.G.

    1999-03-01

    This report presents the results of laboratory studies to develop and evaluate a method for the sampling and analysis of phosgene from stationary sources of air emissions using diethylamine (DEA) in toluene as the collection media. The method extracts stack gas from emission sources and stabilizes the reactive gas for subsequent analysis. DEA was evaluated both in a benchtop study and in a laboratory train spiking study. This report includes results for both the benchtop study and the train spiking study. Benchtop studies to evaluate the suitability of DEA for collecting and analyzing phosgene investigated five variables: storage time, DEAmore » concentration, moisture/pH, phosgene concentration, and sample storage temperature. Prototype sampling train studies were performed to determine if the benchtop chemical studies were transferable to a Modified Method 5 sampling train collecting phosgene in the presence of clean air mixed with typical stack gas components. Four conditions, which varied the moisture and phosgene spike were evaluated in triplicate. In addition to research results, the report includes a detailed draft method for sampling and analysis of phosgene from stationary source emissions.« less

  6. Determination of regulatory ionophore coccidiostat residues in feedstuffs at carry-over levels by liquid chromatography-mass spectrometry

    PubMed Central

    Stramenga, Arianna; Colagrande, Maria Novella; Campana, Guido; Scortichini, Giampiero; Migliorati, Giacomo; Compagnone, Dario

    2017-01-01

    In this study samples of feedstuffs were collected from different feed mills and animal farms located in central Italy and analyzed for ionophore coccidiostat residues at carry-over levels by liquid chromatography-mass spectrometry. Since unavoidable cross-contamination of feedstuffs may occur during their production as well as distribution and storage, the collection of samples covered all these different stages. Residues of lasalocid, monensin, salinomycin and maduramicin were detected in 32.4% of samples, both at production and storage level. The maximum content for unavoidable carry-over set by Regulation (EU) No 574/2011 was exceeded in 11.3% of samples. The variability of the results highlighted the different approach of each investigated feed business operator to avoid any cross-contamination in non-target feed. The method developed in this study can be able to detect ionophore coccidiostats at low concentrations consequent to carry-over. PMID:28792977

  7. Impact of temperature and time storage on the microbial detection of oral samples by Checkerboard DNA-DNA hybridization method.

    PubMed

    do Nascimento, Cássio; dos Santos, Janine Navarro; Pedrazzi, Vinícius; Pita, Murillo Sucena; Monesi, Nadia; Ribeiro, Ricardo Faria; de Albuquerque, Rubens Ferreira

    2014-01-01

    Molecular diagnosis methods have been largely used in epidemiological or clinical studies to detect and quantify microbial species that may colonize the oral cavity in healthy or disease. The preservation of genetic material from samples remains the major challenge to ensure the feasibility of these methodologies. Long-term storage may compromise the final result. The aim of this study was to evaluate the effect of temperature and time storage on the microbial detection of oral samples by Checkerboard DNA-DNA hybridization. Saliva and supragingival biofilm were taken from 10 healthy subjects, aliquoted (n=364) and processed according to proposed protocols: immediate processing and processed after 2 or 4 weeks, and 6 or 12 months of storage at 4°C, -20°C and -80°C. Either total or individual microbial counts were recorded in lower values for samples processed after 12 months of storage, irrespective of temperatures tested. Samples stored up to 6 months at cold temperatures showed similar counts to those immediately processed. The microbial incidence was also significantly reduced in samples stored during 12 months in all temperatures. Temperature and time of oral samples storage have relevant impact in the detection and quantification of bacterial and fungal species by Checkerboard DNA-DNA hybridization method. Samples should be processed immediately after collection or up to 6 months if conserved at cold temperatures to avoid false-negative results. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. Collection and storage of urine specimens for measurement of urolithiasis risk factors.

    PubMed

    Wu, Wenqi; Yang, Dong; Tiselius, Hans-Goran; Ou, Lili; Mai, Zanlin; Chen, Kang; Zhu, Hanliang; Xu, Shaohong; Zhao, Zhijian; Zeng, Guohua

    2015-02-01

    To evaluate how different methods for storage and preservation of urine samples affected the outcome of analysis of risk factors for stone formation. Spot urine samples were collected from 21 healthy volunteers. Each fresh urine sample was divided into ten 10-mL aliquots: 2 without preservative, 2 with thymol, 2 with toluene, 2 with hydrochloric acid (HCl), and 2 with sodium azide. One sample of each pair was stored at 4 °C and the other at room temperature. The concentrations of calcium, magnesium, sodium, phosphate, urate, oxalate, citrate, and pH in each urine sample were analyzed immediately after collection (0 hour) and after 24 and 48 hours. There were no significant differences in calcium, oxalate, magnesium, phosphate, sodium, urate or pH (without acidification) between samples with different preservation methods (P >.05). Urinary citrate, however, was significantly lower in the urine collected with HCl than when other preservatives were used, both at room temperature and at 4 °C. Urine pH was significantly higher after 48 hours than after 24 hours, whether the samples were stored at room temperature or at 4 °C. Antibacterial preservatives (eg, thymol or toluene) can be recommended as preservatives for 24-hour urine collections. Ideally, the samples should be stored at 4 °C. When HCl is used as a preservative, it seems essential to neutralize the samples before analysis. This is particularly obvious with the chromatographic method used for analysis of citrate that was used in this study. Copyright © 2015 Elsevier Inc. All rights reserved.

  9. Effect of short-term room temperature storage on the microbial community in infant fecal samples.

    PubMed

    Guo, Yong; Li, Sheng-Hui; Kuang, Ya-Shu; He, Jian-Rong; Lu, Jin-Hua; Luo, Bei-Jun; Jiang, Feng-Ju; Liu, Yao-Zhong; Papasian, Christopher J; Xia, Hui-Min; Deng, Hong-Wen; Qiu, Xiu

    2016-05-26

    Sample storage conditions are important for unbiased analysis of microbial communities in metagenomic studies. Specifically, for infant gut microbiota studies, stool specimens are often exposed to room temperature (RT) conditions prior to analysis. This could lead to variations in structural and quantitative assessment of bacterial communities. To estimate such effects of RT storage, we collected feces from 29 healthy infants (0-3 months) and partitioned each sample into 5 portions to be stored for different lengths of time at RT before freezing at -80 °C. Alpha diversity did not differ between samples with storage time from 0 to 2 hours. The UniFrac distances and microbial composition analysis showed significant differences by testing among individuals, but not by testing between different time points at RT. Changes in the relative abundance of some specific (less common, minor) taxa were still found during storage at room temperature. Our results support previous studies in children and adults, and provided useful information for accurate characterization of infant gut microbiomes. In particular, our study furnished a solid foundation and justification for using fecal samples exposed to RT for less than 2 hours for comparative analyses between various medical conditions.

  10. Effect of sample storage temperature and buffer formulation on faecal immunochemical test haemoglobin measurements.

    PubMed

    Symonds, Erin L; Cole, Stephen R; Bastin, Dawn; Fraser, Robert Jl; Young, Graeme P

    2017-12-01

    Objectives Faecal immunochemical test accuracy may be adversely affected when samples are exposed to high temperatures. This study evaluated the effect of two sample collection buffer formulations (OC-Sensor, Eiken) and storage temperatures on faecal haemoglobin readings. Methods Faecal immunochemical test samples returned in a screening programme and with ≥10 µg Hb/g faeces in either the original or new formulation haemoglobin stabilizing buffer were stored in the freezer, refrigerator, or at room temperature (22℃-24℃), and reanalysed after 1-14 days. Samples in the new buffer were also reanalysed after storage at 35℃ and 50℃. Results were expressed as percentage of the initial concentration, and the number of days that levels were maintained to at least 80% was calculated. Results Haemoglobin concentrations were maintained above 80% of their initial concentration with both freezer and refrigerator storage, regardless of buffer formulation or storage duration. Stability at room temperature was significantly better in the new buffer, with haemoglobin remaining above 80% for 20 days compared with six days in the original buffer. Storage at 35℃ or 50℃ in the new buffer maintained haemoglobin above 80% for eight and two days, respectively. Conclusion The new formulation buffer has enhanced haemoglobin stabilizing properties when samples are exposed to temperatures greater than 22℃.

  11. The effect of extraction, storage, and analysis techniques on the measurement of airborne endotoxin from a large dairy

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to fill in additional knowledge gaps with respect to the extraction, storage, and analysis of airborne endotoxin, with a specific focus on samples from a dairy production facility. We utilized polycarbonate filters to collect total airborne endotoxins, sonication as ...

  12. Biobanking human endometrial tissue and blood specimens: standard operating procedure and importance to reproductive biology research and diagnostic development.

    PubMed

    Sheldon, Elizabeth; Vo, Kim Chi; McIntire, Ramsey A; Aghajanova, Lusine; Zelenko, Zara; Irwin, Juan C; Giudice, Linda C

    2011-05-01

    To develop a standard operating procedure (SOP) for collection, transport, storage of human endometrial tissue and blood samples, subject and specimen annotation, and establishing sample priorities. The SOP synthesizes sound scientific procedures, the literature on ischemia research, sample collection and gene expression profiling, good laboratory practices, and the authors' experience of workflow and sample quality. The National Institutes of Health, University of California, San Francisco, Human Endometrial Tissue and DNA Bank. Women undergoing endometrial biopsy or hysterectomy for nonmalignant indications. Collecting, processing, storing, distributing endometrial tissue and blood samples under approved institutional review board protocols and written informed consent from participating subjects. Standard operating procedure. The SOP addresses rigorous and consistent subject annotation, specimen processing and characterization, strict regulatory compliance, and a reference for researchers to track collection and storage times that may influence their research. The comprehensive and systematic approach to the procurement of human blood and endometrial tissue in this SOP ensures the high quality, reliability, and scientific usefulness of biospecimens made available to investigators by the National Institutes of Health, University of California, San Francisco, Human Endometrial Tissue and DNA Bank. The detail and perspective in this SOP also provides a blueprint for implementation of similar collection programs at other institutions. Copyright © 2011 American Society for Reproductive Medicine. Published by Elsevier Inc. All rights reserved.

  13. Standard operating procedures for pre-analytical handling of blood and urine for metabolomic studies and biobanks.

    PubMed

    Bernini, Patrizia; Bertini, Ivano; Luchinat, Claudio; Nincheri, Paola; Staderini, Samuele; Turano, Paola

    2011-04-01

    (1)H NMR metabolic profiling of urine, serum and plasma has been used to monitor the impact of the pre-analytical steps on the sample quality and stability in order to propose standard operating procedures (SOPs) for deposition in biobanks. We analyzed the quality of serum and plasma samples as a function of the elapsed time (t = 0-4 h) between blood collection and processing and of the time from processing to freezing (up to 24 h). The stability of the urine metabolic profile over time (up to 24 h) at various storage temperatures was monitored as a function of the different pre-analytical treatments like pre-storage centrifugation, filtration, and addition of the bacteriostatic preservative sodium azide. Appreciable changes in the profiles, reflecting changes in the concentration of a number of metabolites, were detected and discussed in terms of chemical and enzymatic reactions for both blood and urine samples. Appropriate procedures for blood derivatives collection and urine preservation/storage that allow maintaining as much as possible the original metabolic profile of the fresh samples emerge, and are proposed as SOPs for biobanking.

  14. Evaluation of epididymis storage temperature and cryopreservation conditions for improved mitochondrial membrane potential, membrane integrity, sperm motility and in vitro fertilization in bovine epididymal sperm.

    PubMed

    Nichi, M; Rijsselaere, T; Losano, Jda; Angrimani, Dsr; Kawai, Gkv; Goovaerts, Igf; Van Soom, A; Barnabe, V H; De Clercq, Jbp; Bols, Pej

    2017-04-01

    The maintaining of the epididymis at lower temperatures during storage and transport improves sperm quality. Our study aimed to test whether epididymis storage temperature (post-mortem) and sperm cryopreservation affect sperm kinetics, membrane integrity, mitochondrial potential and fertility capacity. Thirty-six epididymides were collected from 18 bulls after slaughter and divided into two groups: at 4 or 34°C for 2-3 hr. The sperm was collected from the epididymis cauda. The evaluation consisted of computer-assisted sperm analysis (CASA), SYBR14/PI/JC1 to evaluate membrane integrity, mitochondrial membrane potential (MMP) and measurement of lipid peroxidation (TBARS). The sperm was then frozen using an automatic device. After thawing, sperm samples were evaluated by the same variables and further in vitro fertilization rates. Cryopreservation negatively affected sperm motility in samples stored at 4 and 34°C. Nevertheless, the 4°C samples yielded higher rates of blastocyst formation. Pre-freeze sperm motility, progressive motility and velocity were higher in sperm from epididymis stored at 4°C while post-thaw sperm motility, progressive motility and velocity remained the same among samples from epididymis stored at 4 or 34°C. However, with regard to the kinetic patterns, samples collected from epididymis stored at 34°C had lower values when compared to those stored at 4°C prior the cryopreservation process. Our results indicate that epididymis handling conditions after cryopreservation may affect sperm quality after thawing, especially due to compromised MMP in sperm collected from epididymis stored at higher temperatures. © 2016 Blackwell Verlag GmbH.

  15. Individualization of pubic hair bacterial communities and the effects of storage time and temperature.

    PubMed

    Williams, Diana W; Gibson, Greg

    2017-01-01

    A potential application of microbial genetics in forensic science is detection of transfer of the pubic hair microbiome between individuals during sexual intercourse using high-throughput sequencing. In addition to the primary need to show whether the pubic hair microbiome is individualizing, one aspect that must be addressed before using the microbiome in criminal casework involves the impact of storage on the microbiome of samples recovered for forensic testing. To test the effects of short-term storage, pubic hair samples were collected from volunteers and stored at room temperature (∼20°C), refrigerated (4°C), and frozen (-20°C) for 1 week, 2 weeks, 4 weeks, and 6 weeks along with a baseline sample. Individual microbial profiles (R 2 =0.69) and gender (R 2 =0.17) were the greatest sources of variation between samples. Because of this variation, individual and gender could be predicted using Random Forests supervised classification in this sample set with an overall error rate of 2.7%± 5.8% and 1.7%±5.2%, respectively. There was no statistically significant difference attributable to time of sampling or temperature of storage within individuals. Further work on larger sample sets will quantify the temporal consistency of individual profiles and define whether it is plausible to detect transfer between sexual partners. For short-term storage (≤6 weeks), recovery of the microbiome was not affected significantly by either storage time or temperature, suggesting that investigators and crime laboratories can use existing evidence storage methods. Published by Elsevier Ireland Ltd.

  16. Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping.

    PubMed

    Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A; Debelius, Justine; Morton, James T; Hyde, Embriette; Robbins-Pianka, Adam; Knight, Rob

    2017-01-01

    The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria , grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., "bloomed") at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called "Deblur" to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has been shown not to obscure physiologically relevant microbiome differences between comparison groups, there are still changes in specific bacterial taxa, notably, in members of the class Gammaproteobacteria , that can make microbiome profiles difficult to interpret. Here we identify the most problematic taxa and show that removing sequences from just a few fast-growing taxa is sufficient to correct microbiome profiles.

  17. Correcting for Microbial Blooms in Fecal Samples during Room-Temperature Shipping

    PubMed Central

    Amir, Amnon; McDonald, Daniel; Navas-Molina, Jose A.; Debelius, Justine; Morton, James T.; Hyde, Embriette; Robbins-Pianka, Adam

    2017-01-01

    ABSTRACT The use of sterile swabs is a convenient and common way to collect microbiome samples, and many studies have shown that the effects of room-temperature storage are smaller than physiologically relevant differences between subjects. However, several bacterial taxa, notably members of the class Gammaproteobacteria, grow at room temperature, sometimes confusing microbiome results, particularly when stability is assumed. Although comparative benchmarking has shown that several preservation methods, including the use of 95% ethanol, fecal occult blood test (FOBT) and FTA cards, and Omnigene-GUT kits, reduce changes in taxon abundance during room-temperature storage, these techniques all have drawbacks and cannot be applied retrospectively to samples that have already been collected. Here we performed a meta-analysis using several different microbiome sample storage condition studies, showing consistent trends in which specific bacteria grew (i.e., “bloomed”) at room temperature, and introduce a procedure for removing the sequences that most distort analyses. In contrast to similarity-based clustering using operational taxonomic units (OTUs), we use a new technique called “Deblur” to identify the exact sequences corresponding to blooming taxa, greatly reducing false positives and also dramatically decreasing runtime. We show that applying this technique to samples collected for the American Gut Project (AGP), for which participants simply mail samples back without the use of ice packs or other preservatives, yields results consistent with published microbiome studies performed with frozen or otherwise preserved samples. IMPORTANCE In many microbiome studies, the necessity to store samples at room temperature (i.e., remote fieldwork) and the ability to ship samples without hazardous materials that require special handling training, such as ethanol (i.e., citizen science efforts), is paramount. However, although room-temperature storage for a few days has been shown not to obscure physiologically relevant microbiome differences between comparison groups, there are still changes in specific bacterial taxa, notably, in members of the class Gammaproteobacteria, that can make microbiome profiles difficult to interpret. Here we identify the most problematic taxa and show that removing sequences from just a few fast-growing taxa is sufficient to correct microbiome profiles. PMID:28289733

  18. Optimization of HPV DNA detection in urine by improving collection, storage, and extraction.

    PubMed

    Vorsters, A; Van den Bergh, J; Micalessi, I; Biesmans, S; Bogers, J; Hens, A; De Coster, I; Ieven, M; Van Damme, P

    2014-11-01

    The benefits of using urine for the detection of human papillomavirus (HPV) DNA have been evaluated in disease surveillance, epidemiological studies, and screening for cervical cancers in specific subgroups. HPV DNA testing in urine is being considered for important purposes, notably the monitoring of HPV vaccination in adolescent girls and young women who do not wish to have a vaginal examination. The need to optimize and standardize sampling, storage, and processing has been reported.In this paper, we examined the impact of a DNA-conservation buffer, the extraction method, and urine sampling on the detection of HPV DNA and human DNA in urine provided by 44 women with a cytologically normal but HPV DNA-positive cervical sample. Ten women provided first-void and midstream urine samples. DNA analysis was performed using real-time PCR to allow quantification of HPV and human DNA.The results showed that an optimized method for HPV DNA detection in urine should (a) prevent DNA degradation during extraction and storage, (b) recover cell-free HPV DNA in addition to cell-associated DNA, (c) process a sufficient volume of urine, and (d) use a first-void sample.In addition, we found that detectable human DNA in urine may not be a good internal control for sample validity. HPV prevalence data that are based on urine samples collected, stored, and/or processed under suboptimal conditions may underestimate infection rates.

  19. Hydrologic and Geochemical Evaluation of Aquifer Storage Recovery in the Santee Limestone/Black Mingo Aquifer, Charleston, South Carolina, 1998-2002

    USGS Publications Warehouse

    Petkewich, Matthew D.; Parkhurst, David L.; Conlon, Kevin J.; Campbell, Bruce G.; Mirecki, June E.

    2004-01-01

    The hydrologic and geochemical effects of aquifer storage recovery were evaluated to determine the potential for supplying the city of Charleston, South Carolina, with large quantities of potable water during emergencies, such as earthquakes, hurricanes, or hard freezes. An aquifer storage recovery system, including a production well and three observation wells, was installed at a site located on the Charleston peninsula. The focus of this study was the 23.2-meter thick Tertiary-age carbonate and sand aquifer of the Santee Limestone and the Black Mingo Group, the northernmost equivalent of the Floridan aquifer system. Four cycles of injection, storage, and recovery were conducted between October 1999 and February 2002. Each cycle consisted of injecting between 6.90 and 7.19 million liters of water for storage periods of 1, 3, or 6 months. The volume of recovered water that did not exceed the U.S. Environmental Protection Agency secondary standard for chloride (250 milligrams per liter) varied from 1.48 to 2.46 million liters, which is equivalent to 21 and 34 percent of the total volume injected for the individual tests. Aquifer storage recovery testing occurred within two productive zones of the brackish Santee Limestone/Black Mingo aquifer. The individual productive zones were determined to be approximately 2 to 4 meters thick, based on borehole geophysical logs, electromagnetic flow-meter testing, and specific-conductance profiles collected within the observation wells. A transmissivity and storage coefficient of 37 meters squared per day and 3 x 10-5, respectively, were determined for the Santee Limestone/Black Mingo aquifer. Water-quality and sediment samples collected during this investigation documented baseline aquifer and injected water quality, aquifer matrix composition, and changes in injected/aquifer water quality during injection, storage, and recovery. A total of 193 water-quality samples were collected and analyzed for physical properties, major and minor ions, and nutrients. The aquifer and treated surface water were sodiumchloride and calcium/sodium-bicarbonate water types, respectively. Forty-five samples were collected and analyzed for total trihalomethane. Total trihalomethane data collected during aquifer storage recovery cycle 4 indicated that this constituent would not restrict the use of recovered water for drinking-water purposes. Analysis of six sediment samples collected from a cored well located near the aquifer storage recovery site showed that quartz and calcite were the dominant minerals in the Santee Limestone/Black Mingo aquifer. Estimated cation exchange capacity ranged from 12 to 36 milliequivalents per 100 grams in the lower section of the aquifer. A reactive transport model was developed that included two 2-meter thick layers to describe each of the production zones. The four layers composing the production zones were assigned porosities ranging from 0.1 to 0.3 and hydraulic conductivities ranging from 1 to 8.4 meters per day. Specific storage of the aquifer and confining units was estimated to be 1.5 x 10-5 meter-1. Longitudinal dispersivity of all layers was specified to be 0.5 meter. Leakage through the confining unit was estimated to be minimal and, therefore, not used in the reactive transport modeling. Inverse geochemical modeling indicates that mixing, cation exchange, and calcite dissolution are the dominant reactions that occur during aquifer storage recovery testing in the Santee Limestone/Black Mingo aquifer. Potable water injected into the Santee Limestone/Black Mingo aquifer evolved chemically by mixing with brackish background water and reaction with calcite and cation exchangers in the sediment. Reactive-transport model simulations indicated that the calcite and exchange reactions could be treated as equilibrium processes. Simulations with the calibrated reactive transport model indicated that approximately one-fourth of the total volume of water injected into

  20. Report of Operation FITZWILLIAM. Volume 1, Design of Operation and Summary of Results (REDACTED)

    DTIC Science & Technology

    1948-01-01

    storage tanks {400 lbs/sq in) to pel’mit the collection o£ sampleD or gas in the vicinity of the radio- active clouc1. P~dicective LUlC.lysis of -~he gas...Corps Furnish ground dust sampling units and wrap-around countera. 4. Navy Naval :Research Lab. (a) FW:-nish ground dust sampling units...direct as necessar,r the collection or air• craft filters and gaseou.s samples trca aircraft based at Km.falem. (6) Vector Destro,.er-M:lne...Swee

  1. The Organization of a Computer Software Collection Using an Information Storage and Retrieval Software Package.

    ERIC Educational Resources Information Center

    Davies, Denise M.

    1985-01-01

    Discusses design, development, and use of a database to provide organization and access to a computer software collection at the University of Hawaii School of Library Studies. Field specifications, samples of report forms, and a description of the physical organization of the software collection are included. (MBR)

  2. Storage mite contamination of commercial dry dog food in south-eastern Australia.

    PubMed

    Hibberson, C E; Vogelnest, L J

    2014-06-01

    To evaluate contamination of unopened and opened stored sources of commercial dry dog food by viable storage mites. Prospective laboratory and field study. Samples were collected from nine brands of previously unopened bags (new bags) of dry food and 20 field sources of stored dry food in homes in Sydney and Canberra, Australia. All samples were initially examined for the presence of mites using a stereo-binocular microscope and then placed in separate filter-paper-sealed containers. Field samples were incubated at an average temperature of 29°C and 78% relative humidity (RH) for 5 weeks and then at average 26°C/83% RH for 8 weeks. Paired new-bag samples were stored under room conditions (average 23°C/47% RH) and controlled incubator conditions (average 26°C/80% RH) for 6 weeks. All samples were thoroughly examined for mites, mite eggs and visible mould once weekly using a stereo-binocular microscope. Storage mites were not visualised in any of the field samples or in new-bag samples stored at room temperature. Storage mites, identified as Tyrophagus putrescentiae, were visualised in increasing numbers in seven of nine new-bag samples after incubation, with first mites and then eggs evident after 3 weeks of incubation. We confirmed the presence of viable storage mites in a range of previously unopened commercial dry dog foods in Australia and confirmed the possibility of heavy storage mite contamination for dry food stored under conditions of moderate temperature and high humidity. These findings have relevance to storage mite and/or dust mite sensitivity in canine atopic dermatitis. © 2014 Australian Veterinary Association.

  3. NHEXAS PHASE I ARIZONA STUDY--STANDARD OPERATING PROCEDURE FOR COLLECTION, STORAGE AND SHIPMENT OF DRINKING AND TAP WATER SAMPLES FOR PESTICIDES (EPA 225.2) AND CARBARYL (EPA 531.1) (UA-F-17.1)

    EPA Science Inventory

    The purpose of this SOP is to describe how to collect, store, and ship tap and drinking water samples for analysis by EPA Method 525.2 (revision 1.0) and EPA method 531.1 (revision 3) for the NHEXAS Arizona project. This SOP provides a brief description of the sample containers,...

  4. Effect of room temperature transport vials on DNA quality and phylogenetic composition of faecal microbiota of elderly adults and infants.

    PubMed

    Hill, Cian J; Brown, Jillian R M; Lynch, Denise B; Jeffery, Ian B; Ryan, C Anthony; Ross, R Paul; Stanton, Catherine; O'Toole, Paul W

    2016-05-10

    Alterations in intestinal microbiota have been correlated with a growing number of diseases. Investigating the faecal microbiota is widely used as a non-invasive and ethically simple proxy for intestinal biopsies. There is an urgent need for collection and transport media that would allow faecal sampling at distance from the processing laboratory, obviating the need for same-day DNA extraction recommended by previous studies of freezing and processing methods for stool. We compared the faecal bacterial DNA quality and apparent phylogenetic composition derived using a commercial kit for stool storage and transport (DNA Genotek OMNIgene GUT) with that of freshly extracted samples, 22 from infants and 20 from older adults. Use of the storage vials increased the quality of extracted bacterial DNA by reduction of DNA shearing. When infant and elderly datasets were examined separately, no differences in microbiota composition were observed due to storage. When the two datasets were combined, there was a difference according to a Wilcoxon test in the relative proportions of Faecalibacterium, Sporobacter, Clostridium XVIII, and Clostridium XlVa after 1 week's storage compared to immediately extracted samples. After 2 weeks' storage, Bacteroides abundance was also significantly different, showing an apparent increase from week 1 to week 2. The microbiota composition of infant samples was more affected than that of elderly samples by storage, with significantly higher Spearman distances between paired freshly extracted and stored samples (p < 0.001). When the microbiota profiles were analysed at the operational taxonomic unit (OTU) level, three infant datasets in the study did not cluster together, while only one elderly dataset did not. The lower microbiota diversity of the infant gut microbiota compared to the elderly gut microbiota (p < 0.001) means that any alteration in the infant datasets has a proportionally larger effect. The commercial storage vials appear to be suitable for high diversity microbiota samples, but may be less appropriate for lower diversity samples. Differences between fresh and stored samples mean that where storage is unavoidable, a consistent storage regime should be used. We would recommend extraction ideally within the first week of storage.

  5. An Information Storage and Retrieval System for Biological and Geological Data. Interim Report.

    ERIC Educational Resources Information Center

    Squires, Donald F.

    A project is being conducted to test the feasibility of an information storage and retrieval system for museum specimen data, particularly for natural history museums. A pilot data processing system has been developed, with the specimen records from the national collections of birds, marine crustaceans, and rocks used as sample data. The research…

  6. Monitoring of occupational exposure to methylene chloride: sampling protocol and stability of urine samples.

    PubMed

    Hoffer, Erica; Tabak, Arek; Shcherb, Inna; Wiener, Avi; Bentur, Yedidia

    2005-01-01

    A sampling protocol for biomonitoring of the volatile solvent methylene chloride (MeCl(2)) by analysis of urine from exposed workers was established. Storage temperature, sample volume in headspace vial (HSV), and time to sealing HSV on determination of MeCl(2) in urine were evaluated. MeCl(2) was analyzed by a solid-phase microextraction technique combined with gas chromatography. Volume of urine in HSV has no effect on MeCl(2) analysis. Delays of 30 and 60 min from collection of urine until sealing the HSV caused 14.47 +/- 6.98% and 26.17 +/- 9.57% decreases from baseline concentration, respectively. MeCl(2) concentration in spiked urine samples stored in sealed HSVs decreased on day 2 and then remained stable for 2 weeks. Refrigeration did not improve recovery although it seems to be associated with less variability. MeCl(2) in urine samples of seven exposed workers was in the range of 0.02-0.06 mg/L. Sampling of MeCl(2)-containing urine should include collection of urine in closed plastic bottles, transfer to HSV within 15 min, sealing and clamping of HSV within 15 s, and storage of HSV in refrigeration until analysis, but no longer than 2 weeks. Standard samples should be prepared on the day of test sample collection and handled under the same conditions.

  7. Working Up a Good Sweat – The Challenges of Standardising Sweat Collection for Metabolomics Analysis

    PubMed Central

    Hussain, Joy N; Mantri, Nitin; Cohen, Marc M

    2017-01-01

    Introduction Human sweat is a complex biofluid of interest to diverse scientific fields. Metabolomics analysis of sweat promises to improve screening, diagnosis and self-monitoring of numerous conditions through new applications and greater personalisation of medical interventions. Before these applications can be fully developed, existing methods for the collection, handling, processing and storage of human sweat need to be revised. This review presents a cross-disciplinary overview of the origins, composition, physical characteristics and functional roles of human sweat, and explores the factors involved in standardising sweat collection for metabolomics analysis. Methods A literature review of human sweat analysis over the past 10 years (2006–2016) was performed to identify studies with metabolomics or similarly applicable ‘omics’ analysis. These studies were reviewed with attention to sweat induction and sampling techniques, timing of sweat collection, sweat storage conditions, laboratory derivation, processing and analytical platforms. Results Comparative analysis of 20 studies revealed numerous factors that can significantly impact the validity, reliability and reproducibility of sweat analysis including: anatomical site of sweat sampling, skin integrity and preparation; temperature and humidity at the sweat collection sites; timing and nature of sweat collection; metabolic quenching; transport and storage; qualitative and quantitative measurements of the skin microbiota at sweat collection sites; and individual variables such as diet, emotional state, metabolic conditions, pharmaceutical, recreational drug and supplement use. Conclusion Further development of standard operating protocols for human sweat collection can open the way for sweat metabolomics to significantly add to our understanding of human physiology in health and disease. PMID:28798503

  8. The NASA Ames Life Sciences Data Archive: Biobanking for the Final Frontier

    NASA Technical Reports Server (NTRS)

    Rask, Jon; Chakravarty, Kaushik; French, Alison J.; Choi, Sungshin; Stewart, Helen J.

    2017-01-01

    The NASA Ames Institutional Scientific Collection involves the Ames Life Sciences Data Archive (ALSDA) and a biospecimen repository, which are responsible for archiving information and non-human biospecimens collected from spaceflight and matching ground control experiments. The ALSDA also manages a biospecimen sharing program, performs curation and long-term storage operations, and facilitates distribution of biospecimens for research purposes via a public website (https:lsda.jsc.nasa.gov). As part of our best practices, a tissue viability testing plan has been developed for the repository, which will assess the quality of samples subjected to long-term storage. We expect that the test results will confirm usability of the samples, enable broader science community interest, and verify operational efficiency of the archives. This work will also support NASA open science initiatives and guides development of NASA directives and policy for curation of biological collections.

  9. Laboratory evaluation of sample collection methods (organs vs swabs) for Tasmanian salmon reovirus detection in farmed Atlantic salmon, Salmo salar L.

    PubMed

    Zainathan, S C; Carson, J; Crane, M St J; Nowak, B F

    2013-04-01

    The use of swabs relative to organs as a sample collection method for the detection of Tasmanian salmon reovirus (TSRV) in farmed Tasmanian Atlantic salmon, Salmo salar L., was evaluated by RT-qPCR. Evaluation of individual and pooled sample collection (organs vs swabs) was carried out to determine the sensitivity of the collection methods and the effect of pooling of samples for the detection of TSRV. Detection of TSRV in individual samples was as sensitive when organs were sampled compared to swabs, and in pooled samples, organs demonstrated a sensitivity of one 10-fold dilution higher than sampling of pooled swabs. Storage of swabs at 4 °C for t = 24 h demonstrated results similar to those at t = 0. Advantages of using swabs as a preferred sample collection method for the detection of TSRV compared to organ samples are evident from these experimental trials. © 2012 Blackwell Publishing Ltd.

  10. Efficacy of sanitized ice in reducing bacterial load on fish fillet and in the water collected from the melted ice.

    PubMed

    Feliciano, Lizanel; Lee, Jaesung; Lopes, John A; Pascall, Melvin A

    2010-05-01

    This study investigated the efficacy of sanitized ice for the reduction of bacteria in the water collected from the ice that melted during storage of whole and filleted Tilapia fish. Also, bacterial reductions on the fish fillets were investigated. The sanitized ice was prepared by freezing solutions of PRO-SAN (an organic acid formulation) and neutral electrolyzed water (NEW). For the whole fish study, the survival of the natural microflora was determined from the water of the melted ice prepared with PRO-SAN and tap water. These water samples were collected during an 8 h storage period. For the fish fillet study, samples were inoculated with Escherichia coli K12, Listeria innocua, and Pseudomonas putida then stored on crushed sanitized ice. The efficacies of these were tested by enumerating each bacterial species on the fish fillet and in the water samples at 12 and 24 h intervals for 72 h, respectively. Results showed that each bacterial population was reduced during the test. However, a bacterial reduction of < 1 log CFU was obtained for the fillet samples. A maximum of approximately 2 log CFU and > 3 log CFU reductions were obtained in the waters sampled after the storage of whole fish and the fillets, respectively. These reductions were significantly (P < 0.05) higher in the water from sanitized ice when compared with the water from the unsanitized melted ice. These results showed that the organic acid formulation and NEW considerably reduced the bacterial numbers in the melted ice and thus reduced the potential for cross-contamination.

  11. Handling and storage of human body fluids for analysis of extracellular vesicles

    PubMed Central

    Yuana, Yuana; Böing, Anita N.; Grootemaat, Anita E.; van der Pol, Edwin; Hau, Chi M.; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk

    2015-01-01

    Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze–thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze–thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100–200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze–thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories. PMID:26563735

  12. Handling and storage of human body fluids for analysis of extracellular vesicles.

    PubMed

    Yuana, Yuana; Böing, Anita N; Grootemaat, Anita E; van der Pol, Edwin; Hau, Chi M; Cizmar, Petr; Buhr, Egbert; Sturk, Auguste; Nieuwland, Rienk

    2015-01-01

    Because procedures of handling and storage of body fluids affect numbers and composition of extracellular vesicles (EVs), standardization is important to ensure reliable and comparable measurements of EVs in a clinical environment. We aimed to develop standard protocols for handling and storage of human body fluids for EV analysis. Conditions such as centrifugation, single freeze-thaw cycle, effect of time delay between blood collection and plasma preparation and storage were investigated. Plasma is the most commonly studied body fluid in EV research. We mainly focused on EVs originating from platelets and erythrocytes and investigated the behaviour of these 2 types of EVs independently as well as in plasma samples of healthy subjects. EVs in urine and saliva were also studied for comparison. All samples were analysed simultaneously before and after freeze-thawing by resistive pulse sensing, nanoparticle tracking analysis, conventional flow cytometry (FCM) and transmission (scanning) electron microscopy. Our main finding is that the effect of centrifugation markedly depends on the cellular origin of EVs. Whereas erythrocyte EVs remain present as single EVs after centrifugation, platelet EVs form aggregates, which affect their measured concentration in plasma. Single erythrocyte and platelet EVs are present mainly in the range of 100-200 nm, far below the lower limit of what can be measured by conventional FCM. Furthermore, the effects of single freeze-thaw cycle, time delay between blood collection and plasma preparation up to 1 hour and storage up to 1 year are insignificant (p>0.05) on the measured concentration and diameter of EVs from erythrocyte and platelet concentrates and EVs in plasma, urine and saliva. In conclusion, in standard protocols for EV studies, centrifugation to isolate EVs from collected body fluids should be avoided. Freezing and storage of collected body fluids, albeit their insignificant effects, should be performed identically for comparative EV studies and to create reliable biorepositories.

  13. Quality of groundwater at and near an aquifer storage and recovery site, Bexar, Atascosa, and Wilson Counties, Texas, June 2004-August 2008

    USGS Publications Warehouse

    Otero, Cassi L.; Petri, Brian L.

    2010-01-01

    The U.S. Geological Survey, in cooperation with the San Antonio Water System, did a study during 2004-08 to characterize the quality of native groundwater from the Edwards aquifer and pre- and post-injection water from the Carrizo aquifer at and near an aquifer storage and recovery (ASR) site in Bexar, Atascosa, and Wilson Counties, Texas. Groundwater samples were collected and analyzed for selected physical properties and constituents to characterize the quality of native groundwater from the Edwards aquifer and pre- and post-injection water from the Carrizo aquifer at and near the ASR site. Geochemical and isotope data indicated no substantial changes in major-ion, trace-element, and isotope chemistry occurred as the water from the Edwards aquifer was transferred through a 38-mile pipeline to the aquifer storage and recovery site. The samples collected from the four ASR recovery wells were similar in major-ion and stable isotope chemistry compared to the samples collected from the Edwards aquifer source wells and the ASR injection well. The similarity could indicate that as Edwards aquifer water was injected, it displaced native Carrizo aquifer water, or, alternatively, if mixing of Edwards and Carrizo aquifer waters was occurring, the major-ion and stable isotope signatures for the Carrizo aquifer water might have been obscured by the signatures of the injected Edwards aquifer water. Differences in the dissolved iron and dissolved manganese concentrations indicate that either minor amounts of mixing occurred between the waters from the two aquifers, or as Edwards aquifer water displaced Carrizo aquifer water it dissolved the iron and manganese directly from the Carrizo Sand. Concentrations of radium-226 in the samples collected at the ASR recovery wells were smaller than the concentrations in samples collected from the Edwards aquifer source wells and from the ASR injection well. The smaller radium-226 concentrations in the samples collected from the ASR recovery wells likely indicate some degree of mixing of the two waters occurred rather than continued decay of radium-226 in the injected water. Geochemical and isotope data measured in samples collected in May 2005 from two Carrizo aquifer monitoring wells and in July 2008 from the three ASR production-only wells in the northern section of the ASR site indicate that injected Edwards aquifer water had not migrated to these five sites. Geochemical and isotope data measured in samples collected from Carrizo aquifer wells in 2004, 2005, and 2008 were graphically analyzed to determine if changes in chemistry could be detected. Major-ion, trace element, and isotope chemistry varied spatially in the samples collected from the Carrizo aquifer. With the exception of a few samples, major-ion concentrations measured in samples collected in Carrizo aquifer wells in 2004, 2005, and 2008 were similar. A slightly larger sulfate con-centration and a slightly smaller bicarbonate concentration were measured in samples collected in 2005 and 2008 from well NC1 compared to samples collected at well NC1 in 2004. Larger sodium concentrations and smaller calcium, magnesium, bicarbonate, and sulfate concentrations were measured in samples collected in 2008 from well WC1 than in samples collected at this well in 2004 and 2005. Larger calcium and magnesium concentrations and a smaller sodium concentration were measured in the samples collected in 2008 at well EC2 compared to samples collected at this well in 2004 and 2005. While in some cases the computed percent differences (compared to concentrations from June 2004) in dissolved iron and dissolved manganese concentrations in 11 wells sampled in the Carrizo aquifer in 2005 and 2008 were quite large, no trends that might have been caused by migration of injected Edwards aquifer water were observed. Because of the natural variation in geochemical data in the Carrizo aquifer and the small data set collected for this study, differences in major-ion and

  14. Vintage venoms: proteomic and pharmacological stability of snake venoms stored for up to eight decades.

    PubMed

    Jesupret, Clémence; Baumann, Kate; Jackson, Timothy N W; Ali, Syed Abid; Yang, Daryl C; Greisman, Laura; Kern, Larissa; Steuten, Jessica; Jouiaei, Mahdokht; Casewell, Nicholas R; Undheim, Eivind A B; Koludarov, Ivan; Debono, Jordan; Low, Dolyce H W; Rossi, Sarah; Panagides, Nadya; Winter, Kelly; Ignjatovic, Vera; Summerhayes, Robyn; Jones, Alun; Nouwens, Amanda; Dunstan, Nathan; Hodgson, Wayne C; Winkel, Kenneth D; Monagle, Paul; Fry, Bryan Grieg

    2014-06-13

    For over a century, venom samples from wild snakes have been collected and stored around the world. However, the quality of storage conditions for "vintage" venoms has rarely been assessed. The goal of this study was to determine whether such historical venom samples are still biochemically and pharmacologically viable for research purposes, or if new sample efforts are needed. In total, 52 samples spanning 5 genera and 13 species with regional variants of some species (e.g., 14 different populations of Notechis scutatus) were analysed by a combined proteomic and pharmacological approach to determine protein structural stability and bioactivity. When venoms were not exposed to air during storage, the proteomic results were virtually indistinguishable from that of fresh venom and bioactivity was equivalent or only slightly reduced. By contrast, a sample of Acanthophis antarcticus venom that was exposed to air (due to a loss of integrity of the rubber stopper) suffered significant degradation as evidenced by the proteomics profile. Interestingly, the neurotoxicity of this sample was nearly the same as fresh venom, indicating that degradation may have occurred in the free N- or C-terminus chains of the proteins, rather than at the tips of loops where the functional residues are located. These results suggest that these and other vintage venom collections may be of continuing value in toxin research. This is particularly important as many snake species worldwide are declining due to habitat destruction or modification. For some venoms (such as N. scutatus from Babel Island, Flinders Island, King Island and St. Francis Island) these were the first analyses ever conducted and these vintage samples may represent the only venom ever collected from these unique island forms of tiger snakes. Such vintage venoms may therefore represent the last remaining stocks of some local populations and thus are precious resources. These venoms also have significant historical value as the Oxyuranus venoms analysed include samples from the first coastal taipan (Oxyuranus scutellatus) collected for antivenom production (the snake that killed the collector Kevin Budden), as well as samples from the first Oxyuranus microlepidotus specimen collected after the species' rediscovery in 1976. These results demonstrate that with proper storage techniques, venom samples can retain structural and pharmacological stability. This article is part of a Special Issue entitled: Proteomics of non-model organisms. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Variability in, variability out: best practice recommendations to standardize pre-analytical variables in the detection of circulating and tissue microRNAs.

    PubMed

    Khan, Jenna; Lieberman, Joshua A; Lockwood, Christina M

    2017-05-01

    microRNAs (miRNAs) hold promise as biomarkers for a variety of disease processes and for determining cell differentiation. These short RNA species are robust, survive harsh treatment and storage conditions and may be extracted from blood and tissue. Pre-analytical variables are critical confounders in the analysis of miRNAs: we elucidate these and identify best practices for minimizing sample variation in blood and tissue specimens. Pre-analytical variables addressed include patient-intrinsic variation, time and temperature from sample collection to storage or processing, processing methods, contamination by cells and blood components, RNA extraction method, normalization, and storage time/conditions. For circulating miRNAs, hemolysis and blood cell contamination significantly affect profiles; samples should be processed within 2 h of collection; ethylene diamine tetraacetic acid (EDTA) is preferred while heparin should be avoided; samples should be "double spun" or filtered; room temperature or 4 °C storage for up to 24 h is preferred; miRNAs are stable for at least 1 year at -20 °C or -80 °C. For tissue-based analysis, warm ischemic time should be <1 h; cold ischemic time (4 °C) <24 h; common fixative used for all specimens; formalin fix up to 72 h prior to processing; enrich for cells of interest; validate candidate biomarkers with in situ visualization. Most importantly, all specimen types should have standard and common workflows with careful documentation of relevant pre-analytical variables.

  16. The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots.

    PubMed

    Schwartz, Alanna; Baidjoe, Amrish; Rosenthal, Philip J; Dorsey, Grant; Bousema, Teun; Greenhouse, Bryan

    2015-05-01

    Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We investigated the effects of storage conditions, length of storage, and DNA extraction methods on amplification via three PCR-based assays using field samples and laboratory controls. Samples stored as DBS for 2 or more years at ambient temperature showed a significant loss of sensitivity that increased with time; after 10 years only 10% samples with parasite densities > 1,000 parasites/μL were detectable by nested polymerase chain reaction (PCR). Conversely, DBS and extracted DNA stored at -20°C showed no loss of sensitivity with time. Samples with low parasite densities amplified more successfully with saponin/Chelex compared with spin-column-based extraction, though the latter method performed better on samples with higher parasite densities stored for 2 years at ambient temperature. DNA extracted via both methods was stable after 20 freeze-thaw cycles. Our results suggest that DBS should be stored at -20°C or extracted immediately, especially if anticipating 2 or more years of storage. © The American Society of Tropical Medicine and Hygiene.

  17. The Effect of Storage and Extraction Methods on Amplification of Plasmodium falciparum DNA from Dried Blood Spots

    PubMed Central

    Schwartz, Alanna; Baidjoe, Amrish; Rosenthal, Philip J.; Dorsey, Grant; Bousema, Teun; Greenhouse, Bryan

    2015-01-01

    Extraction and amplification of DNA from dried blood spots (DBS) collected in field studies is commonly used for detection of Plasmodium falciparum. However, there have been few systematic efforts to determine the effects of storage and extraction methods on the sensitivity of DNA amplification. We investigated the effects of storage conditions, length of storage, and DNA extraction methods on amplification via three PCR-based assays using field samples and laboratory controls. Samples stored as DBS for 2 or more years at ambient temperature showed a significant loss of sensitivity that increased with time; after 10 years only 10% samples with parasite densities > 1,000 parasites/μL were detectable by nested polymerase chain reaction (PCR). Conversely, DBS and extracted DNA stored at −20°C showed no loss of sensitivity with time. Samples with low parasite densities amplified more successfully with saponin/Chelex compared with spin-column-based extraction, though the latter method performed better on samples with higher parasite densities stored for 2 years at ambient temperature. DNA extracted via both methods was stable after 20 freeze-thaw cycles. Our results suggest that DBS should be stored at −20°C or extracted immediately, especially if anticipating 2 or more years of storage. PMID:25758652

  18. Apparatus and process for collection of gas and vapor samples

    DOEpatents

    Jackson, Dennis G [Augusta, GA; Peterson, Kurt D [Aiken, SC; Riha, Brian D [Augusta, GA

    2008-04-01

    A gas sampling apparatus and process is provided in which a standard crimping tool is modified by an attached collar. The collar permits operation of the crimping tool while also facilitating the introduction of a supply of gas to be introduced into a storage vial. The introduced gas supply is used to purge ambient air from a collection chamber and an interior of the sample vial. Upon completion of the purging operation, the vial is sealed using the crimping tool.

  19. Evaluation of FTA cards as a laboratory and field sampling device for the detection of foot-and-mouth disease virus and serotyping by RT-PCR and real-time RT-PCR.

    PubMed

    Muthukrishnan, Madhanmohan; Singanallur, Nagendrakumar B; Ralla, Kumar; Villuppanoor, Srinivasan A

    2008-08-01

    Foot-and-mouth disease virus (FMDV) samples transported to the laboratory from far and inaccessible areas for serodiagnosis pose a major problem in a tropical country like India, where there is maximum temperature fluctuation. Inadequate storage methods lead to spoilage of FMDV samples collected from clinically positive animals in the field. Such samples are declared as non-typeable by the typing laboratories with the consequent loss of valuable epidemiological data. The present study evaluated the usefulness of FTA Classic Cards for the collection, shipment, storage and identification of the FMDV genome by RT-PCR and real-time RT-PCR. The stability of the viral RNA, the absence of infectivity and ease of processing the sample for molecular methods make the FTA cards a useful option for transport of FMDV genome for identification and serotyping. The method can be used routinely for FMDV research as it is economical and the cards can be transported easily in envelopes by regular document transport methods. Live virus cannot be isolated from samples collected in FTA cards, which is a limitation. This property can be viewed as an advantage as it limits the risk of transmission of live virus.

  20. Preservation of Multiple Mammalian Tissues to Maximize Science Return from Ground Based and Spaceflight Experiments.

    PubMed

    Choi, Sungshin; Ray, Hami E; Lai, San-Huei; Alwood, Joshua S; Globus, Ruth K

    2016-01-01

    Even with recent scientific advancements, challenges posed by limited resources and capabilities at the time of sample dissection continue to limit the collection of high quality tissues from experiments that can be conducted only infrequently and at high cost, such as in space. The resources and time it takes to harvest tissues post-euthanasia, and the methods and duration of long duration storage, potentially have negative impacts on sample quantity and quality, thereby limiting the scientific outcome that can be achieved. The goals of this study were to optimize methods for both sample recovery and science return from rodent experiments, with possible relevance to both ground based and spaceflight studies. The first objective was to determine the impacts of tissue harvest time post-euthanasia, preservation methods, and storage duration, focusing on RNA quality and enzyme activities in liver and spleen as indices of sample quality. The second objective was to develop methods that will maximize science return by dissecting multiple tissues after long duration storage in situ at -80°C. Tissues of C57Bl/6J mice were dissected and preserved at various time points post-euthanasia and stored at -80°C for up to 11 months. In some experiments, tissues were recovered from frozen carcasses which had been stored at -80°C up to 7 months. RNA quantity and quality was assessed by measuring RNA Integrity Number (RIN) values using an Agilent Bioanalyzer. Additionally, the quality of tissues was assessed by measuring activities of hepatic enzymes (catalase, glutathione reductase and GAPDH). Fresh tissues were collected up to one hour post-euthanasia, and stored up to 11 months at -80°C, with minimal adverse effects on the RNA quality of either livers or RNAlater-preserved spleens. Liver enzyme activities were similar to those of positive controls, with no significant effect observed at any time point. Tissues dissected from frozen carcasses that had been stored for up to 7 months at -80°C had variable results, depending on the specific tissue analyzed. RNA quality of liver, heart, and kidneys were minimally affected after 6-7 months of storage at -80°C, whereas RNA degradation was evident in tissues such as small intestine, bone, and bone marrow when they were collected from the carcasses frozen for 2.5 months. These results demonstrate that 1) the protocols developed for spaceflight experiments with on-orbit dissections support the retrieval of high quality samples for RNA expression and some protein analyses, despite delayed preservation post-euthanasia or prolonged storage, and 2) many additional tissues for gene expression analysis can be obtained by dissection even following prolonged storage of the tissue in situ at -80°C. These findings have relevance both to high value, ground-based experiments when sample collection capability is severely constrained, and to spaceflight experiments that entail on-orbit sample recovery by astronauts.

  1. Effect of storage time at -20°C on markers used for assessment of renal damage in children: albumin, γ-glutamyl transpeptidase, N-acetyl-β-D-glucosaminidase and α1-microglobulin.

    PubMed

    Trachtenberg, Felicia; Barregard, Lars

    2010-11-01

    The objective of this study is to examine the influence of storage time at -20°C on the concentration of albumin, γ-glutamyl transpeptidase (γ-GT), N-acetyl-β-D-glucosaminidase (NAG), α(1)-microglobulin (A1M) and creatinine in a large sample of healthy children. The New England Children's Amalgam Trial followed 534 children, aged 6-10 at baseline, for 5 years, with annual urine collections. Urine samples were analysed for creatinine, albumin, γ-GT, NAG and A1M concentrations. Repeated measures analysis of covariance was used to model the effect of storage time on these concentrations. The γ-GT concentration decreased significantly with storage time at -20°C. There was also a limited decrease in NAG. Albumin, A1M and creatinine concentrations did not appear to be affected by storage time at -20°C. If it is necessary to interpret results from samples stored for a long time at -20°C, it is advisable to account for storage time in statistical models.

  2. Evaluation of a New Technique for iFOBT Utilising a New Sample Collection Device with Increased Buffer Stability.

    PubMed

    Bruns-Toepler, Markus; Hardt, Philip

    2017-07-01

    The aims of the present study were: (i) Evaluate specificity and sensitivity of Hb Smart enzyme-linked immunosorbent assay (ELISA) (ScheBo Biotech) compared to colonoscopy results and (ii) assess stability of a new sample collection device containing a newly formulated buffer to extract haemoglobin using buffer and stool samples spiked with defined concentrations of haemoglobin. Stool samples were quantified with the ELISA method. The stability of haemoglobin in the extraction buffer and in native stool samples, respectively, was determined daily by ELISA during storage for 5 days at 4°C and at room temperature after addition of haemoglobin. Haemoglobin ELISA had a sensitivity of 78.4% for detection of CRC with a specificity of 98%. Haemoglobin extracted in corresponding extraction buffer demonstrated stability throughout storage for 5 days at 4°C and at room temperature. Hb Smart represents a very promising tool for large-scale screening of CRC with regard to sample handling, stability and analysis of haemoglobin in faeces. Copyright© 2017, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.

  3. Maternal–Child Microbiome: Specimen Collection, Storage and Implications for Research and Practice

    PubMed Central

    Jordan, Sheila; Baker, Brenda; Dunn, Alexis; Edwards, Sara; Ferranti, Erin; Mutic, Abby D.; Yang, Irene; Rodriguez, Jeannie

    2017-01-01

    Background The maternal microbiome is a key contributor to the development and outcomes of pregnancy and the health status of both mother and infant. Significant advances are occurring in the science of the maternal and child microbiome and hold promise in improving outcomes related to pregnancy complications, child development, and chronic health conditions of mother and child. Objectives The purpose of the paper is to review site-specific considerations in the collection and storage of maternal and child microbiome samples and its implications for nursing research and practice. Approach Microbiome sampling protocols were reviewed and synthesized. Precautions across sampling protocols were also noted. Results Oral, vaginal, gut, placental, and breastmilk are viable sources for sampling the maternal and/or child microbiome. Prior to sampling special considerations need to be addressed related to various factors including current medications, health status, and hygiene practices. Proper storage of samples will avoid degradation of cellular and DNA structures vital for analysis. Discussion Changes in the microbiome throughout the perinatal, postpartum and childhood periods are dramatic and significant to outcomes of the pregnancy and the long-term health of mother and child. Proper sampling techniques are required to produce reliable results from which evidence-based practice recommendations will be built. Ethical and practical issues surrounding study design and protocol development must also be considered when researching vulnerable groups such as pregnant women and infants. Nurses hold the responsibility to both perform the research and to translate findings from microbiome investigations for clinical use. PMID:28252577

  4. [Study on content of ethyl carbamate in yellow rice wine and its change in Zhejiang province].

    PubMed

    Shen, Xiang-hong; Wu, Ping-gu; Wang, Li-yuan; Zhang, Jing; Tan, Ying; Ying, Ying; Ma, Bing-jie

    2013-11-01

    To understand the content status of ethyl carbamate (EC) in yellow rice wine and the changes in storage period and shelf life in Zhejiang province. A total of 475 samples of yellow rice wine purchased randomly from supermarkets and food stores in Zhejiang province during 2008-2012, and 49 samples collected from manufacturers were measured for EC content. The sample collected from manufacturers by filter sterilization was placed at 4 °C, room temperature and 37 °C for 400 d, respectively;a bottled wine and a wine in bag were bought from market were placed for 400 d in room temperature to conduct shelf life storage test, and measure the content in every point in 2011. The EC of the samples was determinated by gas chromatography-mass spectrometry after the samples were diluted with D5-EC isotope dilution technique, and purified by alkaline diatomite solid phase extraction column. The overall detection rate of EC was 99% (472/475) in yellow rice wine of Zhejiang province in 2008-2012, the median value was 70-112 µg/kg, the 90th percentile was 190-333 µg/kg, the 95th percentile was 214-393 µg/kg, and the maximum value was 430-515 µg/kg. The content of EC was increased gradually along with the increasing of storage age in commercially yellow rice wine, and the average content of EC were positively correlated with storage age(r = 0.988). The contents of EC in yellow rice wine after sterilization increased from 74 µg/kg to 86 µg/kg, 127 µg/kg and 509 µg/kg at 4 °C, room temperature and 37°C, respectively for 400 d storage, the differences had statistical significance (F = 14.73, P < 0.01). The content of EC in yellow rice wines in shelf life, which stored in room temperature with bottle and bag package, was decreased slightly with increasing storage time in the beginning, from 215 to 184 µg/kg and 196 to 158 µg/kg, respectively, and increased again with increasing storage time after 250 d, with 252 µg/kg and 210 µg/kg in bottle and bag package after 400 d, respectively, the differences had statistical significance (Z = 2.37, P < 0.05). EC is widespread in rice wine, the content of EC was correlated with storage time and temperature.

  5. Microbial contamination of water intended for milk container washing in smallholder dairy farming and milk retailing houses in southern Ethiopia.

    PubMed

    Amenu, Kebede; Shitu, Desalew; Abera, Mesele

    2016-01-01

    The water used during handling and processing of milk products can be potential sources of microbial contamination with possible negative consequences on food safety. Especially, the water used in keeping the hygiene of milking and milk storage utensils is crucial to keep the quality and safety of the products. Therefore, the current study was designed to assess the bacteriological quality of water used for cleaning milking and milk storage equipment in smallholder dairy production in Hawassa and its surroundings. A total of 79 water samples were collected: 26 from milk collecting houses in Hawassa and 53 from selected smallholder dairy farms (Hawassa = 14, Arsi Negele = 29 and Yirgalem = 10). Out of the total samples, 18 samples were collected directly from pipe and 61 from storage containers (46 from narrow opening and 15 from wide opening containers). The overall prevalence of E. coli exceeding zero CFU/ml was 39.2 %. From analyzed samples, high prevalence of positive samples for E. coli was found in water samples taken from wide opening containers (66.7 %). A number of bacteria were isolated and presumptively identified which include Bacillus sp. 6.3 % (n = 5), Citrobacter sp. 1.3 %(n = 1), E. coli 39.2 % (n = 31), Enterobacter sp. 2.5 % (n = 2), Klebisella sp. 7.6 % (n = 6), Micrococcus sp. 6.3 % (n = 5), Pseudomonas sp. 6.3 % (n = 5), Staphylococcus aureus 6.3 % (n = 5), Staphylococcus epidermidis 13.9 % (n = 11) and Streptococcus sp. 1.3 % (n = 1). The bacteriological quality of water especially, water stored in household storage containers in present study area was found to be contaminated with different bacteria indicating potential food safety problem and health risk to the society. In this respect, people handling water should be educated on its proper handling and the risk of contamination during storage. To minimize contamination, materials with narrow mouth and lid should be used. Further study is recommended on the relationship between the bacteriological quality of water and the behavior of water users.

  6. Evaluation of BBL™ Sensi-Discs™ and FTA® cards as sampling devices for detection of rotavirus in stool samples

    PubMed Central

    Tam, Ka Ian; Esona, Mathew D.; Williams, Alice; Ndze, Valentine N.; Boula, Angeline; Bowen, Michael D.

    2015-01-01

    Rotavirus is the most important cause of severe childhood gastroenteritis worldwide. Rotavirus vaccines are available and rotavirus surveillance is carried out to assess vaccination impact. In surveillance studies, stool samples are stored typically at 4°C or frozen to maintain sample quality. Uninterrupted cold storage is a problem in developing countries because of power interruptions. Cold-chain transportation of samples from collection sites to testing laboratories is costly. In this study, we evaluated the use of BBL™ Sensi-Discs™ and FTA® cards for storage and transportation of samples for virus isolation, EIA, and RT-PCR testing. Infectious rotavirus was recovered after 30 days of storage on Sensi-Discs™ at room temperature. We were able to genotype 98–99% of samples stored on Sensi-Discs™ and FTA® cards at temperatures ranging from −80°C to 37°C up to 180 days. A field sampling test using samples prepared and shipped from Cameroon, showed that both matrices yielded 100% genotyping success compared with whole stool and Sensi-Discs™ demonstrated 95% concordance with whole stool in EIA testing. The utilization of BBL™ Sensi-Discs™ and FTA® cards for stool sample storage and shipment has the potential to have great impact on global public health by facilitating surveillance and epidemiological investigations of rotavirus strains worldwide at a reduced cost. PMID:26022083

  7. Evaluation of BBL™ Sensi-Discs™ and FTA® cards as sampling devices for detection of rotavirus in stool samples.

    PubMed

    Tam, Ka Ian; Esona, Mathew D; Williams, Alice; Ndze, Valantine N; Boula, Angeline; Bowen, Michael D

    2015-09-15

    Rotavirus is the most important cause of severe childhood gastroenteritis worldwide. Rotavirus vaccines are available and rotavirus surveillance is carried out to assess vaccination impact. In surveillance studies, stool samples are stored typically at 4°C or frozen to maintain sample quality. Uninterrupted cold storage is a problem in developing countries because of power interruptions. Cold-chain transportation of samples from collection sites to testing laboratories is costly. In this study, we evaluated the use of BBL™ Sensi-Discs™ and FTA(®) cards for storage and transportation of samples for virus isolation, EIA, and RT-PCR testing. Infectious rotavirus was recovered after 30 days of storage on Sensi-Discs™ at room temperature. We were able to genotype 98-99% of samples stored on Sensi-Discs™ and FTA(®) cards at temperatures ranging from -80°C to 37°C up to 180 days. A field sampling test using samples prepared and shipped from Cameroon, showed that both matrices yielded 100% genotyping success compared with whole stool and Sensi-Discs™ demonstrated 95% concordance with whole stool in EIA testing. The utilization of BBL™ Sensi-Discs™ and FTA(®) cards for stool sample storage and shipment has the potential to have great impact on global public health by facilitating surveillance and epidemiological investigations of rotavirus strains worldwide at a reduced cost. Published by Elsevier B.V.

  8. Variation in Gas and Volatile Compound Emissions from Human Urine as It Ages, Measured by an Electronic Nose.

    PubMed

    Esfahani, Siavash; Sagar, Nidhi M; Kyrou, Ioannis; Mozdiak, Ella; O'Connell, Nicola; Nwokolo, Chuka; Bardhan, Karna D; Arasaradnam, Ramesh P; Covington, James A

    2016-01-25

    The medical profession is becoming ever more interested in the use of gas-phase biomarkers for disease identification and monitoring. This is due in part to its rapid analysis time and low test cost, which makes it attractive for many different clinical arenas. One technology that is showing promise for analyzing these gas-phase biomarkers is the electronic nose--an instrument designed to replicate the biological olfactory system. Of the possible biological media available to "sniff", urine is becoming ever more important as it is easy to collect and to store for batch testing. However, this raises the question of sample storage shelf-life, even at -80 °C. Here we investigated the effect of storage time (years) on stability and reproducibility of total gas/vapour emissions from urine samples. Urine samples from 87 patients with Type 2 Diabetes Mellitus were collected over a four-year period and stored at -80 °C. These samples were then analyzed using FAIMS (field-asymmetric ion mobility spectrometry--a type of electronic nose). It was discovered that gas emissions (concentration and diversity) reduced over time. However, there was less variation in the initial nine months of storage with greater uniformity and stability of concentrations together with tighter clustering of the total number of chemicals released. This suggests that nine months could be considered a general guide to a sample shelf-life.

  9. Evaluation of reactive oxygen metabolites in frozen serum samples. Effect of storage and repeated thawing.

    PubMed

    Cavalleri, A; Colombo, C; Venturelli, E; Miceli, R; Mariani, L; Cornelli, U; Pala, V; Berrino, F; Secreto, G

    2004-01-01

    Measuring the free radical activity in serum samples from prospective studies is the best way to investigate the association between oxidative stress and human diseases. Prospective studies require the analysis of serum samples that have often been stored for a long time. Our study was designed to determine the effect of storage at -30 degrees C and -80 degrees C for two years on free radical activity. We analyzed the free radical activity by measuring circulating hydroperoxides in a pool of sera at baseline and after one day, one week, one month and 25 months of storage, using a photometric method (d-ROMs test). Measurements were performed in aliquots thawed only once at each time point and in aliquots frozen and thawed repeatedly over the study period. After two years we observed a small but statistically significant 4% decrease in the hydroperoxide concentration, which was substantially unaffected by storage temperatures and repeated freeze-thaw cycles. We also carried out the d-ROMs test in sera from ten apparently healthy volunteers at 2, 8, 24, and 48 hours after collection and storage at 4 degrees C and did not observe any significant variation. In conclusion, the d-ROMs test is a simple method suitable to evaluate the free radical activity in frozen serum samples after long-term storage.

  10. About Rubella (German Measles, Three-Day Measles)

    MedlinePlus

    ... Professionals Pregnancy and Rubella Rubella Vaccination Travelers’ Health Information on Rubella Laboratory Testing CDC Laboratory Testing & Procedures Serology RNA Detection Genetic Analysis Specimen Collection, Storage, & Shipment Sample Submission Q&A ...

  11. Rubella (German Measles, Three-Day Measles) Photos

    MedlinePlus

    ... Professionals Pregnancy and Rubella Rubella Vaccination Travelers’ Health Information on Rubella Laboratory Testing CDC Laboratory Testing & Procedures Serology RNA Detection Genetic Analysis Specimen Collection, Storage, & Shipment Sample Submission Q&A ...

  12. Sample handling factors affecting the enumeration of lactobacilli and cellulolytic bacteria in equine feces

    USDA-ARS?s Scientific Manuscript database

    The objectives were to compare media types and evaluate the effects of fecal storage time and temperature on the enumeration of cellulolytic bacteria and lactobacilli from horses. Fecal samples were collected from horses (n = 3) and transported to the lab (CO2, 37 ºC, 0.5 h). The samples were assign...

  13. Identification of control parameters for the sulfur gas storability with bag sampling methods

    USDA-ARS?s Scientific Manuscript database

    Air samples containing sulfur compounds are often collected and stored in sample bags prior to analyses. The storage stability of six gaseous sulfur compounds (H2S, CH3SH, DMS, CS2, DMDS and SO2) was compared between two different bag materials (polyvinyl fluoride (PVF) and polyester aluminum (PEA))...

  14. Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection

    PubMed Central

    Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J.; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

    2015-01-01

    Background In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Methodology Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Principal Findings Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic “gold standard”, the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). Conclusions The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments PMID:26360049

  15. Use of Occult Blood Detection Cards for Real-Time PCR-Based Diagnosis of Schistosoma Mansoni Infection.

    PubMed

    Schunk, Mirjam; Kebede Mekonnen, Seleshi; Wondafrash, Beyene; Mengele, Carolin; Fleischmann, Erna; Herbinger, Karl-Heinz; Verweij, Jaco J; Geldmacher, Christof; Bretzel, Gisela; Löscher, Thomas; Zeynudin, Ahmed

    2015-01-01

    In Schistosoma mansoni infection, diagnosis and control after treatment mainly rely on parasitological stool investigations which are laborious and have limited sensitivity. PCR methods have shown equal or superior sensitivity but preservation and storage methods limit their use in the field. Therefore, the use of occult blood detection cards (fecal cards) for easy sampling and storage of fecal samples for further PCR testing was evaluated in a pilot study. Stool specimens were collected in a highly endemic area for S. mansoni in Ethiopia and submitted in an investigator-blinded fashion to microscopic examination by Kato-Katz thick smear as well as to real-time PCR using either fresh frozen stool samples or stool smears on fecal cards which have been stored at ambient temperature for up to ten months. Out of 55 stool samples, 35 were positive by microscopy, 33 and 32 were positive by PCR of frozen samples and of fecal card samples, respectively. When microscopy was used as diagnostic "gold standard", the sensitivity of PCR on fresh stool was 94.3% (95%-CI: 86.6; 100) and on fecal cards 91.4% (95%-CI: 82.2; 100). The use of fecal cards proved to be a simple and useful method for stool collection and prolonged storage prior to PCR based diagnosis of S. mansoni infection. This technique may be a valuable approach for large scale surveillance and post treatment assessments.

  16. Ergovaline Stability in Tall Fescue Based on Sample Handling and Storage Methods

    NASA Astrophysics Data System (ADS)

    Lea, Krista; Smith, Lori; Gaskill, Cynthia; Coleman, Robert; Smith, S.

    2014-09-01

    Ergovaline is an ergot alkaloid produced by the endophyte Neotyphodium coenophialum (Morgan-Jones and Gams) found in tall fescue (Schedonorus arundinacea (Schreb.) Dumort.) and blamed for a multitude of livestock disorders. Ergovaline is known to be unstable and affected by many variables. The objective of this study was to determine the effect of sample handling and storage on the stability of ergovaline in tall fescue samples. Fresh tall fescue was collected from a horse farm in central Kentucky at three harvest dates and transported on ice to the University of Kentucky Veterinary Diagnostic Laboratory. Plant material was frozen in liquid nitrogen, milled and mixed before being allocated into different sub-samples. Three sub-samples were assigned to each of 14 sample handling or storage treatments. Sample handling included increased heat and UV light to simulate transportation in a vehicle and on ice in a cooler per standard transportation recommendations. Storage conditions included storage at 22oC, 5oC and -20oC for up to 28 days. Each sub-sample was then analyzed for ergovaline concentration using HPLC with fluorescence detection and this experiment was repeated for each harvest date. Sub-samples exposed to UV light and heat lost a significant fraction of ergovaline in 2 hours, while sub-samples stored on ice in a cooler showed no change in ergovaline in 2 hours. All sub-samples stored at 22oC, 5oC and -20oC lost a significant fraction of ergovaline in the first 24 hours of storage. There was little change in ergovaline in the freezer (-20oC) after the first 24 hours up to 28 days of storage but intermittent losses were observed at 22oC and 5oC. To obtain results that most closely represent levels in the field, all samples should be transported on ice to the laboratory immediately after harvest for same day analysis. If immediate testing is not possible, samples should be stored at -20oC until analysis.

  17. Evaluation of a simple method for storage of blood samples that enables isolation of circulating tumor cells 96 h after sample collection.

    PubMed

    Apostolou, Panagiotis; Ntanovasilis, Dimitrios-Athanasios; Papasotiriou, Ioannis

    2017-12-01

    Minimizing the effects of transportation on the properties of biological material is a major challenge for the scientific community. The viability of cells is important in cases where their study is urgent for evaluation of treatment response or for the study of cancer progression. Circulating tumor cells (CTCs) constitute a cell subpopulation with great importance for oncologists, because of their prognostic value. Detection and isolation of CTCs from blood samples is a routine activity in many laboratories, but concerns exist with regard to the maintenance of the cells during transportation. In this study, experiments were conducted to determine the stability of gene and protein expression in CTCs over a period of 96 h. Blood samples collected from healthy individuals and patients with cancer were each divided into five aliquots, which were stored at 2-8 °C and analyzed after 0, 24, 48, 72 and 96 h of storage. CTCs from patients and CD45-negative cells from healthy individuals were isolated each day using enrichment protocols, and qPCR was performed to determine expression levels of genes encoding specific biological markers. In addition, cells from breast and colon cancer cell lines were spiked into blood samples from healthy individuals, and these samples were stored and analyzed over a period of 96 h by PCR and by flow cytometry. The markers that were studied included housekeeping genes and genes associated with the response to chemotherapy, as well as genes encoding transcription factors. The results demonstrated that the expression profiles of specific genes and proteins in CTCs were not significantly affected by 72 h of storage. After 96 h of storage, expression of some genes was altered. The transportation of blood at low temperature (2-8 °C) in the presence of the anticoagulant EDTA can protect CTCs from alteration of gene and protein expression for at least 72 h. Furthermore, under these conditions, CTCs can be detected and isolated 96 h after blood collection.

  18. Effects of preservation method on canine (Canis lupus familiaris) fecal microbiota.

    PubMed

    Horng, Katti R; Ganz, Holly H; Eisen, Jonathan A; Marks, Stanley L

    2018-01-01

    Studies involving gut microbiome analysis play an increasing role in the evaluation of health and disease in humans and animals alike. Fecal sampling methods for DNA preservation in laboratory, clinical, and field settings can greatly influence inferences of microbial composition and diversity, but are often inconsistent and under-investigated between studies. Many laboratories have utilized either temperature control or preservation buffers for optimization of DNA preservation, but few studies have evaluated the effects of combining both methods to preserve fecal microbiota. To determine the optimal method for fecal DNA preservation, we collected fecal samples from one canine donor and stored aliquots in RNAlater, 70% ethanol, 50:50 glycerol:PBS, or without buffer at 25 °C, 4 °C, and -80 °C. Fecal DNA was extracted, quantified, and 16S rRNA gene analysis performed on Days 0, 7, 14, and 56 to evaluate changes in DNA concentration, purity, and bacterial diversity and composition over time. We detected overall effects on bacterial community of storage buffer ( F -value = 6.87, DF  = 3, P  < 0.001), storage temperature ( F -value=1.77, DF  = 3, P  = 0.037), and duration of sample storage ( F -value = 3.68, DF  = 3, P  < 0.001). Changes in bacterial composition were observed in samples stored in -80 °C without buffer, a commonly used method for fecal DNA storage, suggesting that simply freezing samples may be suboptimal for bacterial analysis. Fecal preservation with 70% ethanol and RNAlater closely resembled that of fresh samples, though RNAlater yielded significantly lower DNA concentrations ( DF  = 8.57, P  < 0.001). Although bacterial composition varied with temperature and buffer storage, 70% ethanol was the best method for preserving bacterial DNA in canine feces, yielding the highest DNA concentration and minimal changes in bacterial diversity and composition. The differences observed between samples highlight the need to consider optimized post-collection methods in microbiome research.

  19. “It’s my blood”: ethical complexities in the use, storage and export of biological samples: perspectives from South African research participants

    PubMed Central

    2014-01-01

    Background The use of biological samples in research raises a number of ethical issues in relation to consent, storage, export, benefit sharing and re-use of samples. Participant perspectives have been explored in North America and Europe, with only a few studies reported in Africa. The amount of research being conducted in Africa is growing exponentially with volumes of biological samples being exported from the African continent. In order to investigate the perspectives of African research participants, we conducted a study at research sites in the Western Cape and Gauteng, South Africa. Methods Data were collected using a semi-structured questionnaire that captured both quantitative and qualitative information at 6 research sites in South Africa. Interviews were conducted in English and Afrikaans. Data were analysed both quantitatively and qualitatively. Results Our study indicates that while the majority of participants were supportive of providing samples for research, serious concerns were voiced about future use, benefit sharing and export of samples. While researchers view the provision of biosamples as a donation, participants believe that they still have ownership rights and are therefore in favour of benefit sharing. Almost half of the participants expressed a desire to be re-contacted for consent for future use of their samples. Interesting opinions were expressed with respect to export of samples. Conclusions Eliciting participant perspectives is an important part of community engagement in research involving biological sample collection, export, storage and future use. A tiered consent process appears to be more acceptable to participants in this study. Eliciting opinions of researchers and research ethics committee (REC) members would contribute multiple perspectives. Further research is required to interrogate the concept of ownership and the consent process in research involving biological samples. PMID:24447822

  20. Sample handling for mass spectrometric proteomic investigations of human sera.

    PubMed

    West-Nielsen, Mikkel; Høgdall, Estrid V; Marchiori, Elena; Høgdall, Claus K; Schou, Christian; Heegaard, Niels H H

    2005-08-15

    Proteomic investigations of sera are potentially of value for diagnosis, prognosis, choice of therapy, and disease activity assessment by virtue of discovering new biomarkers and biomarker patterns. Much debate focuses on the biological relevance and the need for identification of such biomarkers while less effort has been invested in devising standard procedures for sample preparation and storage in relation to model building based on complex sets of mass spectrometric (MS) data. Thus, development of standardized methods for collection and storage of patient samples together with standards for transportation and handling of samples are needed. This requires knowledge about how sample processing affects MS-based proteome analyses and thereby how nonbiological biased classification errors are avoided. In this study, we characterize the effects of sample handling, including clotting conditions, storage temperature, storage time, and freeze/thaw cycles, on MS-based proteomics of human serum by using principal components analysis, support vector machine learning, and clustering methods based on genetic algorithms as class modeling and prediction methods. Using spiking to artificially create differentiable sample groups, this integrated approach yields data that--even when working with sample groups that differ more than may be expected in biological studies--clearly demonstrate the need for comparable sampling conditions for samples used for modeling and for the samples that are going into the test set group. Also, the study emphasizes the difference between class prediction and class comparison studies as well as the advantages and disadvantages of different modeling methods.

  1. Effects of Dry Storage and Resubmersion of Oysters on Total Vibrio vulnificus and Total and Pathogenic (tdh+/trh+) Vibrio parahaemolyticus Levels.

    PubMed

    Kinsey, Thomas P; Lydon, Keri A; Bowers, John C; Jones, Jessica L

    2015-08-01

    Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are the two leading causes of bacterial illnesses associated with raw shellfish consumption. Levels of these pathogens in oysters can increase during routine antifouling aquaculture practices involving dry storage in ambient air conditions. After storage, common practice is to resubmerge these stored oysters to reduce elevated Vv and Vp levels, but evidence proving the effectiveness of this practice is lacking. This study examined the changes in Vv and in total and pathogenic (thermostable direct hemolysin gene and the tdh-related hemolysin gene, tdh+ and trh+) Vp levels in oysters after 5 or 24 h of dry storage (28 to 32°C), followed by resubmersion (27 to 32°C) for 14 days. For each trial, replicate oyster samples were collected at initial harvest, after dry storage, after 7 days, and after 14 days of resubmersion. Oysters not subjected to dry storage were collected and analyzed to determine natural undisturbed vibrio levels (background control). Vibrio levels were measured using a most-probable-number enrichment followed by real-time PCR. After storage, vibrio levels (excluding tdh+ and trh+ Vp during 5-h storage) increased significantly (P < 0.001) from initial levels. After 7 days of resubmersion, Vv and total Vp levels (excluding total Vp in oysters stored for 5 h) were not significantly different (P < 0.1) from levels in background oysters. Vv and total and pathogenic Vp levels were not significantly different (P > 0.1) from levels in background oysters after 14 days of resubmersion, regardless of dry storage time. These data demonstrate that oyster resubmersion after dry storage at elevated ambient temperatures allows vibrio levels to return to those of background control samples. These results can be used to help minimize the risk of Vv and Vp illnesses and to inform the oyster industry on the effectiveness of routine storing and resubmerging of aquaculture oysters.

  2. Experimental and analytical variation in human urine in 1H NMR spectroscopy-based metabolic phenotyping studies.

    PubMed

    Maher, Anthony D; Zirah, Séverine F M; Holmes, Elaine; Nicholson, Jeremy K

    2007-07-15

    1H NMR spectroscopy potentially provides a robust approach for high-throughput metabolic screening of biofluids such as urine and plasma, but sample handling and preparation need careful optimization to ensure that spectra accurately report biological status or disease state. We have investigated the effects of storage temperature and time on the 1H NMR spectral profiles of human urine from two participants, collected three times a day on four different days. These were analyzed using modern chemometric methods. Analytical and preparation variation (tested between -40 degrees C and room temperature) and time of storage (to 24 h) were found to be much less influential than biological variation in sample classification. Statistical total correlation spectroscopy and discriminant function methods were used to identify the specific metabolites that were hypervariable due to preparation and biology. Significant intraindividual variation in metabolite profiles were observed even for urine collected on the same day and after at least 6 h fasting. The effect of long-term storage at different temperatures was also investigated, showing urine is stable if frozen for at least 3 months and that storage at room temperature for long periods (1-3 months) results in a metabolic profile explained by bacterial activity. Presampling (e.g., previous day) intake of food and medicine can also strongly influence the urinary metabolic profiles indicating that collective detailed participant historical meta data are important for interpretation of metabolic phenotypes and for avoiding false biomarker discovery.

  3. Evaluation of diagnostic procedures for subclinical mastitis in meat-producing sheep.

    PubMed

    Clements, Archie C A; Taylor, David J; Fitzpatrick, Julie L

    2003-05-01

    Samples of foremilk were collected from 261 clinically normal glands of 150 ewes, and tested using the California mastitis test (CMT). Further samples were collected from 195 of these glands for determination of automated somatic cell counts (SCC), and from 60 of these glands for bacteriological assessment. The sensitivity and specificity of CMT for detecting samples with SCC above different threshold levels and for CMT and SCC in determining bacteriological status were evaluated using two-graph receiver operating characteristics (TG-ROC). Milk samples were obtained subsequently from ten CMT positive, and five CMT negative first- and second-lactation ewes. Samples were cultured using a variety of media, incubation temperatures and atmospheric conditions, immediately after collection, and 1 week after storage at 4 degrees C and -21 degrees C. Results suggested that CMT is best used as a diagnostic test for ovine subclinical mastitis (SCM) with a cut-off of 3 (distinct gel formation), and that automated SCC thresholds of > 1200 x 10(3) cells/ml are appropriate, especially where low prevalences are expected (e.g. < 5%). Additionally, this study showed that routine bacteriological methods were appropriate for isolation of most species of pathogen responsible for ovine SCM, but storage of samples prior to culture, either at 4 degrees C or -21 degrees C, was detrimental to the isolation of several of these organisms.

  4. Use of FTA sampling cards for molecular detection of avian influenza virus in wild birds.

    PubMed

    Keeler, Shamus P; Ferro, Pamela J; Brown, Justin D; Fang, Xingwang; El-Attrache, John; Poulson, Rebecca; Jackwood, Mark W; Stallknecht, David E

    2012-03-01

    Current avian influenza (AI) virus surveillance programs involving wild birds rely on sample collection methods that require refrigeration or low temperature freezing to maintain sample integrity for virus isolation and/or reverse-transcriptase (RT) PCR. Maintaining the cold chain is critical for the success of these diagnostic assays but is not always possible under field conditions. The aim of this study was to test the utility of Finders Technology Associates (FTA) cards for reliable detection of AI virus from cloacal and oropharyngeal swabs of wild birds. The minimum detectable titer was determined, and the effect of room temperature storage was evaluated experimentally using multiple egg-propagated stock viruses (n = 6). Using real time RT-PCR, we compared results from paired cloacal swab and samples collected on FTA cards from both experimentally infected mallards (Anasplatyrhynchos) and hunter-harvested waterfowl sampled along the Texas Gulf Coast. Based on the laboratory trials, the average minimal detectable viral titer was determined to be 1 x 10(4.7) median embryo infectious dose (EID50)/ml (range: 1 x 10(4.3) to 1 x 10(5.4) EID50/ml), and viral RNA was consistently detectable on the FTA cards for a minimum of 20 days and up to 30 days for most subtypes at room temperature (23 C) storage. Real-time RT-PCR of samples collected using the FTA cards showed fair to good agreement in live birds when compared with both real-time RT-PCR and virus isolation of swabs. AI virus detection rates in samples from several wild bird species were higher when samples were collected using the FTA cards compared with cloacal swabs. These results suggest that FTA cards can be used as an alternative sample collection method when traditional surveillance methods are not possible, especially in avian populations that have historically received limited testing or situations in which field conditions limit the ability to properly store or ship swab samples.

  5. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature.

    PubMed

    Tuorto, Steven J; Brown, Chris M; Bidle, Kay D; McGuinness, Lora R; Kerkhof, Lee J

    2015-01-01

    This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry "field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.

  6. BioDry: An Inexpensive, Low-Power Method to Preserve Aquatic Microbial Biomass at Room Temperature

    PubMed Central

    Tuorto, Steven J.; Brown, Chris M.; Bidle, Kay D.; McGuinness, Lora R.; Kerkhof, Lee J.

    2015-01-01

    This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry “field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited. PMID:26710122

  7. Analysis of the Storage Methods for Raw Human Milk from Mothers with Infants Admitted to a Neonatal Intensive Care Unit, According to Brazilian Regulations.

    PubMed

    Grazziotin, Maria Celestina Bonzanini; Grazziotin, Ana Laura; Vidal, Newton Medeiros; Freire, Marcia Helena de Souza; da Silva, Regina Paula Guimarães Vieira Cavalcante

    2016-08-01

    Milk safety is an important concern in neonatal units and human milk banks. Therefore, evidence-based recommendations regarding raw milk handling and storage are needed to safely promote supplying hospitalized infants with their mother's own milk. To evaluate raw human milk storage methods according to Brazilian milk management regulations by investigating the effects of refrigeration (5°C) for 12 hours and freezing (-20°C) for 15 days on the acidity and energy content in a large number of raw milk samples. Expressed milk samples from 100 distinct donors were collected in glass bottles. Each sample was separated into 3 equal portions that were analyzed at room temperature and after either 12 hours of refrigeration or 15 days of freezing. Milk acidity and energy content were determined by Dornic titration and creamatocrit technique, respectively. All samples showed Dornic acidity values within the established acceptable limit (≤ 8°D), as required by Brazilian regulations. In addition, energy content did not significantly differ among fresh, refrigerated and frozen milk samples (median of ~50 kcal/100 mL for each). Most samples tested (> 80%) were considered top quality milk (< 4°D) based on acidity values, and milk energy content was preserved after storage. We conclude that the storage methods required by Brazilian regulations are suitable to ensure milk safety and energy content of stored milk when supplied to neonates. © The Author(s) 2016.

  8. Effects of Storage and Type of Blood Collection Tubes on Hepatitis C Virus Level in Whole Blood Samples

    PubMed Central

    Kessler, Harald H.; Stelzl, Evelyn; Raggam, Reinhard B.; Haas, Josef; Kirchmeir, Franz; Hegenbarth, Karin; Daghofer, Elisabeth; Santner, Brigitte I.; Marth, Egon; Stauber, Rudolf E.

    2001-01-01

    In this study, we compared serum hepatitis C virus (HCV) RNA concentrations with HCV RNA concentrations in whole blood collection tubes, including two different types of EDTA tubes and nucleic acid stabilization tubes (NASTs). We also investigated the impact of a processing delay on HCV RNA concentration in these tubes. In NASTs, the mean HCV RNA concentration was comparable to the mean serum HCV RNA concentration at “date zero.” In EDTA tubes, mean baseline HCV RNA concentrations were higher. Storage at room temperature up to 96 h did not result in a decline of HCV RNA concentration in any of the whole blood collection tubes. In NASTs, HCV RNA concentrations remained stable during the whole study period, whereas a significant increase of HCV RNA was observed in both types of EDTA tubes at 96 h compared to date zero. We concluded that HCV RNA remains stable in NASTs at room temperature for at least 96 h, allowing greater flexibility in sample collection and transport. PMID:11325991

  9. Assessment of the stability of DNA in specimens collected under conditions for drug testing-A pilot study.

    PubMed

    White, Robert M; Mitchell, John M; Hart, E Dale; Evans, Amy; Meaders, Meredith; Norsworthy, Sarah E; Hayes, Eugene D; Flegel, Ron; Maha, George C; Shaffer, Megan D; Hall, Erin M; Rogers, Kelley

    2018-02-01

    For forensic biological sample collections, the specimen donor is linked solidly to his or her specimen through a chain of custody (CoC) sometimes referenced as a chain of evidence. Rarely, a donor may deny that a urine or oral fluid (OF) specimen is his or her specimen even with a patent CoC. The goal of this pilot study was to determine the potential effects of short-term storage on the quality and quantity of DNA in both types of specimen under conditions that may be encountered with employment-related drug testing specimens. Fresh urine and freshly collected oral fluid all produced complete STR profiles. For the "pad" type OF collectors, acceptable DNA was extractable both from the buffer/preservative and the pad. Although fresh urine and OF produced complete STR profiles, partial profiles were obtained after storage for most samples. An exception was the DNA in the Quantisal OF collector, from which a complete profile was obtained for both freshly collected OF and stored OF. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Evaluating Protocols for Porcine Faecal Microbiome Recollection, Storage and DNA Extraction: from the Farm to the Lab.

    PubMed

    Muiños-Bühl, Anixa; González-Recio, Oscar; Muñoz, María; Óvilo, Cristina; García-Casco, Juan; Fernández, Ana I

    2018-06-01

    There is a growing interest in understanding the role of the gut microbiome on productive and meat quality-related traits in livestock species in order to develop new useful tools for improving pig production systems and industry. Faecal samples are analysed as a proxy of gut microbiota and here the selection of suitable protocols for faecal sampling and DNA isolation is a critical first step in order to obtain reliable results, even more to compare results obtained from different studies. The aim of the current study was to establish in a cost-effective way, using automated ribosomal intergenic spacer analysis technique, a protocol for porcine faecal sampling and storage at farm and slaughterhouse and to determine the most efficient microbiota DNA isolation kit among those most widely used. Operational Taxonomic Unit profiles were compared from Iberian pig faecal samples collected from rectum or ground, stored with liquid N 2 , room temperature or RNAlater, and processed with QIAamp DNA Stool (Qiagen), PowerFecal DNA Isolation (Mobio) or SpeedTools Tissue DNA extraction (Biotools) commercial kits. The results, focused on prokaryote sampling, based on DNA yield and quality, OTU number and Sørensen similarity Indexes, indicate that the recommended protocol for porcine faecal microbiome sampling at farm should include: the collection from porcine rectum to avoid contamination; the storage in liquid N 2 or even at room temperature, but not in RNAlater; and the isolation of microbiota DNA using PowerFecal DNA Isolation kit. These conditions provide more reliable DNA samples for further microbiome analysis.

  11. Effects of Chicken Litter Storage Time and Ammonia Content on Thermal Resistance of Desiccation-Adapted Salmonella spp.

    PubMed

    Chen, Zhao; Wang, Hongye; Ionita, Claudia; Luo, Feng; Jiang, Xiuping

    2015-10-01

    Broiler chicken litter was kept as a stacked heap on a poultry farm, and samples were collected up to 9 months of storage. Chicken litter inoculated with desiccation-adapted Salmonella cells was heat-treated at 75, 80, 85, and 150°C. Salmonella populations decreased in all these samples during heat treatment, and the inactivation rates became lower in chicken litter when storage time was extended from 0 to 6 months. There was no significant difference (P > 0.05) in thermal resistance of Salmonella in 6- and 9-month litter samples, indicating that a threshold for thermal resistance was reached after 6 months. Overall, the thermal resistance of Salmonella in chicken litter was affected by the storage time of the litter. The changes in some chemical, physical, and microbiological properties during storage could possibly contribute to this difference. Moisture and ammonia could be two of the most significant factors influencing the thermal resistance of Salmonella cells in chicken litter. Our results emphasize the importance of adjusting time and temperature conditions for heat processing chicken litter when it is removed from the chicken house at different time intervals. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  12. Effects of Chicken Litter Storage Time and Ammonia Content on Thermal Resistance of Desiccation-Adapted Salmonella spp.

    PubMed Central

    Chen, Zhao; Wang, Hongye; Ionita, Claudia; Luo, Feng

    2015-01-01

    Broiler chicken litter was kept as a stacked heap on a poultry farm, and samples were collected up to 9 months of storage. Chicken litter inoculated with desiccation-adapted Salmonella cells was heat-treated at 75, 80, 85, and 150°C. Salmonella populations decreased in all these samples during heat treatment, and the inactivation rates became lower in chicken litter when storage time was extended from 0 to 6 months. There was no significant difference (P > 0.05) in thermal resistance of Salmonella in 6- and 9-month litter samples, indicating that a threshold for thermal resistance was reached after 6 months. Overall, the thermal resistance of Salmonella in chicken litter was affected by the storage time of the litter. The changes in some chemical, physical, and microbiological properties during storage could possibly contribute to this difference. Moisture and ammonia could be two of the most significant factors influencing the thermal resistance of Salmonella cells in chicken litter. Our results emphasize the importance of adjusting time and temperature conditions for heat processing chicken litter when it is removed from the chicken house at different time intervals. PMID:26209673

  13. Storage conditions for stability of offline measurement of fractional exhaled nitric oxide after collection for epidemiologic research

    PubMed Central

    2012-01-01

    Background The measurement of fractional concentration of nitric oxide in exhaled air (FeNO) is valuable for the assessment of airway inflammation. Offline measurement of FeNO has been used in some epidemiologic studies. However, the time course of the changes in FeNO after collection has not been fully clarified. In this study, the effects of storage conditions on the stability of FeNO measurement in exhaled air after collection for epidemiologic research were examined. Methods Exhaled air samples were collected from 48 healthy adults (mean age 43.4 ± 12.1 years) in Mylar bags. FeNO levels in the bags were measured immediately after collection. The bags were then stored at 4°C or room temperature to measure FeNO levels repeatedly for up to 168 hours. Results In the bags stored at room temperature after collection, FeNO levels were stable for 9 hours, but increased starting at 24 hours. FeNO levels remained stable for a long time at 4°C, and they were 99.7% ± 7.7% and 101.3% ± 15.0% relative to the baseline values at 24 and 96 hours, respectively. When the samples were stored at 4°C, FeNO levels gradually decreased with time among the subjects with FeNO ≥ 51 ppb immediately after collection, although there were almost no changes among the other subjects. FeNO levels among current smokers increased even at 4°C, although the values among ex-smokers decreased gradually, and those among nonsmokers remained stable. The rate of increase was significantly higher among current smokers than among nonsmokers and ex-smokers from 9 hours after collection onwards. Conclusions Storage at 4°C could prolong the stability of FeNO levels after collection. This result suggests that valid measurements can be performed within several days if the samples are stored at 4°C. However, the time course of the changes in FeNO levels differed in relation to initial FeNO values and cigarette smoking. PMID:23116255

  14. Long-term room temperature preservation of corpse soft tissue: an approach for tissue sample storage

    PubMed Central

    2011-01-01

    Background Disaster victim identification (DVI) represents one of the most difficult challenges in forensic sciences, and subsequent DNA typing is essential. Collected samples for DNA-based human identification are usually stored at low temperature to halt the degradation processes of human remains. We have developed a simple and reliable procedure for soft tissue storage and preservation for DNA extraction. It ensures high quality DNA suitable for PCR-based DNA typing after at least 1 year of room temperature storage. Methods Fragments of human psoas muscle were exposed to three different environmental conditions for diverse time periods at room temperature. Storage conditions included: (a) a preserving medium consisting of solid sodium chloride (salt), (b) no additional substances and (c) garden soil. DNA was extracted with proteinase K/SDS followed by organic solvent treatment and concentration by centrifugal filter devices. Quantification was carried out by real-time PCR using commercial kits. Short tandem repeat (STR) typing profiles were analysed with 'expert software'. Results DNA quantities recovered from samples stored in salt were similar up to the complete storage time and underscored the effectiveness of the preservation method. It was possible to reliably and accurately type different genetic systems including autosomal STRs and mitochondrial and Y-chromosome haplogroups. Autosomal STR typing quality was evaluated by expert software, denoting high quality profiles from DNA samples obtained from corpse tissue stored in salt for up to 365 days. Conclusions The procedure proposed herein is a cost efficient alternative for storage of human remains in challenging environmental areas, such as mass disaster locations, mass graves and exhumations. This technique should be considered as an additional method for sample storage when preservation of DNA integrity is required for PCR-based DNA typing. PMID:21846338

  15. Gamma-hydroxybutyric acid endogenous production and post-mortem behaviour - the importance of different biological matrices, cut-off reference values, sample collection and storage conditions.

    PubMed

    Castro, André L; Dias, Mário; Reis, Flávio; Teixeira, Helena M

    2014-10-01

    Gamma-Hydroxybutyric Acid (GHB) is an endogenous compound with a story of clinical use, since the 1960's. However, due to its secondary effects, it has become a controlled substance, entering the illicit market for recreational and "dance club scene" use, muscle enhancement purposes and drug-facilitated sexual assaults. Its endogenous context can bring some difficulties when interpreting, in a forensic context, the analytical values achieved in biological samples. This manuscript reviewed several crucial aspects related to GHB forensic toxicology evaluation, such as its post-mortem behaviour in biological samples; endogenous production values, whether in in vivo and in post-mortem samples; sampling and storage conditions (including stability tests); and cut-off reference values evaluation for different biological samples, such as whole blood, plasma, serum, urine, saliva, bile, vitreous humour and hair. This revision highlights the need of specific sampling care, storage conditions, and cut-off reference values interpretation in different biological samples, essential for proper practical application in forensic toxicology. Copyright © 2014 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

  16. Method and Apparatus for the Collection, Storage, and Real Time Analysis of Blood and Other Bodily Fluids

    NASA Technical Reports Server (NTRS)

    Whiitson, Peggy A. (Inventor); Clift, Vaughan L. (Inventor)

    1999-01-01

    The present invention provides a method and apparatus for separating a blood sample having a volume of up to about 20 milliliters into cellular and acellular fractions. The apparatus includes a housing divided by a fibrous filter into a blood sample collection chamber having a volume of at least about 1 milliliter and a serum sample collection chamber. The fibrous filter has a pore size of less than about 3 microns, and is coated with a mixture including between about 1-40 wt/vol % mannitol and between about 0.1-15 wt/vol % of plasma fraction protein (or an animal or vegetable equivalent thereof). The coating causes the cellular fraction to be trapped by the small pores, leaving the cellular fraction intact on the fibrous filter while the acellular fraction passes through the filter for collection in unaltered form from the serum sample collection chamber.

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

    DOE Data Explorer

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

    2003-01-01

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

  18. Quantitation of major allergens in dust samples from urban populations collected in different seasons in two climatic areas of the Basque region (Spain).

    PubMed

    Echechipía, S; Ventas, P; Audícana, M; Urrutia, I; Gastaminza, G; Polo, F; Fernández de Corres, L

    1995-06-01

    We present the results of allergen content evaluation in 80 dust samples from 31 homes of atopic patients from two climatic areas (humid and subhumid), collected in two seasons of the year (autumn and winter). Monoclonal antibody-based immunoassays were used to quantify Der p 1, Der f 1, Der 2, Lep d 1, and Fel d 1. The results were compared according to climate, season, and the type of sensitization (Pyroglyphidae mites, storage mites, or grass pollens). We underline the predominance of Dermatophagoides pteronyssinus (89% of samples) over D. farinae (16% of samples) in our environment. Der p 1 rates were higher in the humid area (Mann-Whitney P < 0.001), especially in the autumn (Wilcoxon P < 0.05). Lep d 1 was detected in 23% of samples and Lep d 1 levels were higher in the homes of patients sensitized to storage mites (Mann-Whitney P < 0.05), whereas this allergen was not detected in the homes of pollen-allergic patients. Fel d 1 was detected in nine of the 31 homes (16% of samples) although there was a cat in only one home.

  19. Methodology challenges in studying human gut microbiota - effects of collection, storage, DNA extraction and next generation sequencing technologies.

    PubMed

    Panek, Marina; Čipčić Paljetak, Hana; Barešić, Anja; Perić, Mihaela; Matijašić, Mario; Lojkić, Ivana; Vranešić Bender, Darija; Krznarić, Željko; Verbanac, Donatella

    2018-03-23

    The information on microbiota composition in the human gastrointestinal tract predominantly originates from the analyses of human faeces by application of next generation sequencing (NGS). However, the detected composition of the faecal bacterial community can be affected by various factors including experimental design and procedures. This study evaluated the performance of different protocols for collection and storage of faecal samples (native and OMNIgene.GUT system) and bacterial DNA extraction (MP Biomedicals, QIAGEN and MO BIO kits), using two NGS platforms for 16S rRNA gene sequencing (Ilumina MiSeq and Ion Torrent PGM). OMNIgene.GUT proved as a reliable and convenient system for collection and storage of faecal samples although favouring Sutterella genus. MP provided superior DNA yield and quality, MO BIO depleted Gram positive organisms while using QIAGEN with OMNIgene.GUT resulted in greatest variability compared to other two kits. MiSeq and IT platforms in their supplier recommended setups provided comparable reproducibility of donor faecal microbiota. The differences included higher diversity observed with MiSeq and increased capacity of MiSeq to detect Akkermansia muciniphila, [Odoribacteraceae], Erysipelotrichaceae and Ruminococcaceae (primarily Faecalibacterium prausnitzii). The results of our study could assist the investigators using NGS technologies to make informed decisions on appropriate tools for their experimental pipelines.

  20. Optimizing blood collection, transport and storage conditions for cell free DNA increases access to prenatal testing.

    PubMed

    Wong, David; Moturi, Sharmili; Angkachatchai, Vach; Mueller, Reinhold; DeSantis, Grace; van den Boom, Dirk; Ehrich, Mathias

    2013-08-01

    Fetal mutations and fetal chromosomal abnormalities can be detected by molecular analysis of circulating cell free fetal DNA (ccffDNA) from maternal plasma. This comprehensive study was aimed to investigate and verify blood collection and blood shipping conditions that enable Noninvasive Prenatal Testing. Specifically, the impact of shipping and storage on the stability and concentration of circulating cell-free DNA (ccfDNA) in Streck® Cell-Free DNA™ Blood Collection Tubes (Streck BCTs, Streck, Omaha NE). These BCTs were designed to minimize cellular degradation, and thus effectively prevent dilution of fetal ccf DNA by maternal genomic DNA, was evaluated. Peripheral venous maternal blood was collected into Streck BCTs to investigate four aspects of handling and processing conditions: (1) time from blood draw to plasma processing; (2) storage temperature; (3) mechanical stress; and (4) lot-to-lot tube variations. Maternal blood stored in Streck BCTs for up to 7 days at ambient temperature provides stable concentrations of ccffDNA. The amount of fetal DNA did not change over a broad range of storage temperatures (4°C, 23°C, 37°C, 40°C), but the amount of total (largely maternal) DNA increased in samples stored at 23°C and above, indicating maternal cell degradation and genomic DNA release at elevated temperatures. Shipping maternal blood in Streck BCTs, did not affect sample quality. Maternal plasma DNA stabilized for 0 to 7 days in Streck BCTs can be used for non-invasive prenatal molecular applications, when temperatures are maintained within the broad parameters assessed in this study. Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  1. Analysis of dust samples collected from spent nuclear fuel interim storage containers at Hope Creek, Delaware, and Diablo Canyon, California

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

    Bryan, Charles R.; Enos, David George

    2014-07-01

    Potentially corrosive environments may form on the surface of spent nuclear fuel dry storage canisters by deliquescence of deposited dusts. To assess this, samples of dust were collected from in-service dry storage canisters at two near-marine sites, the Hope Creek and Diablo Canyon storage installations, and have been characterized with respect to mineralogy, chemistry, and texture. At both sites, terrestrially-derived silicate minerals, including quartz, feldspars, micas, and clays, comprise the largest fraction of the dust. Also significant at both sites were particles of iron and iron-chromium metal and oxides generated by the manufacturing process. Soluble salt phases were minor componentmore » of the Hope Creek dusts, and were compositionally similar to inland salt aerosols, rich in calcium, sulfate, and nitrate. At Diablo Canyon, however, sea-salt aerosols, occurring as aggregates of NaCl and Mg-sulfate, were a major component of the dust samples. The seasalt aerosols commonly occurred as hollow spheres, which may have formed by evaporation of suspended aerosol seawater droplets, possibly while rising through the heated annulus between the canister and the overpack. The differences in salt composition and abundance for the two sites are attributed to differences in proximity to the open ocean and wave action. The Diablo Canyon facility is on the shores of the Pacific Ocean, while the Hope Creek facility is on the shores of the Delaware River, several miles from the open ocean.« less

  2. Minimizing microbial contamination of sperm samples

    USGS Publications Warehouse

    Jenkins, Jill A.; Tiersch, Terrence R.; Green, Christopher C.

    2011-01-01

    Taken from the Methods section: With the collection and translocation of gametes from aquatic species, a potential hazard exists for microbial transfer. Contamination of semen can occur during collection, processing, storage, and transport. Some preventative measures are described below for limiting the spread and amplification of microorganisms such as bacteria, viruses, fungi, mycoplasmas, and parasites. Generally, sanitation during collection is essential. Materials and equipment used to freeze semen should be sterile. Following good practice guidelines for handling and processing samples collected for freezing is especially important for non-domestic animals where disease-free status cannot be guaranteed and unsophisticated technology is used (Russell et al. 1977).

  3. Variation in Gas and Volatile Compound Emissions from Human Urine as It Ages, Measured by an Electronic Nose

    PubMed Central

    Esfahani, Siavash; Sagar, Nidhi M.; Kyrou, Ioannis; Mozdiak, Ella; O’Connell, Nicola; Nwokolo, Chuka; Bardhan, Karna D.; Arasaradnam, Ramesh P.; Covington, James A.

    2016-01-01

    The medical profession is becoming ever more interested in the use of gas-phase biomarkers for disease identification and monitoring. This is due in part to its rapid analysis time and low test cost, which makes it attractive for many different clinical arenas. One technology that is showing promise for analyzing these gas-phase biomarkers is the electronic nose—an instrument designed to replicate the biological olfactory system. Of the possible biological media available to “sniff”, urine is becoming ever more important as it is easy to collect and to store for batch testing. However, this raises the question of sample storage shelf-life, even at −80 °C. Here we investigated the effect of storage time (years) on stability and reproducibility of total gas/vapour emissions from urine samples. Urine samples from 87 patients with Type 2 Diabetes Mellitus were collected over a four-year period and stored at −80 °C. These samples were then analyzed using FAIMS (field-asymmetric ion mobility spectrometry—a type of electronic nose). It was discovered that gas emissions (concentration and diversity) reduced over time. However, there was less variation in the initial nine months of storage with greater uniformity and stability of concentrations together with tighter clustering of the total number of chemicals released. This suggests that nine months could be considered a general guide to a sample shelf-life. PMID:26821055

  4. Quantitative assessment of anthrax vaccine immunogenicity using the dried blood spot matrix.

    PubMed

    Schiffer, Jarad M; Maniatis, Panagiotis; Garza, Ilana; Steward-Clark, Evelene; Korman, Lawrence T; Pittman, Phillip R; Mei, Joanne V; Quinn, Conrad P

    2013-03-01

    The collection, processing and transportation to a testing laboratory of large numbers of clinical samples during an emergency response situation present significant cost and logistical issues. Blood and serum are common clinical samples for diagnosis of disease. Serum preparation requires significant on-site equipment and facilities for immediate processing and cold storage, and significant costs for cold-chain transport to testing facilities. The dried blood spot (DBS) matrix offers an alternative to serum for rapid and efficient sample collection with fewer on-site equipment requirements and considerably lower storage and transport costs. We have developed and validated assay methods for using DBS in the quantitative anti-protective antigen IgG enzyme-linked immunosorbent assay (ELISA), one of the primary assays for assessing immunogenicity of anthrax vaccine and for confirmatory diagnosis of Bacillus anthracis infection in humans. We have also developed and validated high-throughput data analysis software to facilitate data handling for large clinical trials and emergency response. Published by Elsevier Ltd.

  5. Sampling of Atmospheric Precipitation and Deposits for Analysis of Atmospheric Pollution

    PubMed Central

    Skarżyńska, K.; Polkowska, Ż; Namieśnik, J.

    2006-01-01

    This paper reviews techniques and equipment for collecting precipitation samples from the atmosphere (fog and cloud water) and from atmospheric deposits (dew, hoarfrost, and rime) that are suitable for the evaluation of atmospheric pollution. It discusses the storage and preparation of samples for analysis and also presents bibliographic information on the concentration ranges of inorganic and organic compounds in the precipitation and atmospheric deposit samples. PMID:17671615

  6. Reuse of samples: ethical issues encountered by two institutional ethics review committees in Kenya.

    PubMed

    Langat, Simon K

    2005-10-01

    There is growing concern about the reuse and exploitation of biological materials (human tissues) for use in research worldwide. Most discussions about samples have taken place in developed countries, where genetic manipulation techniques have greatly advanced in recent years. There is very little discussion in developing countries, although collaborative research with institutions from developed countries is on the increase. The study sought to identify and describe ethical issues arising in the storage, reuse and exportation of samples in a developing country. Research protocols presented to two Ethics Review Committees in Kenya during a period of two years were reviewed. A record was made of the protocol title, sample collected, request for storage, reuse or exportation and whether or not subject consent was sought. The findings indicated that about 25% out of the 388 protocols sought permission for reuse and only half of those actually informed subjects of the contemplated re-use. Less than 20% requested storage and again, about half of them sought consent from subjects. There is an indication that investigators do not see the need to seek consent for storage, reuse and exportation of samples. It is proposed that these issues should be addressed through policy interventions at both the national and global levels.

  7. Influence of Freezing and Storage Procedure on Human Urine Samples in NMR-Based Metabolomics

    PubMed Central

    Rist, Manuela J.; Muhle-Goll, Claudia; Görling, Benjamin; Bub, Achim; Heissler, Stefan; Watzl, Bernhard; Luy, Burkhard

    2013-01-01

    It is consensus in the metabolomics community that standardized protocols should be followed for sample handling, storage and analysis, as it is of utmost importance to maintain constant measurement conditions to identify subtle biological differences. The aim of this work, therefore, was to systematically investigate the influence of freezing procedures and storage temperatures and their effect on NMR spectra as a potentially disturbing aspect for NMR-based metabolomics studies. Urine samples were collected from two healthy volunteers, centrifuged and divided into aliquots. Urine aliquots were frozen either at −20 °C, on dry ice, at −80 °C or in liquid nitrogen and then stored at −20 °C, −80 °C or in liquid nitrogen vapor phase for 1–5 weeks before NMR analysis. Results show spectral changes depending on the freezing procedure, with samples frozen on dry ice showing the largest deviations. The effect was found to be based on pH differences, which were caused by variations in CO2 concentrations introduced by the freezing procedure. Thus, we recommend that urine samples should be frozen at −20 °C and transferred to lower storage temperatures within one week and that freezing procedures should be part of the publication protocol. PMID:24957990

  8. Influence of Freezing and Storage Procedure on Human Urine Samples in NMR-Based Metabolomics.

    PubMed

    Rist, Manuela J; Muhle-Goll, Claudia; Görling, Benjamin; Bub, Achim; Heissler, Stefan; Watzl, Bernhard; Luy, Burkhard

    2013-04-09

    It is consensus in the metabolomics community that standardized protocols should be followed for sample handling, storage and analysis, as it is of utmost importance to maintain constant measurement conditions to identify subtle biological differences. The aim of this work, therefore, was to systematically investigate the influence of freezing procedures and storage temperatures and their effect on NMR spectra as a potentially disturbing aspect for NMR-based metabolomics studies. Urine samples were collected from two healthy volunteers, centrifuged and divided into aliquots. Urine aliquots were frozen either at -20 °C, on dry ice, at -80 °C or in liquid nitrogen and then stored at -20 °C, -80 °C or in liquid nitrogen vapor phase for 1-5 weeks before NMR analysis. Results show spectral changes depending on the freezing procedure, with samples frozen on dry ice showing the largest deviations. The effect was found to be based on pH differences, which were caused by variations in CO2 concentrations introduced by the freezing procedure. Thus, we recommend that urine samples should be frozen at -20 °C and transferred to lower storage temperatures within one week and that freezing procedures should be part of the publication protocol.

  9. QUALITY SYSTEMS AND IMPLEMENTATION PLAN FOR A PILOT STUDY OF CHILDREN'S TOTAL EXPOSURE TO PERSISTENT PESTICIDES AND OTHER PERSISTENT ORGANIC PESTICIDES (CTEPP)

    EPA Science Inventory

    The Quality System Implementation Plan (QSIP) describes the quality assurance and quality control procedures developed for the CTEPP study. It provides the QA/QC procedures used in recruitment of subjects, sample field collection, sample extraction and analysis, data storage, and...

  10. PROCEEDINGS OF THE WORKSHOP ON SAMPLING GEOTHERMAL EFFLUENTS (2ND) HELD AT LAS VEGAS, NEVADA ON FEBRUARY 15-17, 1977

    EPA Science Inventory

    A partial listing of contents includes: The use of gas sampling bags for the collection and storage of hydrothermal gases; Heavy metal emissions from geothermal power plants; The dynamic measurement of ambient airborne gases near geothermal areas; Analysis of radon in geothermal ...

  11. Isotopic (d18O/d2H) integrity of water samples collected and stored by automatic samplers

    USDA-ARS?s Scientific Manuscript database

    Stable water isotopes are increasingly becoming part of routine monitoring programs that utilize automatic samplers. The objectives of this study were to quantify the uncertainty in isotope signatures due to the length of sample storage (1-24 d) inside autosamplers over a range of air temperatures (...

  12. Shelf-life extension of vacuum-packaged meat from pheasant (Phasianus colchicus) by lactic acid treatment.

    PubMed

    Pfeifer, Agathe; Smulders, Frans J M; Paulsen, Peter

    2014-07-01

    We investigated the influence of lactic acid treatment of pheasant meat before vacuum-packaged storage of 3, 7, and 10 d at +6°C on microbiota and pH. Breast muscle samples were collected from carcasses of slaughtered as well as from hunted (shot) wild pheasants. Immersion of meat samples in 3% (wt/wt) lactic acid for 60 s effectuated a significant drop in pH of approximately 0.5 to 0.7 units, which remained during the entire storage period. In parallel, total aerobic counts of such treated and stored samples were on an average 1.5 to 1.7 log units lower than in non-acid-treated samples. Similar results were found for Enterobacteriaceae. A significant decrease in pH was measured at d 7 and 10 in the acid-treated samples in comparison with the untreated ones. In summary, the immersion of pheasant breast meat cuts in dilute lactic acid significantly reduced microbiota during vacuum-packed storage, even at slight temperature abuse conditions. © 2014 Poultry Science Association Inc.

  13. Modular biowaste monitoring system

    NASA Technical Reports Server (NTRS)

    Fogal, G. L.

    1975-01-01

    The objective of the Modular Biowaste Monitoring System Program was to generate and evaluate hardware for supporting shuttle life science experimental and diagnostic programs. An initial conceptual design effort established requirements and defined an overall modular system for the collection, measurement, sampling and storage of urine and feces biowastes. This conceptual design effort was followed by the design, fabrication and performance evaluation of a flight prototype model urine collection, volume measurement and sampling capability. No operational or performance deficiencies were uncovered as a result of the performance evaluation tests.

  14. Uncertainty in monitoring E. coli concentrations in streams and stormwater runoff

    NASA Astrophysics Data System (ADS)

    Harmel, R. D.; Hathaway, J. M.; Wagner, K. L.; Wolfe, J. E.; Karthikeyan, R.; Francesconi, W.; McCarthy, D. T.

    2016-03-01

    Microbial contamination of surface waters, a substantial public health concern throughout the world, is typically identified by fecal indicator bacteria such as Escherichia coli. Thus, monitoring E. coli concentrations is critical to evaluate current conditions, determine restoration effectiveness, and inform model development and calibration. An often overlooked component of these monitoring and modeling activities is understanding the inherent random and systematic uncertainty present in measured data. In this research, a review and subsequent analysis was performed to identify, document, and analyze measurement uncertainty of E. coli data collected in stream flow and stormwater runoff as individual discrete samples or throughout a single runoff event. Data on the uncertainty contributed by sample collection, sample preservation/storage, and laboratory analysis in measured E. coli concentrations were compiled and analyzed, and differences in sampling method and data quality scenarios were compared. The analysis showed that: (1) manual integrated sampling produced the lowest random and systematic uncertainty in individual samples, but automated sampling typically produced the lowest uncertainty when sampling throughout runoff events; (2) sample collection procedures often contributed the highest amount of uncertainty, although laboratory analysis introduced substantial random uncertainty and preservation/storage introduced substantial systematic uncertainty under some scenarios; and (3) the uncertainty in measured E. coli concentrations was greater than that of sediment and nutrients, but the difference was not as great as may be assumed. This comprehensive analysis of uncertainty in E. coli concentrations measured in streamflow and runoff should provide valuable insight for designing E. coli monitoring projects, reducing uncertainty in quality assurance efforts, regulatory and policy decision making, and fate and transport modeling.

  15. Insect Population Dynamics in Commercial Grain Elevators

    USDA-ARS?s Scientific Manuscript database

    Data were collected in 1998-2002 from wheat stored in commercial grain elevators in south-central Kansas. Storage bins at these elevators had concrete walls and were typically 6-9 m in diameter and 30-35 m tall. A vacuum-probe sampler was used to collect ten 3-kg grain samples in the top 12 m of the...

  16. Apparatus and method for monitoring of gas having stable isotopes

    DOEpatents

    Clegg, Samuel M; Fessenden-Rahn, Julianna E

    2013-03-05

    Gas having stable isotopes is monitored continuously by using a system that sends a modulated laser beam to the gas and collects and transmits the light not absorbed by the gas to a detector. Gas from geological storage, or from the atmosphere can be monitored continuously without collecting samples and transporting them to a lab.

  17. Collection of biological samples in forensic toxicology.

    PubMed

    Dinis-Oliveira, R J; Carvalho, F; Duarte, J A; Remião, F; Marques, A; Santos, A; Magalhães, T

    2010-09-01

    Forensic toxicology is the study and practice of the application of toxicology to the purposes of the law. The relevance of any finding is determined, in the first instance, by the nature and integrity of the specimen(s) submitted for analysis. This means that there are several specific challenges to select and collect specimens for ante-mortem and post-mortem toxicology investigation. Post-mortem specimens may be numerous and can endow some special difficulties compared to clinical specimens, namely those resulting from autolytic and putrefactive changes. Storage stability is also an important issue to be considered during the pre-analytic phase, since its consideration should facilitate the assessment of sample quality and the analytical result obtained from that sample. The knowledge on degradation mechanisms and methods to increase storage stability may enable the forensic toxicologist to circumvent possible difficulties. Therefore, advantages and limitations of specimen preservation procedures are thoroughfully discussed in this review. Presently, harmonized protocols for sampling in suspected intoxications would have obvious utility. In the present article an overview is given on sampling procedures for routinely collected specimens as well as on alternative specimens that may provide additional information on the route and timing of exposure to a specific xenobiotic. Last, but not least, a discussion on possible bias that can influence the interpretation of toxicological results is provided. This comprehensive review article is intented as a significant help for forensic toxicologists to accomplish their frequently overwhelming mission.

  18. Method and Apparatus for the Collection Storage and Real Time Analysis of Blood and Other Bodily Fluids

    NASA Technical Reports Server (NTRS)

    Whitson, Peggy A. (Inventor); Clift, Vaughan L. (Inventor)

    1997-01-01

    The present invention provides an apparatus for separating a relatively large volume of blood into cellular and acellular fractions without the need for centrifugation. The apparatus comprises a housing divided by a fibrous filter into a blood sample collection chamber having a volume of at least about 1 milliliter and a serum sample collection chamber. The fibrous filter has a pore size of less than about 3 microns, and is coated with a mixture of mannitol and plasma fraction protein (or an animal or vegetable equivalent thereof). The coating causes the cellular fraction to be trapped by the small pores, leaving the cellular fraction intact on the fibrous filter while the acellular fraction passes through the filter for collection in unaltered form from the serum sample collection chamber.

  19. Stability of selected serum hormones and lipids after long-term storage in the Janus Serum Bank.

    PubMed

    Gislefoss, Randi E; Grimsrud, Tom K; Mørkrid, Lars

    2015-04-01

    The potential value of a biobank depends on the quality of the samples, i.e. how well they reflect the biological or biochemical state of the donors at the time of sampling. Documentation of sample quality has become a particularly important issue for researchers and users of biobank studies. The aim of this study was to investigate the long-term stability of selected components: cholesterol, high density cholesterol (HDLC), low density cholesterol (LDLC), apolipoprotein A1 (apo-A1), apolipoprotein B (apo B), follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), thyroid stimulating hormone (TSH) and free thyroxin (FT4). Samples, stored at -25°C, from 520 men aged 40-49 years at blood sampling distributed in equally sized groups (n=130) according to length of storage, 0, 4, 17 and 29 years, respectively, were used in a cross sectional design. The freshly collected serum samples were used as a reference group to calculate storage related changes. The differences between fresh samples and samples stored for 29 years were substantial for apo-A1 (+12%), apo-B (+22.3%), HDLC (-69.2%), LDLC (+31.3%), and PRL (-33.5%), while total cholesterol, FSH, LH, TSH and FT4 did not show any significant difference. The study showed large differences in serum level of the selected components. The lipids and apolipoproteins were all changed except for total cholesterol. Most hormones investigated (FSH, LH, TSH and FT4) proved to be stable after 29 years of storage while PRL showed sign of degradation. The observed differences are probably due to long-term storage effects and/or external factors (i.e. diet and smoking). Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  20. Effect of ambient temperature storage on potable water coliform population estimations.

    PubMed Central

    Standridge, J H; Delfino, J J

    1983-01-01

    The effect of the length of time between sampling potable water and performing coliform analyses has been a long-standing controversial issue in environmental microbiology. The issue is of practical importance since reducing the sample-to-analysis time may substantially increase costs for water analysis programs. Randomly selected samples (from those routinely collected throughout the State of Wisconsin) were analyzed for total coliforms after being held at room temperature (20 +/- 2 degrees C) for 24 and 48 h. Differences in results for the two holding times were compared with differences predicted by probability calculations. The study showed that storage of the potable water for up to 48 h had little effect on the public health significance of most samples containing more than two coliforms per 100 ml. PMID:6651296

  1. Analytical results from an environmental investigation of six sites on Kirtland Air Force Base, New Mexico, 1993-94

    USGS Publications Warehouse

    Wilcox, Ralph

    1995-01-01

    The six sites investigated include silver recovery units; a buried caustic drain line; a neutralization pit; an evaporation/infiltration pond; the Manzano fire training area; and a waste oil underground storage tank. Environmental samples of soil, pond sediment, soil gas, and water and gas in floor drains were collected and analyzed. Field quality-control samples were also collected and analyzed in association with the environmental samples. The six sites were investigated because past or current activities could have resulted in contamination of soil, pond sediment, or water and sediment in drains.

  2. Capture and exploration of sample quality data to inform and improve the management of a screening collection.

    PubMed

    Charles, Isabel; Sinclair, Ian; Addison, Daniel H

    2014-04-01

    A new approach to the storage, processing, and interrogation of the quality data for screening samples has improved analytical throughput and confidence and enhanced the opportunities for learning from the accumulating records. The approach has entailed the design, development, and implementation of a database-oriented system, capturing information from the liquid chromatography-mass spectrometry capabilities used for assessing the integrity of samples in AstraZeneca's screening collection. A Web application has been developed to enable the visualization and interactive annotation of the analytical data, monitor the current sample queue, and report the throughput rate. Sample purity and identity are certified automatically on the chromatographic peaks of interest if predetermined thresholds are reached on key parameters. Using information extracted in parallel from the compound registration and container inventory databases, the chromatographic and spectroscopic profiles for each vessel are linked to the sample structures and storage histories. A search engine facilitates the direct comparison of results for multiple vessels of the same or similar compounds, for single vessels analyzed at different time points, or for vessels related by their origin or process flow. Access to this network of information has provided a deeper understanding of the multiple factors contributing to sample quality assurance.

  3. Bias of Tedlar bags in the measurement of agricultural odorants.

    PubMed

    Trabue, Steven L; Anhalt, Jennifer C; Zahn, James A

    2006-01-01

    Odor regulations typically specify the use of dynamic dilution olfactometery (DDO) as a method to quantify odor emissions, and Tedlar bags are the preferred holding container for grab samples. This study was conducted to determine if Tedlar bags affect the integrity of sampled air from animal operations. Air samples were collected simultaneously in both Tedlar bags and Tenax thermal desorption tubes. Sample sources originated from either a hydrocarbon-free air tank, dynamic headspace chamber (DHC), or swine-production facility, and were analyzed by gas chromatography-mass spectrometry-olfactometry (GC-MS-O). Several background contaminants were identified from Tedlar bags, which included the odorous compounds N,N-dimethyl acetamide (DMAC), acetic acid, and phenol. Samples from the DHC demonstrated that recovery of malodor compounds was dependent on residence time in the Tedlar bag with longer residence time leading to lower recovery. After 24 h of storage, recovery of C3-C6 volatile fatty acids (VFA) averaged 64%, 4-methylphenol and 4-ethylphenol averaged 10%, and indole and 3-methylindole were below the detection limits of GC-MS-O. The odor activity value (OAV) of grab samples collected in Tedlar bags were 33 to 65% lower following 24 h of storage. These results indicate that significant odorant bias occurs when using Tedlar bags for the sampling of odors from animal production facilities.

  4. CTC-mRNA (AR-V7) Analysis from Blood Samples—Impact of Blood Collection Tube and Storage Time

    PubMed Central

    Luk, Alison W. S.; Ma, Yafeng; Ding, Pei N.; Young, Francis P.; Chua, Wei; Balakrishnar, Bavanthi; Dransfield, Daniel T.; de Souza, Paul; Becker, Therese M.

    2017-01-01

    Circulating tumour cells (CTCs) are an emerging resource for monitoring cancer biomarkers. New technologies for CTC isolation and biomarker detection are increasingly sensitive, however, the ideal blood storage conditions to preserve CTC-specific mRNA biomarkers remains undetermined. Here we tested the preservation of tumour cells and CTC-mRNA over time in common anticoagulant ethylene-diamine-tetra-acetic acid (EDTA) and acid citrate dextrose solution B (Citrate) blood tubes compared to preservative-containing blood tubes. Blood samples spiked with prostate cancer cells were processed after 0, 24, 30, and 48 h storage at room temperature. The tumour cell isolation efficiency and the mRNA levels of the prostate cancer biomarkers androgen receptor variant 7 (AR-V7) and total AR, as well as epithelial cell adhesion molecule (EpCAM) were measured. Spiked cells were recovered across all storage tube types and times. Surprisingly, tumour mRNA biomarkers were readily detectable after 48 h storage in EDTA and Citrate tubes, but not in preservative-containing tubes. Notably, AR-V7 expression was detected in prostate cancer patient blood samples after 48 h storage in EDTA tubes at room temperature. This important finding presents opportunities for measuring AR-V7 expression from clinical trial patient samples processed within 48 h—a much more feasible timeframe compared to previous recommendations. PMID:28498319

  5. Osmotic and aging effects in caviar oocytes throughout water and lipid changes assessed by 1H NMR T1 and T2 relaxation and MRI.

    PubMed

    Gussoni, Maristella; Greco, Fulvia; Vezzoli, Alessandra; Paleari, Maria Antonietta; Moretti, Vittorio Maria; Lanza, Barbara; Zetta, Lucia

    2007-01-01

    By combining NMR relaxation spectroscopy and magnetic resonance imaging techniques, unsalted (us) and salted (s) caviar (Acipenser transmontanus) oocytes were characterized over a storage period of up to 90 days. The aging and the salting effects on the two major cell constituents, water and lipids, were separately assessed. T1 and T2 decays were interpreted by assuming a two-site exchange model. At Day 0, two water compartments that were not in fast exchange were identified by the T1 relaxation measurements on the us oocytes. In the s samples, T1 decay was monoexponential. During the time of storage, an increment of the free water amount was found for the us oocytes, ascribed to an increased metabolism. T1 and T2 of the s oocytes shortened as a consequence of the osmotic stress produced by salting. Selective images showed the presence of water endowed with different regional mobility that severely changed during the storage. Lipid T1 relaxation decays collected on us and s samples were found to be biexponential, and the T1 values lengthened during storage. In us and s oocytes, the increased lipid mobility with the storage was ascribed to lipolysis. Selective images of us samples showed lipids that were confined to the cytoplasm for up to 60 days of storage.

  6. Impact of low intensity summer rainfall on E. coli-discharge event dynamics with reference to sample acquisition and storage.

    PubMed

    Oliver, David M; Porter, Kenneth D H; Heathwaite, A Louise; Zhang, Ting; Quilliam, Richard S

    2015-07-01

    Understanding the role of different rainfall scenarios on faecal indicator organism (FIO) dynamics under variable field conditions is important to strengthen the evidence base on which regulators and land managers can base informed decisions regarding diffuse microbial pollution risks. We sought to investigate the impact of low intensity summer rainfall on Escherichia coli-discharge (Q) patterns at the headwater catchment scale in order to provide new empirical data on FIO concentrations observed during baseflow conditions. In addition, we evaluated the potential impact of using automatic samplers to collect and store freshwater samples for subsequent microbial analysis during summer storm sampling campaigns. The temporal variation of E. coli concentrations with Q was captured during six events throughout a relatively dry summer in central Scotland. The relationship between E. coli concentration and Q was complex with no discernible patterns of cell emergence with Q that were repeated across all events. On several occasions, an order of magnitude increase in E. coli concentrations occurred even with slight increases in Q, but responses were not consistent and highlighted the challenges of attempting to characterise temporal responses of E. coli concentrations relative to Q during low intensity rainfall. Cross-comparison of E. coli concentrations determined in water samples using simultaneous manual grab and automated sample collection was undertaken with no difference in concentrations observed between methods. However, the duration of sample storage within the autosampler unit was found to be more problematic in terms of impacting on the representativeness of microbial water quality, with unrefrigerated autosamplers exhibiting significantly different concentrations of E. coli relative to initial samples after 12-h storage. The findings from this study provide important empirical contributions to the growing evidence base in the field of catchment microbial dynamics.

  7. Improvement in the stability of serum samples stored in an automated refrigerated module.

    PubMed

    Parra-Robert, Marina; Rico-Santana, Naira; Alcaraz-Quiles, José; Sandalinas, Silvia; Fernández, Esther; Falcón, Isabel; Pérez-Riedweg, Margarita; Bedini, Josep Lluís

    2016-12-01

    In clinical laboratories it is necessary to know for how long the analytes are stable in the samples with specific storage conditions. Our laboratory has implemented the new Aptio Automation System (AAS) (Siemens Healthcare Diagnostics) where the analyzed samples are stored in a refrigerated storage module (RSM) after being sealed. The aim of the study was to evaluate the stability of serum samples with the AAS and comparing the results with a previous study using a conventional refrigerated system. Serum samples from a total of 50 patients were collected and for each of them 27 biochemical analytes were analyzed. The samples were divided in 5 sets of 10 samples. Each set was re-analyzed at one of the following times: 24, 48, 72, 96 and 120h. Stability was evaluated according to the Total Limit of Change (TLC) criteria, which combine both analytical and biologic variation. A total of 26 out of 27 analytes were stable at the end of the study according to TLC criteria. Lactate dehydrogenase was not stable at 48h observing a decrease in its concentration until the end of the study. In the previous study (conventional storage system) 9 biochemical analytes were not stable with an increase of their levels due to the evaporation process. The RSM connected to the AAS improves the stability of serum samples. This system avoids the evaporation process due to the sealing of samples and allows better control of the samples during their storage. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  8. Long-term frozen storage of urine samples: a trouble to get PCR results in Schistosoma spp. DNA detection?

    PubMed

    Fernández-Soto, Pedro; Velasco Tirado, Virginia; Carranza Rodríguez, Cristina; Pérez-Arellano, José Luis; Muro, Antonio

    2013-01-01

    Human schistosomiasis remains a serious worldwide public health problem. At present, a sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. The potential for detecting schistosome-derived DNA by PCR-based methods in human clinical samples is currently being investigated as a diagnostic tool with potential application in routine schistosomiasis diagnosis. Collection of diagnostic samples such as stool or blood is usually difficult in some populations. However, urine is a biological sample that can be collected in a non-invasive method, easy to get from people of all ages and easy in management, but as a sample for PCR diagnosis is still not widely used. This could be due to the high variability in the reported efficiency of detection as a result of the high variation in urine samples' storage or conditions for handling and DNA preservation and extraction methods. We evaluate different commercial DNA extraction methods from a series of long-term frozen storage human urine samples from patients with parasitological confirmed schistosomiasis in order to assess the PCR effectiveness for Schistosoma spp. detection. Patients urine samples were frozen for 18 months up to 7 years until use. Results were compared with those obtained in PCR assays using fresh healthy human urine artificially contaminated with Schistosoma mansoni DNA and urine samples from mice experimentally infected with S. mansoni cercariae stored frozen for at least 12 months before use. PCR results in fresh human artificial urine samples using different DNA based extraction methods were much more effective than those obtained when long-term frozen human urine samples were used as the source of DNA template. Long-term frozen human urine samples are probably not a good source for DNA extraction for use as a template in PCR detection of Schistosoma spp., regardless of the DNA method of extraction used.

  9. Comparison of storage stability of odorous VOCs in polyester aluminum and polyvinyl fluoride tedlar bags

    USDA-ARS?s Scientific Manuscript database

    Whole air sampling using containers such as flexible bags or rigid canisters is commonly used to collect samples of volatile organic compounds (VOC) in air. The objective of this study was to compare the stability of polyester aluminum (PEA) and polyvinyl fluoride (PVF, brand name Tedlar®) bags for ...

  10. Collection, Storage, and Preparation of Human Blood Cells

    PubMed Central

    Dagur, Pradeep K.; McCoy, J. Philip

    2015-01-01

    Human peripheral blood is often studied by flow cytometry in both the research and clinical laboratories. The methods for collection, storage, and preparation of peripheral blood will vary depending on the cell lineage to be examined as well as the type of assay to be performed. This unit presents protocols for collection of blood, separation of leukocytes from whole blood by lysis of erythrocytes, isolating mononuclear cells by density gradient separation, and assorted non-flow sorting methods, such as magnetic bead separations, for enriching specific cell populations, including monocytes, T lymphocytes, B lymphocytes, neutrophils,, , and platelets prior to flow cytometric analysis. A protocol is also offered for cryopreservation of cells since clinical research often involves retrospective flow cytometric analysis of samples stored over a period of months or years. PMID:26132177

  11. Validity and Reliability of Perinatal Biomarkers after Storage as Dry Blood Spots on Paper

    PubMed Central

    Mihalopoulos, Nicole L.; Phillips, Terry M.; Slater, Hillarie; Thomson, J. Anne; Varner, Michael W.; Moyer-Mileur, Laurie J.

    2013-01-01

    Ojective To validate use of chip-based immunoaffinity capillary electrophoresis on dry blood spot samples (DBSS) to measure obesity-related cytokines. Methods Chip-based immunoaffinity capillary electrophoresis was used to measure adiponectin, leptin and insulin in serum and DBSS in pregnant women, cord blood, and infant heelstick at birth and 6 weeks. Concordance of measurements was determined with Pearson's correlation. Results We report high concordance between results obtained from serum and DBSS with the exception of cord blood specimens. Conclusions Ease of sample collection and storage makes DBSS an optimal method for use in studies involving neonates and young children. PMID:21735507

  12. Maximizing Science Return from Future Rodent Experiments on the International Space Station (ISS): Tissue Preservation

    NASA Technical Reports Server (NTRS)

    Choi, S. Y.; Lai, S.; Klotz, R.; Popova, Y.; Chakravarty, K.; Beegle, J. E.; Wigley, C. L.; Globus, R. K.

    2014-01-01

    To better understand how mammals adapt to long duration habitation in space, a system for performing rodent experiments on the ISS is under development. Rodent Research-1 is the first flight and will include validation of both on-orbit animal support and tissue preservation. To evaluate plans for on-orbit sample dissection and preservation, we simulated conditions for euthanasia, tissue dissection, and prolonged sample storage on the ISS, and we also developed methods for post-flight dissection and recovery of high quality RNA from multiple tissues following prolonged storage in situ for future science return. Livers and spleens from mice were harvested under conditions that simulated nominal, on-orbit euthanasia and dissection procedures including storage at minus 80 degrees Centigrade for 4 months. The RNA recovered was of high quality (RNA Integrity Number, RNA Integrity Number (RIN) greater than 8) and quantity, and the liver enzyme contents and activities (catalase, glutathione reductase, GAPDH) were similar to positive controls, which were collected under standard laboratory conditions. We also assessed the impact of possible delayed on-orbit dissection scenarios (off-nominal) by dissecting and preserving the spleen (RNA, later) and liver (fast-freezing) at various time points post-euthanasia (from 5 minutes up to 105 minutes). The RNA recovered was of high quality (spleen, RIN greater than 8; liver, RIN greater than 6) and liver enzyme activities were similar to positive controls at all time points, although an apparent decline in select enzyme activities was evident at 105 minutes. Additionally, various tissues were harvested from either intact or partially dissected, frozen carcasses after storage for approximately 2 months; most of the tissues (brain, heart, kidney, eye, adrenal glands and muscle) were of acceptable RNA quality for science return, whereas some tissues (small intestine, bone marrow and bones) were not. These data demonstrate: 1) The protocols developed for future flight experiments will support science return despite delayed preservation post-euthanasia or prolonged storage, and 2) High-quality RNA samples from many different tissues can be recovered by dissection following prolonged storage of the tissue in situ at minus 80 degrees Centigrade. These findings have relevance both to high-value, ground-based experiments when sample collection capability is severely constrained, and to future spaceflight experiments that entail on-orbit sample recovery by the ISS crew.

  13. Long-Term Frozen Storage of Urine Samples: A Trouble to Get PCR Results in Schistosoma spp. DNA Detection?

    PubMed Central

    Fernández-Soto, Pedro; Velasco Tirado, Virginia; Carranza Rodríguez, Cristina; Pérez-Arellano, José Luis; Muro, Antonio

    2013-01-01

    Background Human schistosomiasis remains a serious worldwide public health problem. At present, a sensitive and specific assay for routine diagnosis of schistosome infection is not yet available. The potential for detecting schistosome-derived DNA by PCR-based methods in human clinical samples is currently being investigated as a diagnostic tool with potential application in routine schistosomiasis diagnosis. Collection of diagnostic samples such as stool or blood is usually difficult in some populations. However, urine is a biological sample that can be collected in a non-invasive method, easy to get from people of all ages and easy in management, but as a sample for PCR diagnosis is still not widely used. This could be due to the high variability in the reported efficiency of detection as a result of the high variation in urine samples’ storage or conditions for handling and DNA preservation and extraction methods. Methodology/Principal Findings We evaluate different commercial DNA extraction methods from a series of long-term frozen storage human urine samples from patients with parasitological confirmed schistosomiasis in order to assess the PCR effectiveness for Schistosoma spp. detection. Patientś urine samples were frozen for 18 months up to 7 years until use. Results were compared with those obtained in PCR assays using fresh healthy human urine artificially contaminated with Schistosoma mansoni DNA and urine samples from mice experimentally infected with S. mansoni cercariae stored frozen for at least 12 months before use. PCR results in fresh human artificial urine samples using different DNA based extraction methods were much more effective than those obtained when long-term frozen human urine samples were used as the source of DNA template. Conclusions/Significance Long-term frozen human urine samples are probably not a good source for DNA extraction for use as a template in PCR detection of Schistosoma spp., regardless of the DNA method of extraction used. PMID:23613907

  14. Ethics, Human Use, and the Department of Defense Serum Repository.

    PubMed

    Pavlin, Julie A; Welch, Robert A

    2015-10-01

    The Department of Defense Serum Repository (DoDSR) contains a growing archive of sera from service members collected to perform medical surveillance, clinical diagnosis, and epidemiologic studies to identify, prevent, and control diseases associated with military service. The specimens are a mandatory collection under DoD and U.S. regulations and do not include informed consent for uses beyond force health protection. Any use of the specimens for research requires deidentification of the samples and must be approved by Institutional Review Boards. However, as expansion of the DoDSR is contemplated, ethical considerations of sample collection, storage, and use must be carefully reconsidered. Other similar programs for research use of specimens collected for public health purpose are also undergoing similar reviews. It is recommended that at a minimum, service members are informed of the potential storage and use of their specimens and are allowed to opt out of additional use, or a broad informed consent is provided. The DoDSR provides a tremendous resource to the DoD and global health community, and to ensure its continued existence and improvement, the DoD must stay consistent with all principles of research ethics. Reprint & Copyright © 2015 Association of Military Surgeons of the U.S.

  15. 40 CFR 63.694 - Testing methods and procedures.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... determine treatment process required HAP biodegradation efficiency (Rbio) for compliance with standards... procedures to minimize the loss of compounds due to volatilization, biodegradation, reaction, or sorption... compounds due to volatilization, biodegradation, reaction, or sorption during the sample collection, storage...

  16. 40 CFR 63.694 - Testing methods and procedures.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... determine treatment process required HAP biodegradation efficiency (Rbio) for compliance with standards... procedures to minimize the loss of compounds due to volatilization, biodegradation, reaction, or sorption... compounds due to volatilization, biodegradation, reaction, or sorption during the sample collection, storage...

  17. 40 CFR 63.694 - Testing methods and procedures.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... determine treatment process required HAP biodegradation efficiency (Rbio) for compliance with standards... procedures to minimize the loss of compounds due to volatilization, biodegradation, reaction, or sorption... compounds due to volatilization, biodegradation, reaction, or sorption during the sample collection, storage...

  18. Bisphenol A exposure assessment from olive oil consumption.

    PubMed

    Abou Omar, Tarek F; Sukhn, Carol; Fares, Souha A; Abiad, Mohamad G; Habib, Rima R; Dhaini, Hassan R

    2017-07-01

    The use of bisphenol A (BPA) in packaging has grown over the past 50 years despite concerns of its migration into packaged food and beverages, resulting in human exposure. Many studies have reported tumorigenic effects and endocrine alterations associated with BPA in animal models. This study aims at assessing human exposure to BPA from olive oil. A total of 27 olive oil samples were collected from mills and local villagers in the Hasbaya District, a major olive oil harvesting region in Lebanon. Information on storage conditions was also collected. BPA was extracted and quantified by HPLC. Results showed significantly higher BPA levels in olive oil samples stored in plastic vs. non-plastic packaging (mean = 333 vs. 150 μg/kg, p value = 0.006), samples with a plastic storage duration of >1 year compared to those with a storage duration of <1 year (mean = 452 vs. 288 μg/kg, p value = 0.008), and oil samples sourced from locals compared to oil mills (mean = 376 vs. 228 μg/kg, p value = 0.022). Statistically significant higher BPA levels remained for samples stored in plastic vs. non-plastic packaging in the bootstrap multivariable linear regression (B = 121.56, 95% CI 53.44-194.39, p value = 0.009). This is the first report on BPA levels in Mediterranean olive oil. The estimated exposure was 1.38% of the EFSA tolerable daily intake, hence there are no concerns about potential health risks from olive oil consumption.

  19. Apollo Lunar Sample Photographs: Digitizing the Moon Rock Collection

    NASA Technical Reports Server (NTRS)

    Lofgren, Gary E.; Todd, Nancy S.; Runco, S. K.; Stefanov, W. L.

    2011-01-01

    The Acquisition and Curation Office at JSC has undertaken a 4-year data restoration project effort for the lunar science community funded by the LASER program (Lunar Advanced Science and Exploration Research) to digitize photographs of the Apollo lunar rock samples and create high resolution digital images. These sample photographs are not easily accessible outside of JSC, and currently exist only on degradable film in the Curation Data Storage Facility

  20. Environmental investigation for property transfer, Fort Holabird Crime Records Center. Environmental investigation and alternatives assessment. Final report

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

    Finger, F.J.; Todd, Q.R.

    An Environmental Investigation and Alternatives Assessment was conducted for the Crime Records Center at Fort Holabird in Baltimore, Maryland. Groundwater sampling, underground storage tank tightness testing, radon sampling, asbestos sampling, soil observations, and a review of previously collected data were used to develop remedial alternatives and recommendations. Removal and record keeping were recommended for asbestos. The recommendation for groundwater was no action.

  1. Feasibility and quality development of biomaterials in the pretest studies of the German National Cohort.

    PubMed

    Kühn, A; Nieters, A; Köttgen, A; Goek, O N; Michels, K; Nöthlings, U; Jacobs, G; Meisinger, C; Pessler, F; Akmatov, M F; Kühnisch, J; Moebus, S; Glocker, E; Naus, S; Keimling, M; Leitzmann, M; Linseisen, J; Sarioglu, H; von Toerne, C; Hauck, S M; Wallaschofski, H; Wichmann, H E; Illig, Thomas

    2014-11-01

    The German National Cohort (GNC) is designed to address research questions concerning a wide range of possible causes of major chronic diseases (e.g. cancer, diabetes, infectious, allergic, neurologic and cardiovascular diseases) as well as to identify risk factors and prognostic biomarkers for early diagnosis and prevention of these diseases. The collection of biomaterials in combination with extensive information from questionnaires and medical examinations represents one of the central study components. In two pretest studies of the German National Cohort conducted between 2011 and 2013, a range of biomaterials from a defined number of participants was collected. Ten study centres were involved in pretest 1 and 18 study centres were involved in pretest 2. Standard operation procedures (SOP) were developed and evaluated to minimize pre-analytical artefacts during biosample collection. Within the pretest studies different aspects concerning feasibility of sample collection/preparation [pretest 1 (a)] and quality control of biomarkers and proteome analyses were investigated [pretest 1 (b), (c)]. Additionally, recruitment of study participants for specific projects and examination procedures of all study centres in a defined time period according to common standards as well as transportation and decentralized storage of biological samples were tested (pretest 2). These analyses will serve as the basis for the biomaterial collection in the main study of the GNC starting in 2014. Participants, randomly chosen from the population (n = 1000 subjects recruited at ten study sites in pretest 1) were asked to donate blood, urine, saliva and stool samples. Additionally, nasal and oropharyngeal swabs were collected at the study sites and nasal swabs were collected by the participants at home. SOPs for sample collection, preparation, storage and transportation were developed and adopted for pretest 2. In pretest 2, 18 study sites (n = 599 subjects) collected biomaterials mostly identical to pretest 1. Biomarker analyses to test the quality of the biomaterials were performed. In pretest 1 and 2, it was feasible to collect all biomaterials from nearly all invited participants without major problems. The mean response rate of the subjects was 95 %. As one important result we found for example that after blood draw the cellular fraction should be separated from the plasma and serum fractions during the first hour with no significant variation for up to 6 h at 4 ℃ for all analysed biomarkers. Moreover, quality control of samples using a proteomics approach showed no significant clustering of proteins according to different storage conditions. All developed SOPs were validated for use in the main study after some adaptation and modification. Additionally, electronic and paper documentation sheets were developed and tested to record time stamps, volumes, freezing times, and aliquot numbers of the collected biomaterials. The collection of the biomaterials was feasible without major problems at all participating study sites. However, the processing times were in some cases too long. To avoid pre-analytical artefacts in sample collection, appropriate standardisation among the study sites is necessary. To achieve this, blood and urine collection will have to be adapted to specific conditions of usage of liquid handling robots, which will be available at all participating study centres in the main study of the GNC. Strict compliance with the SOPs, thorough training of the staff and accurate documentation are mandatory to obtain high sample quality for later analyses. The so obtained biomaterials represent a valuable resource for research on infectious and other common complex diseases in the GNC.

  2. High-throughput metataxonomic characterization of the raw milk microbiota identifies changes reflecting lactation stage and storage conditions.

    PubMed

    Doyle, Conor J; Gleeson, David; O'Toole, Paul W; Cotter, Paul D

    2017-08-16

    Low temperature is used to control the growth of bacteria in milk, both pre- and post-pasteurization. As the duration of refrigerated storage extends, psychrotrophs dominate the milk microbiota, that can produce heat stable lipases which negatively impact the organoleptic qualities of milk. Here we examine the influence that refrigeration temperature (2°C, 4°C and 6°C) and storage duration (96h) have on the microbiota composition (16S profiling) of raw bulk tank milk (BTM). To reflect a proposed change to current farming practices, raw milk was blended after each milking (8 milkings) and stored for five consecutive days in each temperature-specific tank. Here 16S rRNA-based microbiota compositional analysis was performed after milk was collected on day 1 and again after the final addition of milk at day 5. In addition to assessing the impact of the duration and temperature of storage, the influence of lactation stage, i.e. mid- versus late-lactation, on the microbiota of the blended BTM was also examined. Overall, both temperature and length of storage had surprisingly little influence on the raw milk microbiota, other than an increase in proportions of Gammaproteobacteria in the blended milk samples collected after pooling on day 5, and in samples stored at 6°C. However, lactation stage had a considerable influence on microbiota composition, with milk from mid-lactation containing higher proportions of Bacteroides, Faecalibacterium, Campylobacter and Rhodanobacter, and late-lactation milk containing higher proportions of Actinobacteria. Overall, the study demonstrates that current temperature and storage duration practises impact the microbiota of raw milk, but these impacts are modest relative to the more considerable differences between mid and late-lactation milk. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. National Aeronautics and Space Administration Biological Specimen Repository

    NASA Technical Reports Server (NTRS)

    McMonigal, Kathleen A.; Pietrzyk, Robert a.; Johnson, Mary Anne

    2008-01-01

    The National Aeronautics and Space Administration Biological Specimen Repository (Repository) is a storage bank that is used to maintain biological specimens over extended periods of time and under well-controlled conditions. Samples from the International Space Station (ISS), including blood and urine, will be collected, processed and archived during the preflight, inflight and postflight phases of ISS missions. This investigation has been developed to archive biosamples for use as a resource for future space flight related research. The International Space Station (ISS) provides a platform to investigate the effects of microgravity on human physiology prior to lunar and exploration class missions. The storage of crewmember samples from many different ISS flights in a single repository will be a valuable resource with which researchers can study space flight related changes and investigate physiological markers. The development of the National Aeronautics and Space Administration Biological Specimen Repository will allow for the collection, processing, storage, maintenance, and ethical distribution of biosamples to meet goals of scientific and programmatic relevance to the space program. Archiving of the biosamples will provide future research opportunities including investigating patterns of physiological changes, analysis of components unknown at this time or analyses performed by new methodologies.

  4. Quantitative evaluation of plasma after methylene blue and white light treatment in four Chinese blood centers.

    PubMed

    Chunhui, Yang; Guohui, Bian; Hong, Yang; Xiaopu, Xiao; Zherong, Bai; Mingyuan, Wang; Xinsheng, Zhang; Juanjuan, Wang; Changqing, Li; Wuping, Li

    2013-12-01

    Pathogen reduction technology is an important process in the blood safety system, including solvent/detergent treatment, filtration and methylene blue-photochemical technology (MB-PCT). To investigate the quality of MB-PCT-treated plasma, plasma samples from four Chinese blood centers were analyzed over 12 months of storage to determine the recovery of activities and levels of various plasma proteins. Ten plasma units each from the Suzhou, Yancheng, Chongqing and Shandong blood centers were divided into two aliquots. One was subjected to treatment with one of two methylene blue-photochemical technology instruments and the other was used as control. The treated and untreated sample pairs were stored at -30°C. The recovery rates of coagulation factors, inhibitor proteins, total protein, immunoglobulins, and complement proteins were measured at different time points after storage. The mean recovery of most proteins exceeded 80% after MB treatment. The exceptions were factor XI and fibrinogen, of which only 71.3-74% and 69.0-92.3% were retained during storage. The two equipment types differed in terms of residual MB concentration in the plasma samples (0.18 μM and 0.29 μM, respectively). They had similar protein recovery rates at 0.5 month but differed at later time points. The four blood centers differed significantly with regard to factor II, V, VIII and fibrinogen activities. Only the samples from the Shandong blood center met the methylene blue treated fresh frozen plasma requirement. The major factor that influenced the quality of the MB-FFP samples was the time taken between blood collection and storage. Methylene blue treated plasma showed reduced coagulation factor (CF) activity and protein levels. The MB treatment-induced damage to the proteins was acceptable at the four Chinese blood centers, but the quality of the MB-treated plasma in general was not satisfactory. The main factor affecting plasma quality may be the duration of the collection and processing. Copyright © 2013 Elsevier Ltd. All rights reserved.

  5. Fluid transport container

    DOEpatents

    DeRoos, Bradley G.; Downing, Jr., John P.; Neal, Michael P.

    1995-01-01

    An improved fluid container for the transport, collection, and dispensing of a sample fluid that maintains the fluid integrity relative to the conditions of the location at which it is taken. More specifically, the invention is a fluid sample transport container that utilizes a fitment for both penetrating and sealing a storage container under controlled conditions. Additionally, the invention allows for the periodic withdrawal of portions of the sample fluid without contamination or intermixing from the environment surrounding the sample container.

  6. Assessment of petroleum-hydrocarbon contamination in the surficial sediments and ground water at three former underground storage tank locations, Fort Jackson, South Carolina, 1995

    USGS Publications Warehouse

    Robertson, J.F.

    1996-01-01

    Ground-water and sediment contamination by petroleum hydrocarbons resulting from leaks and overfills was detected during tank removal activities at three former underground storage tank locations at Fort Jackson, near Columbia, South Carolina. Investigations were initiated to assess the effect of contamination to the surficial aquifer at Sites 1062, 2438, and 2444. These investigations involved the installation of permanent monitoring wells and the collection and analysis of sediment and ground-water samples at the three sites. Water-level data were collected at all sites to determine hydraulic gradients and the direction of ground-water flow. In addition, aquifer tests were made at Site 1062 to determine the hydraulic conductivity of the surficial aquifer at that site. Sediment borings were made at the three sites to collect subsurface-sediment samples for lithologic description and laboratory analyses, and for the installation of ground-water monitoring wells. Laboratory analyses of sediment samples collected from boreholes at Site 1062 indicated elevated concentrations of petroleum hydrocarbons at three locations. Total Petroleum Hydrocarbons - Diesel Range Organics were detected at one borehole at a concentration of 388,000 micrograms per kilogram. Total benzene, toluene, ethylbenzene, and xylene concentrations in sediment from the site ranged from less than 350 to over 100,000 micrograms per kilogram. Total lead was detected at concentrations ranging from 2,900 to 5,900 micrograms per kilogram. Petroleum hydrocarbons were detected at Site 2438 in one borehole at a trace concentration of 112 micrograms per kilogram of para- and meta-xylenes. No concentrations exceeding the detection limits were reported for petroleum hydrocarbons in sediment samples collected from Site 2444; however, total lead was detected in sediment samples from two boreholes, each at concentrations of 600 micrograms per kilogram. Ground-water samples were collected from each site for laboratory analysis and field-property determinations. Petroleum hydrocarbons and lead were detected at concentrations exceeding regulatory limits for drinking water in ground water from Site 1062 only. Petroleum hydrocarbons were detected in ground water from three wells at Site 1062, with the highest concentrations occurring in the area of the former underground storage tanks. Benzene was detected at concentrations as much as 28 micrograms per liter; toluene as much as 558 micrograms per liter; para- and meta-xylenes as much as 993 micrograms per liter; and naphthalene as much as 236 micrograms per liter. Ethylbenzene and ortho-xylene were detected in one well at concentrations of 70 and 6 micrograms per liter, respectively. Dissolved lead was detected in ground water from four wells at concentrations from 5 to 152 micrograms per liter. Analysis of ground-water samples collected from Sites 2438 and 2444 showed little evidence of petroleum-hydrocarbon contamination. Petroleum hydrocarbons were not detected in any of the ground-water samples collected from Site 2438. With the exception of a low concentration of naphthalene (11 micrograms per liter) detected in ground water from one well, petroleum hydrocarbons and lead were not detected in ground water collected from Site 2444.

  7. Human DNA extraction from whole saliva that was fresh or stored for 3, 6 or 12 months using five different protocols

    PubMed Central

    GARBIERI, Thais Francini; BROZOSKI, Daniel Thomas; DIONÍSIO, Thiago José; SANTOS, Carlos Ferreira; NEVES, Lucimara Teixeira das

    2017-01-01

    Abstract Saliva when compared to blood collection has the following advantages: it requires no specialized personnel for collection, allows for remote collection by the patient, is painless, well accepted by participants, has decreased risks of disease transmission, does not clot, can be frozen before DNA extraction and possibly has a longer storage time. Objective and Material and Methods This study aimed to compare the quantity and quality of human DNA extracted from saliva that was fresh or frozen for three, six and twelve months using five different DNA extraction protocols: protocol 1 – Oragene™ commercial kit, protocol 2 – QIAamp DNA mini kit, protocol 3 – DNA extraction using ammonium acetate, protocol 4 – Instagene™ Matrix and protocol 5 – Instagene™ Matrix diluted 1:1 using proteinase K and 1% SDS. Briefly, DNA was analyzed using spectrophotometry, electrophoresis and PCR. Results Results indicated that time spent in storage typically decreased the DNA quantity with the exception of protocol 1. The purity of DNA was generally not affected by storage times for the commercial based protocols, while the purity of the DNA samples extracted by the noncommercial protocols typically decreased when the saliva was stored longer. Only protocol 1 consistently extracted unfragmented DNA samples. In general, DNA samples extracted through protocols 1, 2, 3 and 4, regardless of storage time, were amplified by human specific primers whereas protocol 5 produced almost no samples that were able to be amplified by human specific primers. Depending on the protocol used, it was possible to extract DNA in high quantities and of good quality using whole saliva, and furthermore, for the purposes of DNA extraction, saliva can be reliably stored for relatively long time periods. Conclusions In summary, a complicated picture emerges when taking into account the extracted DNA’s quantity, purity and quality; depending on a given researchers needs, one protocol’s particular strengths and costs might be the deciding factor for its employment. PMID:28403355

  8. The effect of pre-storage cooling on 2,3-DPG levels in red cells stored in SAG-M.

    PubMed

    Llohn, Abid Hussain; Vetlesen, Annette; Fagerhol, Magne Kristoffer; Kjeldsen-Kragh, Jens

    2005-10-01

    The concentration of red cell 2,3-DPG (2,3-diphosphoglycerate) rapidly decreases during storage. A favourable effect on red cell 2,3-DPG has been demonstrated by rapid cooling of whole blood prior to storage. In our study we have investigated how different methods of cooling whole blood immediately after donation effect 2,3-DPG levels during storage. Thirty-six whole blood units (in 6 groups) of 450 ml were collected in 63 ml CPD. SAG-M was used as preservative solution for red cell concentrates (RCC). The units in one group were cooled down at ambient temperature, while units in the other groups were cooled down rapidly by different ways immediately after bleeding. Samples from the whole blood units were collected at various days during storage for 2,3-DPG measurements. The decline in 2,3-DPG during the first two weeks of storage was significantly slower in the groups which were cooled down rapidly to 17-18 degrees C within 1h after bleeding (all p

  9. Effect of storage time on gene expression data acquired from unfrozen archived newborn blood spots.

    PubMed

    Ho, Nhan T; Busik, Julia V; Resau, James H; Paneth, Nigel; Khoo, Sok Kean

    2016-11-01

    Unfrozen archived newborn blood spots (NBS) have been shown to retain sufficient messenger RNA (mRNA) for gene expression profiling. However, the effect of storage time at ambient temperature for NBS samples in relation to the quality of gene expression data is relatively unknown. Here, we evaluated mRNA expression from quantitative real-time PCR (qRT-PCR) and microarray data obtained from NBS samples stored at ambient temperature to determine the effect of storage time on the quality of gene expression. These data were generated in a previous case-control study examining NBS in 53 children with cerebral palsy (CP) and 53 matched controls. NBS sample storage period ranged from 3 to 16years at ambient temperature. We found persistently low RNA integrity numbers (RIN=2.3±0.71) and 28S/18S rRNA ratios (~0) across NBS samples for all storage periods. In both qRT-PCR and microarray data, the expression of three common housekeeping genes-beta cytoskeletal actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and peptidylprolyl isomerase A (PPIA)-decreased with increased storage time. Median values of each microarray probe intensity at log 2 scale also decreased over time. After eight years of storage, probe intensity values were largely reduced to background intensity levels. Of 21,500 genes tested, 89% significantly decreased in signal intensity, with 13,551, 10,730, and 9925 genes detected within 5years, > 5 to <10years, and >10years of storage, respectively. We also examined the expression of two gender-specific genes (X inactivation-specific transcript, XIST and lysine-specific demethylase 5D, KDM5D) and seven gene sets representing the inflammatory, hypoxic, coagulative, and thyroidal pathways hypothesized to be related to CP risk to determine the effect of storage time on the detection of these biologically relevant genes. We found the gender-specific genes and CP-related gene sets detectable in all storage periods, but exhibited differential expression (between male vs. female or CP vs. control) only within the first six years of storage. We concluded that gene expression data quality deteriorates in unfrozen archived NBS over time and that differential gene expression profiling and analysis is recommended for those NBS samples collected and stored within six years at ambient temperature. Copyright © 2016 Elsevier Inc. All rights reserved.

  10. Pre-analytic evaluation of volumetric absorptive microsampling and integration in a mass spectrometry-based metabolomics workflow.

    PubMed

    Volani, Chiara; Caprioli, Giulia; Calderisi, Giovanni; Sigurdsson, Baldur B; Rainer, Johannes; Gentilini, Ivo; Hicks, Andrew A; Pramstaller, Peter P; Weiss, Guenter; Smarason, Sigurdur V; Paglia, Giuseppe

    2017-10-01

    Volumetric absorptive microsampling (VAMS) is a novel approach that allows single-drop (10 μL) blood collection. Integration of VAMS with mass spectrometry (MS)-based untargeted metabolomics is an attractive solution for both human and animal studies. However, to boost the use of VAMS in metabolomics, key pre-analytical questions need to be addressed. Therefore, in this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. We first evaluated the best extraction procedure for the polar metabolome and found that the highest number and amount of metabolites were recovered upon extraction with acetonitrile/water (70:30). In contrast, basic conditions (pH 9) resulted in divergent metabolite profiles mainly resulting from the extraction of intracellular metabolites originating from red blood cells. In addition, the prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but once the VAMS devices were stored at - 80 °C, the metabolome remained stable for up to 6 months. The time used for drying the sample did also affect the metabolome. In fact, some metabolites were rapidly degraded or accumulated in the sample during the first 48 h at room temperature, indicating that a longer drying step will significantly change the concentration in the sample. Graphical abstract Volumetric absorptive microsampling (VAMS) is a novel technology that allows single-drop blood collection and, in combination with mass spectrometry (MS)-based untargeted metabolomics, represents an attractive solution for both human and animal studies. In this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. The latter revealed that prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but if VAMS devices were stored at - 80 °C, the metabolome remained stable for up to 6 months.

  11. A Systematic Evaluation of Blood Serum and Plasma Pre-Analytics for Metabolomics Cohort Studies

    PubMed Central

    Jobard, Elodie; Trédan, Olivier; Postoly, Déborah; André, Fabrice; Martin, Anne-Laure; Elena-Herrmann, Bénédicte; Boyault, Sandrine

    2016-01-01

    The recent thriving development of biobanks and associated high-throughput phenotyping studies requires the elaboration of large-scale approaches for monitoring biological sample quality and compliance with standard protocols. We present a metabolomic investigation of human blood samples that delineates pitfalls and guidelines for the collection, storage and handling procedures for serum and plasma. A series of eight pre-processing technical parameters is systematically investigated along variable ranges commonly encountered across clinical studies. While metabolic fingerprints, as assessed by nuclear magnetic resonance, are not significantly affected by altered centrifugation parameters or delays between sample pre-processing (blood centrifugation) and storage, our metabolomic investigation highlights that both the delay and storage temperature between blood draw and centrifugation are the primary parameters impacting serum and plasma metabolic profiles. Storing the blood drawn at 4 °C is shown to be a reliable routine to confine variability associated with idle time prior to sample pre-processing. Based on their fine sensitivity to pre-analytical parameters and protocol variations, metabolic fingerprints could be exploited as valuable ways to determine compliance with standard procedures and quality assessment of blood samples within large multi-omic clinical and translational cohort studies. PMID:27929400

  12. Hematopoietic Lineage Transcriptome Stability and Representation in PAXgene™ Collected Peripheral Blood Utilising SPIA Single-Stranded cDNA Probes for Microarray

    PubMed Central

    Kennedy, Laura; Vass, J. Keith; Haggart, D. Ross; Moore, Steve; Burczynski, Michael E.; Crowther, Dan; Miele, Gino

    2008-01-01

    Peripheral blood as a surrogate tissue for transcriptome profiling holds great promise for the discovery of diagnostic and prognostic disease biomarkers, particularly when target tissues of disease are not readily available. To maximize the reliability of gene expression data generated from clinical blood samples, both the sample collection and the microarray probe generation methods should be optimized to provide stabilized, reproducible and representative gene expression profiles faithfully representing the transcriptional profiles of the constituent blood cell types present in the circulation. Given the increasing innovation in this field in recent years, we investigated a combination of methodological advances in both RNA stabilisation and microarray probe generation with the goal of achieving robust, reliable and representative transcriptional profiles from whole blood. To assess the whole blood profiles, the transcriptomes of purified blood cell types were measured and compared with the global transcriptomes measured in whole blood. The results demonstrate that a combination of PAXgene™ RNA stabilising technology and single-stranded cDNA probe generation afforded by the NuGEN Ovation RNA amplification system V2™ enables an approach that yields faithful representation of specific hematopoietic cell lineage transcriptomes in whole blood without the necessity for prior sample fractionation, cell enrichment or globin reduction. Storage stability assessments of the PAXgene™ blood samples also advocate a short, fixed room temperature storage time for all PAXgene™ blood samples collected for the purposes of global transcriptional profiling in clinical studies. PMID:19578521

  13. Mars rover 1988 concepts

    NASA Technical Reports Server (NTRS)

    Pivirotto, Donna Shirley; Penn, Thomas J.; Dias, William C.

    1989-01-01

    Results of FY88 studies of a sample-collecting Mars rover are presented. A variety of rover concepts are discussed which include different technical approaches to rover functions. The performance of rovers with different levels of automation is described and compared to the science requirement for 20 to 40 km to be traversed on the Martian surface and for 100 rock and soil samples to be collected. The analysis shows that a considerable amount of automation in roving and sampling is required to meet this requirement. Additional performance evaluation shows that advanced RTG's producing 500 W and 350 WHr of battery storage are needed to supply the rover.

  14. Method development for the analysis of organophosphate and pyrethroid insecticides at low parts per trillion levels in water.

    PubMed

    Wang, Dongli; Weston, Donald P; Lydy, Michael J

    2009-06-15

    In the current study, organophosphate and pyrethroid insecticides including diazinon, chlorpyrifos, bifenthrin, fenpropathrin, permethrin, lambda-cyhalothrin, cyfluthrin, cypermethrin, esfenvalerate and deltamethrin were analyzed in laboratory and field-collected water samples. Water samples were extracted and analyzed by gas chromatography/electron capture detector (GC/ECD) and gas chromatography/nitrogen-phosphorous detector (GC/NPD). Comparison of results from liquid-liquid extraction and subsequent normal phase solid-phase extraction cleanup (LLE-NPSPE), and reversed phase solid-phase extraction (RPSPE) showed that LLE-NPSPE was the better choice to extract trace amounts of pesticides from water. Pesticide recoveries from four spiked water samples using LLE-NPSPE ranged from 63.2 to 148.8% at four spiking concentrations. Method detection limits were 0.72-1.69 ng/L using four different water sources. The stability of the target pesticides in lake water was investigated at 4 degrees C for 1h, 1d, 4d, and 7d under three conditions: (1) water samples only; (2) with 20 mL hexane used as a keeper solvent; and (3) with acidification to pH 2 with HCl. Results showed that water storage without treatment resulted in slow degradation of some pesticides with storage time, storage using water acidification led to significant degradation and loss of diazinon and chlorpyrifos, while water storage with hexane as a keeper solvent showed good stability for all of the target pesticides over the 7d storage period.

  15. Pre-analytical issues in the haemostasis laboratory: guidance for the clinical laboratories.

    PubMed

    Magnette, A; Chatelain, M; Chatelain, B; Ten Cate, H; Mullier, F

    2016-01-01

    Ensuring quality has become a daily requirement in laboratories. In haemostasis, even more than in other disciplines of biology, quality is determined by a pre-analytical step that encompasses all procedures, starting with the formulation of the medical question, and includes patient preparation, sample collection, handling, transportation, processing, and storage until time of analysis. This step, based on a variety of manual activities, is the most vulnerable part of the total testing process and is a major component of the reliability and validity of results in haemostasis and constitutes the most important source of erroneous or un-interpretable results. Pre-analytical errors may occur throughout the testing process and arise from unsuitable, inappropriate or wrongly handled procedures. Problems may arise during the collection of blood specimens such as misidentification of the sample, use of inadequate devices or needles, incorrect order of draw, prolonged tourniquet placing, unsuccessful attempts to locate the vein, incorrect use of additive tubes, collection of unsuitable samples for quality or quantity, inappropriate mixing of a sample, etc. Some factors can alter the result of a sample constituent after collection during transportation, preparation and storage. Laboratory errors can often have serious adverse consequences. Lack of standardized procedures for sample collection accounts for most of the errors encountered within the total testing process. They can also have clinical consequences as well as a significant impact on patient care, especially those related to specialized tests as these are often considered as "diagnostic". Controlling pre-analytical variables is critical since this has a direct influence on the quality of results and on their clinical reliability. The accurate standardization of the pre-analytical phase is of pivotal importance for achieving reliable results of coagulation tests and should reduce the side effects of the influence factors. This review is a summary of the most important recommendations regarding the importance of pre-analytical factors for coagulation testing and should be a tool to increase awareness about the importance of pre-analytical factors for coagulation testing.

  16. Characterization of nutrients and fecal indicator bacteria at a concentrated swine feeding operation in Wake County, North Carolina, 2009-2011

    USGS Publications Warehouse

    Harden, Stephen L.; Rogers, Shane W.; Jahne, Michael A.; Shaffer, Carrie E.; Smith, Douglas G.

    2012-01-01

    Study sites were sampled for laboratory analysis of nutrients, total suspended solids (TSS), and (or) fecal indicator bacteria (FIB). Nutrient analyses included measurement of dissolved ammonia, total and dissolved ammonia + organic nitrogen, dissolved nitrate + nitrite, dissolved orthophosphate, and total phosphorus. The FIB analyses included measurement of Escherichia coli and enterococci. Samples of wastewater at the swine facility were collected from a pipe outfall from the swine housing units, two storage lagoons, and the spray fields for analysis of nutrients, TSS, and FIB. Soil samples collected from a spray field were analyzed for FIB. Monitoring locations were established for collecting discharge and water-quality data during storm events at three in-field runoff sites and two sites on the headwater stream (one upstream and one downstream) next to the swine facility. Stormflow samples at the five monitoring locations were collected for four storm events during 2009 to 2010 and analyzed for nutrients, TSS, and FIB. Monthly water samples also were collected during base-flow conditions at all four stream sites for laboratory analysis of nutrients, TSS, and (or) FIB.

  17. Characterization of seed storage protein patterns of Heliotropium digynum.

    PubMed

    Alwhibi, Mona Soliman

    2017-09-01

    Heliotropium digynum , is a shrub that has ecological importance. The height of the plant differs from one population to another and the difference in length of the inflorescence can be attributed to environmental factors, such as rainfall or type of soil and temperature. To date, no study has shed light on estimation in seed samples of H. digynum in Saudi Arabia. So, the aim is to evaluate and characterize the protein patterns of seed storage proteins of H. digynum to be used as fingerprint of this plant in Saudi Arabia. It is collected from different locations in the central region of Saudi Arabia and total protein extraction from plant was compared in SDS-PAGE. The genetic relationships among all cultivars were analyzed using UPGMA and NJ using Total Lab TL and in the same way using Jaccard Similarity Coefficient dendrogram using STATISTICA (ver.8) software. Results, our data show that amounts of protein are different, although they are of the same type or from the same geographical region. Amounts ranged between 22 and 1.5 mg/g of dry weight. Less amount of protein was obtained from the group of samples collected from Dir'iyah area, and the highest amount of protein was from the group of samples collected from Dyrab area in general.

  18. During cooled storage the extender influences processed autophagy marker light chain 3 (LC3B) of stallion spermatozoa.

    PubMed

    Bolaños, J M Gallardo; Morán, A Miró; da Silva, C M Balao; Dávila, M Plaza; Muñoz, P Martín; Aparicio, I M; Tapia, J A; Ferrusola, C Ortega; Peña, F J

    2014-02-01

    To investigate the role of the processed autophagy marker light chain 3 (LC3B) protein in sperm survival in stallion semen processing during cooled storage, split ejaculates were diluted in two different extenders, KMT and INRA 96, and LC3B processing and sperm quality evaluated during incubation at 5°C for five days. After 3 days of incubation there was a drop in total motility in both extenders, although the percentage of progressive motile sperm was greater (P<0.05) in samples extended in INRA96. On Day 5 of cooled storage all sperm parameters decreased significantly independent of the extender, however, samples extended in INRA 96 maintained motility values while those extended in KMT had a further decrease in motility compared with data collected on Day 3 of incubation. The percentage of live sperm decreased over the time of incubation, but only in samples incubated in KMT. The extender had a marked effect in LC3B processing during cooled storage. Spermatozoa maintained in KMT extender did not exhibit LC3B processing, while in spermatozoa incubated in INRA96 there was an increase (P<0.01) in LC3B processing after 5 days of cooled storage. Stallion spermatozoa experience LC3B turnover during cooled storage, however, the extent depends on the extender used. Apparently LC3B turnover is associated with enhanced survival. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Final work plan : supplemental upward vapor intrusion investigation at the former CCC/USDA grain storage facility in Hanover, Kansas.

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

    LaFreniere, L. M.; Environmental Science Division

    The Commodity Credit Corporation (CCC), an agency of the U.S. Department of Agriculture (USDA), operated a grain storage facility at the northeastern edge of the city of Hanover, Kansas, from 1950 until the early 1970s. During this time, commercial grain fumigants containing carbon tetrachloride were in common use by the grain storage industry to preserve grain in their facilities. In February 1998, trace to low levels of carbon tetrachloride (below the maximum contaminant level [MCL] of 5.0 {micro}g/L) were detected in two private wells near the former grain storage facility at Hanover, as part of a statewide USDA private wellmore » sampling program that was implemented by the Kansas Department of Health and Environment (KDHE) near former CCC/USDA facilities. In 2007, the CCC/USDA conducted near-surface soil sampling at 61 locations and also sampled indoor air at nine residences on or adjacent to its former Hanover facility to address the residents concerns regarding vapor intrusion. Low levels of carbon tetrachloride were detected at four of the nine homes. The results were submitted to the KDHE in October 2007 (Argonne 2007). On the basis of the results, the KDHE requested sub-slab sampling and/or indoor air sampling (KDHE 2007). This Work Plan describes, in detail, the proposed additional scope of work requested by the KDHE and has been developed as a supplement to the comprehensive site investigation work plan that is pending (Argonne 2008). Indoor air samples collected previously from four homes at Hanover were shown to contain the carbon tetrachloride at low concentrations (Table 2.1). It cannot be concluded from these previous data that the source of the detected carbon tetrachloride is vapor intrusion attributable to former grain storage operations of the CCC/USDA at Hanover. The technical objective of the vapor intrusion investigation described here is to assess the risk to human health due to the potential for upward migration of carbon tetrachloride and chloroform into four homes located on or adjacent to the former CCC/USDA facility. The technical objective will be accomplished by collecting sub-slab vapor samples. The preliminary data collected during the July 2007 investigation did not fully address the source of or migration pathway for the carbon tetrachloride detected in the four homes. The scope of work proposed here will generate additional data needed to help evaluate whether the source of the detected carbon tetrachloride is vapor intrusion attributable to activities of the CCC/USDA. The additional vapor sampling at Hanover will be performed, on behalf of the CCC/USDA, by the Environmental Science Division of Argonne National Laboratory and H&P Mobile Geochemistry of San Diego (http://www.handpmg.com). Argonne is a nonprofit, multidisciplinary research center operated by UChicago Argonne, LLC, for the U.S. Department of Energy (DOE). The CCC/USDA has entered into an interagency agreement with DOE, under which Argonne provides technical assistance to the CCC/USDA with environmental site characterization and remediation at its former grain storage facilities. The professional staff members of H&P Mobile Geochemistry are nationally leading experts in soil gas sampling and vapor intrusion investigations.« less

  20. Lipolytic and proteolytic activity of Pseudomonas spp. isolated during milking and storage of refrigerated raw milk.

    PubMed

    Capodifoglio, Eduardo; Vidal, Ana Maria Centola; Lima, Joyce Aparecida Santos; Bortoletto, Fernanda; D'Abreu, Léa Furlan; Gonçalves, Ana Carolina Siqueira; Vaz, Andreia Cristina Nakashima; Balieiro, Julio Cesar de Carvalho; Netto, Arlindo Saran

    2016-07-01

    The aim of this study was to verify the presence of lipolytic and proteolytic Pseudomonas spp. during milking and storage of refrigerated raw milk. We also intended to compare samples collected during rainy and dry seasons, from farms with manual and mechanical milking systems. For this, samples of milkers' hands, cows' teats, water, expansion tanks, equipment, and utensils used during milking were analyzed regarding Pseudomonas spp. Positive samples were tested for the production of lipolytic and proteolytic enzymes. Microorganisms of the genus Pseudomonas were isolated from all sampling points. A higher isolation rate of the bacterium was found in the rainy season except for 6 sampling points, with all of these associated with mechanical milking systems. Pseudomonas spp. exhibiting lipolytic activity were found to be predominant during the dry season, since no activity was detected during the rainy season in 26 of the 29 sampling sites. The highest number of lipolytic Pseudomonas isolates was obtained from water. Presence of lipase-producing Pseudomonas spp. was verified in 7 and 36% of the samples collected from farms with manual and mechanical milking, respectively. When analyzing raw milk collected from expansion tanks immediately (0 h) and 24h after milking, we observed that for dairy properties with manual milking process, 10% of the Pseudomonas isolates were positive for lipolytic activity. The percentage increased to 12% 48h after milking. Mean averages were 32, 33, and 39% immediately after, 24 and 48h after milking, respectively, for farms with mechanical milking. All sampling points showed the presence of proteolytic strains of Pseudomonas. The highest proteolytic activity was found during the rainy season, except for the samples collected from milkers' hands before milking, buckets, and teat cup inner surfaces after milking and from the water in dairy farms with mechanical milking system. Of these samples, 72, 56, and 50%, respectively, were positive for proteolysis during the dry season. For the water samples, a statistical difference was observed between mechanical (50%) and manual (7%) milking systems in the percentage of proteolytic activity. No production of proteolytic enzyme was detected in the samples from milkers' hands taken after milking and no statistically significant difference was found among manual (19.91%) and mechanical (47.85%) milking. During the rainy months, no proteolysis was detected in the samples taken from cows' teats after the predipping. It is evident, therefore, that preventive measures capable of minimizing the contamination with Pseudomonas spp. during milking and storage of refrigerated raw milk are needed, regardless of season. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  1. Influence of storage and preservation on microbiological quality of silo ovine milk.

    PubMed

    de Garnica, M L; Santos, J A; Gonzalo, C

    2011-04-01

    This study was designed to analyze the effects of the storage and preservation conditions on counts of mesophilic, thermoduric, psychotrophic, coliform, Escherichia coli, Streptococcus agalactiae, and Staphylococcus aureus organisms in silo ovine milk. A total of 910 analytical determinations were conducted from aliquots of 10 silo ovine milks. The conditions tested were unpreserved and azidiol-preserved milk stored at 4°C, and unpreserved milk stored at -20°C. Milk aged 2, 24, 48, 72, and 96 h post-collection for refrigerated aliquots, and 7, 15, and 30 d post-collection for frozen aliquots. The factors silo and storage conditions significantly contributed to variation of all microbiological variables, although milk age effect within storage was only significant for mesophilic, psychrotrophic, and coliform bacteria counts. In refrigerated raw milk, mesophile, psychrotroph, and coliform counts significantly increased over 96 h post-collection, whereas the other groups and bacteria species tested maintained their initial concentration. In all cases, azidiol preservation maintained the initial bacterial concentration in raw sheep milk under refrigeration throughout 96 h. Thus, azidiol was a suitable preservative for microbiological studies in sheep milk. Smallest counts were registered for frozen samples, particularly for coliforms, E. coli, Strep. agalactiae and Staph. aureus. Estimates of mesophilic, thermoduric and psychrotrophic organisms showed similar values on both azidiol-preserved and frozen milk samples. Coliforms and E. coli counts significantly decrease over time after freezing. Consequently, freezing at -20°C could also be appropriate for analysis of mesophilic, thermoduric, and psychrotrophic bacterial groups, but not for coliforms or mammary pathogens. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  2. Geomorphic controls on floodplain organic carbon storage in sediment along five rivers in interior Alaska

    NASA Astrophysics Data System (ADS)

    Lininger, K.; Wohl, E.; Rose, J. R.

    2016-12-01

    High latitude permafrost regions contain large amounts of organic carbon (OC) in the subsurface, but little work has quantified OC storage in floodplain sediment in the high latitudes. Floodplains influence the export of OC to the ocean by temporarily storing OC at timescales of 101 to 103 years. To fully understand terrestrial carbon cycling, the storage and residence time of OC in floodplains, and the geomorphic controls on OC storage, must be taken into account. Small-scale spatial variations in OC storage within floodplains likely reflect geomorphic processes of deposition and floodplain development. We present results of floodplain OC storage and residence time in sediment along 5 rivers in the Yukon Flats National Wildlife Refuge in interior Alaska, a region with discontinuous permafrost. We collected sediment samples within the active layer along tributaries to the Yukon River and the mainstem Yukon River and analyzed the sediment samples for OC content. We classified sample locations by geomorphic type (filled secondary channels, levees, point bars) and vegetation type (herbaceous, deciduous/shrub, white spruce, and black spruce wetlands), and found that both geomorphology and vegetation influence OC concentration and OC mass per area. Preliminary results suggest that filled secondary channels contain more OC per area compared to other geomorphic types. We present results of radiocarbon dates from river cutbanks associated with our sampling sites, which give a maximum age for residence times of OC in sediment before erosion and transport. The radiocarbon dates also provide estimates of long-term OC accretion within the Yukon Flats floodplains. Small-scale variations within floodplains as a result of floodplain depositional processes and vegetation communities shed light on the geomorphic controls on OC storage. This work will help constrain the spatial variation in OC storage and OC residence time across the landscape in a region experiencing rapid climate change and permafrost thaw.

  3. The biobank of the Norwegian mother and child cohort Study: A resource for the next 100 years

    PubMed Central

    Rønningen, Kjersti S.; Paltiel, Liv; Meltzer, Helle M.; Nordhagen, Rannveig; Lie, Kari K.; Hovengen, Ragnhild; Haugen, Margaretha; Nystad, Wenche; Magnus, Per; Hoppin, Jane A.

    2007-01-01

    Introduction Long-term storage of biological materials is a critical component of any epidemiological study. In designing specimen repositories, efforts need to balance future needs for samples with logistical constraints necessary to process and store samples in a timely fashion. Objectives In the Norwegian Mother and Child Cohort Study (MoBa), the Biobank was charged with long-term storage of more than 380,000 biological samples from pregnant women, their partners and their children for up to 100 years. Methods Biological specimens include whole blood, plasma, DNA and urine; samples are collected at 50 hospitals in Norway. All samples are sent via ordinary mail to the Biobank in Oslo where the samples are registered, aliquoted and DNA extracted. DNA is stored at −20 °C while whole blood, urine and plasma are stored at − 80 °C. Results As of July 2006, over 227,000 sample sets have been collected, processed and stored at the Biobank. Currently 250–300 sets are received daily. An important part of the Biobank is the quality control program. Conclusion With the unique combination of biological specimens and questionnaire data, the MoBa Study will constitute a resource for many future investigations of the separate and combined effects of genetic, environmental factors on pregnancy outcome and on human morbidity, mortality and health in general. PMID:17031521

  4. Tank vapor characterization project. Headspace vapor characterization of Hanford waste tank 241-BY-108: Second comparison study results from samples collected on 3/28/96

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

    Thomas, B.L.; Pool, K.H.; Evans, J.C.

    1997-01-01

    This report describes the analytical results of vapor samples taken from the headspace of waste storage tank 241-BY-108 (Tank BY-108) at the Hanford Site in Washington State. The results described in this report is the second in a series comparing vapor sampling of the tank headspace using the Vapor Sampling System (VSS) and In Situ Vapor Sampling (ISVS) system without high efficiency particulate air (HEPA) prefiltration. The results include air concentrations of water (H{sub 2}O) and ammonia (NH{sub 3}), permanent gases, total non-methane organic compounds (TO-12), and individual organic analytes collected in SUMMA{trademark} canisters and on triple sorbent traps (TSTs).more » Samples were collected by Westinghouse Hanford Company (WHC) and analyzed by Pacific Northwest National Laboratory (PNNL). Analyses were performed by the Vapor Analytical Laboratory (VAL) at PNNL. Analyte concentrations were based on analytical results and, where appropriate, sample volume measurements provided by WHC.« less

  5. Collection, storage, inspection and quality control of platelet concentrates obtained by apheresis: The situation in Spain.

    PubMed

    Castrillo, Azucena; Jimenez-Marco, Teresa; Arroyo, José L; Jurado, María L; Larrea, Luis; Maymo, Rosa M; Monge, Jorge; Muñoz, Carmen; Pajares, Ángel; Yáñez, Marta

    2017-06-01

    Diverse variables are involved in apheresis platelet collection, processing and storage. This survey shows how these are realized in Spain. An analysis of collected data was performed in a questionnaire completed by ten Transfusion Centers (TC) which perform between 50 and 520 apheresis procedures per month. This information comprises the procedures used to collect, inspect and store apheresis platelet concentrates (PC), and quality control data. Macroscopic inspection of PC is performed in all TC, especially during the first few hours post-collection and before distribution. The type of processor, duration of post-collection resting periods and temperature from the time of collection until distribution are similar in all TC. In 80% of TC, PC with small and scarce aggregates are distributed to transfusion services. The presence of clumps is influenced by type of processor, female donor, cold ambient temperature and collection of hyperconcentrated platelets, and is often recurrent in the same donor, although some TC have not found any influential variables. Overall, no objective inspection methods are followed, although there are exceptions. The degree of compliance with quality control parameters, such as the number of units studied, mean platelet yield, residual leukocyte counts and pH at expiry date, is acceptable in all TC. Compliance in terms of number of microbiological culture samples is variable. The usual practice in Spanish TC with respect to the collection, post-collection handling and storage of apheresis PC can be considered uniform, although some specific aspects of analyses should follow more objective methods. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. The precision of wet atmospheric deposition data from national atmospheric deposition program/national trends network sites determined with collocated samplers

    USGS Publications Warehouse

    Nilles, M.A.; Gordon, J.D.; Schroder, L.J.

    1994-01-01

    A collocated, wet-deposition sampler program has been operated since October 1988 by the U.S. Geological Survey to estimate the overall sampling precision of wet atmospheric deposition data collected at selected sites in the National Atmospheric Deposition Program and National Trends Network (NADP/NTN). A duplicate set of wet-deposition sampling instruments was installed adjacent to existing sampling instruments at four different NADP/NTN sites for each year of the study. Wet-deposition samples from collocated sites were collected and analysed using standard NADP/NTN procedures. Laboratory analyses included determinations of pH, specific conductance, and concentrations of major cations and anions. The estimates of precision included all variability in the data-collection system, from the point of sample collection through storage in the NADP/NTN database. Sampling precision was determined from the absolute value of differences in the analytical results for the paired samples in terms of median relative and absolute difference. The median relative difference for Mg2+, Na+, K+ and NH4+ concentration and deposition was quite variable between sites and exceeded 10% at most sites. Relative error for analytes whose concentrations typically approached laboratory method detection limits were greater than for analytes that did not typically approach detection limits. The median relative difference for SO42- and NO3- concentration, specific conductance, and sample volume at all sites was less than 7%. Precision for H+ concentration and deposition ranged from less than 10% at sites with typically high levels of H+ concentration to greater than 30% at sites with low H+ concentration. Median difference for analyte concentration and deposition was typically 1.5-2-times greater for samples collected during the winter than during other seasons at two northern sites. Likewise, the median relative difference in sample volume for winter samples was more than double the annual median relative difference at the two northern sites. Bias accounted for less than 25% of the collocated variability in analyte concentration and deposition from weekly collocated precipitation samples at most sites.A collocated, wet-deposition sampler program has been operated since OCtober 1988 by the U.S Geological Survey to estimate the overall sampling precision of wet atmospheric deposition data collected at selected sites in the National Atmospheric Deposition Program and National Trends Network (NADP/NTN). A duplicate set of wet-deposition sampling instruments was installed adjacent to existing sampling instruments four different NADP/NTN sites for each year of the study. Wet-deposition samples from collocated sites were collected and analysed using standard NADP/NTN procedures. Laboratory analyses included determinations of pH, specific conductance, and concentrations of major cations and anions. The estimates of precision included all variability in the data-collection system, from the point of sample collection through storage in the NADP/NTN database.

  7. DNA quality and quantity from up to 16 years old post-mortem blood stored on FTA cards.

    PubMed

    Rahikainen, Anna-Liina; Palo, Jukka U; de Leeuw, Wiljo; Budowle, Bruce; Sajantila, Antti

    2016-04-01

    Blood samples preserved on FTA cards offer unique opportunities for genetic research. DNA recovered from these cards should be stable for long periods of time. However, it is not well established as how well the DNA stored on FTA card for substantial time periods meets the demands of forensic or genomic DNA analyses and especially so for from post-mortem (PM) samples in which the quality can vary upon initial collection. The aim of this study was to evaluate the time-dependent degradation on DNA quality and quantity extracted from up to 16 years old post-mortem bloodstained FTA cards. Four random FTA samples from eight time points spanning 1998 to 2013 (n=32) were collected and extracted in triplicate. The quantity and quality of the extracted DNA samples were determined with Quantifiler(®) Human Plus (HP) Quantification kit. Internal sample and sample-to-sample variation were evaluated by comparing recovered DNA yields. The DNA from the triplicate samplings were subsequently combined and normalized for further analysis. The practical effect of degradation on DNA quality was evaluated from normalized samples both with forensic and pharmacogenetic target markers. Our results suggest that (1) a PM change, e.g. blood clotting prior to sampling, affects the recovered DNA yield, creating both internal and sample-to-sample variation; (2) a negative correlation between the FTA card storage time and DNA quantity (r=-0.836 at the 0.01 level) was observed; (3) a positive correlation (r=0.738 at the level 0.01) was found between FTA card storage time and degradation levels. However, no inhibition was observed with the method used. The effect of degradation was manifested clearly with functional applications. Although complete STR-profiles were obtained for all samples, there was evidence of degradation manifested as decreased peak heights in the larger-sized amplicons. Lower amplification success was notable with the large 5.1 kb CYP2D6 gene fragment which strongly supports degradation of the stored samples. According to our results, DNA stored on FTA cards is rather stable over a long time period. DNA extracted from this storage medium can be used as human identification purposes as the method used is sufficiently sensitive and amplicon sizes tend to be <400 bp. However, DNA integrity was affected during storage. This effect should be taken into account depending on the intended application especially if high quality DNA and long PCR amplicons are required. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  8. The storage stability of biogenic volatile organic compounds (BVOCs) in polyester aluminum bags

    NASA Astrophysics Data System (ADS)

    Ahn, Jeong-Hyeon; Deep, Akash; Kim, Ki-Hyun

    2016-09-01

    In this study, the sorptive loss properties of biogenic volatile organic compounds (BVOCs) in polyester aluminum bags were investigated as a function of storage duration. To this end, the relative recovery of gas phase standards of BVOCs, obtained via vaporization of liquid phase standards, was computed by calibrating their standards (response factors: RF) represnting each phase. Accordingly, the results indicated either slight loss (-5.59% (isoprene), -2.39% (camphene), -1.69% ((R)-(+)-limonene), -0.88% (p-cymene)) or gain (1.47% (γ-terpinene), 2.27% (α-terpinene), 2.63% (α-phellandrene), 2.73% ((+)-3-carene), 3.93% ((+)-β-pinene), and 5.98% ((+)-α-pinene)). Through comparison of the calibration results across storage time, the temporal stability of BVOCs was assessed. Longer BVOC storage time in polyester aluminum (PEA) bags lowered the relative recovery of BVOCs. The relative loss of BVOCs, if calculated in terms of mean bag standard RF ratios (relative to liquid standard) across elapsed time, decreased systematically: 0.99 ± 0.05 (0 h), 0.88 ± 0.06 (24 h), 0.66 ± 0.11 (72 h), and 0.62 ± 0.14 (120 h). It is thus recommended to complete the analysis of BVOC in PEA bags within 24 h of sample acquisition. As such, it is important to apply appropriate sampling approaches with a proper storage plan when measuring ambient BVOCs collected by bag sampling methods.

  9. Field methods for sampling and storing nectar from flowers with low nectar volumes.

    PubMed

    Morrant, D S; Schumann, R; Petit, S

    2009-02-01

    Although several methods of sampling and storing floral nectar are available, little information exists on sampling and storing nectar from flowers with low nectar volumes. Methods for sampling and storing nectar from the flowers of species with low floral nectar volumes (<1 microL) were investigated using the flowers of Eucalyptus species. Sampling with microcapillary tubes, blotting up with filter paper, washing and rinsing were compared to determine masses of sugars recovered and differences in sugar ratios. Storage methods included room temperature, refrigeration and freezing treatments; the addition of antimicrobial agents benzyl alcohol or methanol to some of these treatments was also evaluated. Nectar samples were analysed using high-performance liquid chromatography and the masses of sucrose, glucose and fructose in each sample were determined. Masses of sugars varied significantly among sampling treatments, but the highest yielding methods, rinsing and washing, were not significantly different. A washing time of 1 min was as effective as one of 20 min. Storage trials showed that the sugar concentration measurements of nectar solutions changed rapidly, with the best results achieved for refrigeration with no additive (sucrose and fructose were stable for at least 2 weeks). Sugar ratios, however, remained relatively stable in most treatments and did not change significantly across 4 weeks for the methanol plus refrigerator and freezing treatments, and 2 weeks for the refrigeration treatment with no additive. Washing is recommended for nectar collection from flowers with low nectar volumes in the field (with the understanding that one wash underestimates the amounts of sugars present in a flower), as is immediate analysis of sugar mass. In view of the great variation in results depending on nectar collection and storage methods, caution should be exercised in their choice, and their accuracy should be evaluated. The use of pulsed amperometric detection, more specific than refractive index detection, may improve the accuracy of nectar sugar analysis.

  10. Occurrence of Penicillium verrucosum, ochratoxin A, ochratoxin B and citrinin in on-farm stored winter wheat from the Canadian Great Lakes Region

    PubMed Central

    Miller, J. David; Schaafsma, Arthur W.

    2017-01-01

    The occurrence of P. verrucosum and ochratoxin A (OTA) were surveyed for 3 and 4 years, respectively. A total of 250 samples was collected from an average of 30 farms during the 2011, 2012, 2013 and 2014 winter seasons. Most storage bins surveyed were typically 11 m high round bins made of corrugated, galvanized steel, with flat-bottoms and conical roofs. Samples of clumped grain contained the most P. verrucosum (p<0.05, n = 10) followed by samples taken from the first load (n = 24, mean = 147±87 CFU/g) and last load (n = 17, mean = 101±77 CFU/g). Five grain samples (2.2%) tested positive for OTA, citrinin and OTB at concentrations of 14.7±7.9, 4.9±1.9 and 1.2±0.7 ng/g, with only three samples exceeding 5 ng/g. Grain samples positive for OTA were related to moisture resulting from either condensation or migrating moist warm air in the bin or areas where precipitation including snow entered the bin. Bins containing grain and clumps contaminated with OTA were studied in detail. A number of statistically-significant risk factors for OTA contamination were identified. These included 1) grain clumps accumulated around or directly under manhole openings, 2) debris and residue of old grain or grain clumps collected from the bin walls or left on storage floor and augers and 3) grain clumps accumulated around side doors. Even when grain enters storage below the 14.5% threshold of moisture, condensation and moisture migration occurs in hotspots in modern corrugated steel storage bins. Hot spots of OTA contamination were most often in areas affected by moisture migration due to inadequate aeration and exposure to moisture from precipitation or condensation. Further, we found that the nature of the condensation affects the nature and distribution of small and isolated areas with high incidence of toxin contamination and/or P. verrucosum prevalence in the grain bins examined. PMID:28749978

  11. Occurrence of Penicillium verrucosum, ochratoxin A, ochratoxin B and citrinin in on-farm stored winter wheat from the Canadian Great Lakes Region.

    PubMed

    Limay-Rios, Victor; Miller, J David; Schaafsma, Arthur W

    2017-01-01

    The occurrence of P. verrucosum and ochratoxin A (OTA) were surveyed for 3 and 4 years, respectively. A total of 250 samples was collected from an average of 30 farms during the 2011, 2012, 2013 and 2014 winter seasons. Most storage bins surveyed were typically 11 m high round bins made of corrugated, galvanized steel, with flat-bottoms and conical roofs. Samples of clumped grain contained the most P. verrucosum (p<0.05, n = 10) followed by samples taken from the first load (n = 24, mean = 147±87 CFU/g) and last load (n = 17, mean = 101±77 CFU/g). Five grain samples (2.2%) tested positive for OTA, citrinin and OTB at concentrations of 14.7±7.9, 4.9±1.9 and 1.2±0.7 ng/g, with only three samples exceeding 5 ng/g. Grain samples positive for OTA were related to moisture resulting from either condensation or migrating moist warm air in the bin or areas where precipitation including snow entered the bin. Bins containing grain and clumps contaminated with OTA were studied in detail. A number of statistically-significant risk factors for OTA contamination were identified. These included 1) grain clumps accumulated around or directly under manhole openings, 2) debris and residue of old grain or grain clumps collected from the bin walls or left on storage floor and augers and 3) grain clumps accumulated around side doors. Even when grain enters storage below the 14.5% threshold of moisture, condensation and moisture migration occurs in hotspots in modern corrugated steel storage bins. Hot spots of OTA contamination were most often in areas affected by moisture migration due to inadequate aeration and exposure to moisture from precipitation or condensation. Further, we found that the nature of the condensation affects the nature and distribution of small and isolated areas with high incidence of toxin contamination and/or P. verrucosum prevalence in the grain bins examined.

  12. Maximizing Science Return from Future Rodent Experiments on the International Space Station (ISS): Tissue Preservation

    NASA Technical Reports Server (NTRS)

    Choi, S. Y.; Lai, S.; Klotz, R.; Popova, Y.; Chakravarty, K.; Beegle, J. E.; Wigley, C. L.; Globus, R. K.

    2014-01-01

    To better understand how mammals adapt to long duration habitation in space, a system for performing rodent experiments on the ISS is under development; Rodent Research-1 is the first flight and will include validation of both on-orbit animal support and tissue preservation. To evaluate plans for on-orbit sample dissection and preservation, we simulated conditions for euthanasia, tissue dissection, and prolonged sample storage on the ISS, and we also developed methods for post-flight dissection and recovery of high quality RNA from multiple tissues following prolonged storage in situ for future science. Mouse livers and spleens were harvested under conditions that simulated nominal, on-orbit euthanasia and dissection operations including storage at -80 C for 4 months. The RNA recovered was of high quality (RNA Integrity Number, RIN(is) greater than 8) and quantity, and the liver enzyme contents and activities (catalase, glutathione reductase, GAPDH) were similar to positive controls, which were collected under standard laboratory conditions. We also assessed the impact of possible delayed on-orbit dissection scenarios (off-nominal) by dissecting and preserving the spleen (RNAlater) and liver (fast-freezing) at various time points post-euthanasia (from 5 min up to 105 min). The RNA recovered was of high quality (spleen, RIN (is) greater than 8; liver, RIN (is) greater than 6) and liver enzyme activities were similar to positive controls at all time points, although an apparent decline in select enzyme activities was evident at the latest time (105 min). Additionally, various tissues were harvested from either intact or partially dissected, frozen carcasses after storage for approximately 2 months; most of the tissues (brain, heart, kidney, eye, adrenal glands and muscle) were of acceptable RNA quality for science return, whereas some tissues (small intestine, bone marrow and bones) were not. These data demonstrate: 1) The protocols developed for future flight experiments will support science return despite delayed preservation post-euthanasia or prolonged storage, and 2) Many additional tissues for gene expression analysis can be obtained by dissection following prolonged storage of the tissue in situ at -80 C. These findings have relevance both to high value, ground-based experiments when sample collection capability is severely constrained, and to all future spaceflight experiments that entail on-orbit sample recovery by the ISS crew.

  13. Filtration Device for On-Site Collection, Storage and Shipment of Cells from Urine and Its Application to DNA-Based Detection of Bladder Cancer.

    PubMed

    Andersson, Elin; Dahmcke, Christina M; Steven, Kenneth; Larsen, Louise K; Guldberg, Per

    2015-01-01

    Molecular analysis of cells from urine provides a convenient approach to non-invasive detection of bladder cancer. The practical use of urinary cell-based tests is often hampered by difficulties in handling and analyzing large sample volumes, the need for rapid sample processing to avoid degradation of cellular content, and low sensitivity due to a high background of normal cells. We present a filtration device, designed for home or point-of-care use, which enables collection, storage and shipment of urinary cells. A special feature of this device is a removable cartridge housing a membrane filter, which after filtration of urine can be transferred to a storage unit containing an appropriate preserving solution. In spiking experiments, the use of this device provided efficient recovery of bladder cancer cells with elimination of >99% of excess smaller-sized cells. The performance of the device was further evaluated by DNA-based analysis of urinary cells collected from 57 patients subjected to transurethral resection following flexible cystoscopy indicating the presence of a tumor. All samples were tested for FGFR3 mutations and seven DNA methylation markers (BCL2, CCNA1, EOMES, HOXA9, POU4F2, SALL3 and VIM). In the group of patients where a transitional cell tumor was confirmed at histopathological evaluation, urine DNA was positive for one or more markers in 29 out of 31 cases (94%), including 19 with FGFR3 mutation (61%). In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%). Only one patient in this group was positive for a FGFR3 mutation. This patient had a stage Ta tumor resected 6 months later. The ability to easily collect, store and ship diagnostic cells from urine using the presented device may facilitate non-invasive testing for bladder cancer.

  14. Filtration Device for On-Site Collection, Storage and Shipment of Cells from Urine and Its Application to DNA-Based Detection of Bladder Cancer

    PubMed Central

    Andersson, Elin; Dahmcke, Christina M.; Steven, Kenneth; Larsen, Louise K.; Guldberg, Per

    2015-01-01

    Molecular analysis of cells from urine provides a convenient approach to non-invasive detection of bladder cancer. The practical use of urinary cell-based tests is often hampered by difficulties in handling and analyzing large sample volumes, the need for rapid sample processing to avoid degradation of cellular content, and low sensitivity due to a high background of normal cells. We present a filtration device, designed for home or point-of-care use, which enables collection, storage and shipment of urinary cells. A special feature of this device is a removable cartridge housing a membrane filter, which after filtration of urine can be transferred to a storage unit containing an appropriate preserving solution. In spiking experiments, the use of this device provided efficient recovery of bladder cancer cells with elimination of >99% of excess smaller-sized cells. The performance of the device was further evaluated by DNA-based analysis of urinary cells collected from 57 patients subjected to transurethral resection following flexible cystoscopy indicating the presence of a tumor. All samples were tested for FGFR3 mutations and seven DNA methylation markers (BCL2, CCNA1, EOMES, HOXA9, POU4F2, SALL3 and VIM). In the group of patients where a transitional cell tumor was confirmed at histopathological evaluation, urine DNA was positive for one or more markers in 29 out of 31 cases (94%), including 19 with FGFR3 mutation (61%). In the group of patients with benign histopathology, urine DNA was positive for methylation markers in 13 out of 26 cases (50%). Only one patient in this group was positive for a FGFR3 mutation. This patient had a stage Ta tumor resected 6 months later. The ability to easily collect, store and ship diagnostic cells from urine using the presented device may facilitate non-invasive testing for bladder cancer. PMID:26151138

  15. Vaginal Prostate Specific Antigen (PSA) Is a Useful Biomarker of Semen Exposure Among HIV-Infected Ugandan Women.

    PubMed

    Woolf-King, Sarah E; Muyindike, Winnie; Hobbs, Marcia M; Kusasira, Adrine; Fatch, Robin; Emenyonu, Nneka; Johnson, Mallory O; Hahn, Judith A

    2017-07-01

    The practical feasibility of using prostate specific antigen (PSA) as a biomarker of semen exposure was examined among HIV-infected Ugandan women. Vaginal fluids were obtained with self-collected swabs and a qualitative rapid test (ABAcard ® p30) was used to detect PSA. Trained laboratory technicians processed samples on-site and positive PSA tests were compared to self-reported unprotected vaginal sex (UVS) in the last 48 h. A total of 77 women submitted 126 samples for PSA testing at up to three study visits. Of these samples, 31 % (n = 39/126) were PSA positive, and 64 % (n = 25/39) of the positive PSA samples were accompanied by self-report of no UVS at the study visit the PSA was collected. There were no reported difficulties with specimen collection, storage, or processing. These findings provide preliminary data on high levels of misreported UVS among HIV-infected Ugandan women using practically feasible methods for PSA collection and processing.

  16. Sample selection, preparation methods, and the apparent tensile properties of silkworm (B. mori) cocoon silk.

    PubMed

    Reed, Emily J; Bianchini, Lindsay L; Viney, Christopher

    2012-06-01

    Reported literature values of the tensile properties of natural silk cover a wide range. While much of this inconsistency is the result of variability that is intrinsic to silk, some is also a consequence of differences in the way that silk is prepared for tensile tests. Here we explore how measured mechanical properties of Bombyx mori cocoon silk are affected by two intrinsic factors (the location from which the silk is collected within the cocoon, and the color of the silk), and two extrinsic factors (the storage conditions prior to testing, and different styles of reeling the fiber). We find that extrinsic and therefore controllable factors can affect the properties more than the intrinsic ones studied. Our results suggest that enhanced inter-laboratory collaborations, that lead to standardized sample collection, handling, and storage protocols prior to mechanical testing, would help to decrease unnecessary (and complicating) variation in reported tensile properties. Copyright © 2011 Wiley Periodicals, Inc.

  17. Implications of the pH and temperature of diluted, cooled boar semen on fresh and frozen-thawed sperm motility characteristics.

    PubMed

    Purdy, P H; Tharp, N; Stewart, T; Spiller, S F; Blackburn, H D

    2010-10-15

    Boar semen is typically collected, diluted and cooled for AI use over numerous days, or frozen immediately after shipping to capable laboratories. The storage temperature and pH of the diluted, cooled boar semen could influence the fertility of boar sperm. Therefore, the purpose of this study was to determine the effects of pH and storage temperature on fresh and frozen-thawed boar sperm motility end points. Semen samples (n = 199) were collected, diluted, cooled and shipped overnight to the National Animal Germplasm Program laboratory for freezing and analysis from four boar stud facilities. The temperature, pH and motility characteristics, determined using computer automated semen analysis, were measured at arrival. Samples were then cryopreserved and post-thaw motility determined. The commercial stud was a significant source of variation for mean semen temperature and pH, as well as total and progressive motility, and numerous other sperm motility characteristics. Based on multiple regression analysis, pH was not a significant source of variation for fresh or frozen-thawed boar sperm motility end points. However, significant models were derived which demonstrated that storage temperature, boar, and the commercial stud influenced sperm motility end points and the potential success for surviving cryopreservation. We inferred that maintaining cooled boar semen at approximately 16 °C during storage will result in higher fresh and frozen-thawed boar sperm quality, which should result in greater fertility. Copyright © 2010 Elsevier Inc. All rights reserved.

  18. Effect of delayed serum separation and storage temperature on serum glucose concentration in horse, dog, alpaca, and sturgeon.

    PubMed

    Collicutt, Nancy B; Garner, Bridget; Berghaus, Roy D; Camus, Melinda S; Hart, Kelsey

    2015-03-01

    Although delays between blood sample collection and analysis are common in veterinary medicine, the effect of prolonged serum-clot contact time on serum glucose concentration is not well established and species differences have not been elucidated. The objective was to investigate the effect of storage time and temperature on serum glucose concentration in stored whole blood samples from horse, dog, alpaca, and sturgeon. Whole blood specimens were divided into 7 no-additive tubes and serum was separated from one sample within one hour, serving as the reference sample. The remaining samples were stored at 4°C and 25°C, then centrifuged and serum glucose measured by automated analysis at 2, 4, and 8 hours postcollection. Glucose concentrations were compared using linear mixed models. The decline in serum glucose concentration for all samples stored at 4°C was not statistically significant, except for the 8-hour samples from sturgeon and dog. At 25°C, serum glucose concentration was comparable to reference values at 2 hours in sturgeon and alpaca, but significantly lower at 4 and 8 hours in those species, and at all time points in equine and canine specimens, being most prominent after 8 hours of storage in canine specimens. Storage at 4°C limits serum glucose decline for at least 4 hours in all species tested and up to 8 hours in specimens of horse and alpaca. At 25°C, serum-clot contact time should not exceed 1 hour in equine and canine samples, and 2 hours in specimens from alpaca and sturgeon. © 2014 American Society for Veterinary Clinical Pathology.

  19. The effect of anaerobic digestion and storage on indicator microorganisms in swine and dairy manure.

    PubMed

    Costa, Annamaria; Gusmara, Claudia; Gardoni, Davide; Zaninelli, Mauro; Tambone, Fulvia; Sala, Vittorio; Guarino, Marcella

    2017-11-01

    The aim of this experimental study was to evaluate the influence of anaerobic digestion and storage on indicator microorganisms in swine and dairy excreta. Samples were collected every 90 days for 15 months at eight farms, four pig, and four dairy farms, four of them having a biogas plant. Moreover, to evaluate storage effects on samples, 20 l of manure and slurry taken at each farm (digested manure only in farms with a biogas plant) were stored in a controlled climatic chamber at 18 °C, for 6 months. The bacterial load and the chemical-physical characteristics of excreta were evaluated at each sampling time, stored slurry, and manure were sampled and analyzed every 2 months. A high variability of the concentration of bacteria in the different excreta types was observed during the experiment, mainly depending on the type and time of treatment. No sample revealed either the presence of Escherichia coli O157:H7 or of Salmonella, usually linked to the temporary rearing of infected animals in facilities. Anaerobic digestion and storage affected in a significant way the reduction of indicator bacteria like lactobacilli, coliforms, and streptococci. Anaerobic digestion lowered coliforms in pig slurry (- 2.80 log, P < 0.05), streptococci in dairy manure (- 2.44 log, P < 0.001) and in pig slurry (- 1.43 log, P < 0.05), and lactobacilli in pig slurry (- 3.03 log, P < 0.05). Storage lowered coliforms and the other indicators counts, in particular in fresh wastes, while clostridia did not show a reduction in concentration.

  20. Stability of 35 biochemical and immunological routine tests after 10 hours storage and transport of human whole blood at 21°C.

    PubMed

    Henriksen, Linda O; Faber, Nina R; Moller, Mette F; Nexo, Ebba; Hansen, Annebirthe B

    2014-10-01

    Suitable procedures for transport of blood samples from general practitioners to hospital laboratories are requested. Here we explore routine testing on samples stored and transported as whole blood in lithium-heparin or serum tubes. Blood samples were collected from 106 hospitalized patients, and analyzed on Architect c8000 or Advia Centaur XP for 35 analytes at base line, and after storage and transport of whole blood in lithium-heparin or serum tubes at 21 ± 1°C for 10 h. Bias and imprecision (representing variation from analysis and storage) were calculated from values at baseline and after storage, and differences tested by paired t-tests. Results were compared to goals set by the laboratory. We observed no statistically significant bias and results within the goal for imprecision between baseline samples and 10-h samples for albumin, alkaline phosphatase, antitrypsin, bilirubin, creatinine, free triiodothyronine, γ-glutamyl transferase, haptoglobin, immunoglobulin G, lactate dehydrogenase, prostate specific antigen, total carbon dioxide, and urea. Alanine aminotransferase, amylase, C-reactive protein, calcium, cholesterol, creatine kinase, ferritin, free thyroxine, immunoglobulin A, immunoglobulin M, orosomucoid, sodium, transferrin, and triglycerides met goals for imprecision, though they showed a minor, but statistically significant bias in results after storage. Cobalamin, folate, HDL-cholesterol, iron, phosphate, potassium, thyroid stimulating hormone and urate warranted concern, but only folate and phosphate showed deviations of clinical importance. We conclude that whole blood in lithium-heparin or serum tubes stored for 10 h at 21 ± 1°C, may be used for routine analysis without restrictions for all investigated analytes but folate and phosphate.

  1. Developments of solid materials for UF 6 sampling

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

    Smith, Nicholas; Hebden, Andrew; Savina, Joseph

    2017-11-15

    This project demonstrated that a device using majority Commercial Off the Shelf (COTS) components could be used to collect uranium hexafluoride samples safely from gaseous or solid sources. The device was based on the successful Cristallini method developed by ABACC over the past 10 years. The system was designed to capture and store the UF 6 as an inert fluoride salt to ease transportation regulations. In addition, the method was considerably faster than traditional cryogenic methods, collected enough material to perform analyses without undue waste, and could be used either inside a facility or in the storage yard.

  2. Analysis of street sweepings, Portland, Oregon

    USGS Publications Warehouse

    Miller, Timothy L.; Rinella, Joseph F.; McKenzie, Stuart W.; Parmenter, Jerry

    1977-01-01

    A brief study involving collection and analysis of street sweepings was undertaken to provide the U.S. Army Corps of Engineers with data on physical, chemical, and biological characteristics of dust and dirt accumulating on Portland streets. Most of the analyses selected were based on the pollutant loads predicted by the Storage, Treatment, Overflow, and Runoff Model (STORM). Five different basins were selected for sampling, and samples were collected three times in each basin. Because the literature reports no methodology for analysis of dust and dirt, the analytical methodology is described in detail. Results of the analyses are summarized in table 1.

  3. Value of semen parameters, with special reference to TNF-α, in predicting the quality of boar semen after short-term storage.

    PubMed

    Mrkun, Janko; Kosec, Marjan; Zrimšek, Petra

    2013-06-01

    The aim of this study was to address the question whether changes in boar semen quality after short-term storage could be predicted on the basis of standard semen parameters and TNF-α level determined on the day of semen collection under commercial conditions. Progressive motility showed the highest positive correlation with morphology on day 0 of collection, and progressive motility on day 3 (P < 0.05) showed a negative correlation with acrosome abnormalities (P < 0.05). According to the area under receiver operating characteristics (ROC) curves (AUCs), progressive motility could also be used in predicting semen quality after 3 days of storage (AUC > 0.5; P < 0.05). TNF-α in seminal plasma is the only parameter measured on day 0 to show a significant correlation with the percentage of viable spermatozoa after 3 days of semen storage (r = 0.495, P < 0.05). ROC analysis shows that TNF-α level is helpful in discriminating viability outcome after semen storage (AUC = 0.94, P < 0.001). We can predict with 92.35% certainty that fresh semen samples with more than 150 pg/ml of TNF-α in the seminal plasma will retain more than 85% of viable spermatozoa after 3 days of storage. Thus, TNF-α can contribute to predicting the quality of short-term stored semen.

  4. Bioavailability of dissolved organic nitrogen (DON) in wastewaters from animal feedlots and storage lagoons.

    PubMed

    Sun, Jingyi; Khan, Eakalak; Simsek, Senay; Ohm, Jae-Bom; Simsek, Halis

    2017-11-01

    Dissolved organic nitrogen (DON) from animal wastes can contribute to pollution of surface waters. Bioavailable DON (ABDON) is a portion of DON utilized by algae with or without bacteria. This study determined DON and ABDON levels in animal wastewater collected from two different sources: an animal feedlot wastewater storage tank and a sheep wastewater storage lagoon. Inocula for the ABDON bioassays were comprised of individual species and several combinations involving two algae (Chlamydomonas reinhardtii and Chlorella vulgaris) and a mixed liquor suspended solids (MLSS) bacterial culture. The ratio of initial DON to initial total dissolved nitrogen was 18% in the feedlot wastewater samples and 70% in the lagoon wastewater samples. The results showed that between 1.6 and 4.5 mg-NL-1 DON (45-79% of initial DON) in the feedlot samples and between 3.4 and 7.5 mg-NL-1 DON (36%-79% of initial DON) in the lagoon samples were bioavailable with the inocula tested. These results suggest that when considering eutrophication potential of livestock wastewater, organic nitrogen should be included in addition to the obvious culprits, ammonia and nitrate. Copyright © 2017 Elsevier Ltd. All rights reserved.

  5. Impact of Storage Time on Hepatitis B Virus DNA Stability in Clinical Specimens Determined by Quantitative Real-time PCR.

    PubMed

    Zhang, Xiaolian; Yang, Dongmei; Lu, Yu; Lao, Xianjun; Qin, Xue; Li, Shan

    2016-01-01

    Detecting blood levels of hepatitis B virus (HBV) DNA must be accurate and credible. Shipment and storage conditions of clinical samples affect the quality of nucleic acids and can interfere with HBV DNA analysis. The aim of our study was to compare HBV DNA stability in plasma specimens at 4 degrees C for different storage periods. Blood samples from 30 hepatitis B surface antigen (HBsAg) positive patients were collected in tubes containing EDTA-K2. Each sample was divided into eight aliquots, one of which was measured immediately for the initial viral load. The remaining aliquots were then stored at 4 degrees C and assessed after 1, 2, 3, 7, 14, 21, and 30 days of storage. Quantification of HBV DNA was performed by real-time polymerase chain reaction (RT-PCR), and the difference in HBV DNA concentrations between two different time points was analysed with a paired-samples t-test. HBV DNA was measured in a range of 2.00 - 8.00 IU/mL, with low within-run and between-run coefficients of variation (< 10%). Storing plasma for one month at 4 degrees C revealed no significant decrease in HBV DNA level (p = 0.231), and no trend was evident to indicate continued reduction over a 3-week storage period. Based on the results of this study, storing plasma for up to one month at 4 degrees C does not affect the stability of HBV DNA, regardless of the initial viral load.

  6. The macronutrients in human milk change after storage in various containers.

    PubMed

    Chang, Yu-Chuan; Chen, Chao-Huei; Lin, Ming-Chih

    2012-06-01

    The concentrations of macronutrients in human milk can be influenced by various processes, such as storage, freezing, and thawing, that are performed by lactating working mothers and breast milk banks. We evaluated the impact of various containers on the nutrient concentrations in human milk. A total of 42 breast milk samples from 18 healthy lactating mothers were collected. A baseline macronutrient concentration was determined for each sample. Then, the breast milk samples were divided and stored in nine different commercial milk containers. After freezing at -20°C for 2 days, the milk samples were thawed and analyzed again. A midinfrared human milk analyzer (HMA) was used to measure the protein, fat, and carbohydrate contents. There was a significant decrease in the fat content following the storage, freezing, and thawing processes, ranging from 0.27-0.30 g/dL (p=0.02), but no significant decrease in energy content (p=0.069) was noted in the nine different containers. There were statistically significant increases in protein and carbohydrate concentrations in all containers (p=0.021 and 0.001, respectively), however there were no significant differences between the containers in terms of fat, protein, carbohydrate, or energy contents. Human milk, when subjected to storage, freezing, and thawing processes, demonstrated a significant decrease in fat content (up to 9% reduction) in various containers. It is better for infants to receive milk directly from the mother via breastfeeding. More studies are warranted to evaluate the effects of milk storage on infant growth and development. Copyright © 2012. Published by Elsevier B.V.

  7. Preservation of mithun (Bos frontalis) semen at refrigeration temperature.

    PubMed

    Karunakaran, M; Dhali, A; Mech, A; Khate, K; Rajkhowa, C; Mishra, D P

    2007-10-01

    The objective of the present study was to investigate the possibility of preserving mithun (Bos frontalis) spermatozoa at refrigeration temperature using tris-egg yolk diluent. Semen samples were collected from four adult mithun bulls through rectal massage method. Good quality semen samples (n=30) were preserved at 4 degrees C using tris-egg yolk diluent for 72 h. Progressive motility, live spermatozoa count and morphological abnormalities were evaluated every 12 h until 72 h of preservation. The colour, consistency and mass activity of fresh semen samples were found to be creamy white, medium and 3+ to 4+ (5+ scale), respectively. The average (mean+/-S.E.) volume (ml), pH and spermatozoa concentration (10(6) ml(-1)) of fresh semen samples were found to be 0.6+/-0.01, 6.8+/-0.03 and 425+/-48, respectively. Progressive motility and live spermatozoa count were found to be less than 30% (P<0.01) after 48 h of storage. Head (P<0.05), midpiece (P<0.05), tail (P<0.01) and total (P<0.01) abnormalities were found to be increased significantly over the time of storage. It was observed that progressive motility and live spermatozoa count remained above 30% and 40%, respectively, until 36 h of storage. Simultaneously the percentage of morphologically abnormal spermatozoa was found to be significantly low until 36 h of storage. The results indicate that it is possible to preserve mithun spermatozoa at refrigeration temperature in tris-egg yolk diluent, which can be further used for artificial insemination within 36 h of storage.

  8. Comparison of storage stability of odorous VOCs in polyester aluminum and polyvinyl fluoride Tedlar® bags.

    PubMed

    Kim, Yong-Hyun; Kim, Ki-Hyun; Jo, Sang-Hee; Jeon, Eui-Chan; Sohn, Jong Ryeul; Parker, David B

    2012-01-27

    Whole air sampling using containers such as flexible bags or rigid canisters is commonly used to collect samples of volatile organic compounds (VOC) in air. The objective of this study was to compare the stability of polyester aluminum (PEA) and polyvinyl fluoride (PVF, brand name Tedlar(®)) bags for gaseous VOC sampling. Eight VOC standards (benzene, toluene, p-xylene, styrene, methyl ethyl ketone, methyl isobutyl ketone, butyl acetate, and isobutyl alcohol) were placed into each bag at storage times of 0, 2, and 3 days prior to analyses by gas chromatography/mass spectrometry (GC/MS). From each bag representing each storage day, samples of 3 different mass loadings were withdrawn and analyzed to derive response factors (RF) of each chemical between the slope of the GC response (y-axis) vs. loaded mass (x-axis). The relative recoveries (RR) of VOC, if derived by dividing RF value of a given storage day by that of 0 day, varied by time, bag type, and VOC type. If the RR values after three days are compared, those of methyl isobutyl ketone were the highest with 96 (PVF) and 99% (PEA); however, the results of isobutyl alcohol were highly contrasting between the two bags with 31 and 94%, respectively. Differences in RR values between the two bag types increased with storage time, such that RR of PEA bags (88±10%) were superior to those of PVF bags (73±22%) after three days, demonstrating that VOC in PEA bags were more stable than in PVF bags. Copyright © 2011 Elsevier B.V. All rights reserved.

  9. Illicit Drugs in Oral Fluid: Evaluation of Two Collection Devices.

    PubMed

    Cohier, Camille; Mégarbane, Bruno; Roussel, Olivier

    2017-01-01

    Driving after illicit drug use is a worldwide growing concern requiring rapid and sensitive screening at the roadside. It is noteworthy that the sampling method used to collect oral fluid (OF) may significantly influence drug concentrations in the collected sample and thus alter the accuracy of the measurement. We evaluated two OF collection devices, Quantisal ® and Certus ® collectors, for their suitability for collecting samples to allow laboratory confirmation of driving after illicit drug use. Four parameters were studied including (i) the collected OF volume; (ii) the recovery efficiency using OFs spiked with opiates, cannabinoids, amphetamines, cocaine and its metabolites; (iii) drug stability after storage for 1, 7 and 14 days at -20°C, +4°C and room temperature; and (iv) the impact of mouth cells present in the collected OF on drug stability. Drug concentrations were measured using gas and liquid chromatography-mass spectrometry. Certus ® collector allowed the collection of significantly larger (0.94 ± 0.18 mL vs. 0.84 ± 0.06 mL, P = 0.08) but less reproducible OF volumes (19 vs. 6.7%) compared with Quantisal ® collector. Drug recovery was significantly better with Quantisal ® than with Certus ® collector, especially when used to detect cannabinoids (0.94 vs. 0.54, P < 0.001 for ∆9-tetrahydrocannabinol (THC)). For both OF collectors, storage at 4°C was preferable except for methadone, the stability of which was altered by adherence to the collector device. In the presence of mouth cells in the OF sample, THC concentrations were significantly decreased at Day 7 in comparison with Day 1 with both collection devices (P = 0.001 with Quantisal ® collector and P = 0.01 with Certus ® collector). In conclusion, Quantisal ® collector is more reliable than Certus ® collector although the practicability of both devices remains to be determined at the roadside. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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

    PubMed

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

    2009-12-01

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

  11. The effect of storage conditions on microbial community composition and biomethane potential in a biogas starter culture.

    PubMed

    Hagen, Live Heldal; Vivekanand, Vivekanand; Pope, Phillip B; Eijsink, Vincent G H; Horn, Svein J

    2015-07-01

    A new biogas process is initiated by adding a microbial community, typically in the form of a sample collected from a functional biogas plant. This inoculum has considerable impact on the initial performance of a biogas reactor, affecting parameters such as stability, biogas production yields and the overall efficiency of the anaerobic digestion process. In this study, we have analyzed changes in the microbial composition and performance of an inoculum during storage using barcoded pyrosequencing of bacterial and archaeal 16S ribosomal RNA (rRNA) genes, and determination of the biomethane potential, respectively. The inoculum was stored at room temperature, 4 and -20 °C for up to 11 months and cellulose was used as a standard substrate to test the biomethane potential. Storage up to 1 month resulted in similar final methane yields, but the rate of methane production was reduced by storage at -20 °C. Longer storage times resulted in reduced methane yields and slower production kinetics for all storage conditions, with room temperature and frozen samples consistently giving the best and worst performance, respectively. Both storage time and temperature affected the microbial community composition and methanogenic activity. In particular, fluctuations in the relative abundance of Bacteroidetes were observed. Interestingly, a shift from hydrogenotrophic methanogens to methanogens with the capacity to perform acetoclastic methanogensis was observed upon prolonged storage. In conclusion, this study suggests that biogas inocula may be stored up to 1 month with low loss of methanogenic activity, and identifies bacterial and archaeal species that are affected by the storage.

  12. Effects of field storage method on E. coli concentrations measured in storm water runoff.

    PubMed

    Harmel, Daren; Wagner, Kevin; Martin, Emily; Smith, Doug; Wanjugi, Pauline; Gentry, Terry; Gregory, Lucas; Hendon, Tina

    2016-03-01

    Storm water runoff is increasingly assessed for fecal indicator organisms (e.g., Escherichia coli, E. coli) and its impact on contact recreation. Concurrently, use of autosamplers along with logistic, economic, technical, and personnel barriers is challenging conventional protocols for sample holding times and storage conditions in the field. A common holding time limit for E. coli is 8 h with a 10 °C storage temperature, but several research studies support longer hold time thresholds. The use of autosamplers to collect E. coli water samples has received little field research attention; thus, this study was implemented to compare refrigerated and unrefrigerated autosamplers and evaluate potential E. coli concentration differences due to field storage temperature (storms with holding times ≤24 h) and due to field storage time and temperature (storms >24 h). Data from 85 runoff events on four diverse watersheds showed that field storage times and temperatures had minor effects on mean and median E. coli concentrations. Graphs and error values did, however, indicate a weak tendency for higher concentrations in the refrigerated samplers, but it is unknown to what extent differing die-off and/or regrowth rates, heterogeneity in concentrations within samples, and laboratory analysis uncertainty contributed to the results. The minimal differences in measured E. coli concentrations cast doubt on the need for utilizing the rigid conventional protocols for field holding time and storage temperature. This is not to say that proper quality assurance and quality control is not important but to emphasize the need to consider the balance between data quality and practical constraints related to logistics, funding, travel time, and autosampler use in storm water studies.

  13. Determining the authenticity of athlete urine in doping control by DNA analysis.

    PubMed

    Devesse, Laurence; Syndercombe Court, Denise; Cowan, David

    2015-10-01

    The integrity of urine samples collected from athletes for doping control is essential. The authenticity of samples may be contested, leading to the need for a robust sample identification method. DNA typing using short tandem repeats (STR) can be used for identification purposes, but its application to cellular DNA in urine has so far been limited. Here, a reliable and accurate method is reported for the successful identification of urine samples, using reduced final extraction volumes and the STR multiplex kit, Promega® PowerPlex ESI 17, with capillary electrophoretic characterisation of the alleles. Full DNA profiles were obtained for all samples (n = 20) stored for less than 2 days at 4 °C. The effect of different storage conditions on yield of cellular DNA and probability of obtaining a full profile were also investigated. Storage for 21 days at 4 °C resulted in allelic drop-out in some samples, but the random match probabilities obtained demonstrate the high power of discrimination achieved through targeting a large number of STRs. The best solution for long-term storage was centrifugation and removal of supernatant prior to freezing at -20 °C. The method is robust enough for incorporation into current anti-doping protocols, and was successfully applied to 44 athlete samples for anti-doping testing with 100% concordant typing. Copyright © 2015 John Wiley & Sons, Ltd.

  14. The Use of Chlorhexidine/n-Propyl Gallate (CPG) as an Ambient-Temperature Urine Preservative

    NASA Technical Reports Server (NTRS)

    Nillen, Jeannie L.; Smith, Scott M.

    2003-01-01

    A safe, effective ambient temperature urine preservative, chlorhexidine/n-propyl gallate (CPG), has been formulated for use during spacefli ght that reduces the effects of oxidation and bacterial contamination on sample integrity while maintaining urine pH. The ability of this preservative to maintain stability of nine key analytes was evaluated for a period of one year. CPG effectively maintained stability of a mmonia, total nitrogen, 3-methylhistidine, chloride, sodium, potassiu m, and urea; however, creatinine and osmolality were not preserved by CPG. These data indicate that CPG offers prolonged room-temperature storage for multiple urine analytes, reducing the requirements for f rozen urine storage on future spaceflights. Iii medical applications on Earth, this technology can allow urine samples to be collected in remote settings and eliminate the need to ship frozen samples.

  15. Mycobiota of cocoa: from farm to chocolate.

    PubMed

    Copetti, Marina V; Iamanaka, Beatriz T; Frisvad, Jens C; Pereira, José L; Taniwaki, Marta H

    2011-12-01

    The present work was carried out to study the mycobiota of cocoa beans from farm to chocolate. Four hundred and ninety-four samples were analyzed at various stages of cocoa processing: (i) primary stage at the farm (fermentation, drying, and storage), (ii) secondary stage at processing (testa, nibs, liquor, butter, cake and powder) and (iii) the final chocolate product (dark, milk, white and powdered) collected from retail outlets. Direct plating or dilution plating on Dichloran 18% Glycerol agar were used for cocoa beans and processed product analyses, respectively. Fungi were isolated and identified using different keys of identification. The largest numbers and diversity of fungi were observed in the samples collected at the farm, especially during drying and storage. The species with the highest occurrence among samples were: Absidia corymbifera, Aspergillus sp. nov., A. flavus, Penicillium paneum and yeasts. A total of 1132 potentially toxigenic fungi were isolated from the following species or species groups: A. flavus, Aspergillus parasiticus, Aspergillus nomius, Aspergillus niger group, Aspergillus carbonarius and Aspergillus ochraceus group. The highest percentage of toxigenic fungi was found at the drying and storage stages. The industrial processing reduced the fungal contamination in all fractions and no fungi were found in the final chocolate products. The knowledge of which fungi are dominant at each processing stage of cocoa provides important data about their ecology. This understanding leads to a reduction in fungal spoilage and mycotoxin production in this product. Copyright © 2011 Elsevier Ltd. All rights reserved.

  16. Storage at -80°C preserves the antioxidant capacity of preterm human milk.

    PubMed

    Akdag, Arzu; Sari, Fatma Nur; Dizdar, Evrim Alyamac; Uras, Nurdan; Isikoglu, Semra; Erel, Ozcan; Dilmen, Ugur

    2014-09-01

    It is essential to establish optimum parameters for maintaining the quality of stored milk until the moment of consumption with minimal deterioration of its properties. The aim of the study was to evaluate total antioxidant capacity (TAC) and total oxidation status (TOS) of fresh and freeze-stored samples (at -80°C) of preterm human milk (HM). Samples of colostrum were collected from 98 healthy women within the first 4 days after delivery. The total milk volume collected (6 ml) was divided in two aliquot parts: 3 ml for the fresh analysis which was done immediately after the extraction and 3 ml for storage under freezing conditions at -80°C for three months. The antioxidant status and oxidative stress of the fresh and stored breast milk were assessed via determination of TAC and TOS levels. The mean gestational age and the birth weight of the infants were 31.26 ± 2.93 weeks and 1620 ± 581.91 g; respectively. There were no significant correlations between maternal age, route of delivery and milk oxidative stress. There was no significant difference between the levels of TAC, TOS and the oxidative stress index in fresh and freeze-stored samples of colostrum in preterm HM (p > 0.05). Freeze storage of preterm HM at -80°C for three months preserves the antioxidant capacity without changing oxidative status of HM, which could be noteworthy for the preterm infant nutrition. © 2014 Wiley Periodicals, Inc.

  17. Antioxidative Activity of Colostrum and Human Milk: Effects of Pasteurization and Storage.

    PubMed

    Marinković, Vesna; Ranković-Janevski, Milica; Spasić, Snežana; Nikolić-Kokić, Aleksandra; Lugonja, Nikoleta; Djurović, Dijana; Miletić, Srdjan; Vrvić, Miroslav M; Spasojević, Ivan

    2016-06-01

    Milk banks collect, pasteurize, and freeze/store human milk. The processing may alter redox properties of milk, but the effects have not been fully examined. We collected 10 mature milk and 10 colostrum samples and applied a battery of biochemical assays and electron paramagnetic resonance spectroscopy to inspect changes that milk undergoes with pasteurization and 30 days storage at -20°C. Pasteurization and storage of raw milk did not affect total nonenzymatic antioxidative capacity, but specific components and features were altered. Urate radical and ascorbyl radical emerge as products of exposure of milk to hydroxyl radical-generating system. Processing shifted the load of antioxidative activity from ascorbate to urate and lowered the capacity of milk to diminish hydroxyl radical. Pasteurization caused a significant drop in the activity of 2 major antioxidative enzymes-superoxide dismutase and glutathione peroxidase, whereas freezing/storage of raw milk affected only superoxide dismutase. Colostrum showed drastically higher total nonenzymatic antioxidative capacity, hydroxyl radical scavenging ability, and glutathione reductase activity compared with mature milk. Pasteurization and storage affect nonenzymatic and enzymatic antioxidative agents in human milk. It appears that nonenzymatic antioxidative systems in colostrum and milk are different. The effects of processing may be partially compensated by fortification/spiking with ascorbate before use.

  18. Does laboratory automation for the preanalytical phase improve data quality?

    PubMed

    Lima-Oliveira, Gabriel; Lippi, Giuseppe; Salvagno, Gian Luca; Danese, Elisa; Montagnana, Martina; Brocco, Giorgio; Voi, Monica; Picheth, Geraldo; Guidi, Gian Cesare

    2013-10-01

    Our aim was to evaluate whether automation for the preanalytical phase improves data quality. Blood from 100 volunteers was collected into two vacuum tubes. One sample from each volunteer was respectively assigned to (G1) traditional processing, starting with centrifugation at 1200 g for 10 min, and (G2) the MODULAR PRE-ANALYTICALS EVO-MPA system. The routine clinical chemistry tests were performed in duplicate on the same instrument Cobas 6000 module. G1 samples were uncapped manually and immediately placed into the instrument. G2 samples were directly fed from the MPA system to the instrument without further staff intervention. At the end, (1) the G1 samples were stored for 6 h at 4 °C as prescribed in our accredited laboratory and (2) the G2 samples were stored for 6 h in the MPA output buffer. Results from G1 and G2, before and after storage, were compared. Significant increases were observed in G1 compared with G2 samples as follows: (1) before storage for alkaline phosphatase (ALP), lactate dehydrogenase (LDH), phosphate (P), magnesium (MG), iron (FE), and hemolysis index and (2) after storage for total cholesterol (COL), triglycerides (TG), total protein (TP), albumin (ALB), blood urea nitrogen (BUN), creatinine (CRE), uric acid (UA), ALP, pancreatic amylase, aspartate aminotransferase (AST), alanine aminotransferase (ALT), g-glutamyltransferase (GGT), LDH, creatine kinase (CK), calcium (CA), FE, sodium (NA), potassium (K), and hemolysis index. Moreover, significant increases were observed in (3) G1-after versus G1-before storage samples for COL, high-density lipoprotein cholesterol, TG, TP, ALB, BUN, CRE, UA, AST, ALT, GGT, LDH, P, CA, MG, FE, NA, K, and hemolysis index and (4) G2-after versus G2-before storage only for BUN, AST, LDH, P, and CA. In conclusion, our results show that the MPA system improves the quality of laboratory testing.

  19. Use of External Quality Control Material for HIV-1 RNA Testing To Assess the Comparability of Data Generated in Separate Laboratories and the Stability of HIV-1 RNA in Samples after Prolonged Storage.

    PubMed

    Jennings, Cheryl; Wager, Carrie G; Scianna, Salvatore R; Zaccaro, Daniel J; Couzens, Amy; Mellors, John W; Coombs, Robert W; Bremer, James W

    2018-06-01

    The National Institute of Allergy and Infectious Diseases (NIAID) AIDS Clinical Trials Group (ACTG) stores specimens from its clinical trials in a biorepository and permits the use of these specimens for nonprotocol exploratory studies, once the studies for the original protocol are concluded. We sought to assess the comparability of the data generated from real-time HIV-1 RNA testing during two clinical trials with the data generated from the retesting of different aliquots of the same samples after years of storage at -80°C. Overall, there was 92% agreement in the data generated for 1,570 paired samples (kappa statistic = 0.757; 95% confidence interval [CI], 0.716 to 0.797), where samples were tested in one laboratory using the microwell plate (MWP) version of the Roche HIV-1 Monitor test within 1 to 37 days of collection and retested in another laboratory using the Cobas version of the assay after a median of 6.7 years of storage (range, 5.7 to 8.6 years). Historical external quality control data submitted to the NIAID Virology Quality Assurance program (VQA) by client laboratories using the same two versions of the Monitor assay were used to differentiate between systematic differences in the assays to evaluate the stability of HIV-1 RNA in the stored samples. No significant loss of RNA was noted in samples containing either a low concentration (<50 copies/ml) or a high concentration (≥50 copies/ml) of HIV-1 RNA ( P = 0.10 and P = 0.90, respectively) regardless of the time in storage. These data confirm the quality of the plasma samples in the ACTG biorepository following long-term storage. Copyright © 2018 American Society for Microbiology.

  20. NASA Biological Specimen Repository

    NASA Technical Reports Server (NTRS)

    Pietrzyk, Robert; McMonigal, K. A.; Sams, C. F.; Johnson, M. A.

    2009-01-01

    The NASA Biological Specimen Repository (NBSR) has been established to collect, process, annotate, store, and distribute specimens under the authority of the NASA/JSC Committee for the Protection of Human Subjects. The International Space Station (ISS) provides a platform to investigate the effects of microgravity on human physiology prior to lunar and exploration class missions. The NBSR is a secure controlled storage facility that is used to maintain biological specimens over extended periods of time, under well-controlled conditions, for future use in approved human spaceflight-related research protocols. The repository supports the Human Research Program, which is charged with identifying and investigating physiological changes that occur during human spaceflight, and developing and implementing effective countermeasures when necessary. The storage of crewmember samples from many different ISS flights in a single repository will be a valuable resource with which researchers can validate clinical hypotheses, study space-flight related changes, and investigate physiological markers All samples collected require written informed consent from each long duration crewmember. The NBSR collects blood and urine samples from all participating long duration ISS crewmembers. These biological samples are collected pre-flight at approximately 45 days prior to launch, during flight on flight days 15, 30, 60 120 and within 2 weeks of landing. Postflight sessions are conducted 3 and 30 days following landing. The number of inflight sessions is dependent on the duration of the mission. Operations began in 2007 and as of October 2009, 23 USOS crewmembers have completed or agreed to participate in this project. As currently planned, these human biological samples will be collected from crewmembers covering multiple ISS missions until the end of U.S. presence on the ISS or 2017. The NBSR will establish guidelines for sample distribution that are consistent with ethical principles, protection of crewmember confidentiality, prevailing laws and regulations, intellectual property policies, and consent form language. A NBSR Advisory Board composed of representatives of all participating agencies will be established to evaluate each request by an investigator for use of the samples to ensure the request reflects the mission of the NBSR.

  1. Headspace vapor characterization of Hanford Waste Tank 241-S-102: Results from samples collected on January 26, 1996. Tank Vapor Characterization Project

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

    Evans, J.C.; Thomas, B.L.; Pool, K.H.

    1996-07-01

    This report describes the results of vapor samples obtained to compare vapor sampling of the tank headspace using the Vapor Sampling System (VSS) and In Situ Vapor Sampling System (ISVS) with and without particulate prefiltration. Samples were collected from the headspace of waste storage tank 241-S-102 (Tank S-102) at the Hanford Site in Washington State. Pacific Northwest National Laboratory (PNNL) was contracted by Westinghouse Hanford Company (WHC) to provide sampling devices and analyze samples for water, ammonia, permanent gases, total nonmethane hydrocarbons (TNMHCs, also known as TO-12), and organic analytes in samples collected in SUMMA{trademark} canisters and on triple sorbentmore » traps (TSTs) from the tank headspace. The analytical work was performed by the PNNL Vapor Analytical Laboratory (VAL) by the Tank Vapor Characterization Project. Work performed was based on a sampling and analysis plan (SAP) prepared by WHC. The SAP provided job-specific instructions for samples, analyses, and reporting. The SAP for this sample job was {open_quotes}Sampling and Analysis Plan for Tank Vapor Sampling Comparison Test{close_quote}, and the sample jobs were designated S6007, S6008, and S6009. Samples were collected by WHC on January 26, 1996, using the VSS, a truck-based sampling method using a heated probe; and the ISVS with and without particulate prefiltration.« less

  2. Headspace vapor characterization of Hanford Waste Tank 241-BY-108: Results from samples collected January 23, 1996. Tank Vapor Characterization Project

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

    Pool, K.H.; Evans, J.C.; Thomas, B.L.

    1996-07-01

    This report describes the results of vapor samples obtained to compare vapor sampling of the tank headspace using the Vapor Sampling System (VSS) and In Situ Vapor Sampling System (ISVS) with and without particulate prefiltration. Samples were collected from the headspace of waste storage tank 241-BY-108 (Tank BY-108) at the Hanford Site in Washington State. Pacific Northwest National Laboratory (PNNL) was contracted by Westinghouse Hanford Company (WHC) to provide sampling devices and analyze samples for water, ammonia, permanent gases, total nonmethane hydrocarbons (TNMHCs, also known as TO-12), and organic analytes in samples collected in SUMMA{trademark} canisters and on triple sorbentmore » traps (TSTs) from the tank headspace. The analytical work was performed by the PNNL Vapor Analytical Laboratory (VAL) by the Tank Vapor Characterization Project. Work performed was based on a sampling and analysis plan (SAP) prepared by WHC. The SAP provided job-specific instructions for samples, analyses, and reporting. The SAP for this sample job was {open_quotes}Sampling and Analysis Plan for Tank Vapor Sampling Comparison Test{close_quotes}, and the sample jobs were designated S6004, S6005, and S6006. Samples were collected by WHC on January 23, 1996, using the VSS, a truck-based sampling method using a heated probe; and the ISVS with and without particulate prefiltration.« less

  3. The effect of storage on whole blood chemiluminescence measurement of equine neutrophils.

    PubMed

    Krumrych, Wiesław; Skórzewski, Radosław; Malinowski, Edward

    2013-01-01

    The aim of this study was to determine the effect of duration and temperature of sample storage on whole blood chemiluminescence measurement results. Venous blood from 18 clinically healthy Polish half-bred horses aged 4 to 11 years were used in the study. Luminol dependent chemiluminescence (CL) was used to measure neutrophil oxygen metabolism in whole blood. Blood samples were examined for spontaneous CL and stimulated by a surface receptor stimulus as well as extra-receptor stimulus. The assay was performed in two parallel experimental sets with samples stored at 4 and 22 °C, respectively. Whole blood CL was estimated at 2, 6, 24, 48, 72, 96 and 120 h after collection. The study demonstrated that temperature and duration of sample storage are factors that determine the quality of CL measurements of whole blood in horses. The study concluded that samples should be stored at 4 °C and the assay should be performed as early as possible. It was also shown that the viability period of horse blood for CL assays is relatively long. Material stored at room temperature for 24 h and even up to 48 h at 4 °C did not show any significant decrease in spontaneous or stimulated chemiluminescence. Copyright © 2012 John Wiley & Sons, Ltd.

  4. Mutagen Sensitivity, Apoptosis, and Polymorphism in DNA Repair as Measures of Prostate Cancer Risk

    DTIC Science & Technology

    2005-02-01

    blood tubes are immediately refrigerated and delivered on ice within 6 hours to the GCRC core facility at Georgetown University for processing . Case...is CLIA certified. There is a centrally monitored storage facility with -80’C freezers. The established recruitment and optimized sample processing in...controls. We have established the recruitment procedures, sample collection, processing , repository, and data management. This is a substantial effort

  5. Methods of human body odor sampling: the effect of freezing.

    PubMed

    Lenochova, Pavlina; Roberts, S Craig; Havlicek, Jan

    2009-02-01

    Body odor sampling is an essential tool in human chemical ecology research. However, methodologies of individual studies vary widely in terms of sampling material, length of sampling, and sample processing. Although these differences might have a critical impact on results obtained, almost no studies test validity of current methods. Here, we focused on the effect of freezing samples between collection and use in experiments involving body odor perception. In 2 experiments, we tested whether axillary odors were perceived differently by raters when presented fresh or having been frozen and whether several freeze-thaw cycles affected sample quality. In the first experiment, samples were frozen for 2 weeks, 1 month, or 4 months. We found no differences in ratings of pleasantness, attractiveness, or masculinity between fresh and frozen samples. Similarly, almost no differences between repeatedly thawed and fresh samples were found. We found some variations in intensity; however, this was unrelated to length of storage. The second experiment tested differences between fresh samples and those frozen for 6 months. Again no differences in subjective ratings were observed. These results suggest that freezing has no significant effect on perceived odor hedonicity and that samples can be reliably used after storage for relatively long periods.

  6. Evaluation of the impact of refrigeration on next generation sequencing-based assessment of the canine and feline fecal microbiota.

    PubMed

    Weese, J Scott; Jalali, Mohammad

    2014-09-30

    Evaluation of factors that might impact microbiota assessment is important to avoid spurious results, particularly in field and multicenter studies where sample collection may occur distant from the laboratory. This study evaluated the impact of refrigeration on next generation sequence-based assessment of the canine and feline fecal microbiota. Fecal samples were collected from seven dogs and ten cats, and analysed at baseline and after 3, 7, 10 and 14 days of storage at 4°C. There were no differences in community membership or population structure between timepoints for either dogs or cats, nor were there any differences in richness, diversity and evenness. There were few differences in relative abundance of phyla or predominant genera, with the only differences being significant increases in Actinobacteria between Days 0-14 (P = 0.0184) and 1-14 (P = 0.0182) for canine samples, and a decrease in Erysipelotrichaceae incertae sedis between Day 0 and Day 7 (median 4.9 vs 2.2%, P = 0.046) in feline samples. Storage for at least 14 days at 4°C has limited impact on culture-independent assessment of the canine and feline fecal microbiota, although changes in some individual groups may occur.

  7. The efficacy of protoporphyrin as a predictive biomarker for lead exposure in canvasback ducks: effect of sample storage time

    USGS Publications Warehouse

    Franson, J.C.; Hohman, W.L.; Moore, J.L.; Smith, M.R.

    1996-01-01

    We used 363 blood samples collected from wild canvasback dueks (Aythya valisineria) at Catahoula Lake, Louisiana, U.S.A. to evaluate the effect of sample storage time on the efficacy of erythrocytic protoporphyrin as an indicator of lead exposure. The protoporphyrin concentration of each sample was determined by hematofluorometry within 5 min of blood collection and after refrigeration at 4 °C for 24 and 48 h. All samples were analyzed for lead by atomic absorption spectrophotometry. Based on a blood lead concentration of ≥0.2 ppm wet weight as positive evidence for lead exposure, the protoporphyrin technique resulted in overall error rates of 29%, 20%, and 19% and false negative error rates of 47%, 29% and 25% when hematofluorometric determinations were made on blood at 5 min, 24 h, and 48 h, respectively. False positive error rates were less than 10% for all three measurement times. The accuracy of the 24-h erythrocytic protoporphyrin classification of blood samples as positive or negative for lead exposure was significantly greater than the 5-min classification, but no improvement in accuracy was gained when samples were tested at 48 h. The false negative errors were probably due, at least in part, to the lag time between lead exposure and the increase of blood protoporphyrin concentrations. False negatives resulted in an underestimation of the true number of canvasbacks exposed to lead, indicating that hematofluorometry provides a conservative estimate of lead exposure.

  8. The Generation R Study: Biobank update 2015.

    PubMed

    Kruithof, Claudia J; Kooijman, Marjolein N; van Duijn, Cornelia M; Franco, Oscar H; de Jongste, Johan C; Klaver, Caroline C W; Mackenbach, Johan P; Moll, Henriëtte A; Raat, Hein; Rings, Edmond H H M; Rivadeneira, Fernando; Steegers, Eric A P; Tiemeier, Henning; Uitterlinden, Andre G; Verhulst, Frank C; Wolvius, Eppo B; Hofman, Albert; Jaddoe, Vincent W V

    2014-12-01

    The Generation R Study is a population-based prospective cohort study from fetal life until adulthood. The study is designed to identify early environmental and genetic causes and causal pathways leading to normal and abnormal growth, development and health from fetal life, childhood and young adulthood. In total, 9,778 mothers were enrolled in the study. Data collection in children and their parents include questionnaires, interviews, detailed physical and ultrasound examinations, behavioural observations, Magnetic Resonance Imaging and biological samples. Efforts have been conducted for collecting biological samples including blood, hair, faeces, nasal swabs, saliva and urine samples and generating genomics data on DNA, RNA and microbiome. In this paper, we give an update of the collection, processing and storage of these biological samples and available measures. Together with detailed phenotype measurements, these biological samples provide a unique resource for epidemiological studies focused on environmental exposures, genetic and genomic determinants and their interactions in relation to growth, health and development from fetal life onwards.

  9. Human Milk Fatty Acid Composition: Comparison of Novel Dried Milk Spot Versus Standard Liquid Extraction Methods.

    PubMed

    Rudolph, Michael C; Young, Bridget E; Jackson, Kristina Harris; Krebs, Nancy F; Harris, William S; MacLean, Paul S

    2016-12-01

    Accurate assessment of the long chain polyunsaturated fatty acid (LC-PUFA) content of human milk (HM) provides a powerful means to evaluate the FA nutrient status of breastfed infants. The conventional standard for FA composition analysis of HM is liquid extraction, trans-methylation, and analyte detection resolved by gas chromatography. This standard approach requires fresh or frozen samples, storage in deep freeze, organic solvents, and specialized equipment in processing and analysis. Further, HM collection is often impractical for many studies in the free living environment, particularly for studies in developing countries. In the present study, we compare a novel and more practical approach to sample collection and processing that involves the spotting and drying ~50 μL of HM on a specialized paper stored and transported at ambient temperatures until analysis. Deming regression indicated the two methods aligned very well for all LC-PUFA and the abundant HM FA. Additionally, strong correlations (r > 0.85) were observed for DHA, ARA, EPA, linoleic (LA), and alpha-linolenic acids (ALA), which are of particular interest to the health of the developing infant. Taken together, our data suggest this more practical and inexpensive method of collection, storage, and transport of HM milk samples could dramatically facilitate studies of HM, as well as understanding its lipid composition influences on human health and development.

  10. Results of stainless steel canister corrosion studies and environmental sample investigations

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

    Bryan, Charles R.; Enos, David

    2014-12-01

    This progress report describes work being done at Sandia National Laboratories (SNL) to assess the localized corrosion performance of container/cask materials used in the interim storage of used nuclear fuel. The work involves both characterization of the potential physical and chemical environment on the surface of the storage canisters and how it might evolve through time, and testing to evaluate performance of the canister materials under anticipated storage conditions. To evaluate the potential environment on the surface of the canisters, SNL is working with the Electric Power Research Institute (EPRI) to collect and analyze dust samples from the surface ofmore » in-service SNF storage canisters. In FY 13, SNL analyzed samples from the Calvert Cliffs Independent Spent Fuel Storage Installation (ISFSI); here, results are presented for samples collected from two additional near-marine ISFSI sites, Hope Creek NJ, and Diablo Canyon CA. The Hope Creek site is located on the shores of the Delaware River within the tidal zone; the water is brackish and wave action is normally minor. The Diablo Canyon site is located on a rocky Pacific Ocean shoreline with breaking waves. Two types of samples were collected: SaltSmart™ samples, which leach the soluble salts from a known surface area of the canister, and dry pad samples, which collected a surface salt and dust using a swipe method with a mildly abrasive ScotchBrite™ pad. The dry samples were used to characterize the mineralogy and texture of the soluble and insoluble components in the dust via microanalytical techniques, including mapping X-ray Fluorescence spectroscopy and Scanning Electron Microscopy. For both Hope Creek and Diablo Canyon canisters, dust loadings were much higher on the flat upper surfaces of the canisters than on the vertical sides. Maximum dust sizes collected at both sites were slightly larger than 20 μm, but Phragmites grass seeds ~1 mm in size, were observed on the tops of the Hope Creek canisters. At both sites, the surface dust could be divided into fractions generated by manufacturing processes and by natural processes. The fraction from manufacturing processes consisted of variably-oxidized angular and spherical particles of stainless steel and iron, generated by machining and welding/cutting processes, respectively. Dust from natural sources consisted largely of detrital quartz and aluminosilicates (feldspars and clays) at both sites. At Hope Creek, soluble salts were dominated by sulfates and nitrates, mostly of calcium. Chloride was a trace component and the only chloride mineral observed by SEM was NaCl. Chloride surface loads measured by the Saltsmart™ sensors were very low, less than 60 mg m –2 on the canister top, and less than 10 mg m –2 on the canister sides. At Diablo Canyon, sea-salt aggregates of NaCl and Mg-SO 4, with minor K and Ca, were abundant in the dust, in some cases dominating the observed dust assemblage. Measured Saltsmart™ chloride surface loads were very low (<5 mg m –2); however, high canister surface temperatures damaged the Saltsmart™ sensors, and, in view of the SEM observations of abundant sea-salts on the package surfaces, the measured surface loads may not be valid. Moreover, the more heavily-loaded canister tops at Diablo Canyon were not sampled with the Saltsmart™ sensors. The observed low surface loads do not preclude chloride-induced stress corrosion cracking (CISCC) at either site, because (1) the measured data may not be valid for the Diablo Canyon canisters; (2) the surface coverage was not complete (for instance, the 45º offset between the outlet and inlet vents means that near-inlet areas, likely to have heavier dust and salt loads, were not sampled); and (3) CISCC has been experimentally been observed at salt loads as low as 5-8 mg/m 2. Experimental efforts at SNL to assess corrosion of interim storage canister materials include three tasks in FY14. First, a full-diameter canister mockup, made using materials and techniques identical to those used to make interim storage canisters, was designed and ordered from Ranor Inc., a cask vendor for Areva/TN. The mockup will be delivered prior to the end of FY14, and will be used for evaluating weld residual stresses and degrees of sensitization for typical interim storage canister welds. Following weld characterization, the mockup will be sectioned and provided to participating organizations for corrosion testing purposes. A test plan is being developed for these efforts. In a second task, experimental work was carried out to evaluate crevice corrosion of 304SS in the presence of limited reactants, as would be present on a dustcovered storage canister. This work tests the theory that limited salt loads will limit corrosion penetration over time, and is a continuation of work carried out in FY13. Laser confocal microscopy was utilized to assess the volume and depth of corrosion pits formed during the crevice corrosion tests. Results indicate that for the duration of the current experiments (100 days), no stifling of corrosion occurred due to limitations in the amount of reactants present at three different salt loadings. Finally, work has been carried out this year perfecting an instrument for depositing sea-salts onto metal surfaces for atmospheric corrosion testing purposes. The system uses an X-Y plotter system with a commercial airbrush, and deposition is monitored with a quartz crystal microbalance. The system is capable of depositing very even salt loadings, even at very low total deposition rates.« less

  11. Determination of ochratoxin A in traditional Chinese medicinal plants by HPLC-FLD.

    PubMed

    Yang, Lei; Wang, Linan; Pan, Jianyu; Xiang, Lan; Yang, Meihua; Logrieco, Antonio F

    2010-07-01

    Traditional Chinese medicinal plants (TCMPs), commonly used as spices, additives or foods, are also widely used in China to prevent and cure human disease. Due to their provenance, TCMPs may be contaminated by various fungal species, including ochratoxigenic fungi, during growth, collection, transportation and, especially, storage. A reliable method for the determination of ochratoxin A (OTA) in TCMPs of different origins was developed to monitor OTA levels in China. Analyses involved the extraction of OTA by methanol/water, immunoaffinity column (IAC) clean-up and HPLC quantification with fluorescence detection (FLD). Positive results were further confirmed by LC-ESI-MS/MS. The limit of detection (LOD) was 0.3 microg kg(-1), based on a signal-to-noise ratio of 3 : 1. Among the total of 57 TCMPs samples collected from six different areas, 31 were visibly moldy due to inappropriate storage; 26 sample, purchased from local TCMPs markets, were not visibly moldy. The results showed that 23 of the visibly moldy samples and two of the not visibly moldy were contaminated with OTA at levels ranging 1.2-158.7 and 2.5-5.6 microg kg(-1), respectively. This is the first report of the natural occurrence of OTA in TCMPs.

  12. Impact of collection container material and holding times on sample integrity for mercury and methylmercury in water

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

    Riscassi, Ami L; Miller, Carrie L; Brooks, Scott C

    Mercury (Hg) and methylmercury (MeHg) concentrations in streamwater can vary on short timescales (hourly or less) during storm flow and on a diel cycle; the frequency and timing of sampling required to accurately characterize these dynamics may be difficult to accomplish manually. Automated sampling can assist in sample collection; however use has been limited for Hg and MeHg analysis due to stability concerns of trace concentrations during extended storage times. We examined the viability of using automated samplers with disposable low-density polyethylene (LDPE) sample bags to collect industrially contaminated streamwater for unfiltered and filtered Hg and MeHg analysis. Specifically wemore » investigated the effect of holding times ranging from hours to days on streamwater collected during baseflow and storm flow. Unfiltered and filtered Hg and MeHg concentrations decreased with increases in time prior to sample processing; holding times of 24 hours or less resulted in concentration changes (mean 11 7% different) similar to variability in duplicates collected manually during analogous field conditions (mean 7 10% different). Comparisons of samples collected with manual and automated techniques throughout a year for a wide range of stream conditions were also found to be similar to differences observed between duplicate grab samples. These results demonstrate automated sampling into LDPE bags with holding times of 24 hours or less can be effectively used to collect streamwater for Hg and MeHg analysis, and encourage the testing of these materials and methods for implementation in other aqueous systems where high-frequency sampling is warranted.« less

  13. Development of monitoring techniques for potential seepage of CO2 from sub-seafloor storage sites: Field studies at Sleipner, North Sea

    NASA Astrophysics Data System (ADS)

    James, R. H.; Connelly, D. P.; Bull, J. M.; Lichtschlag, A.; Cevatoglu, M.; Le Bas, T.; Wright, I. C.

    2012-12-01

    Although CO2 has been stored at the Sleipner site in the North Sea for over 15 years, and a number of other sub-seafloor storage sites are now either in operation or planned, almost nothing is known about the effect of potential seepage on marine ecosystems. To address this, we will undertake a comprehensive field campaign to Sleipner (RRS James Cook Cruise 77) in September 2012 that aims to: (i) Constrain the potential pathways of seepage from the storage site. (ii) Test methods for the detection of seepage, including formation fluids, natural gas and CO2, as it passes through the sedimentary overburden and into the water column. (iii) Develop a monitoring strategy suitable for all offshore carbon capture and storage projects. To this end, we will conduct an extensive AUV survey in the vicinity of the sub-seafloor CO2 plume, using our novel, long-range AUTOSUB system. AUTOSUB will be equipped with a variety of instrumentation, including sidescan sonar and an EM2000 multibeam systems, as well as a CHIRP profiler capable of inspecting the architecture of the sedimentary overburden at unprecedented spatial resolution. Other instrumentation will include a series of sensors (including a pH sensor), to detect and monitor the dispersion of potential seepage, and a new colour camera. Areas of interest, revealed by the AUV surveys, will be inspected and sampled using a hybrid remotely operated vehicle, equipped with high resolution video cameras, a grab sampling device, and instrumentation for the collection of precisely-located water samples. Further water samples will be collected using the ship-based CTD system. Fluid and gas seeps will be sampled using a vibrocoring system, and analyses of the porefluid chemistry will be used to quantify fluxes across the sediment-seawater interface, and the source, transformation, and fate of dissolved constituents. Longer-term monitoring will be addressed by deployment of a seafloor lander, that is equipped with a flow meter, a seismometer and a hydrophone as well as other geophysical and geochemical sensors. Preliminary results from the cruise will be presented.

  14. PHOSPHORUS SORPTION DYNAMICS IN SOILS AND COUPLING WITH SURFACE AND PORE WATER IN RIVERINE WETLANDS

    EPA Science Inventory

    Adsorption to soils is one of the dominant mechanisms of P storage in wetlands. We examined P sorption dynamics in soils collected at 12 sample points with diverse hydrology, geomorphic position, mineralogy, and plant communities in two riverine wetlands in northern Minnesota and...

  15. Development and testing of a portable wind sensitive directional air sampler

    NASA Technical Reports Server (NTRS)

    Deyo, J.; Toma, J.; King, R. B.

    1975-01-01

    A portable wind sensitive directional air sampler was developed as part of an air pollution source identification system. The system is designed to identify sources of air pollution based on the directional collection of field air samples and their analysis for TSP and trace element characteristics. Sources can be identified by analyzing the data on the basis of pattern recognition concepts. The unit, designated Air Scout, receives wind direction signals from an associated wind vane. Air samples are collected on filter slides using a standard high volume air sampler drawing air through a porting arrangement which tracks the wind direction and permits collection of discrete samples. A preset timer controls the length of time each filter is in the sampling position. At the conclusion of the sampling period a new filter is automatically moved into sampling position displacing the previous filter to a storage compartment. Thus the Air Scout may be set up at a field location, loaded with up to 12 filter slides, and left to acquire air samples automatically, according to the wind, at any timer interval desired from 1 to 30 hours.

  16. Effect of cryopreservation methods and precryopreservation storage on bottlenose dolphin (Tursiops truncatus) spermatozoa.

    PubMed

    Robeck, T R; O'Brien, J K

    2004-05-01

    Research was conducted to develop an effective method for cryopreserving bottlenose dolphin (Tursiops truncatus) semen processed immediately after collection or after 24-h liquid storage. In each of two experiments, four ejaculates were collected from three males. In experiment 1, three cryopreservation methods (CM1, CM2, and CM3), two straw sizes (0.25 and 0.5 ml), and three thawing rates (slow, medium, and fast) were evaluated. Evaluations were conducted at collection, prefreeze, and 0-, 3-, and 6-h postthaw. A sperm motility index (SMI; total motility [TM] x % progressive motility [PPM] x kinetic rating [KR, scale of 0-5]) was calculated and expressed as a percentage MI of the initial ejaculate. For all ejaculates, initial TM and PPM were greater than 85%, and KR was five. At 0-h postthaw, differences in SMI among cryopreservation methods and thaw rates were observed (P < 0.05), but no effect of straw size was observed. In experiment 2, ejaculates were divided into four aliquots for dilution (1:1) and storage at 4 degrees C with a skim milk- glucose or a N-tris(hydroxymethyl)methyl-2-aminoethane sulfonic acid (TES)-TRIS egg yolk solution and at 21 degrees C with a Hepes-Tyrode balanced salt solution (containing bovine albumin and HEPES) (TALP) medium or no dilution. After 24 h, samples were frozen and thawed (CM3, 0.5-ml straws, fast thawing rate) at 20 x 10(6) spermatozoa ml(-1) (low concentration) or at 100 x 10(6) spermatozoa ml(-1) (standard concentration). The SMI at 0-h postthaw was higher for samples stored at 4 degrees C than for samples stored at 21 degrees C (P < 0.001), and at 6-h postthaw, the SMI was higher for samples frozen at the standard concentration than for samples frozen at the low concentration (P < 0.05). For both experiments, acrosome integrity was similar across treatments. In summary, a semen cryopreservation protocol applied to fresh or liquid-stored semen maintained high levels of initial ejaculate sperm characteristics.

  17. Influence of preservation methods on the quality of colostrum sourced from New Zealand dairy farms.

    PubMed

    Denholm, K S; Hunnam, J C; Cuttance, E L; McDougall, S

    2017-09-01

    To assess the effect of two temperatures (ambient temperature and 4°C), three preservation methods (no preservative, yoghurt and potassium sorbate), and two periods of storage (3 and 7 days) on Brix and total bacterial and coliform counts of colostrum collected from New Zealand dairy farms. One litre of colostrum destined to be fed to newborn calves was collected from 55 New Zealand dairy farms in the spring of 2015. Six aliquots of 150 mL were obtained from each colostrum sample, with two aliquots left untreated, two treated with potassium sorbate and two with yoghurt, and one of each pair of aliquots stored at ambient temperature and the other at 4°C. All samples were tested for Brix, total bacterial counts and coliform counts before treatment (Day 0), and after 3 and 7 days of storage. The effect of preservation method and storage temperature on the change in Brix, bacterial and coliform counts after 3 or 7 days of storage was analysed using multivariable random effects models. For all outcome variables there was a temperature by preservation interaction. For aliquots preserved with potassium sorbate, changes in Brix and bacterial counts did not differ between aliquots stored at ambient temperature or 4°C, but for aliquots preserved with yoghurt or no preservative the decrease in Brix and increase in bacterial counts was greater for aliquots stored at ambient temperature than 4°C (p<0.001). For aliquots preserved with potassium sorbate, coliform counts decreased at both temperatures, but for aliquots preserved with yoghurt or no preservative coliform counts increased for aliquots stored at 4°C, but generally decreased at ambient temperatures (p<0.001). There was also an interaction between duration of storage and temperature for bacterial counts (p<0.001). The difference in the increase in bacterial counts between aliquots stored at 4°C and ambient temperature after 3 days was greater than between aliquots stored at 4°C and ambient temperature after 7 days. Use of potassium sorbate to preserve colostrum for 3 or 7 days resulted in little or no reduction in Brix and a lower increase in total bacterial counts than colostrum stored without preservative or with yoghurt added. Colostrum quality was not affected by storage temperature for samples preserved with potassium sorbate, but storage at 4°C resulted in better quality colostrum than storage at ambient temperatures for colostrum with no preservative or yoghurt added.

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

    Philipp, Hugh T., E-mail: htp2@cornell.edu; Tate, Mark W.; Purohit, Prafull

    Modern storage rings are readily capable of providing intense x-ray pulses, tens of picoseconds in duration, millions of times per second. Exploiting the temporal structure of these x-ray sources opens avenues for studying rapid structural changes in materials. Many processes (e.g. crack propagation, deformation on impact, turbulence, etc.) differ in detail from one sample trial to the next and would benefit from the ability to record successive x-ray images with single x-ray sensitivity while framing at 5 to 10 MHz rates. To this end, we have pursued the development of fast x-ray imaging detectors capable of collecting bursts of imagesmore » that enable the isolation of single synchrotron bunches and/or bunch trains. The detector technology used is the hybrid pixel array detector (PAD) with a charge integrating front-end, and high-speed, in-pixel signal storage elements. A 384×256 pixel version, the Keck-PAD, with 150 µm × 150 µm pixels and 8 dedicated in-pixel storage elements is operational, has been tested at CHESS, and has collected data for compression wave studies. An updated version with 27 dedicated storage capacitors and identical pixel size has been fabricated.« less

  19. 77 FR 26045 - Agency Information Collection Activities; Proposed Collection; Comments Requested: Certification...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-02

    ... Secure Gun Storage or Safety Devices ACTION: 30-Day Notice of information collection. The Department of... approved collection. (2) Title of the Form/Collection: Certification of Secure Gun Storage or Safety... to the availability of secure gun storage or safety devices. (5) An estimate of the total number of...

  20. Hair combing to collect organic gunshot residues (OGSR).

    PubMed

    MacCrehan, William A; Layman, Malinda J; Secl, Janelle D

    2003-08-12

    A protocol is presented for the collection and analysis of gunshot residues (GSR) from hair. A fine-toothed comb is used for collection of the residues. A small zip-closure bag serves as a container for both sample storage and extraction of the characteristic organic powder additives. The success of this residue recovery approach was tested on simulated shooters and victims using mannequin-supported human wig hair as well as on human shooters. Residues were collected from four weapons: a revolver and semi-automatic pistol, rifle and shotgun. One characteristic additive, nitroglycerin, was detected by capillary electrophoresis (CE) in the majority of the collection experiments.

  1. Long term storage of Pleurotus ostreatus and Trametes versicolor isolates using different cryopreservation techniques and its impact on laccase activity.

    PubMed

    Eichlerová, Ivana; Homolka, Ladislav; Tomšovský, Michal; Lisá, Ludmila

    2015-12-01

    The strain Pleurotus ostreatus Florida f6, its 45 basidiospore-derived isolates (both monokaryons and dikaryons prepared in our laboratory), Trametes versicolor strain CCBAS 614 and 22 other T. versicolor isolates obtained from the sporocarps collected in distant localities were successfully preserved for 12 y using perlite and straw cryopreservation protocols. All tested isolates survived a 12-year storage in liquid nitrogen (LN) and their laccase production and Poly B411 decolorization capacity was preserved. Also mycelium extension rate and the types of colony appearance of individual isolates remained unchanged. Different cryopreservation techniques were also tested for the short time (24 h) and the long time (6 m) storage of the culture liquid with extracellular laccase produced by T. versicolor strain CCBAS 614. The results showed that 10 % glycerol was the most suitable cryopreservant. The absence of the cryopreservant did not cause high loss of laccase activity in the samples; the presence of DMSO (5 or 10 %) in LN-stored samples caused mostly a decrease of laccase activity. For the preservation of laccase activity in the liquid culture the storage in the freezer at -80 °C is more convenient than the storage in liquid nitrogen. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  2. Performance and Storage Integrity of Dried Blood Spots for PCB, BFR and Pesticide Measurements

    PubMed Central

    Batterman, Stuart; Chernyak, Sergei

    2014-01-01

    Dried blood spots (DBS) can provide accurate and valuable estimates of exposure to environmental toxicants, and the use of information derived from archived newborn DBS information has enormous potential to open up new research on the impacts of early chemical exposure on disease. Broad application of DBS for the purpose of quantitative exposure estimation requires robust and validated methods. This study investigates the suitability of DBS analyses for population studies of exposure to three chemical groups: polychlorinated biphenyls (PCBs), brominated flame retardants (BFRs), and chlorinated pesticides. It examines background (matrix) contamination, recovery and extraction variability, sensitivity, and storage stability. DBS samples prepared using 50 μL of adult blood were analyzed by GC/MS, and method performance was confirmed by using certified materials and paired DBS-blood samples from six volunteers. Several of the target compounds and their degradation products have not been previously measured in DBS. All target compounds were detected in DBS samples collected from the volunteers. Sample DBS cards showed background contamination of several compounds. When stored at room temperature, target compounds, excluding PBDEs, were stable for up to one month. When refrigerated or frozen, stability was acceptable for all compounds up to one year, and multiyear storage appears acceptable at colder (e.g., −80 °C) temperatures. Multicompartment models may be used to estimate or correct for storage losses. Considering concentrations of contaminants for adults and children reported in the literature, and experimental values of detection limits and background contamination, DBS samples are suitable for quantifying exposures to many PCBs, BFRs and persistent pesticides. PMID:25058892

  3. What a drop can do: dried blood spots as a minimally invasive method for integrating biomarkers into population-based research.

    PubMed

    McDade, Thomas W; Williams, Sharon; Snodgrass, J Josh

    2007-11-01

    Logistical constraints associated with the collection and analysis of biological samples in community-based settings have been a significant impediment to integrative, multilevel bio-demographic and biobehavioral research. However recent methodological developments have overcome many of these constraints and have also expanded the options for incorporating biomarkers into population-based health research in international as well as domestic contexts. In particular using dried blood spot (DBS) samples-drops of whole blood collected on filter paper from a simple finger prick-provides a minimally invasive method for collecting blood samples in nonclinical settings. After a brief discussion of biomarkers more generally, we review procedures for collecting, handling, and analyzing DBS samples. Advantages of using DBS samples-compared with venipuncture include the relative ease and low cost of sample collection, transport, and storage. Disadvantages include requirements for assay development and validation as well as the relatively small volumes of sample. We present the results of a comprehensive literature review of published protocols for analysis of DBS samples, and we provide more detailed analysis of protocols for 45 analytes likely to be of particular relevance to population-level health research. Our objective is to provide investigators with the information they need to make informed decisions regarding the appropriateness of blood spot methods for their research interests.

  4. Factors influencing moisture analysis in the 3013 destructive examination surveillance program

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

    Scogin, J. H.

    Thermogravimetric analysis of a solid sample with mass spectrometry (TGA-MS) of the evolved gas is used in the destructive examination (DE) portion of the Integrated Surveillance Program to quantify the moisture content of the material stored in a 3013 container. As with any measurement determined from a small sample, the collection, storage, transportation, and handling of the sample can affect its ability to represent the properties of the bulk material. During the course of the DE program, questions have periodically arisen concerning the ability of the moisture sample to reflect reliably the actual moisture content of the entire material storedmore » in the 3013 container. Most concerns are related to the ability to collect a representative sample and to preserve the moisture content of the sample between collection and analysis. Recent delays in analysis caused by maintenance issues with the TGA-MS instrument presented a unique opportunity to document and quantify the effects various factors have on the TGA-MS moisture measurement. This report will use recent data to document the effects that current sample collection and handling practices have on the TGA-MS moisture measurement. Some suggestions will be made which could improve the current sample collection and handling practices for the TGA-MS moisture measurement so that the analytical results more accurately reflect the moisture content of the material stored in the 3013 container.« less

  5. Cariogenic potential of stored human milk--an in-vitro study.

    PubMed

    Hegde, Amitha M; Vikyath, Rani

    2007-01-01

    Human milk samples collected from ten lactating mothers in the K. S. Hegde Medical Hospital, Mangalore were divided into five different parts and stored at different temperatures for varying durations. The pH, buffer capacity and growth of Streptococcus mutans were assessed in each of these samples. There was a fall in pH of human milk stored at various temperatures. The buffer capacity of human milk increased with duration of storage. There was an increase in Streptococcus colony count in stored human milk proportional to the duration of storage and it increased more rapidly in case of milk stored at higher temperatures (0 degrees C - 4 degrees C) compared to the milk stored in the freezer (-19 degrees C). Milk samples stored at room temperature for 6 hours and in the freezer at -19 degrees C for 2 weeks were found to be relatively safe.

  6. Evaluation of a solid matrix for collection and ambient storage of RNA from whole blood

    PubMed Central

    2014-01-01

    Background Whole blood gene expression-based molecular diagnostic tests are becoming increasingly available. Conventional tube-based methods for obtaining RNA from whole blood can be limited by phlebotomy, volume requirements, and RNA stability during transport and storage. A dried blood spot matrix for collecting high-quality RNA, called RNA Stabilizing Matrix (RSM), was evaluated against PAXgene® blood collection tubes. Methods Whole blood was collected from 25 individuals and subjected to 3 sample storage conditions: 18 hours at either room temperature (baseline arm) or 37°C, and 6 days at room temperature. RNA was extracted and assessed for integrity by Agilent Bioanalyzer, and gene expression was compared by RT-qPCR across 23 mRNAs comprising a clinical test for obstructive coronary artery disease. Results RSM produced RNA of relatively high integrity across the various tested conditions (mean RIN ± 95% CI: baseline arm, 6.92 ± 0.24; 37°C arm, 5.98 ± 0.48; 6-day arm, 6.72 ± 0.23). PAXgene samples showed comparable RNA integrity in both baseline and 37°C arms (8.42 ± 0.17; 7.92 ± 0.1 respectively) however significant degradation was observed in the 6-day arm (3.19 ± 1.32). Gene expression scores on RSM were highly correlated between the baseline and 37°C and 6-day study arms (median r = 0.96, 0.95 respectively), as was the correlation to PAXgene tubes (median r = 0.95, p < 0.001). Conclusion RNA obtained from RSM shows little degradation and comparable RT-qPCR performance to PAXgene RNA for the 23 genes analyzed. Further development of this technology may provide a convenient method for collecting, shipping, and storing RNA for gene expression assays. PMID:24855452

  7. Stability of (13) C-Urea Breath Test Samples Over Time in the Diagnosis of Helicobacter pylori.

    PubMed

    Perets, Tsachi Tsadok; Shporn, Einav; Boltin, Doron; Dickman, Ram; Niv, Yaron

    2016-05-01

    The accuracy and repeatability of breath test in the diagnosis of Helicobacter pylori infection have not been adequately investigated. Although it has been shown that storage for long periods does not affect the analysis results, no data are available on the effect of repetitive testing. In this study, our aim was to evaluate the repeatability of the analyses of breath samples at room temperature. A total of 202 positive breath samples were collected in duplicates, before and after administration of 75 mg (13) C- urea dissolved in 100 ml of orange juice. Breath test results were expressed as delta (13) CO2 . The cut-off value was 3.5 parts per thousand. Each sample was analyzed in a mass spectrometer 7, 14, 21, and 28 days after collection. The accuracy calculation was based on the comparison of the delta (13) CO2 obtained in the three consecutive weeks following the first test run to the delta (13) CO2 obtained in the first test run. Two hundred (99%), 197 (97.52%), and 196 (97%) of the 202 samples tested positive in the second, third, and fourth test runs, respectively. The accuracy of the delta (13) CO2 was 98.6%, 99.2%, and 96.7% in the three consecutive runs, respectively. Short-term storage of 1 month does not affect sample stability or the results of (13) C-urea breath tests in up to three consecutive repeats. © 2015 Wiley Periodicals, Inc.

  8. Planned Variation in Preanalytical Conditions to Evaluate Biospecimen Stability in the National Children’s Study (NCS)

    PubMed Central

    Mechanic, Leah; Mendez, Armando; Merrill, Lori; Rogers, John; Layton, Marnie; Todd, Deborah; Varanasi, Arti; O’Brien, Barbara; Meyer, William A.; Zhang, Ming; Schleicher, Rosemary L.; Moye, Jack

    2014-01-01

    BACKGROUND Preanalytical conditions encountered during collection, processing, and storage of biospecimens may influence laboratory results. The National Children’s Study (NCS) is a planned prospective cohort study of 100,000 families to examine the influence of a wide variety of exposures on child health. In developing biospecimen collection, processing, and storage procedures for the NCS, we identified several analytes of different biochemical categories for which it was unclear to what extent deviations from NCS procedures could influence measurement results. METHODS A pilot study was performed to examine effects of preanalytic sample handling conditions (delays in centrifugation, freezing delays, delays in separation from cells, additive delay, and tube type) on concentrations of eight different analytes. 2,825 measurements were made to assess 15 unique combinations of analyte and handling conditions in blood collected from 151 women of childbearing age (≥20 individuals per handling condition). RESULTS The majority of analytes were stable under the conditions evaluated. However, levels of plasma interleukin-6 and serum insulin were decreased in response to sample centrifugation delays of up to 5.5 hours post collection (P<0.0001). In addition, delays in freezing centrifuged plasma samples (comparing 24, 48 and 72 hours to immediate freezing) resulted in increased levels of adrenocorticotropic hormone (P=0.0014). CONCLUSIONS Determining stability of proposed analytes in response to preanalytical conditions and handling helps to ensure high-quality specimens for study now and in the future. The results inform development of procedures, plans for measurement of analytes, and interpretation of laboratory results. PMID:23924524

  9. 76 FR 40749 - Agency Information Collection Activities: Records and Supporting Data: Importation, Receipt...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-11

    ...] Agency Information Collection Activities: Records and Supporting Data: Importation, Receipt, Storage, and... collection. (2) Title of the Form/Collection: Records and Supporting Data: Importation, Receipt, Storage, and... importation, manufacture, receipt, storage, and disposition of all explosive materials covered under 18 U.S.C...

  10. Testing odorants recovery from a novel metallized fluorinated ethylene propylene gas sampling bag.

    PubMed

    Zhu, Wenda; Koziel, Jacek A; Cai, Lingshuang; Wright, Donald; Kuhrt, Fred

    2015-12-01

    Industry-standard Tedlar bags for odor sample collection from confined animal feeding operations (CAFOs) have been challenged by the evidence of volatile organic compound (VOC) losses and background interferences. Novel impermeable aluminum foil with a thin layer of fluorinated ethylene propylene (FEP) film on the surface that is in contact with a gas sample was developed to address this challenge. In this research, Tedlar and metallized FEP bags were compared for (a) recoveries of four characteristic CAFO odorous VOCs (ethyl mercaptan, butyric acid, isovaleric acid and p-cresol) after 30 min and 24 hr sample storage time and for (b) chemical background interferences. All air sampling and analyses were performed with solid-phase microextraction (SPME) followed by gas chromatography-mass spectroscopy (GC-MS). Mean target gas sample recoveries from metallized FEP bags were 25.9% and 28.0% higher than those in Tedlar bags, for 30 min and 24 hr, respectively. Metallized FEP bags demonstrated the highest p-cresol recoveries after 30-min and 24-hr storage, 96.1±44.5% and 44.8±10.2%, respectively, among different types of sampling bags reported in previous studies. However, a higher variability was observed for p-cresol recovery with metallized FEP bags. A 0% recovery of ethyl mercaptan was observed with Tedlar bags after 24-hr storage, whereas an 85.7±7.4% recovery was achieved with metallized FEP bags. Recoveries of butyric and isovaleric acids were similar for both bag types. Two major impurities in Tedlar bags' background were identified as N,N-dimethylacetamide and phenol, while backgrounds of metallized FEP bags were significantly cleaner. Reusability of metallized FEP bags was tested. Caution is advised when using polymeric materials for storage of livestock-relevant odorous volatile organic compounds. The odorants loss with storage time confirmed that long-term storage in whole-air form is ill advised. A focused short-term odor sample containment should be biased toward the most inert material available relative to the highest impact target odorant. Metallized FEP was identified as such a material to p-cresol as the highest impact odorant from confined animal feeding operations. Metallized FEP bags have much cleaner background than commercial Tedlar bags do. Significantly higher recoveries of methyl mercaptan and p-cresol were also observed with metallized FEP bags.

  11. Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies

    PubMed Central

    Ostler, Michael W.; Porter, James H.; Buxton, Orfeu M.

    2014-01-01

    Biomarkers are directly-measured biological indicators of disease, health, exposures, or other biological information. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spots meet this need, and can be collected in the field with high response rates. These elements are particularly important in longitudinal study designs including interventions where attrition is critical to avoid, and high response rates improve the interpretation of results. Dried Blood Spot sample collection is simple, quick, relatively painless, less invasive then venipuncture, and requires minimal field storage requirements (i.e. samples do not need to be immediately frozen and can be stored for a long period of time in a stable freezer environment before assay). The samples can be analyzed for a variety of different analytes, including cholesterol, C-reactive protein, glycosylated hemoglobin, numerous cytokines, and other analytes, as well as provide genetic material. DBS collection is depicted as employed in several recent studies. PMID:24513728

  12. The NASA Ames Research Center Institutional Scientific Collection: History, Best Practices and Scientific Opportunities

    NASA Technical Reports Server (NTRS)

    Rask, Jon C.; Chakravarty, Kaushik; French, Alison; Choi, Sungshin; Stewart, Helen

    2017-01-01

    The NASA Ames Life Sciences Institutional Scientific Collection (ISC), which is composed of the Ames Life Sciences Data Archive (ALSDA) and the Biospecimen Storage Facility (BSF), is managed by the Space Biosciences Division and has been operational since 1993. The ALSDA is responsible for archiving information and animal biospecimens collected from life science spaceflight experiments and matching ground control experiments. Both fixed and frozen spaceflight and ground tissues are stored in the BSF within the ISC. The ALSDA also manages a Biospecimen Sharing Program, performs curation and long-term storage operations, and makes biospecimens available to the scientific community for research purposes via the Life Science Data Archive public website (https:lsda.jsc.nasa.gov). As part of our best practices, a viability testing plan has been developed for the ISC, which will assess the quality of archived samples. We expect that results from the viability testing will catalyze sample use, enable broader science community interest, and improve operational efficiency of the ISC. The current viability test plan focuses on generating disposition recommendations and is based on using ribonucleic acid (RNA) integrity number (RIN) scores as a criteria for measurement of biospecimen viablity for downstream functional analysis. The plan includes (1) sorting and identification of candidate samples, (2) conducting a statiscally-based power analysis to generate representaive cohorts from the population of stored biospecimens, (3) completion of RIN analysis on select samples, and (4) development of disposition recommendations based on the RIN scores. Results of this work will also support NASA open science initiatives and guides development of the NASA Scientific Collections Directive (a policy on best practices for curation of biological collections). Our RIN-based methodology for characterizing the quality of tissues stored in the ISC since the 1980s also creates unique scientific opportunities for temporal assessment across historical missions. Support from the NASA Space Biology Program and the NASA Human Research Program is gratefully acknowledged.

  13. Microbiological Safety and Food Handling Practices of Seed Sprout Products in the Australian State of Victoria.

    PubMed

    Symes, Sally; Goldsmith, Paul; Haines, Heather

    2015-07-01

    Seed sprouts have been implicated as vehicles for numerous foodborne outbreaks worldwide. Seed sprouts pose a unique food safety concern because of the ease of microbiological seed contamination, the inherent ability of the sprouting process to support microbial growth, and their consumption either raw or lightly cooked. To examine seed sprout safety in the Australian state of Victoria, a survey was conducted to detect specific microbes in seed sprout samples and to investigate food handling practices relating to seed sprouts. A total of 298 seed sprout samples were collected from across 33 local council areas. Escherichia coli was detected in 14.8%, Listeria spp. in 12.3%, and Listeria monocytogenes in 1.3% of samples analyzed. Salmonella spp. were not detected in any of the samples. A range of seed sprout handling practices were identified as potential food safety issues in some food businesses, including temperature control, washing practices, length of storage, and storage in proximity to unpackaged ready-to-eat potentially hazardous foods.

  14. Stability and reproducibility of a single-sample urinary C-peptide/creatinine ratio and its correlation with 24-h urinary C-peptide.

    PubMed

    McDonald, Tim J; Knight, Bridget A; Shields, Beverley M; Bowman, Pamela; Salzmann, Maurice B; Hattersley, Andrew T

    2009-11-01

    C-peptide measurement in blood or 24-h urine samples provides useful information regarding endogenous insulin secretion, but problems related to the rapid degradation of C-peptide in blood and difficulty of 24-h urine collection have limited widespread routine clinical use of this test. We assessed the feasibility of measuring urinary C-peptide (UCP) with correction for creatinine concentration in single urine samples. We analyzed UCP using a routine electrochemiluminescence immunoassay in samples from 21 healthy volunteers. We investigated the stability of UCP with different preservatives and storage conditions and compared the reproducibility of urinary C-peptide/creatinine ratio (UCPCR) in first- and second-void fasting urines, then assessed correlations with 24-h collections. UCPCR was unchanged at room temperature for 24 h and at 4 degrees C for 72 h even in the absence of preservative. UCPCR collected in boric acid was stable at room temperature for 72 h. UCPCR remained stable after 7 freeze-thaw cycles but decreased with freezer storage time and dropped to 82%-84% of baseline by 90 days at -20 degrees C. Second-void fasting UCPCRs were lower than first-void (median 0.78 vs 1.31, P = 0.0003) and showed less variation (CV 33% vs 52%), as second-void UCPCRs were not influenced by evening food-related insulin secretion. Second-void fasting UCPCR was highly correlated with 24-h UCP (r = 0.8, P = 0.00006). Second-void fasting UCPCR is a reproducible measure that correlates well with 24-h UCP in normal samples. The 3-day stability of UCPCR at room temperature greatly increases its potential clinical utility.

  15. Microbial alteration of normal alkane δ13C and δD in sedimentary archives

    NASA Astrophysics Data System (ADS)

    Brittingham, A.; Hren, M. T.; Hartman, G.

    2016-12-01

    Long-carbon chain normal alkanes (e.g. C25-C33) are produced by a wide range of terrestrial plants and commonly preserved in ancient sediments. These serve as a potential paleoclimate proxy because their hydrogen (δD) and carbon (δ13C) isotope values reflect the combined effect of plant-specific species effects and responses to environmental conditions. While these are commonly believed to remain unaltered at low burial temperatures (e.g. <150°C), there is still uncertainty around the role microbes play during the breakdown of these compounds in stored sediment and the potential risk for isotopic alteration. We analyzed two sets of identical samples to assess the role of microbial and other degradation process on the hydrogen and carbon isotope composition of these compounds. The first set of sediment samples were collected in the summer of 2011 from central Armenia, a region with continental climate, and allowed to sit in sealed bags at room temperature for three years. A second and identical set was collected in 2014 and frozen immediately. Stored samples showed high amounts of medium chain length n-alkanes (C19-C26), produced by microorganisms, which were absent from the samples that were collected in 2014 and frozen immediately after sampling. Along with the presence of medium chain length n-alkanes, the average chain length of n-alkanes from C25-C33 decreased significantly in all 2011 samples. Storage of the samples over three years resulted in altered δD and δ13C values of C29 and C31 n-alkanes. While δD values were heavier relative to the control by 4-25‰, δ13C values were mostly lighter (maximum change of -4.2‰ in C29 and -2.9‰ in C31). DNA analysis of the soil showed Rhodococcus and Aeromicrobium, genera that contain multiple coding regions for alkane degrading enzymes CYP153 and AlkB, increased by an order of magnitude during sample storage (from 0.7% to 7.5% of bacteria present). The proliferation of alkane degrading bacteria, combined with the large changes of long-chain n-alkane isotope values, suggest that bacteria may play a larger role than previously expected in altering the measured δD and δ13C values of long-chain n-alkanes during storage. This poses a potentially significant issue for all manner of samples that are not stored frozen, including a variety of sedimentary cores.

  16. The human milk oligosaccharides are not affected by pasteurization and freeze-drying.

    PubMed

    Hahn, Won-Ho; Kim, Jaehan; Song, Seunghyun; Park, Suyeon; Kang, Nam Mi

    2017-11-06

    Human milk oligosaccharides (HMOs) are known as important factors in neurologic and immunologic development of neonates. Moreover, freeze-drying seems to be a promising storage method to improve the processes of human milk banks. However, the effects of pasteurization and freeze-drying on HMOs were not evaluated yet. The purpose of this study is to analyze and compare the HMOs profiles of human milk collected before and after the pasteurization and freeze-drying. Totally nine fresh human milk samples were collected from three healthy mothers at the first, second, and third week after delivery. The samples were treated with Holder pasteurization and freeze-drying. HMOs profiles were analyzed by matrix-assisted laser desorption/ionization (MALDI) time-of-flight/time-of-flight (TOF/TOF) mass spectrometry and compared between samples collected before and after the treatments. Human milk samples showed significantly different HMO patterns between mothers. However, HMOs were not affected by lactation periods within 3 weeks after delivery (r 2  = 0.972-0.999, p < .001). Moreover, both of pasteurization and freeze-drying were found not to affect HMO patterns in a correlation analysis (r 2  = 0.989-0.999, p < .001). HMO patterns were found not to be affected by pasteurization and freeze-drying of donor milks. We hope that introducing freeze-drying to the human milk banks would be encouraged by the present study. However, the storage length without composition changes of HMOs after freeze-drying needs to be evaluated in the further studies.

  17. Monitoring in inline storage sewers for stormwater treatment to determine efficiencies.

    PubMed

    Frehmann, T; Mietzel, T; Kutzner, R; Spengler, B; Geiger, W F

    2004-01-01

    A special structure of combined sewer overflow tanks is the inline storage sewer with downstream discharge (SKU). This layout has the advantage that besides the sewer system, no other structures are required for storm water treatment. Consequently only very little space is required and compared to combined sewer overflow tanks, there is an enormous potential in reducing costs during construction. To investigate the efficiency of an inline storage sewer, a monitoring station was established in Dortmund-Scharnhorst, Germany. The monitoring station was in operation for a period of 2.5 years. Within this period water samples were taken during a total of 20 discharge events. Besides the complete hydraulic data collection, seven water samplers took more than 5,000 water samples during dry and wet weather. This adds up to a total of more than 20,000 individual lab analyses. The average of the total efficiency for the SKU-West is 86%. 29% of this efficiency can be attributed to the throttle flow. The remaining 57% can be divided into a part of 48% that can be attributed to the process storage and 9% that can be attributed to sedimentation and erosion process.

  18. Comparability of the effect of storage time and temperature on serum anti-Müllerian hormone measurement between original and modified enzyme-linked immunosorbent assay.

    PubMed

    Yue, Chao-Yan; Ying, Chun-Mei

    2017-01-01

    To explore the effect of modified enzyme-linked immunosorbent assay on the AMH results is increased or decreased, and to investigate the effect of storage time and temperature on AMH measurements with and without sample premixing assay buffer using the Kangrun ELISA method. Serum AMH concentration were measured by ELISA, consistency between two kits, and comparability between original and the modified assay under different stored conditions were analyzed by Passing-Bablok regression analysis and Bland-Altman bias evaluation. There was a strong consistency between AMH concentrations measured in Kangrun ELISA and Ansh Labs ultra-sensitive AMH ELISA. Pre-mixing serum specimens with assay buffer gave consistent results compared with original assay. Modified protocol can reduce the amplitude of increase affected by sample aged and give the most consistent results regardless of storage conditions. Pre-mixing protocol did not influence the results of fresh serum or frozen serum incubation <3days at 4°C and -80°C, but when specimens detected after collection and stored in other storage conditions, should be pre-mixed with assay buffer to insure its accuracy. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. 77 FR 10559 - Agency Information Collection Activities: Proposed collection; comments requested: Certification...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-22

    ... Secure Gun Storage or Safety Devices ACTION: 60-Day notice of information collection. The Department of...) Title of the Form/Collection: Certification of Secure Gun Storage or Safety Devices. (3) Agency form... gun storage or safety devices. (5) An estimate of the total number of respondents and the amount of...

  20. Do pharmaceuticals, pathogens, and other organic waste water compounds persist when waste water is used for recharge?

    USGS Publications Warehouse

    Cordy, Gail E.; Duran, Norma L.; Bouwer, Herman; Rice, Robert C.; Furlong, Edward T.; Zaugg, Steven D.; Meyer, Michael T.; Barber, Larry B.; Kolpin, Dana W.

    2004-01-01

    A proof-of-concept experiment was devised to determine if pharmaceuticals and other organic waste water compounds (OWCs), as well as pathogens, found in treated effluent could be transported through a 2.4 m soil column and, thus, potentially reach ground water under recharge conditions similar to those in arid or semiarid climates. Treated effluent was applied at the top of the 2.4 m long, 32.5 cm diameter soil column over 23 days, Samples of the column inflow were collected from the effluent storage tank at the beginning (Tbegin) and end (Tend) of the experiment, and a sample of the soil column drainage at the base of the column (Bend) was collected at the end of the experiment. Samples were analyzed for 131 OWCs including veterinary and human antibiotics, other prescription and nonprescription drugs, widely used household and industrial chemicals, and steroids and reproductive hormones, as well as the pathogens Salmonella and Legionella. Analytical results for the two effluent samples taken at the beginning (Tbegin) and end (Tend) of the experiment indicate that the number of OWCs detected in the column inflow decreased by 25% (eight compounds) and the total concentration of OWCs decreased by 46% while the effluent was in the storage tank during the 23-day experiment. After percolating through the soil column, an additional 18 compounds detected in Tend (67% of OWCs) were no longer detected in the effluent (Bend) and the total concentration of OWCs decreased by more than 70%. These compounds may have been subject to transformation (biotic and abiotic), adsorption, and (or) volatilization in the storage tank and during travel through the soil column. Eight compounds—carbamazapine; sulfamethoxazole; benzophenone; 5-methyl-1H-benzotriazole; N,N-diethyltoluamide; tributylphosphate; tri(2-chloroethyl) phosphate; and cholesterol—were detected in all three samples indicating they have the potential to reach ground water under recharge conditions similar to those in arid and semiarid climates. Results from real-time polymerase chain reactions demonstrated the presence of Legionella in all three samples. Salmonella was detected only in Tbegin, suggesting that the bacteria died off in the effluent storage tank over the period of the experiment. This proof-of-concept experiment demonstrates that, under recharge conditions similar to those in arid or semiarid climates, some pharmaceuticals, pathogens, and other OWCs can persist in treated effluent after soil-aquifer treatment.

  1. 40 CFR 80.1502 - What are the survey requirements related to gasoline-ethanol blends?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... surveys, including sample collection, transportation, storage, and analysis. (4) Survey Plan Design... 40 Protection of Environment 17 2013-07-01 2013-07-01 false What are the survey requirements... Requirements for Gasoline-Ethanol Blends § 80.1502 What are the survey requirements related to gasoline-ethanol...

  2. 40 CFR 80.1502 - What are the survey requirements related to gasoline-ethanol blends?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... surveys, including sample collection, transportation, storage, and analysis. (4) Survey Plan Design... 40 Protection of Environment 17 2012-07-01 2012-07-01 false What are the survey requirements... Requirements for Gasoline-Ethanol Blends § 80.1502 What are the survey requirements related to gasoline-ethanol...

  3. 40 CFR 80.1502 - What are the survey requirements related to gasoline-ethanol blends?

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... surveys, including sample collection, transportation, storage, and analysis. (4) Survey Plan Design... 40 Protection of Environment 17 2014-07-01 2014-07-01 false What are the survey requirements... Requirements for Gasoline-Ethanol Blends § 80.1502 What are the survey requirements related to gasoline-ethanol...

  4. Mars oxygen production system design

    NASA Technical Reports Server (NTRS)

    Cotton, Charles E.; Pillow, Linda K.; Perkinson, Robert C.; Brownlie, R. P.; Chwalowski, P.; Carmona, M. F.; Coopersmith, J. P.; Goff, J. C.; Harvey, L. L.; Kovacs, L. A.

    1989-01-01

    The design and construction phase is summarized of the Mars oxygen demonstration project. The basic hardware required to produce oxygen from simulated Mars atmosphere was assembled and tested. Some design problems still remain with the sample collection and storage system. In addition, design and development of computer compatible data acquisition and control instrumentation is ongoing.

  5. Mars oxygen production system design

    NASA Technical Reports Server (NTRS)

    1988-01-01

    This report summarizes the design and construction of the Mars oxygen demonstration project. The basic hardware required to produce oxygen from simulated Mars atmosphere has been assembled and tested. Some design problems still remain with the sample collection and storage system. In addition, design and development of computer data acquisition and control instrumentation is continuing.

  6. Protocol to Reconstruct Historical Contaminant Loading to Large Lakes: The Lake Michigan Sediment Record of Mercury

    EPA Science Inventory

    Samples of opportunity from Pb-210 dated sediment cores collected from Lake Michigan between 1994 and 1996 were analyzed for mercury. The storage of both anthropogenic and total (post-1850) mercury in the lake was calculated to be 186 and 228 metric tons, respectively. By setti...

  7. Microbiological contamination with moulds in work environment in libraries and archive storage facilities.

    PubMed

    Zielinska-Jankiewicz, Katarzyna; Kozajda, Anna; Piotrowska, Malgorzata; Szadkowska-Stanczyk, Irena

    2008-01-01

    Microbiological contamination with fungi, including moulds, can pose a significant health hazard to those working in archives or museums. The species involved include Aspergillus, Penicillium, Geotrichum, Alternaria, Cladosporium, Mucor, Rhizopus, Trichoderma, Fusarium which are associated mostly with allergic response of different types. The aim of the study was to analyse, both in quantitative and qualitative terms, workplace air samples collected in a library and archive storage facilities. Occupational exposure and the related health hazard from microbiological contamination with moulds were assessed in three archive storage buildings and one library. Air samples (total 60) were collected via impact method before work and at hourly intervals during work performance. Surface samples from the artifacts were collected by pressing a counting (RODAC) plate filled with malt extract agar against the surface of the artifacts. The air sample and surface sample analyses yielded 36 different mould species, classified into 19 genera, of which Cladosporium and Penicillium were the most prevalent. Twelve species were regarded as potentially pathogenic for humans: 8 had allergic and 11 toxic properties, the latter including Aspergillus fumigatus. Quantitative analysis revealed air microbiological contamination with moulds at the level ranging from 1.8 x 10(2)-2.3 x 10(3) cfu/m(3). In surface samples from library and archive artifacts, 11 fungal species were distinguished; the number of species per artifact varying from 1-6 and colony count ranging from 4 x 10(1) to 8-10(1) cfu/100 cm(2). Higher contamination levels were found only for Cladosporium cladosporioides (1.48 x 10(3) cfu/100 cm(2)) and Paecillomyces varioti (1.2 x 10(2) cfu/100 cm(2)). At the workposts examined, although no clearly visible signs of mould contamination could be found, the study revealed abundant micromycetes, with the predominant species of Cladosporium and Penicillium. The detected species included also potentially pathogenic microorganisms which can cause allergic and toxic effects, such as Aspergillus fumigatus, that could be hazardous to workers' health. For some species, the concentration levels exceeded the values considered the proposed hygienic standards for total microscopical fungi in occupational settings. The findings of the study point to unsatisfactory hygienic conditions at the worksites examined, resulting in microbiological contamination with moulds, as well as the necessity for prompt remedial activities on the part of the employers.

  8. Stability of flavoured phytosterol-enriched drinking yogurts during storage as affected by different packaging materials.

    PubMed

    Semeniuc, Cristina Anamaria; Cardenia, Vladimiro; Mandrioli, Mara; Muste, Sevastiţa; Borsari, Andrea; Rodriguez-Estrada, Maria Teresa

    2016-06-01

    The aim of this study was to investigate the influence of different packaging materials on storage stability of flavoured phytosterol-enriched drinking yogurts. White vanilla (WV) and blood orange (BO) phytosterol-enriched drinking yogurts conditioned in mono-layer and triple-layer co-extruded plastic bottles were stored at +6 ± 1 °C for 35 days (under alternating 12 h light and 12 h darkness) to simulate shelf-life conditions. Samples were collected at three different storage times and subjected to determination of total sterol content (TSC), peroxide value (PV) and thiobarbituric acid reactive substances (TBARs). TSC was not significantly affected by packaging material or storage time and met the quantity declared on the label. PV was significantly influenced by yogurt type × packaging material × storage time interaction and TBARs by packaging material × storage time interaction. Between the two packaging materials, the triple-layer plastic mini bottle with black coloured and completely opaque intermediate layer offered the best protection against lipid oxidation. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  9. Impact of Collection and Storage of Lung Tumor Tissue on Whole Genome Expression Profiling

    PubMed Central

    Freidin, Maxim B.; Bhudia, Neesa; Lim, Eric; Nicholson, Andrew G.; Cookson, William O.; Moffatt, Miriam F.

    2012-01-01

    Gene expression profiling could assist in revealing biomarkers of lung cancer prognosis and progression. The handling of biological samples may strongly influence global gene expression, a fact that has not been addressed in many studies. We sought to investigate the changes in gene expression that may occur as a result of sample processing time and conditions. Using Illumina Human WG-6 arrays, we quantified gene expression in lung carcinoma samples from six patients obtained at chest opening before and immediately after lung resection with storage in RNAlater [T1a(CO) and T1b(LR)], after receipt of the sample for histopathology, placed in RNAlater [T2a(HP)]; snap frozen [T2b(HP.SF)]; or snap frozen and stored for 1 week [T2c(HP.SFA)], as well as formalin-fixed, paraffin-embedded (FFPE) block samples. Sampling immediately after resection closely represented the tissue obtained in situ, with only 1% of genes differing more than twofold [T1a(CO) versus T1b(LR)]. Delaying tissue harvest for an average of 30 minutes from the operating theater had a significant impact on gene expression, with approximately 25% of genes differing between T1a(CO) and T2a(HP). Many genes previously identified as lung cancer biomarkers were altered during this period. Examination of FFPE specimens showed minimal correlation with fresh samples. This study shows that tissue collection immediately after lung resection with conservation in RNAlater is an optimal strategy for gene expression profiling. PMID:22240448

  10. Biomass Logistics

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

    J. Richard Hess; Kevin L. Kenney; William A. Smith

    Equipment manufacturers have made rapid improvements in biomass harvesting and handling equipment. These improvements have increased transportation and handling efficiencies due to higher biomass densities and reduced losses. Improvements in grinder efficiencies and capacity have reduced biomass grinding costs. Biomass collection efficiencies (the ratio of biomass collected to the amount available in the field) as high as 75% for crop residues and greater than 90% for perennial energy crops have also been demonstrated. However, as collection rates increase, the fraction of entrained soil in the biomass increases, and high biomass residue removal rates can violate agronomic sustainability limits. Advancements inmore » quantifying multi-factor sustainability limits to increase removal rate as guided by sustainable residue removal plans, and mitigating soil contamination through targeted removal rates based on soil type and residue type/fraction is allowing the use of new high efficiency harvesting equipment and methods. As another consideration, single pass harvesting and other technologies that improve harvesting costs cause biomass storage moisture management challenges, which challenges are further perturbed by annual variability in biomass moisture content. Monitoring, sampling, simulation, and analysis provide basis for moisture, time, and quality relationships in storage, which has allowed the development of moisture tolerant storage systems and best management processes that combine moisture content and time to accommodate baled storage of wet material based upon “shelf-life.” The key to improving biomass supply logistics costs has been developing the associated agronomic sustainability and biomass quality technologies and processes that allow the implementation of equipment engineering solutions.« less

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

    Riot, Vincent J.

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing andmore » storage.« less

  12. Changes in postharvest quality of Swiss chard grown using 3 organic preharvest treatments.

    PubMed

    Daiss, N; Lobo, M G; Gonzalez, M

    2008-08-01

    Using storage conditions recommended for conventional chard (4 degrees C, 90% RH and 7 d), the chard treated with some organic preharvest treatments [effective microorganisms, a fermented mixture of effective microorganisms with organic matter (EM-Bokashi + EM), and an auxiliary soil product] lost considerable water (> 2%) and weight (> 25%). These results indicate that organic methods tested produce a vegetable that can not sustain its quality when commercialized through the conventional supply chain. Nevertheless, respiration, color, pH, and titratable acidity practically remained constant during conservation. Ascorbic acid content was constant in chard treated with the different preharvest treatments and collected at 8 wk after sowing (normal harvest). However, the ascorbic acid content of the control chard decreased 60% after 7 d of storage. This vitamin diminished (35%) in chard collected after 19 wk after sowing (late harvest) during the postharvest conservation. The greatest difference in chard quality was registered between sampling dates since chard collected during the late harvest had higher levels of dry matter, sugars, acids, proteins, and ascorbic acid than chard collected during the normal harvest.

  13. Quantifying the effect of 3D spatial resolution on the accuracy of microstructural distributions

    NASA Astrophysics Data System (ADS)

    Loughnane, Gregory; Groeber, Michael; Uchic, Michael; Riley, Matthew; Shah, Megna; Srinivasan, Raghavan; Grandhi, Ramana

    The choice of spatial resolution for experimentally-collected 3D microstructural data is often governed by general rules of thumb. For example, serial section experiments often strive to collect at least ten sections through the average feature-of-interest. However, the desire to collect high resolution data in 3D is greatly tempered by the exponential growth in collection times and data storage requirements. This paper explores the use of systematic down-sampling of synthetically-generated grain microstructures to examine the effect of resolution on the calculated distributions of microstructural descriptors such as grain size, number of nearest neighbors, aspect ratio, and Ω3.

  14. Environmental Monitoring and the Gas Industry: Program Manager Handbook

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

    Gregory D. Gillispie

    1997-12-01

    This document has been developed for the nontechnical gas industry manager who has the responsibility for the development of waste or potentially contaminated soil and groundwater data or must make decisions based on such data for the management or remediation of these materials. It explores the pse of common analytical chemistry instrumentation and associated techniques for identification of environmentally hazardous materials. Sufficient detail is given to familiarize the nontechnical reader with the principles behind the operation of each technique. The scope and realm of the techniques and their constituent variations are portrayed through a discussion of crucial details and, wheremore » appropriate, the depiction of real-life data. It is the author's intention to provide an easily understood handbook for gas industry management. Techniques which determine the presence, composition, and quantification of gas industry wastes are discussed. Greater focus is given to traditional techniques which have been the mainstay of modem analytical benchwork. However, with the continual advancement of instrumental principles and design, several techniques have been included which are likely to receive greater attention in fiture considerations for waste-related detection. Definitions and concepts inherent to a thorough understanding of the principles common to analytical chemistry are discussed. It is also crucial that gas industry managers understand the effects of the various actions which take place before, during, and after the actual sampling step. When a series of sample collection, storage, and transport activities occur, new or inexperienced project managers may overlook or misunderstand the importance of the sequence. Each step has an impact on the final results of the measurement process; errors in judgment or decision making can be costly. Specific techniques and methodologies for the collection, storage, and transport of environmental media samples are not described or discussed in detail in thk handbook. However, the underlying philosophy regarding the importance of proper collection, storage, and transport practices, as well as pertinent references, are presented.« less

  15. Stability of phenytoin in blood collected in vacuum blood collection tubes.

    PubMed

    Parish, R C; Alexander, T

    1990-01-01

    The stability of phenytoin in blood collected in plain and serum separator tubes (SSTs) was investigated under simulated storage and transport conditions. The drug was generally more stable in plain collection tubes than in SSTs. No degradation occurred in plain red-top tubes or in refrigerated SSTs, but clinically significant degradation was present in SSTs stored at room temperature (25 degrees C) and at elevated temperature (32 degrees C) 24 h after collection. The mean loss was 17.9% at 25 degrees C and 25.9% at 32 degrees C. It is recommended that if blood is to be transported or stored in SSTs, the samples be refrigerated unless assay can be performed within 8 h.

  16. Air Quality Inside Police Drug Safes and Drug Storage Areas.

    PubMed

    Doran, Gregory S; Deans, Ralph; De Filippis, Carlo; Kostakis, Chris; Howitt, Julia A

    2018-06-01

    Storage of drug-based evidence inside sealed safes may allow chemical vapors to accumulate, creating concerns of drug exposure by inhalation, or the possibility of cross-contamination of drug evidence. Air samples were taken from inside eight drug safes and one small storage room at nine city and country police stations, as well as a large centralized drug evidence storage vault, in New South Wales (NSW), Australia. Sorbent tubes containing charcoal were used to determine whether any drug residues could be detected in the air, and to identify the types of chemicals present. Carbon traps were extracted and analyzed by LC-MS-MS for a suite of 22 licit and illicit drug residues and 2 metabolites. Carbon traps and SPME fibers were also analyzed by GC-MS for general volatile organic compound (VOC) residues. No detectable drug residues, either as airborne dust or vapor, were found in the safes, the storage room or the large central repository vault. No drugs were detected in any of the 34 urine samples collected at 8 of the 10 sampling locations, while only one of the five hair samples was positive for cocaine (9 pg/mg) provided by police exhibit officers at 3 of the 10 sampling locations. VOC analysis identified a variety of solvents associated with drug manufacture, plasticisers, personal care products and volatiles associated with plants such as cannabis. The results indicate that strong chemical odours emanating from drug safes are unlikely to be drug residues due to low volatility of drugs, and are more likely VOCs associated with their manufacture or from plant growing operations. Consideration should be given to the quality of air flow in rooms in which safes are housed and the use of air filtering inside safes to reduce the likelihood of VOC accumulation, and therefore the risk of human exposure.

  17. Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips

    USGS Publications Warehouse

    Jara, Rocio F.; Sepúlveda, Carolina; Ip, Hon S.; Samuel, Michael D.

    2015-01-01

    Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

  18. Total protein concentration and diagnostic test results for gray wolf (Canis lupus) serum using Nobuto filter paper strips.

    PubMed

    Jara, Rocío F; Sepúlveda, Carolina; Ip, Hon S; Samuel, Michael D

    2015-04-01

    Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

  19. Corrective Action Investigation Plan for Corrective Action Unit 127: Areas 25 and 26 Storage Tanks, Nevada Test Site, Nevada (Rev. No.: 0, August 2002)

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

    NNSA /NV

    This Corrective Action Investigation Plan (CAIP) contains the U.S. Department of Energy, National Nuclear Security Administration Nevada Operations Offices's approach to collect the data necessary to evaluate corrective action alternatives appropriate for the closure of Corrective Action Unit (CAU) 127 under the Federal Facility Agreement and Consent Order. Corrective Action Unit 127 is located on the Nevada Test Site approximately 65 miles northwest of Las Vegas, Nevada. This CAU is comprised of 12 Corrective Action Sites (CASs) located at Test Cell C; the Engine Maintenance, Assembly, and Disassembly (E-MAD) Facility; the X-Tunnel in Area 25; the Pluto Disassembly Facility; themore » Pluto Check Station; and the Port Gaston Training Facility in Area 26. These CASs include: CAS 25-01-05, Aboveground Storage Tank (AST); CAS 25-02-02, Underground Storage Tank (UST); CAS 25-23-11, Contaminated Materials; CAS 25-12-01, Boiler; CAS 25-01-06, AST; CAS 25-01-07, AST; CAS 25-02-13, UST; CAS 26- 01-01, Filter Tank (Rad) and Piping; CAS 26-01-02, Filter Tank (Rad); CAS 26-99-01, Radioactively Contaminated Filters; CAS 26-02-01, UST; CAS 26-23-01, Contaminated Liquids Spreader. Based on site history, process knowledge, and previous field efforts, contaminants of potential concern for CAU 127 include radionuclides, metals, total petroleum hydrocarbons, volatile organic compounds, asbestos, and polychlorinated biphenyls. Additionally, beryllium may be present at some locations. The sources of potential releases are varied, but releases of contaminated liquids may have occurred and may have migrated into and impacted soil below and surrounding storage vessels at some of the CASs. Also, at several CASs, asbestos-containing materials may be present on the aboveground structures and may be friable. Exposure pathways are limited to ingestion, inhalation, and dermal contact (adsorption) of soils/sediments or liquids, or inhalation of contaminants by site workers due to disturbance of contaminated materials. Future land-use scenarios limit subsequent uses of the CASs to various nonresidential (i.e., industrial) activities. Field activities will consist of radiological walkover and screening surveys, and field-screening and collecting of both tank content and soil samples, and further sample testing as appropriate. A two-step data quality objective strategy will be followed: (1) Phase I will be to collect environmental samples for laboratory analysis to confirm the presence or absence of contaminants at concentrations exceeding preliminary action levels; and (2) Phase II will be to collect additional environmental samples for laboratory analysis to determine the extent of contamination identified in Phase I. The results of this field investigation will support a defensible evaluation of corrective action alternatives in the corrective action decision document.« less

  20. Biomolecular characterization, identification, enzyme activities of molds and physiological changes in sweet potatoes (Ipomea batatas) stored under controlled atmospheric conditions

    PubMed Central

    Oladoye, C. O.; Connerton, I. F.; Kayode, R. M. O.; Omojasola, P. F.; Kayode, I. B.

    2016-01-01

    Microbial attacks during storage are one of the primary causes of product deterioration, and can limit the process of prolonging the shelf-life of harvested food. In this study, sweet potatoes were stored at temperatures of 13, 21, and 29 °C for 4 weeks. Samples were collected during storage and plated on potato dextrose agar, from which axenic mold cultures were obtained and identified using 26S rRNA gene sequences. Physiological changes of potato tubers were assessed with respect to pathogenicity, enzyme activity, and atmospheric storage conditions. Six fungal species were identified, namely Penicillium chrysogenum (P. rubens), P. brevicompactum, Mucor circinelloides, Cladosporium cladosporiodes, P. expansum, and P. crustosum. The following fungal isolates, namely P. expansum, P. brevicompactum, and Rhizopus oryzae, were recovered from the re-infected samples and selected according to their levels of enzyme activity. This study revealed high levels of activity for cellulase and pectinase, which were most notable during the initial three days of testing, and were followed by a steady decrease (P<0.05). Polygalacturonase activity was prominent with values ranging from 12.64 to 56.79 U/mg (P. expansum) and 18.36 to 79.01 U/mg (P. brevicompactum). Spoilage was obvious in the control group, which had a 100% decay at the end of the experimental period compared with samples treated with iprodione and sodium hypochlorite, in which the decay rates were 5% and 55%, respectively. The data for the iprodione- and sodium hypochlorite-treated samples at the end of the 3-month storage period showed that they were significantly different (P=0.041), with the sodium hypochlorite-treated samples producing twice the rate of infection compared to the iprodione-treated samples. The comparative rate of the progression of decay in the treated samples can be expressed as iprodione

  1. Blood gas testing and related measurements: National recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    PubMed

    Dukić, Lora; Kopčinović, Lara Milevoj; Dorotić, Adrijana; Baršić, Ivana

    2016-10-15

    Blood gas analysis (BGA) is exposed to risks of errors caused by improper sampling, transport and storage conditions. The Clinical and Laboratory Standards Institute (CLSI) generated documents with recommendations for avoidance of potential errors caused by sample mishandling. Two main documents related to BGA issued by the CLSI are GP43-A4 (former H11-A4) Procedures for the collection of arterial blood specimens; approved standard - fourth edition, and C46-A2 Blood gas and pH analysis and related measurements; approved guideline - second edition. Practices related to processing of blood gas samples are not standardized in the Republic of Croatia. Each institution has its own protocol for ordering, collection and analysis of blood gases. Although many laboratories use state of the art analyzers, still many preanalytical procedures remain unchanged. The objective of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) is to standardize the procedures for BGA based on CLSI recommendations. The Working Group for Blood Gas Testing as part of the Committee for the Scientific Professional Development of the CSMBLM prepared a set of recommended protocols for sampling, transport, storage and processing of blood gas samples based on relevant CLSI documents, relevant literature search and on the results of Croatian survey study on practices and policies in acid-base testing. Recommendations are intended for laboratory professionals and all healthcare workers involved in blood gas processing.

  2. Microbial health risks associated with exposure to stormwater in a water plaza.

    PubMed

    Sales-Ortells, Helena; Medema, Gertjan

    2015-05-01

    Climate change scenarios predict an increase of intense rainfall events in summer in Western Europe. Current urban drainage systems cannot cope with such intense precipitation events. Cities are constructing stormwater storage facilities to prevent pluvial flooding. Combining storage with other functions, such as recreation, may lead to exposure to contaminants. This study assessed the microbial quality of rainwater collected in a water plaza and the health risks associated with recreational exposure. The water plaza collects street run-off, diverges first flush to the sewer system and stores the rest of the run-off in the plaza as open water. Campylobacter, Cryptosporidium and Legionella pneumophila were the pathogens investigated. Microbial source tracking tools were used to determine the origin (human, animal) of the intestinal pathogens. Cryptosporidium was not found in any sample. Campylobacter was found in all samples, with higher concentrations in samples containing human Bacteroides than in samples with zoonotic contamination (15 vs 3.7 gc (genomic copies)/100 mL). In both cases, the estimated disease risk associated with Campylobacter and recreational exposure was higher than the Dutch national incidence. This indicates that the health risk associated with recreational exposure to the water plaza is significant. L. pneumophila was found only in two out of ten pond samples. Legionnaire's disease risks were lower than the Dutch national incidence. Presence of human Bacteroides indicates possible cross-connections with the CSS that should be identified and removed. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Blood gas testing and related measurements: National recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine

    PubMed Central

    Dukić, Lora; Kopčinović, Lara Milevoj; Dorotić, Adrijana; Baršić, Ivana

    2016-01-01

    Blood gas analysis (BGA) is exposed to risks of errors caused by improper sampling, transport and storage conditions. The Clinical and Laboratory Standards Institute (CLSI) generated documents with recommendations for avoidance of potential errors caused by sample mishandling. Two main documents related to BGA issued by the CLSI are GP43-A4 (former H11-A4) Procedures for the collection of arterial blood specimens; approved standard – fourth edition, and C46-A2 Blood gas and pH analysis and related measurements; approved guideline – second edition. Practices related to processing of blood gas samples are not standardized in the Republic of Croatia. Each institution has its own protocol for ordering, collection and analysis of blood gases. Although many laboratories use state of the art analyzers, still many preanalytical procedures remain unchanged. The objective of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) is to standardize the procedures for BGA based on CLSI recommendations. The Working Group for Blood Gas Testing as part of the Committee for the Scientific Professional Development of the CSMBLM prepared a set of recommended protocols for sampling, transport, storage and processing of blood gas samples based on relevant CLSI documents, relevant literature search and on the results of Croatian survey study on practices and policies in acid-base testing. Recommendations are intended for laboratory professionals and all healthcare workers involved in blood gas processing. PMID:27812301

  4. No filters, no fridges: a method for preservation of water samples for eDNA analysis.

    PubMed

    Williams, Kelly E; Huyvaert, Kathryn P; Piaggio, Antoinette J

    2016-06-08

    Advancements in the detection of environmental DNA (eDNA) for detecting species of interest will likely allow for expanded use of these techniques in the field. One obstacle that continues to hinder applications in the field is the requirement of a cold chain of storage for water samples containing eDNA. While eDNA has been successfully preserved using Longmire's lysis buffer applied to filters, it has yet to be tried with freshwater samples collected for eDNA detection of an invasive species. We tested the utility of Longmire's solution (100 mM Tris, 100 mM EDTA, 10 mM NaCl, 0.5 % SDS, 0.2 % sodium azide) as an additive to freshwater samples for preservation of eDNA. Environmental DNA was effectively preserved in 15 mL water samples with Longmire's solution added; eDNA positive detection was comparable to freezing the samples at -80 °C and occurred out to 56 days at the highest concentration (5 mL Longmire's solution: 15 mL sample water). Medium and low concentrations of Longmire's solution added to 15 mL of sample water generally preserved eDNA out to 56 days but not as well as did freezing or application of the highest concentration of Longmire's lysis buffer. Treatment and degradation time had a significant effect on average DNA concentration of samples, although not the interaction of treatment and time. Perfect detection occurred out to 56 days with the high Longmire's treatment group but DNA concentration was significantly lower at this time point compared to 28 days. We conclude that Longmire's lysis buffer is a viable alternative to cold chain storage that can simplify the collection of eDNA by eliminating the need for filtering and allow more time for sample collection when added at our highest concentration (1 part Longmire's:3 parts water sample), which could translate to an increase in the chances of detecting a rare or elusive species.

  5. System and method for measuring fluorescence of a sample

    DOEpatents

    Riot, Vincent J

    2015-03-24

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  6. Solid sorbent air sampling and analytical procedure for methyl-, dimethyl-, ethyl-, and diethylamine

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

    Elskamp, C.J.; Schultz, G.R.

    1986-01-01

    A sampling and analytical procedure for methyl-, dimethyl-, ethyl-, and diethylamine was developed in order to avoid problems typically encountered in the sampling and analysis of low molecular weight aliphatic amines. Samples are collected with adsorbent tubes containing Amberlite XAD-7 resin coated with the derivatizing reagent, NBD chloride (7-chloro-4-nitrobenzo-2-oxa-1,3-diazole). Analysis is performed by high performance liquid chromatography with the use of a fluorescence and/or UV/visible detector. All four amines can be monitored simultaneously, and neither collection nor storage is affected by humidity. Samples are stable at room temperature for at least two weeks. The methodology has been tested for eachmore » of the four amines at sample loadings equivalent to air concentration ranges of 0.5 to 30 ppm for a sample volume of 10 liters. The method shows promise for determining other airborne primary and secondary low molecular weight aliphatic amines.« less

  7. Reducing sampling error in faecal egg counts from black rhinoceros (Diceros bicornis).

    PubMed

    Stringer, Andrew P; Smith, Diane; Kerley, Graham I H; Linklater, Wayne L

    2014-04-01

    Faecal egg counts (FECs) are commonly used for the non-invasive assessment of parasite load within hosts. Sources of error, however, have been identified in laboratory techniques and sample storage. Here we focus on sampling error. We test whether a delay in sample collection can affect FECs, and estimate the number of samples needed to reliably assess mean parasite abundance within a host population. Two commonly found parasite eggs in black rhinoceros (Diceros bicornis) dung, strongyle-type nematodes and Anoplocephala gigantea, were used. We find that collection of dung from the centre of faecal boluses up to six hours after defecation does not affect FECs. More than nine samples were needed to greatly improve confidence intervals of the estimated mean parasite abundance within a host population. These results should improve the cost-effectiveness and efficiency of sampling regimes, and support the usefulness of FECs when used for the non-invasive assessment of parasite abundance in black rhinoceros populations.

  8. 16S Based Microbiome Analysis from Healthy Subjects’ Skin Swabs Stored for Different Storage Periods Reveal Phylum to Genus Level Changes

    PubMed Central

    Klymiuk, Ingeborg; Bambach, Isabella; Patra, Vijaykumar; Trajanoski, Slave; Wolf, Peter

    2016-01-01

    Microbiome research and improvements in high throughput sequencing technologies revolutionize our current scientific viewpoint. The human associated microbiome is a prominent focus of clinical research. Large cohort studies are often required to investigate the human microbiome composition and its changes in a multitude of human diseases. Reproducible analyses of large cohort samples require standardized protocols in study design, sampling, storage, processing, and data analysis. In particular, the effect of sample storage on actual results is critical for reproducibility. So far, the effect of storage conditions on the results of microbial analysis has been examined for only a few human biological materials (e.g., stool samples). There is a lack of data and information on appropriate storage conditions on other human derived samples, such as skin. Here, we analyzed skin swab samples collected from three different body locations (forearm, V of the chest and back) of eight healthy volunteers. The skin swabs were soaked in sterile buffer and total DNA was isolated after freezing at -80°C for 24 h, 90 or 365 days. Hypervariable regions V1-2 were amplified from total DNA and libraries were sequenced on an Illumina MiSeq desktop sequencer in paired end mode. Data were analyzed using Qiime 1.9.1. Summarizing all body locations per time point, we found no significant differences in alpha diversity and multivariate community analysis among the three time points. Considering body locations separately significant differences in the richness of forearm samples were found between d0 vs. d90 and d90 vs. d365. Significant differences in the relative abundance of major skin genera (Propionibacterium, Streptococcus, Bacteroides, Corynebacterium, and Staphylococcus) were detected in our samples in Bacteroides only among all time points in forearm samples and between d0 vs. d90 and d90 vs. d365 in V of the chest and back samples. Accordingly, significant differences were detected in the ratios of the main phyla Actinobacteria, Firmicutes, and Bacteroidetes: Actinobacteria vs. Bacteroidetes at d0 vs. d90 (p-value = 0.0234), at d0 vs. d365 (p-value = 0.0234) and d90 vs. d365 (p-value = 0.0234) in forearm samples and at d90 vs. d365 in V of the chest (p-value = 0.0234) and back samples (p-value = 0.0234). The ratios of Firmicutes vs. Bacteroidetes showed no significant changes in any of the body locations as well as the ratios of Actinobacteria vs. Firmicutes at any time point. Studies with larger sample sizes are required to verify our results and determine long term storage effects with regard to specific biological questions. PMID:28066342

  9. Cryopreservation in fish: current status and pathways to quality assurance and quality control in repository development

    PubMed Central

    Torres, Leticia; Hu, E.; Tiersch, Terrence R.

    2017-01-01

    Cryopreservation in aquatic species in general has been constrained to research activities for more than 60 years. Although the need for application and commercialisation pathways has become clear, the lack of comprehensive quality assurance and quality control programs has impeded the progress of the field, delaying the establishment of germplasm repositories and commercial-scale applications. In this review we focus on the opportunities for standardisation in the practices involved in the four main stages of the cryopreservation process: (1) source, housing and conditioning of fish; (2) sample collection and preparation; (3) freezing and cryogenic storage of samples; and (4) egg collection and use of thawed sperm samples. In addition, we introduce some key factors that would assist the transition to commercial-scale, high-throughput application. PMID:26739583

  10. Critical evaluation of methodology commonly used in sample collection, storage and preparation for the analysis of pharmaceuticals and illicit drugs in surface water and wastewater by solid phase extraction and liquid chromatography-mass spectrometry.

    PubMed

    Baker, David R; Kasprzyk-Hordern, Barbara

    2011-11-04

    The main aim of this manuscript is to provide a comprehensive and critical verification of methodology commonly used for sample collection, storage and preparation in studies concerning the analysis of pharmaceuticals and illicit drugs in aqueous environmental samples with the usage of SPE-LC/MS techniques. This manuscript reports the results of investigations into several sample preparation parameters that to the authors' knowledge have not been reported or have received very little attention. This includes: (i) effect of evaporation temperature and (ii) solvent with regards to solid phase extraction (SPE) extracts; (iii) effect of silanising glassware; (iv) recovery of analytes during vacuum filtration through glass fibre filters and (v) pre LC-MS filter membranes. All of these parameters are vital to develop efficient and reliable extraction techniques; an essential factor given that target drug residues are often present in the aqueous environment at ng L(-1) levels. Presented is also the first comprehensive review of the stability of illicit drugs and pharmaceuticals in wastewater. Among the parameters studied are: time of storage, temperature and pH. Over 60 analytes were targeted including stimulants, opioid and morphine derivatives, benzodiazepines, antidepressants, dissociative anaesthetics, drug precursors, human urine indicators and their metabolites. The lack of stability of analytes in raw wastewater was found to be significant for many compounds. For instance, 34% of compounds studied reported a stability change >15% after only 12 h in raw wastewater stored at 2 °C; a very important finding given that wastewater is typically collected with the use of 24 h composite samplers. The stability of these compounds is also critical given the recent development of so-called 'sewage forensics' or 'sewage epidemiology' in which concentrations of target drug residues in wastewater are used to back-calculate drug consumption. Without an understanding of stability, under (or over) reporting of consumption estimations may take place. Copyright © 2011 Elsevier B.V. All rights reserved.

  11. Effects of packaging and storage conditions on the quality of amoxicillin-clavulanic acid – an analysis of Cambodian samples

    PubMed Central

    2013-01-01

    Background The use of substandard and degraded medicines is a major public health problem in developing countries such as Cambodia. A collaborative study was conducted to evaluate the quality of amoxicillin–clavulanic acid preparations under tropical conditions in a developing country. Methods Amoxicillin-clavulanic acid tablets were obtained from outlets in Cambodia. Packaging condition, printed information, and other sources of information were examined. The samples were tested for quantity, content uniformity, and dissolution. Authenticity was verified with manufacturers and regulatory authorities. Results A total of 59 samples were collected from 48 medicine outlets. Most (93.2%) of the samples were of foreign origin. Using predetermined acceptance criteria, 12 samples (20.3%) were non-compliant. Eight (13.6%), 10 (16.9%), and 20 (33.9%) samples failed quantity, content uniformity, and dissolution tests, respectively. Samples that violated our observational acceptance criteria were significantly more likely to fail the quality tests (Fisher’s exact test, p < 0.05). Conclusions Improper packaging and storage conditions may reduce the quality of amoxicillin–clavulanic acid preparations at community pharmacies. Strict quality control measures are urgently needed to maintain the quality of amoxicillin–clavulanic acid in tropical countries. PMID:23773420

  12. FTA Cards for Preservation of Nucleic Acids for Molecular Assays: A Review on the Use of Cytologic/Tissue Samples.

    PubMed

    da Cunha Santos, Gilda

    2018-03-01

    - Traditional methods for storing histologic and cytologic specimens for future use in molecular assays have consisted of either snap-freezing with cryopreservation or formalin-fixing, paraffin-embedding the samples. Although snap-freezing with cryopreservation is recommended for better preservation of nucleic acids, the infrastructure and space required for archiving impose challenges for high-volume pathology laboratories. Cost-effective, long-term storage at room temperature; relatively easy shipment; and standardized handling can be achieved with formalin-fixed, paraffin-embedded samples, but formalin fixation induces fragmentation and chemical modification of nucleic acids. Advances in next-generation sequencing platforms, coupled with an increase in diagnostic, prognostic, and predictive molecular biomarkers have created a demand for high-quality nucleic acids. To address issues of the quality of nucleic acid and logistics in sample acquisition, alternatives for specimen preservation and long-term storage have been described and include novel universal tissue fixatives, stabilizers, and technologies. - To collect, retrieve, and review information from studies describing the use of nucleic acids recovered from cytologic/tissue specimens stored on Flinders Technology Associates (FTA, GE Whatman, Maidstone, Kent, United Kingdom) cards for downstream molecular applications. - An electronic literature search in the PubMed (National Center for Biotechnology Information, Bethesda, Maryland) database allowed the selection of manuscripts addressing the use of FTA cards for storage of cytologic samples for molecular analysis. Only articles published in English were retrieved. - The use of FTA cards is a versatile method for fostering multicenter, international collaborations and clinical trials that require centralized testing, long-distance shipment, and high-quality nucleic acids for molecular techniques. Studies with controlled temperature are required to test the quality of recovered RNA after long-term storage.

  13. Effects of Agitation and Storage Temperature on Measurements of Hydration Status.

    PubMed

    Adams, Heather M; Eberman, Lindsey E; Yeargin, Susan W; Niemann, Andrew J; Mata, Heather L; Dziedzicki, David J

    2015-12-01

    Hypohydration can have significant implications on normal physiological functions of the body. This study aimed to determine the impact of agitation, storage temperature, and storage time on urine osmolality compared to the criterion control. We used a descriptive diagnostic validity test design. To investigate agitation, we recruited 75 healthy individuals (males = 41, females = 34; mean age = 22 ± 5 years; mean self-reported height = 172 ± 23 cm and mass = 77 ± 17 kg) who provided one or more samples (total = 81). The independent variables were agitation (vortex, hand shaken, no agitation) and temperature (room temperature, freezer, and refrigerator) type. Participants completed informed consent, a health questionnaire and were asked to provide a urine sample, which was split and labeled according to agitation type or storage temperature. Urine osmolality was used to determine hydration status at two time points (within 2 hours [control], 48 hours). We used t-tests to determine the difference between each condition and the control and calculated percent error for each condition. No significant differences for no agitation (t79 = -0.079, P = 0.937), hand shaken (t79 = 1.395, P = 0.167) or vortex mixed (t79 = -0.753, P = 0.453) were identified when compared to the criterion control. No significant differences for room temperature (t82 = -0.720, P = 0.474), refrigerator (t82 = -2.697, P = 0.008) or freezer (t82 = 2.576, P = 0.012) were identified when compared to the criterion control. Our findings suggest agitation of urine specimen is not necessary and samples do not require refrigeration or freezing if assessed within 48 hours. Analysis within two hours of collection is not necessary and samples can be stored for up to 48 hours without impacting the hydration status of the sample.

  14. Effects of freezing on the bactericidal activity of human milk.

    PubMed

    Takci, Sahin; Gulmez, Dolunay; Yigit, Sule; Dogan, Ozlem; Dik, Kezban; Hascelik, Gulsen

    2012-08-01

    Storage of human milk by freezing has been recommended for long-term storage. The present study analyzed the bactericidal activity of human milk on Escherichia coli and Pseudomonas aeruginosa and determined the changes in bactericidal activity following freezing at -20°C and -80°C for 1 month and 3 months. Forty-eight milk samples were collected from 48 lactating mothers. Each sample was divided into 10 aliquots. Two of the samples were processed immediately and the others were stored at both -20°C and -80°C until analysis after 1 month and 3 months of freezing. All of the fresh milk samples showed bactericidal activity against E coli and P aeruginosa. Freezing at -20°C for 1 month did not cause statistically significant alteration in bactericidal activity (P > 0.017), whereas storage for 3 months lowered the degree of bactericidal activity significantly (P < 0.017) against E coli. Bactericidal activity was protected when the samples were stored at -80°C. There was no statistically significant difference in the bactericidal activity of human milk against E coli between freezing at -20°C and -80°C for 1 month (P > 0.017); however, when milk was stored for 3 months, -80°C was significantly more protective (P < 0.017). Freezing at -20°C and -80°C for 1 month and 3 months did not cause any significant change in bactericidal activity against P aeruginosa (P > 0.05). Storage by freezing at -80°C is more appropriate to keep bactericidal capacity of stored human milk >1 month if affordable and available, especially in intensive care settings.

  15. DNA cards: determinants of DNA yield and quality in collecting genetic samples for pharmacogenetic studies.

    PubMed

    Mas, Sergi; Crescenti, Anna; Gassó, Patricia; Vidal-Taboada, Jose M; Lafuente, Amalia

    2007-08-01

    As pharmacogenetic studies frequently require establishment of DNA banks containing large cohorts with multi-centric designs, inexpensive methods for collecting and storing high-quality DNA are needed. The aims of this study were two-fold: to compare the amount and quality of DNA obtained from two different DNA cards (IsoCode Cards or FTA Classic Cards, Whatman plc, Brentford, Middlesex, UK); and to evaluate the effects of time and storage temperature, as well as the influence of anticoagulant ethylenediaminetetraacetic acid on the DNA elution procedure. The samples were genotyped by several methods typically used in pharmacogenetic studies: multiplex PCR, PCR-restriction fragment length polymorphism, single nucleotide primer extension, and allelic discrimination assay. In addition, they were amplified by whole genome amplification to increase genomic DNA mass. Time, storage temperature and ethylenediaminetetraacetic acid had no significant effects on either DNA card. This study reveals the importance of drying blood spots prior to isolation to avoid haemoglobin interference. Moreover, our results demonstrate that re-isolation protocols could be applied to increase the amount of DNA recovered. The samples analysed were accurately genotyped with all the methods examined herein. In conclusion, our study shows that both DNA cards, IsoCode Cards and FTA Classic Cards, facilitate genetic and pharmacogenetic testing for routine clinical practice.

  16. Reduction of hexavalent chromium collected on PVC filters.

    PubMed

    Shin, Y C; Paik, N W

    2000-01-01

    Chromium exists at various valences, including elemental, trivalent, and hexavalent chromium, and undergoes reduction-oxidation reactions in the environment. Since hexavalent chromium is known as a human carcinogen, it is most important to evaluate the oxidation-reduction characteristics of the hexavalent chromium species. Although hexavalent chromium can be reduced to trivalent state, the detailed information on this in workplace environments is limited. The purpose of this study was to investigate hexavalent chromium reduction in time in various conditions. A pilot chrome plating operation was prepared and operated in a laboratory for this study. There was evidence that the hexavalent chromium was reduced by time after mist generation. The percentage ratio (with 95% confidence intervals in parentheses) of hexavalent chromium to total chromium was almost 100% (99.1 approximately 102.3) immediately after mist generation, and was reduced to 87.4% (84.8 approximately 89.9) at 1 hour and 81.0% (78.3 approximately 83.5) at 2 hours, respectively. Another test indicated that hexavalent chromium collected on PVC filters was also reduced by time after sampling. Hexavalent chromium was reduced to 90.8% (88.2 approximately 93.3) at 2 hours after sampling. It also was found that hexavalent chromium was reduced during storage in air. It is recommended that air samples of hexavalent chromium be protected against reduction during storage.

  17. 8 CFR 103.16 - Collection, use and storage of biometric information.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... REGULATIONS IMMIGRATION BENEFITS; BIOMETRIC REQUIREMENTS; AVAILABILITY OF RECORDS Biometric Requirements § 103.16 Collection, use and storage of biometric information. (a) Use of biometric information. Any... 8 Aliens and Nationality 1 2012-01-01 2012-01-01 false Collection, use and storage of biometric...

  18. 8 CFR 103.16 - Collection, use and storage of biometric information.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... REGULATIONS IMMIGRATION BENEFITS; BIOMETRIC REQUIREMENTS; AVAILABILITY OF RECORDS Biometric Requirements § 103.16 Collection, use and storage of biometric information. (a) Use of biometric information. Any... 8 Aliens and Nationality 1 2014-01-01 2014-01-01 false Collection, use and storage of biometric...

  19. 8 CFR 103.16 - Collection, use and storage of biometric information.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... REGULATIONS IMMIGRATION BENEFITS; BIOMETRIC REQUIREMENTS; AVAILABILITY OF RECORDS Biometric Requirements § 103.16 Collection, use and storage of biometric information. (a) Use of biometric information. Any... 8 Aliens and Nationality 1 2013-01-01 2013-01-01 false Collection, use and storage of biometric...

  20. Stability of procalcitonin at room temperature.

    PubMed

    Milcent, Karen; Poulalhon, Claire; Fellous, Christelle Vauloup; Petit, François; Bouyer, Jean; Gajdos, Vincent

    2014-01-01

    The aim was to assess procalcitonin (PCT) stability after two days of storage at room temperature. Samples were collected from febrile children aged 7 to 92 days and were rapidly frozen after sampling. PCT levels were measured twice after thawing: immediately (named y) and 48 hours later after storage at room temperature (named x). PCT values were described with medians and interquartile ranges or by categorizing them into classes with thresholds 0.25, 0.5, and 2 ng/mL. The relationship between x and y PCT levels was analyzed using fractional polynomials in order to predict the PCT value immediately after thawing (named y') from x. A significant decrease in PCT values was observed after 48 hours of storage at room temperature, either in median, 30% lowering (p < 0.001), or as categorical variable (p < 0.001). The relationship between x and y can be accurately modeled with a simple linear model: y = 1.37 x (R2 = 0.99). The median of the predicted PCT values y' was quantitatively very close to the median of y and the distributions of y and y' across categories were very similar and not statistically different. PCT levels noticeably decrease after 48 hours of storage at room temperature. It is possible to pre- dict accurately effective PCT values from the values after 48 hours of storage at room temperature with a simple statistical model.

  1. Apparatus for transporting hazardous materials

    DOEpatents

    Osterman, Robert A.; Cox, Robert

    1992-01-01

    An apparatus and method are provided for selectively receiving, transporting, and releasing one or more radioactive or other hazardous samples for analysis on a differential thermal analysis (DTA) apparatus. The apparatus includes a portable sample transporting apparatus for storing and transporting the samples and includes a support assembly for supporting the transporting apparatus when a sample is transferred to the DTA apparatus. The transporting apparatus includes a storage member which includes a plurality of storage chambers arrayed circumferentially with respect to a central axis. An adjustable top door is located on the top side of the storage member, and the top door includes a channel capable of being selectively placed in registration with the respective storage chambers thereby permitting the samples to selectively enter the respective storage chambers. The top door, when closed, isolates the respective samples within the storage chambers. A plurality of spring-biased bottom doors are located on the bottom sides of the respective storage chambers. The bottom doors isolate the samples in the respective storage chambers when the bottom doors are in the closed position. The bottom doors permit the samples to leave the respective storage chambers from the bottom side when the respective bottom doors are in respective open positions. The bottom doors permit the samples to be loaded into the respective storage chambers after the analysis for storage and transport to a permanent storage location.

  2. Urine Bacterial Community Convergence through Fertilizer Production: Storage, Pasteurization, and Struvite Precipitation.

    PubMed

    Lahr, Rebecca H; Goetsch, Heather E; Haig, Sarah J; Noe-Hays, Abraham; Love, Nancy G; Aga, Diana S; Bott, Charles B; Foxman, Betsy; Jimenez, Jose; Luo, Ting; Nace, Kim; Ramadugu, Kirtana; Wigginton, Krista R

    2016-11-01

    Source-separated human urine was collected from six public events to study the impact of urine processing and storage on bacterial community composition and viability. Illumina 16S rRNA gene sequencing revealed a complex community of bacteria in fresh urine that differed across collection events. Despite the harsh chemical conditions of stored urine (pH > 9 and total ammonia nitrogen > 4000 mg N/L), bacteria consistently grew to 5 ± 2 × 10 8 cells/mL. Storing hydrolyzed urine for any amount of time significantly reduced the number of operational taxonomic units (OTUs) to 130 ± 70, increased Pielou evenness to 0.60 ± 0.06, and produced communities dominated by Clostridiales and Lactobacillales. After 80 days of storage, all six urine samples from different starting materials converged to these characteristics. Urine pasteurization or struvite precipitation did not change the microbial community, even when pasteurized urine was stored for an additional 70 days. Pasteurization decreased metabolic activity by 50 ± 10% and additional storage after pasteurization did not lead to recovery of metabolic activity. Urine-derived fertilizers consistently contained 16S rRNA genes belonging to Tissierella, Erysipelothrix, Atopostipes, Bacteroides, and many Clostridiales OTUs; additional experiments must determine whether pathogenic species are present, responsible for observed metabolic activity, or regrow when applied.

  3. Biopolymers for sample collection, protection, and preservation.

    PubMed

    Sorokulova, Iryna; Olsen, Eric; Vodyanoy, Vitaly

    2015-07-01

    One of the principal challenges in the collection of biological samples from air, water, and soil matrices is that the target agents are not stable enough to be transferred from the collection point to the laboratory of choice without experiencing significant degradation and loss of viability. At present, there is no method to transport biological samples over considerable distances safely, efficiently, and cost-effectively without the use of ice or refrigeration. Current techniques of protection and preservation of biological materials have serious drawbacks. Many known techniques of preservation cause structural damages, so that biological materials lose their structural integrity and viability. We review applications of a novel bacterial preservation process, which is nontoxic and water soluble and allows for the storage of samples without refrigeration. The method is capable of protecting the biological sample from the effects of environment for extended periods of time and then allows for the easy release of these collected biological materials from the protective medium without structural or DNA damage. Strategies for sample collection, preservation, and shipment of bacterial, viral samples are described. The water-soluble polymer is used to immobilize the biological material by replacing the water molecules within the sample with molecules of the biopolymer. The cured polymer results in a solid protective film that is stable to many organic solvents, but quickly removed by the application of the water-based solution. The process of immobilization does not require the use of any additives, accelerators, or plastifiers and does not involve high temperature or radiation to promote polymerization.

  4. Sample integrity evaluation and EPA method 325B interlaboratory comparison for select volatile organic compounds collected diffusively on Carbopack X sorbent tubes

    NASA Astrophysics Data System (ADS)

    Oliver, Karen D.; Cousett, Tamira A.; Whitaker, Donald A.; Smith, Luther A.; Mukerjee, Shaibal; Stallings, Casson; Thoma, Eben D.; Alston, Lillian; Colon, Maribel; Wu, Tai; Henkle, Stacy

    2017-08-01

    A sample integrity evaluation and an interlaboratory comparison were conducted in application of U.S. Environmental Protection Agency (EPA) Methods 325A and 325B for diffusively monitoring benzene and other selected volatile organic compounds (VOCs) using Carbopack X sorbent tubes. To evaluate sample integrity, VOC samples were refrigerated for up to 240 days and analyzed using thermal desorption/gas chromatography-mass spectrometry at the EPA Office of Research and Development laboratory in Research Triangle Park, NC, USA. For the interlaboratory comparison, three commercial analytical laboratories were asked to follow Method 325B when analyzing samples of VOCs that were collected in field and laboratory settings for EPA studies. Overall results indicate that the selected VOCs collected diffusively on sorbent tubes generally were stable for 6 months or longer when samples were refrigerated. This suggests the specified maximum 30-day storage time of VOCs collected diffusively on Carbopack X passive samplers and analyzed using Method 325B might be able to be relaxed. Interlaboratory comparison results were in agreement for the challenge samples collected diffusively in an exposure chamber in the laboratory, with most measurements within ±25% of the theoretical concentration. Statistically significant differences among laboratories for ambient challenge samples were small, less than 1 part per billion by volume (ppbv). Results from all laboratories exhibited good precision and generally agreed well with each other.

  5. Clinical evaluation of high-risk HPV detection on self-samples using the indicating FTA-elute solid-carrier cartridge.

    PubMed

    Geraets, D T; van Baars, R; Alonso, I; Ordi, J; Torné, A; Melchers, W J G; Meijer, C J L M; Quint, W G V

    2013-06-01

    High-risk human papillomavirus (hrHPV) testing in cervical screening is usually performed on physician-taken cervical smears in liquid-based medium. However, solid-state specimen carriers allow easy, non-hazardous storage and transportation and might be suitable for self-collection by non-responders in screening and in low-resource settings. We evaluated the adequacy of self-collected cervicovaginal (c/v) samples using a Viba-brush stored on an Indicating FTA-elute cartridge (FTA-based self-sampling) for hrHPV testing in women referred to a gynecology clinic due to an abnormal smear. 182 women accepted to self-collect a c/v sample. After self-sampling, a physician obtained a conventional liquid-based cervical smear. Finally, women were examined by colposcopy and a biopsy was taken when clinically indicated. Self-samples required only simple DNA elution, and DNA was extracted from physician-obtained samples. Both samples were tested for 14 hrHPVs by GP5+/6+-EIA-LQ Test and SPF(10)-DEIA-LiPA(25). Both assays detected significantly more hrHPV in physician-collected specimens than in self-collected samples (75.3% and 67.6% by SPF(10); 63.3% and 53.3% by GP5+/6+, respectively). The combination of physician-collected specimen and GP5+/6+ testing demonstrated the optimal balance in sensitivity (98.0%) and specificity (48.1%) for CIN2+ detection in this referral population. A test system of FTA-based self-collection and SPF(10) hrHPV detection approached this sensitivity (95.9%) and specificity (42.9%). These results show that the clinical performance of hrHPV detection is determined by both the sample collection system and the test method. FTA-based self-collection with SPF(10) testing might be valuable when a liquid-based medium cannot be used, but requires further investigation in screening populations. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. 40 CFR 80.613 - What defenses apply to persons deemed liable for a violation of a prohibited act under this subpart?

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ..., including sample collection, transportation, storage, and analysis. (4) Survey plan design requirements. The... contractual agreement between the branded refiner or importer and the person designed to prevent such action... conditions and limitations set forth in this paragraph (e): (1) Independent survey association. To comply...

  7. 40 CFR 80.613 - What defenses apply to persons deemed liable for a violation of a prohibited act under this subpart?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ..., including sample collection, transportation, storage, and analysis. (4) Survey plan design requirements. The... contractual agreement between the branded refiner or importer and the person designed to prevent such action... subject to the conditions and limitations set forth in this paragraph (e): (1) Independent survey...

  8. 40 CFR 80.613 - What defenses apply to persons deemed liable for a violation of a prohibited act under this subpart?

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ..., including sample collection, transportation, storage, and analysis. (4) Survey plan design requirements. The... contractual agreement between the branded refiner or importer and the person designed to prevent such action... conditions and limitations set forth in this paragraph (e): (1) Independent survey association. To comply...

  9. Observation of D+ --> etae + nue.

    PubMed

    Mitchell, R E; Shepherd, M R; Besson, D; Pedlar, T K; Cronin-Hennessy, D; Gao, K Y; Hietala, J; Kubota, Y; Klein, T; Lang, B W; Poling, R; Scott, A W; Smith, A; Zweber, P; Dobbs, S; Metreveli, Z; Seth, K K; Tomaradze, A; Ernst, J; Ecklund, K M; Severini, H; Love, W; Savinov, V; Aquines, O; Lopez, A; Mehrabyan, S; Mendez, H; Ramirez, J; Huang, G S; Miller, D H; Pavlunin, V; Sanghi, B; Shipsey, I P J; Xin, B; Adams, G S; Anderson, M; Cummings, J P; Danko, I; Hu, D; Moziak, B; Napolitano, J; He, Q; Insler, J; Muramatsu, H; Park, C S; Thorndike, E H; Yang, F; Artuso, M; Blusk, S; Butt, J; Li, J; Menaa, N; Mountain, R; Nisar, S; Randrianarivony, K; Sia, R; Skwarnicki, T; Stone, S; Wang, J C; Zhang, K; Bonvicini, G; Cinabro, D; Dubrovin, M; Lincoln, A; Asner, D M; Edwards, K W; Naik, P; Briere, R A; Ferguson, T; Tatishvili, G; Vogel, H; Watkins, M E; Rosner, J L; Adam, N E; Alexander, J P; Cassel, D G; Duboscq, J E; Ehrlich, R; Fields, L; Galik, R S; Gibbons, L; Gray, R; Gray, S W; Hartill, D L; Heltsley, B K; Hertz, D; Jones, C D; Kandaswamy, J; Kreinick, D L; Kuznetsov, V E; Mahlke-Krüger, H; Mohapatra, D; Onyisi, P U E; Patterson, J R; Peterson, D; Pivarski, J; Riley, D; Ryd, A; Sadoff, A J; Schwarthoff, H; Shi, X; Stroiney, S; Sun, W M; Wilksen, T; Athar, S B; Patel, R; Potlia, V; Yelton, J; Rubin, P; Cawlfield, C; Eisenstein, B I; Karliner, I; Kim, D; Lowrey, N; Selen, M; White, E J; Wiss, J

    2009-02-27

    Using a 281 pb-1 data sample collected at the psi(3770) resonance with the CLEO-c detector at the Cornell Electron Storage Ring, we report the first observation of D+ --> etae + nue. We also set upper limits for D+ --> eta'e + nue and D + --> varphie + nue that are about 2 orders of magnitude more restrictive than those obtained by previous experiments.

  10. Evaluation of yeasts obtained from Antarctic soil samples as biocontrol agents for the management of postharvest diseases of apple (Malus x domestica)

    USDA-ARS?s Scientific Manuscript database

    Psychrotrophic yeasts isolated from soils collected in Antarctica and selected by its capacity of growing in apple juice at low temperatures were evaluated for their potential as biocontrol agents for the management of postharvest diseases of apple during cold storage. Among the species recovered, ...

  11. Red cell storage in E-Sol 5 and Adsol additive solutions: paired comparison using mixed and non-mixed study designs.

    PubMed

    Radwanski, K; Thill, M; Min, K

    2014-05-01

    If transfusion of older stored red cells is found to negatively affect clinical outcome, one possible alternative to shortened outdate is the use of new additive solutions (AS) that ameliorate the storage lesion. Erythro-Sol (E-Sol), a previously developed next-generation AS, has been reformulated into E-Sol 5, which is compatible with current anticoagulants and AS volumes. The effect of E-Sol 5 on red cells during storage compared to current AS has not been reported. Paired, ABO-matched whole-blood units were collected into CPD anticoagulant, pooled, split and processed into plasma and red cell units with either 110 ml of Adsol or 105 ml of E-Sol 5 within 8 h of collection. In Study 1, paired units in E-Sol 5 and Adsol were sampled on Day 0 and every 7 days up to Day 42 (n = 10). In Study 2, paired units in E-Sol 5 and Adsol were sampled only on Day 0 and Day 42 (n = 10). In Study 1, 2,3 DPG levels were maintained until Day 28 in E-Sol 5 units and Day 14 in Adsol units. ATP levels were higher in E-Sol 5 units until Day 21, after which they were comparable between the two groups. In both studies, metabolic activity was greater in E-Sol 5 units with respect to glucose consumption and lactate production. Morphology scores were higher, and haemolysis and microparticles generated were lower in E-Sol 5 vs. Adsol units. Weekly mixing of units lowered haemolysis and microparticle levels and increased potassium content on Day 42 in both additive solutions. Regardless of whether units are mixed weekly or are stored non-mixed, E-Sol 5 slows the progression of the red cell storage lesion and improves the overall in vitro quality of RBC throughout storage. © 2013 International Society of Blood Transfusion.

  12. Impact of different storage times at room temperature of unspun citrated blood samples on routine coagulation tests results. Results of a bicenter study and review of the literature.

    PubMed

    Toulon, P; Metge, S; Hangard, M; Zwahlen, S; Piaulenne, S; Besson, V

    2017-10-01

    A maximum delay between blood collection and coagulation testing of 4 hours is recommended by most guidelines. As information on optimal storage times is limited, we investigated the potential effect of different storage times of unspun tubes, that is, ≤2, 4, 6, and 8 hours, on routine coagulation test results. Four evacuated polymer tubes containing 0.109 mol/L tri-Na citrate were drawn from 144 patients, including 39 patients on vitamin K-antagonists. Except for storage time, all tubes underwent the same preanalytical process. Prothrombin time (PT)/international normalized ratio (INR), activated partial thromboplastin time (aPTT), fibrinogen, factor V (FV), FVIII, and D-dimer were evaluated in two centers using the same technical conditions. Analytical comparison of aPTT, fibrinogen, FV, and FVIII results evaluated after prolonged storage times vs a <2-hours storage demonstrated significant difference, whereas PT/INR and D-dimer remained unchanged up to 8 hours. Mean bias between test results obtained after prolonged storage times remained below the desirable values for all studied parameters except for FVIII evaluated after 6- and 8-hours storages, but only in patients with FVIII above 100 IU/dL. Even though the corresponding bias of -5.2% and -8.5%, respectively, remained within the GEHT recommended limits of variation, its evaluation after an 8-hours storage could lead to significant underestimation of FVIII. These results suggest that, in the studied technical conditions, PT/INR, aPTT, fibrinogen, FV, and D-dimer can be reliably evaluated in tubes stored unspun at room temperature for up to 8 hours after blood collection. That optimal delay should be of 6 hours for FVIII. © 2017 The Authors. International Journal of Laboratory Hematology Published by John Wiley & Sons Ltd.

  13. Use of FTA card for dry collection, transportation and storage of cervical cell specimen to detect high-risk HPV.

    PubMed

    Gustavsson, Inger; Lindell, Monica; Wilander, Erik; Strand, Anders; Gyllensten, Ulf

    2009-10-01

    The FTA elute micro card, which enable the collection, transport, and archiving of DNA could be an attractive alternative to a liquid based collection system for detection of human papillomavirus (HPV). To develop a method based on the FTA elute micro card for dry collection of cervical epithelial cell samples, suitable for subsequent PCR-based HPV testing. The method was evaluated by a comparison of the DNA collected by cytobrush and the regular FTA elute micro card from 50 cervical cell samples. The method was then used to estimate the DNA amount in 1040 samples applied to the indicating FTA elute micro card. The agreement in HPV positivity between the cytobrush and FTA samples (94%) was excellent (kappa=0.88, 95% CI 0.748-1). All the 1040 samples on the indicating FTA card had sufficient amounts of genomic DNA (>10 copies of a single copy gene) to be suitable for HPV typing. In 53 of the 1040 women the day in the menstrual cycle was noted, and the copy number during follicular phase day 9-13 was found to be statistically significantly lower than for the other three stages in the menstrual cycle (day 4-8, 14, >14) and during menopause. The indicating FTA elute micro card represents a suitable medium for collection of cervical cell samples, although follow-up studies are needed to verify the detection of low frequency HPV types.

  14. Hydrologic and water-quality data for U.S. Coast Guard Support Center Kodiak, Alaska, 1987-89

    USGS Publications Warehouse

    Glass, R.L.

    1996-01-01

    Hydrologic and water-quality data were collected at the U.S. Coast Guard Support Center Kodiak on Kodiak Island, Alaska, to determine regional ground-water conditions and if contamination of soils, ground water, or surface water has occurred. Eighteen areas of possible contamination were identified. Ground-water levels, surface- water stages, surface-water discharges, and results of field and laboratory analyses of soil and water samples are presented in tabular form. Many quality-assurance samples had detectable concentrations of methylene chloride and 1,2-dichloroethane, which may be due to sampling or laboratory contamination. Concentrations were as great as 5.9 micrograms per liter for methylene chloride and 2.6 micrograms per liter for 1,2-dichloroethane. Excluding 1,2-dichloroethane, most soil, ground-water, and surface-water samples contained no detectable concentrations of the organic constituents that were analyzed. Chemical analyses were performed on two lake-bed-material samples and more than 100 soil samples. The median lead concentration was 9.8 milligrams per kilogram. Concentrations of tetrachloroethene were as great as 1.1 milligram per kilogram in soils near a laundry. Water samples were collected from 101 wells. The maximum benzene concentration detected in ground water was 78 micrograms per liter from a well at the air station near a site where aviation fuel was spilled. Wells near a laundry yielded water having concentrations of tetrachloroethene as great as 3,000 micrograms per liter, and vinyl chloride as great as 440 micrograms per liter. A well in a former aviation gasoline storage area yielded water with a concentration of trichloroethene as great as 66 micrograms per liter. Water samples were collected from 59 sites on streams, lakes, or ponds. Surface-water samples had much lower concen- trations of organic compounds; the highest concentration of benzene was 2.2 micrograms per liter in a stream near a former aviation-fuel storage area and the maximum vinyl chloride concentration was 15 micrograms per liter in a stream near a former landfill. Tetrachloroethene and trichloroethene were not detected in any surface-water samples.

  15. Recovery and Stability of Δ9-Tetrahydrocannabinol Using the Oral-Eze® Oral Fluid Collection System and Intercept® Oral Specimen Collection Device.

    PubMed

    Samano, Kimberly L; Anne, Lakshmi; Johnson, Ted; Tang, Kenneth; Sample, R H Barry

    2015-10-01

    Oral fluid (OF) is increasingly used for clinical, forensic and workplace drug testing as an alternative to urine. Uncertainties surrounding OF collection device performance, drug stability and testing reproducibility may be partially responsible for delays in the implementation of OF testing in regulated drug testing programs. Stability of Δ(9)-tetrahydrocannabinol (THC) fortified and authentic specimens was examined after routine collection, transport and laboratory testing. Acceptable recovery and stability were observed when THC-fortified OF (1.5 and 4.5 ng/mL) was applied to Oral-Eze devices. Neat OF samples collected with Oral-Eze, processed per the package insert, and fortified with THC (3 and 6 ng/mL) were stable (±20%) at room temperature (21-25°C), refrigerated (2-8°C) and frozen (-25 to -15°C) conditions up to 1 month, while samples collected with Intercept devices showed decreases at refrigerated and room temperatures. After long-term refrigerated or frozen storage, maximum reductions in THC concentrations were 42% for Oral-Eze and 69% for Intercept. After ≥1 year frozen storage, 80.7% of laboratory specimens positive for THC (3 ng/mL cut-off) by GC-MS were reconfirmed positive (within 25%), with an average THC decrease of 4.2%. Specimens (n = 47) processed with Oral-Eze (diluted) and tested via enzyme immunoassay were concordant with LC-MS-MS results and showed 100% sensitivity and 95% specificity. Paired specimens collected with Oral-Eze and Intercept exhibited 98% overall agreement between the immunoassay test systems. Collectively, these data demonstrate consistent and reproducible recovery and stability of THC in OF after collection, transport and laboratory testing using the Oral-Eze OF Collection System. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  16. FT Raman microscopy of untreated natural plant fibres

    NASA Astrophysics Data System (ADS)

    Edwards, H. G. M.; Farwell, D. W.; Webster, D.

    1997-11-01

    The application of FT-Raman microscopy to the non-destructive analysis of natural plant fibres is demonstrated with samples of flax, jute, ramie, cotton, kapok, sisal and coconut fibre. Vibrational assignments are proposed and characteristic features of each material are presented. Samples were not pre-treated chemically before analysis and were used directly from their respective storage collection; the adaptation of the Raman microscopic technique to the identification of specimens of natural fibres in archaeological burial sites is explored for its forensic potential.

  17. US Geological Survey nutrient preservation experiment : experimental design, statistical analysis, and interpretation of analytical results

    USGS Publications Warehouse

    Patton, Charles J.; Gilroy, Edward J.

    1999-01-01

    Data on which this report is based, including nutrient concentrations in synthetic reference samples determined concurrently with those in real samples, are extensive (greater than 20,000 determinations) and have been published separately. In addition to confirming the well-documented instability of nitrite in acidified samples, this study also demonstrates that when biota are removed from samples at collection sites by 0.45-micrometer membrane filtration, subsequent preservation with sulfuric acid or mercury (II) provides no statistically significant improvement in nutrient concentration stability during storage at 4 degrees Celsius for 30 days. Biocide preservation had no statistically significant effect on the 30-day stability of phosphorus concentrations in whole-water splits from any of the 15 stations, but did stabilize Kjeldahl nitrogen concentrations in whole-water splits from three data-collection stations where ammonium accounted for at least half of the measured Kjeldahl nitrogen.

  18. Stability of mercury concentration measurements in archived soil and peat samples

    USGS Publications Warehouse

    Navrátil, Tomáš; Burns, Douglas; Nováková, Tereza; Kaňa, Jiří; Rohovec, Jan; Roll, Michal; Ettler, Vojtěch

    2018-01-01

    Archived soil samples can provide important information on the history of environmental contamination and by comparison with recently collected samples, temporal trends can be inferred. Little previous work has addressed whether mercury (Hg) concentrations in soil samples are stable with long-term storage under standard laboratory conditions. In this study, we have re-analyzed using cold vapor atomic adsorption spectroscopy a set of archived soil samples that ranged from relatively pristine mountainous sites to a polluted site near a non-ferrous metal smelter with a wide range of Hg concentrations (6 - 6485 µg kg-1). Samples included organic and mineral soils and peats with a carbon content that ranged from 0.2 to 47.7%. Soil samples were stored in polyethylene bags or bottles and held in laboratory rooms where temperature was not kept to a constant value. Mercury concentrations in four subsets of samples were originally measured in 2000, 2005, 2006 and 2007, and re-analyzed in 2017, i.e. after 17, 12, 11 and 10 years of storage. Statistical analyses of either separated or lumped data yielded no significant differences between the original and current Hg concentrations. Based on these analyses, we show that archived soil and peat samples can be used to evaluate historical soil mercury contamination.

  19. Optimization and evaluation of multi-bed adsorbent tube method in collection of volatile organic compounds

    NASA Astrophysics Data System (ADS)

    Ho, Steven Sai Hang; Wang, Liqin; Chow, Judith C.; Watson, John G.; Xue, Yonggang; Huang, Yu; Qu, Linli; Li, Bowei; Dai, Wenting; Li, Lijuan; Cao, Junji

    2018-04-01

    The feasibility of using adsorbent tubes to collect volatile organic compounds (VOCs) has been demonstrated since the 1990's and standardized as Compendium Method TO-17 by the U.S. Environmental Protection Agency (U.S EPA). This paper investigates sampling and analytical variables on concentrations of 57 ozone (O3) precursors (C2-C12 aliphatic and aromatic VOCs) specified for the Photochemical Assessment Monitoring Station (PAMS). Laboratory and field tests examined multi-bed adsorbent tubes containing a sorbate combination of Tenax TA, Carbograph 1 TD, and Carboxen 1003. Analyte stabilities were influenced by both collection tube temperature and ambient O3 concentrations. Analytes degraded during storage, while blank levels were elevated by passive adsorption. Adsorbent tube storage under cold temperatures (- 10 °C) in a preservation container filled with solid silica gel and anhydrous calcium sulfate (CaSO4) ensured sample integrity. A high efficiency (> 99%) O3 scrubber (i.e., copper coil tube filled with saturated potassium iodide [KI]) removed O3 (i.e., < 200 ppbv) from the air stream with a sampling capacity of 30 h. Water vapor scrubbers interfered with VOC measurements. The optimal thermal desorption-gas chromatography/mass spectrometry (TD-GC/MS) desorption time of 8 min was found at 330 °C. Good linearity (R2 > 0.995) was achieved for individual analyte calibrations (with the exception of acetylene) for mixing ratios of 0.08-1.96 ppbv. The method detection limits (MDLs) were below 0.055 ppbv for a 3 L sample volume. Replicate analyses showed relative standard deviations (RSDs) of < 10%, with the majority of the analytes within < 5%.

  20. Investigation of 2,4-dinitrophenylhydrazine impregnated adsorbent tubes for the collection of airborne aldehydes

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

    Holdren, M.W.; Smith, D.L.; Russell, N.K.

    1988-06-01

    The objective of the study was to investigate the use of 2,4-dinitrophenylhydrazine (DNPH) impregnated adsorbents for sampling airborne aldehydes. Experimental results using a 17 cu m environmental chamber and various spiked amounts of aldehyde material (low ppb levels) showed that the DNPH coated cartridge and the DNPH/acetonitrile impinger methods gave equivalent results. Blank levels of the DNPH-coated cartridges were studied as a function of storage time using various containers and temperature conditions. Canisters pressurized with zero-grade nitrogen provided the best storage device. Lower blank levels were also obtained when the cartridges were stored at lower temperatures. Blank levels appear tomore » equilibrate after six days of storage. To assure that quality data will be obtained, cartridges should be grouped according to batch number and blank levels should be determined prior to any field monitoring effort. Blank cartridge levels should be an order of magnitude lower than sample cartridge level. High performance liquid chromatography with UV detection proved to be a sensitive and stable analytical method for the DNPH derivatives.« less

  1. Alteration of municipal and industrial slags under atmospheric conditions

    NASA Astrophysics Data System (ADS)

    Rafał Kowalski, Piotr; Michalik, Marek

    2014-05-01

    The Waste Management System in Poland is being consequently built since 1998. After important changes in legislation, local governments have taken over the duty of waste collection. New points of selective collection of wastes have been opened and new sorting and composting plants were built. The last stage of introducing the Waste Management System is construction of waste incineration power plants. From nine installations which were planned, six are now under construction and they will start operating within the next two years. It is assumed that the consumption of raw wastes for these installations will reach 974 thousand tons per year. These investments will result in increased slags and ashes production. Now in Poland several local waste incinerators are operating and predominant amount of produced incineration residues is landfilled. These materials are exposed to atmospheric conditions in time of short term storage (just after incineration) and afterwards for a longer period of time on the landfill site. During the storage of slags low temperature mineral transformations and chemical changes may occur and also some components can be washed out. These materials are stored wet because of the technological processes. The aim of this study is to investigate the influence of storage in atmospheric conditions on slags from incineration of industrial and municipal wastes. The experiment started in January 2013. During this period slag samples from incineration of industrial and municipal wastes were exposed to atmospheric conditions. Samples were collected after 6 and 12 months. Within this time the pH value was measured monthly, and during the experimental period remained constant on the level of 9.5. After 6 months of exposure only slight changes in mineral compositions were observed in slags. The results of XRD analysis of municipal slags showed increase in content of carbonate minerals in comparison to the raw slag samples. In industrial slags, a decrease in content of soluble minerals, like halite, in comparison to the output samples was noted. These phases where probably dissolved and washed out from the samples. After 12 months of atmospheric exposure in municipal slags only slight changes in weight (1 wt%) were observed, whereas in industrial slags slightly above 10 wt% of the material was removed. After 12 months of atmospheric exposure more significant changes are expected such as changes in chemical and mineral compositions and changes in heavy metals and toxic elements concentrations due to leaching.

  2. Characterization of Crew Refuse Returned from Shuttle Missions with Permanent Gas, Volatile Organic Compound, and Microbial Analyses

    NASA Astrophysics Data System (ADS)

    Peterson, B.; Hummerick, M.; Roberts, M.; Krummins, V.; Kish, A.; Garland, J.; Maxwell, S.; Mills, A.

    In addition to the mass and energy costs associated with bioregenerative systems for advanced life support, the storage and processing of waste on spacecraft requires both atmospheric and biological management. Risks to crew health may arise from the presence of potential human pathogens in waste or from decay processes during waste storage and/or processing. This study reports on the permanent gas, trace volatile organic and microbiological analyses of crew refuse returned from shuttle missions STS-105, 109 and 110. The research objective is to characterize the biological stability of the waste stream, to assess the risks associated with its storage, and to provide baseline measures for the evaluation of waste processing technologies. Microbiological samples were collected from packaging material, food waste, bathroom waste, and bulk liquid collected from the volume F waste container. The number of culturable bacteria and total bacteria were determined by plating on R2A media and by Acridine Orange direct count, respectively. Samples of the trash were analyzed for the presence of fecal and total coliforms and other human-associated bacteria. Dry and ash weights were determined to estimate both water and organic content of the materials. The aerobic and anaerobic bio-stability of stored waste was determined by on-line monitoring of CO2 and by laboratory analysis of off-gas samples for hydrogen sulfide and methane. Volatile organic compounds and permanent gases were analyzed using EPA method TO15 with gas chromatography/mass spectrometry and by gas chromatography with selective detectors . This study establishes a baseline measure of waste composition, labile organics, and microbial load for this material.

  3. Effect of soil and water environment on typeability of PowerPlex Y (Promega) in selected tissue samples.

    PubMed

    Niemcunowicz-Janica, Anna; Pepinski, Witold; Janica, Jacek Robert; Skawronska, Malgorzata; Janica, Jerzy; Koc-Zorawska, Ewa; Stolyszewski, Ireneusz

    2007-01-01

    In cases of decomposed bodies Y chromosomal STR markers may be useful in identification of a male relative. The authors assessed typeability PowerPlex Y (Promega) loci in tissue material stored in water and soil environment. Tissue material was collected during autopsies of five persons aged 20-30 years with time of death determined within the limit of 14 hours. Heart muscle, liver and lung specimens were stored in pond water, sea water, sand and peat soil. DNA was extracted by organic method from tissue samples collected in 7-day intervals. Liver specimens were typeable in all PowerPlex Y loci within 100 days of storage in pond water with gradual decline at DYS392 in sea water. Heart muscle specimens stored in pond water exhibited allelic loss at DYS19, DYS385, DYS389II and DYS392, while all loci were typeable in sea water stored samples. For lung specimens allelic loss was noted throughout the profile. Storage of liver specimens in peat soil for more than 14 days resulted in allelic drop-out, and after 21 days no profiles were typeable. Heart muscle specimens were typeable in all PowerPlex Y systems after 35-day storage in sand, while allelic drop-out and subsequent lack of profiles were noted after 14 and 35 days respectively. Lung specimens stored in garden soil exhibited allelic drop-out and subsequent lack of profiles after 7 and 21 days, respectively. All PowerPlex Y loci were typeable in the latter material in sand up to day 35 with gradual decline of longer amplicons (DYS19, DYS385, DYS389II and DYS392).

  4. Effect of storage temperature on endogenous GHB levels in urine.

    PubMed

    LeBeau, M A; Miller, M L; Levine, B

    2001-06-15

    Because gamma-hydroxybutyrate (GHB) is an endogenous substance present in the body and is rapidly eliminated after ingestion, toxicologists investigating drug-facilitated sexual assault cases are often asked to differentiate between endogenous and exogenous levels of GHB in urine samples. This study was designed to determine the effects of storage temperature on endogenous GHB levels in urine. Specifically, it was designed to ascertain whether endogenous levels can be elevated to a range considered indicative of GHB ingestion. Urine specimens from two subjects that had not been administered exogenous GHB were collected during a 24h period and individually pooled. The pooled specimens were separated into standard sample cups and divided into three storage groups: room temperature ( approximately 25 degrees C), refrigerated (5 degrees C), and frozen (-10 degrees C). Additionally, some specimens were put through numerous freeze/thaw cycles to mimic situations that may occur if multiple laboratories analyze the same specimen. Periodic analysis of the samples revealed increases in the levels of endogenous GHB over a 6-month period. The greatest increase (up to 404%) was observed in the samples maintained at room temperature. The refrigerated specimens showed increases of 140-208%, while the frozen specimens showed smaller changes (88-116%). The specimens subjected to multiple freeze/thaw cycles mirrored specimens that had been thawed only once. None of the stored urine specimens demonstrated increases in GHB concentrations that would be consistent with exogenous GHB ingestion.

  5. Sanitary evaluation of domestic water supply facilities with storage tanks and detection of Aeromonas, enteric and related bacteria in domestic water facilities in Okinawa Prefecture of Japan.

    PubMed

    Miyagi, Kazufumi; Sano, Kouichi; Hirai, Itaru

    2017-08-01

    To provide for temporary restrictions of the public water supply system, storage tanks are commonly installed in the domestic water systems of houses and apartment buildings in Okinawa Prefecture of Japan. To learn more about the sanitary condition and management of these water supply facilities with storage tanks (hereafter called "storage tank water systems") and the extent of bacterial contamination of water from these facilities, we investigated their usage and the existence of Aeromonas, enteric and related bacteria. Verbal interviews concerning the use and management of the storage tank water systems were carried out in each randomly sampled household. A total of 54 water samples were collected for bacteriological and physicochemical examinations. Conventional methods were used for total viable count, fecal coliforms, identification of bacteria such as Aeromonas, Enterobacteriaceae and non-fermentative Gram-negative rods (NF-GNR), and measurement of residual chlorine. On Aeromonas species, tests for putative virulence factor and an identification using 16S rRNA and rpoB genes were also performed. Water from the water storage systems was reported to be consumed directly without boiling in 22 of the 54 houses (40.7%). 31 of the sampled houses had installed water storage tanks of more than 1 cubic meter (m 3 ) per inhabitant, and in 21 of the sampled houses, the tank had never been cleaned. In all samples, the total viable count and fecal coliforms did not exceed quality levels prescribed by Japanese waterworks law. Although the quantity of bacteria detected was not high, 23 NF-GNR, 14 Enterobacteriaceae and 5 Aeromonas were isolated in 42.6%, 7.4% and 3.7% of samples respectively. One isolated A. hydrophila and four A. caviae possessed various putative virulence factors, especially A. hydrophila which had diverse putative pathogenic genes such as aer, hlyA, act, alt, ast, ser, and dam. Many bacteria were isolated when the concentration of residual chlorine was below 0.1 mg/l and the water temperature was above 20 °C. These results suggest that elevated water temperature and mismatch between tank size and water demand lead to loss of residual chlorine in tap water. Therefore, to minimize growth of aquatic bacteria such as Aeromonas spp. and Pseudomonas spp., we recommend that an appropriate size tank and/or volume of stored water is always used, and also suggest installation of some means of reducing water temperature such as shading. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Fit for purpose frozen tissue collections by RNA integrity number-based quality control assurance at the Erasmus MC tissue bank.

    PubMed

    Kap, Marcel; Oomen, Monique; Arshad, Shazia; de Jong, Bas; Riegman, Peter

    2014-04-01

    About 5000 frozen tissue samples are collected each year by the Erasmus Medical Center tissue bank. Two percent of these samples are randomly selected annually for RNA isolation and RNA Integrity Number (RIN) measurement. A similar quality assessment was conducted during centralization of a 20-year-old tissue collection from the cancer institute, a 15-year-old liver sample archive (-80°C), and a 13-year-old clinical pathology frozen biopsy archive (Liquid Nitrogen). Samples were divided into either high-quality (RIN ≥6.5) or low-quality overall categories, or into four "fit-for-purpose" quality groups: RIN <5: not reliable for demanding downstream analysis; 5 ≤RIN <6: suitable for RT-qPCR; 6 ≤RIN <8: suitable for gene array analysis; and RIN ≥8: suitable for all downstream techniques. In general, low RIN values were correlated with fatty, fibrous, pancreatic, or necrotic tissue. When the percentage of samples with RIN ≥6.5 is higher than 90%, the tissue bank performance is adequate. The annual 2011 quality control assessment showed that 90.3% (n=93) of all samples had acceptable RIN values; 97.4% (n=39) of the cancer institute collection had RIN values above 6.5; and 88.6% (n=123) of samples from the liver sample archive collection had RIN values higher than 6.5. As the clinical pathology biopsy collection contained only 58.8% (n=24) acceptable samples, the procurement protocols used for these samples needed immediate evaluation. When the distribution of RIN values of the different collections were compared, no significant differences were found, despite differences in average storage time and temperature. According to the principle of "fit-for-purpose" distribution, the vast majority of samples are considered good enough for most downstream techniques. In conclusion, an annual tissue bank quality control procedure provides useful information on tissue sample quality and sheds light on where and if improvements need to be made.

  7. Dye-dispersion study at proposed pumped-storage project on Hudson River at Cornwall, New York

    USGS Publications Warehouse

    Dunn, Bernard; Gravlee, George C.

    1978-01-01

    Data were collected during a dye-dispersion study on a 6-mile, tide-affected reach of the Hudson River near the proposed Cornwall Pumped Storage Project on September 21-22, 1977. The results indicated that complete mixing did not occur during the first tidal cycle but was complete after two or more cycles. The fluorometric dye-tracing procedure was used to determine the dispersion characteristics of the water mass. Rhodamine WT dye, 20-percent solution, was continuously injected on the west side of the river throughout an ebb tide, and its movement was monitored during a 30-hour period. Samples were collected both individually and continuously. Automatic dye samplers were used at selected cross sections near each bank. Bathymetric measurements were made at eight cross sections between Newburgh and West Point to determine the depths. (Woodard-USGS)

  8. Volatile substance abuse--post-mortem diagnosis.

    PubMed

    Wille, Sarah M R; Lambert, Willy E E

    2004-06-10

    A substantial number of children and adolescents world-wide abuse volatile substances with the intention to experience an euphoric state of consciousness. Although the ratio of deaths to nonfatal inhalation escapades is low, it is an important and preventable cause of death in young people. In the analytical investigation of volatile substances proper sample collection, storage and handling are important in view of the volatile nature of the compounds. Volatile organic compounds in post-mortem matrices such as blood, urine and tissues are generally determined by gas chromatography after extracting the compounds with methods such as static and dynamic headspace or even with pulse-heating and solvent extraction. In post-mortem cases, metabolites in urine seem less relevant, however, trichloroethanol and trichloroacetic acid were determined in several cases. When interpreting qualitative and quantitative results, researchers should be aware of false conclusions. The main reason why scepticism is necessary is the occurrence of losses of analytes during sampling, sample handling and storage, which results in false quantitation.

  9. Comparison of Tillandsia usneoides (Spanish moss) water and leachate dynamics between urban and pristine barrier island maritime oak forests

    NASA Astrophysics Data System (ADS)

    Van Stan, J. T.; Stubbins, A.; Reichard, J. S.; Wright, K.; Jenkins, R. B.

    2013-12-01

    Epiphyte coverage on forest canopies can drastically alter the volume and chemical composition of rainwater reaching soils. Along subtropical and tropical coastlines Tillandisa usneoides L. (Spanish moss), in particular, can envelop urban and natural tree crowns. Several cities actively manage their 'moss' covered forest to enhance aesthetics in the most active tourist areas (e.g., Savannah GA, St. Augustine FL, Charleston SC). Since T. usneoides survives through atmospheric water and solute exchange from specialized trichomes (scales), we hypothesized that T. usneoides water storage dynamics and leachate chemistry may be altered by exposure to this active urban atmosphere. 30 samples of T. usneoides from managed forests around the tourist center of Savannah, Georgia, USA were collected to compare with 30 samples from the pristine maritime live oak (Quercus virginiana Mill.) forests of a nearby undeveloped barrier island (St. Catherines Island, Georgia, USA). Maximum water storage capacities were determined via submersion (for all 60 samples) along with dissolved ion (DI) and organic matter (DOM) concentrations (for 15 samples each) after simulated throughfall generation using milliQ ultrapurified water. Further, DOM quality was evaluated (for 15 samples each) using absorbance and fluorescence spectroscopy (EEMS). Results show significant alterations to water storage dynamics, DI, DOM, and DOM quality metrics under urban atmospheric conditions, suggesting modified C and water cycling in urban forest canopies that may, in turn, influence intrasystem nutrient cycles in urban catchment soils or streams via runoff.

  10. Impact of Nisin-Activated Packaging on Microbiota of Beef Burgers during Storage

    PubMed Central

    Ferrocino, Ilario; Greppi, Anna; La Storia, Antonietta; Rantsiou, Kalliopi; Ercolini, Danilo

    2015-01-01

    Beef burgers were stored at 4°C in a vacuum in nisin-activated antimicrobial packaging. Microbial ecology analyses were performed on samples collected between days 0 and 21 of storage to discover the population diversity. Two batches were analyzed using RNA-based denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. The active packaging retarded the growth of the total viable bacteria and lactic acid bacteria. Culture-independent analysis by pyrosequencing of RNA extracted directly from meat showed that Photobacterium phosphoreum, Lactococcus piscium, Lactobacillus sakei, and Leuconostoc carnosum were the major operational taxonomic units (OTUs) shared between control and treated samples. Beta diversity analysis of the 16S rRNA sequence data and RNA-DGGE showed a clear separation between two batches based on the microbiota. Control samples from batch B showed a significant high abundance of some taxa sensitive to nisin, such as Kocuria rhizophila, Staphylococcus xylosus, Leuconostoc carnosum, and Carnobacterium divergens, compared to control samples from batch A. However, only from batch B was it possible to find a significant difference between controls and treated samples during storage due to the active packaging. Predicted metagenomes confirmed differences between the two batches and indicated that the use of nisin-based antimicrobial packaging can determine a reduction in the abundance of specific metabolic pathways related to spoilage. The present study aimed to assess the viable bacterial communities in beef burgers stored in nisin-based antimicrobial packaging, and it highlights the efficacy of this strategy to prolong beef burger shelf life. PMID:26546424

  11. SELDI-TOF MS of quadruplicate urine and serum samples to evaluate changes related to storage conditions.

    PubMed

    Traum, Avram Z; Wells, Meghan P; Aivado, Manuel; Libermann, Towia A; Ramoni, Marco F; Schachter, Asher D

    2006-03-01

    Proteomic profiling with SELDI-TOF MS has facilitated the discovery of disease-specific protein profiles. However, multicenter studies are often hindered by the logistics required for prompt deep-freezing of samples in liquid nitrogen or dry ice within the clinic setting prior to shipping. We report high concordance between MS profiles within sets of quadruplicate split urine and serum samples deep-frozen at 0, 2, 6, and 24 h after sample collection. Gage R&R results confirm that deep-freezing times are not a statistically significant source of SELDI-TOF MS variability for either blood or urine.

  12. Short communication: Evaluation of sampling socks for detection of Mycobacterium avium ssp. paratuberculosis on dairy farms.

    PubMed

    Wolf, R; Orsel, K; De Buck, J; Kanevets, U; Barkema, H W

    2016-04-01

    Mycobacterium avium ssp. paratuberculosis (MAP) causes Johne's disease, a production-limiting disease in cattle. Detection of infected herds is often done using environmental samples (ES) of manure, which are collected in cattle pens and manure storage areas. Disadvantages of the method are that sample accuracy is affected by cattle housing and type of manure storage area. Furthermore, some sampling locations (e.g., manure lagoons) are frequently not readily accessible. However, sampling socks (SO), as used for Salmonella spp. testing in chicken flocks, might be an easy to use and accurate alternative to ES. The objective of the study was to assess accuracy of SO for detection of MAP in dairy herds. At each of 102 participating herds, 6 ES and 2 SO were collected. In total, 45 herds had only negative samples in both methods and 29 herds had ≥1 positive ES and ≥1 positive SO. Furthermore, 27 herds with ≥1 positive ES had no positive SO, and 1 herd with no positive ES had 1 positive SO. Bayesian simulation with informative priors on sensitivity of ES and MAP herd prevalence provided a posterior sensitivity for SO of 43.5% (95% probability interval=33-58), and 78.5% (95% probability interval=62-93) for ES. Although SO were easy to use, accuracy was lower than for ES. Therefore, with improvements in the sampling protocol (e.g., more SO per farm and more frequent herd visits), as well as improvements in the laboratory protocol, perhaps SO would be a useful alternative for ES. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  13. Storage duration and white blood cell content of red blood cell (RBC) products increases adhesion of stored RBCs to endothelium under flow conditions.

    PubMed

    Anniss, Angela M; Sparrow, Rosemary L

    2006-09-01

    Adherence of red blood cells (RBCs) to vascular endothelium impairs blood flow and decreases oxygen delivery. Although RBCs may be stored for up to 42 days before transfusion under current blood banking guidelines, little is known of how changes to RBCs during storage may affect their adherence properties. The influence of RBC product storage time and white blood cell (WBC) burden on the adherence of RBCs for transfusion to vascular endothelium under conditions of continuous flow was investigated in this study. RBC samples were collected from nonleukoreduced (S-RBC), buffy coat-poor (BCP-RBC), and leukofiltered (LF-RBC) products at fixed time points during storage. Samples were perfused, at controlled shear stress and temperature, across a confluent endothelial cell (EC) monolayer with a parallel-flow chamber mounted to an inverted microscope. RBC-EC interactions were recorded with a digital camera attached to the microscope. The number of RBCs adhering to the EC layer increased significantly with storage time in all RBC products; however, WBC reduction delayed this increase. LF-RBCs were also significantly less adherent than S-RBC or BCP-RBC products on Day 1 of storage (p < 0.05). The strength of RBC attachment to vascular endothelium was significantly stronger in S-RBC products compared to BCP-RBC and LF-RBC products. Our findings indicate that product storage time and WBC burden increase the number and strength of adhesion of RBCs to vascular endothelium. These results may lead to greater understanding of the interaction of transfused RBCs with recipient endothelium and the biologic consequences of this adherence.

  14. Development of a passive sampler for gaseous mercury

    NASA Astrophysics Data System (ADS)

    Gustin, M. S.; Lyman, S. N.; Kilner, P.; Prestbo, E.

    2011-10-01

    Here we describe work toward development of the components of a cost effective passive sampling system for gaseous Hg that could be broadly deployed by nontechnical staff. The passive sampling system included an external shield to reduce turbulence and exposure to precipitation and dust, a diffusive housing that directly protects the collection surface during deployment and handling, and a collection surface. A protocol for cleaning and deploying the sampler and an analytical method were developed. Our final design consisted of a polycarbonate external shield enclosing a custom diffusive housing made from expanded PTFE tubing. Two collection surfaces were investigated, gold sputter-coated quartz plates and silver wires. Research showed the former would require extensive quality control for use, while the latter had interferences with other atmosphere constituents. Although the gold surface exhibited the best performance over space and time, gradual passivation would limit reuse. For both surfaces lack of contamination during shipping, deployment and storage indicated that the handling protocols developed worked well with nontechnical staff. We suggest that the basis for this passive sampling system is sound, but further exploration and development of a reliable collection surface is needed.

  15. IMPROVING BIOMASS LOGISTICS COST WITHIN AGRONOMIC SUSTAINABILITY CONSTRAINTS AND BIOMASS QUALITY TARGETS

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

    J. Richard Hess; Kevin L. Kenney; Christopher T. Wright

    Equipment manufacturers have made rapid improvements in biomass harvesting and handling equipment. These improvements have increased transportation and handling efficiencies due to higher biomass densities and reduced losses. Improvements in grinder efficiencies and capacity have reduced biomass grinding costs. Biomass collection efficiencies (the ratio of biomass collected to the amount available in the field) as high as 75% for crop residues and greater than 90% for perennial energy crops have also been demonstrated. However, as collection rates increase, the fraction of entrained soil in the biomass increases, and high biomass residue removal rates can violate agronomic sustainability limits. Advancements inmore » quantifying multi-factor sustainability limits to increase removal rate as guided by sustainable residue removal plans, and mitigating soil contamination through targeted removal rates based on soil type and residue type/fraction is allowing the use of new high efficiency harvesting equipment and methods. As another consideration, single pass harvesting and other technologies that improve harvesting costs cause biomass storage moisture management challenges, which challenges are further perturbed by annual variability in biomass moisture content. Monitoring, sampling, simulation, and analysis provide basis for moisture, time, and quality relationships in storage, which has allowed the development of moisture tolerant storage systems and best management processes that combine moisture content and time to accommodate baled storage of wet material based upon “shelf-life.” The key to improving biomass supply logistics costs has been developing the associated agronomic sustainability and biomass quality technologies and processes that allow the implementation of equipment engineering solutions.« less

  16. The microbiological quality of pasteurized milk sold by automatic vending machines.

    PubMed

    Angelidis, A S; Tsiota, S; Pexara, A; Govaris, A

    2016-06-01

    The microbiological quality of pasteurized milk samples (n = 39) collected during 13 weekly intervals from three automatic vending machines (AVM) in Greece was investigated. Microbiological counts (total aerobic (TAC), total psychrotrophic (TPC), Enterobacteriaceae (EC), and psychrotrophic aerobic bacterial spore counts (PABSC)) were obtained at the time of sampling and at the end of shelf-life (3 days) after storage of the samples at 4 or 8°C. TAC were found to be below the 10(7 ) CFU ml(-1) limit of pasteurized milk spoilage both during sampling as well as when milk samples were stored at either storage temperature for 3 days. Enterobacteriaceae populations were below 1 CFU ml(-1) in 69·2% of the samples tested at the time of sampling, whereas the remaining samples contained low numbers, typically less than 10 CFU ml(-1) . All samples tested negative for the presence of Listeria monocytogenes. Analogous microbiological data were also obtained by sampling and testing prepackaged, retail samples of pasteurized milk from two dairy companies in Greece (n = 26). From a microbiological standpoint, the data indicate that the AVM milk samples meet the quality standards of pasteurized milk. However, the prepackaged, retail milk samples yielded better results in terms of TAC, TPC and EC, compared to the AVM samples at the end of shelf-life. Recently, Greek dairy farmers organized in cooperatives launched the sale of pasteurized milk via AVM and this study reports on the microbiological quality of this product. The data show that AVM milk is sold at proper refrigeration temperatures and meets the quality standards of pasteurized milk throughout the manufacturer's specified shelf-life. However, based on the microbiological indicators tested, the keeping quality of the tested prepackaged, retail samples of pasteurized milk at the end of shelf-life upon storage under suboptimal refrigeration temperature (8°C) was better. © 2016 The Society for Applied Microbiology.

  17. Influence of storage conditions on aluminum concentrations in serum, dialysis fluid, urine, and tap water.

    PubMed

    Wilhelm, M; Ohnesorge, F K

    1990-01-01

    The influence of storage temperature, vessel type, and treatment on alterations of aluminum (Al) concentrations in serum, urine, and dialysis fluid samples was studied at three different concentrations for each sample over an 18-month period. Furthermore, the influence of acidification on Al levels in tap water, urine, and dialysis fluid samples was studied over a four-month period. Al was measured by atomic absorption spectrometry. Sample storage in glass vessels was unsuitable, whereas only minor alterations of Al levels were observed with storage in polypropylene tubes, polystyrene tubes, and Monovettes. By using appropriate plastic containers, acid washing of the vessels showed no improvement. Frozen storage was superior compared with 4 degrees C, whereas storage at -80 degrees C offered no advantage compared with storage at -20 degrees C. Acidification of tap water samples was necessary to stabilize Al levels during storage. No striking effect of acidification on Al levels in urine and dialysis fluid samples was found. It is concluded that longterm storage of serum, urine, tap water, and dialysis fluid samples is possible if appropriate conditions are used.

  18. Maggot development during morgue storage and its effect on estimating the post-mortem interval.

    PubMed

    Huntington, Timothy E; Higley, Leon G; Baxendale, Frederick P

    2007-03-01

    When insect evidence is obtained during autopsy, forensic entomologists make decisions regarding the effects of low-temperature (-1 degrees C to 4 degrees C) storage of the body and associated insects when estimating the post-mortem interval (PMI). To determine the effects of storage in a morgue cooler on the temperature of maggot masses, temperatures inside and outside of body bags containing a human cadaver and porcine cadavers (seven replicates) were measured during storage. Temperatures remained significantly higher (p<0.05) inside of the body bags relative to the cooler, and remained at levels sufficient for maggot feeding and development. If the assumption that no insect development takes place during preautopsy refrigeration is made, potential error rates in PMI estimation of 8.6-12.8% occur. The potential for blow fly larvae to undergo significant development while being stored in the morgue is a possibility that forensic entomologists should consider during an investigation involving samples collected from autopsy. Case and experimental evidence also demonstrate that substantial tissue loss can occur from maggot feeding during morgue storage.

  19. Preliminary study of a potential CO2 reservoir area in Hungary

    NASA Astrophysics Data System (ADS)

    Sendula, Eszter; Király, Csilla; Szabó, Zsuzsanna; Falus, György; Szabó, Csaba; Kovács, István; Füri, Judit; Kónya, Péter; Páles, Mariann; Forray, Viktória

    2014-05-01

    Since the first international agreement in 1997 (the Kyoto Protocol) the reduction of greenhouse gas emission has a key role in the European Union's energy and climate change policy. Following the Directive 2009/31/EC we are experiencing a significant change in the Hungarian national activity. Since the harmonization procedure, which was completed in May 2012, the national regulation obligates the competent authority to collect and regularly update all geological complexes that are potential for CO2 geological storage. In Hungary the most abundant potential storage formations are mostly saline aquifers of the Great Hungarian Plain (SE-Hungary), with sandstone reservoir and clayey caprock. The Neogene basin of the Great Hungarian Plain was subsided and then filled by a prograding delta system from NW and NE during the Late Miocene, mostly in the Pannonian time. The most potential storage rock was formed as a fine-grained sandy turbidite interlayered by thin argillaceous beds in the deepest part of the basin. It has relatively high porosity, depth and more than 1000 m thickness. Providing a regional coverage for the sandy turbidite, a 400-500 m thick argillaceous succession was formed in the slope environment. The composition, thickness and low permeability is expected to make it a suitable, leakage-safe caprock of the storage system. This succession is underlain by argillaceous rocks that were formed in the basin, far from sediment input and overlain by interfingering siltstone, sandstone and claystone succession formed in delta and shoreline environments and in the alluvial plain. Core samples have been collected from the potential reservoir rock and its cap rock in the Great Hungarian Plain's succession. The water compositions of the studied depth were known from well-log database. Using the information, acquired from these archive documents, we have constructed input data for geochemical modeling in order to to study the effect of pCO2 injection in the potential CO2 storage environment. From the potential reservoir rock samples (sandstone) thin sections were prepared to determine the mineral composition, pore distribution, pore geometry and grain size. The volume ratio of the minerals was calculated using pixel counter. To have more accurate mineral composition, petrographic observation and SEM analyzes have been carried out. The caprock samples involved in the study can be divided into mudstone and aleurolite samples. To determine the mineral composition of these samples, XRD, DTA, FTIR, SEM analysis has been carried out. To obtain a picture about the geochemical behavior of the potential CO2 storage system, geochemical models were made for the reservoir rocks. For the equilibrium geochemical model, PHREEQC 3.0 was used applying LLNL database. The data used in the model are real pore water compositions from the studied area and an average mineral composition based on petrographic microscope and SEM images. In the model we considered the cation-anion ratio (<10%) and the partial pressure of CO2. First of all, we were interested in the direction of the geochemical reactions during an injection process. Present work is focused on the mineralogy of the most potential storage rock and its caprock, and their expectable geochemical reactions for the effect of scCO2.

  20. Molecular epidemiology biomarkers-Sample collection and processing considerations

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

    Holland, Nina T.; Pfleger, Laura; Berger, Eileen

    2005-08-07

    Biomarker studies require processing and storage of numerous biological samples with the goals of obtaining a large amount of information and minimizing future research costs. An efficient study design includes provisions for processing of the original samples, such as cryopreservation, DNA isolation, and preparation of specimens for exposure assessment. Use of standard, two-dimensional and nanobarcodes and customized electronic databases assure efficient management of large sample collections and tracking results of data analyses. Standard operating procedures and quality control plans help to protect sample quality and to assure validity of the biomarker data. Specific state, federal and international regulations are inmore » place regarding research with human samples, governing areas including custody, safety of handling, and transport of human samples. Appropriate informed consent must be obtained from the study subjects prior to sample collection and confidentiality of results maintained. Finally, examples of three biorepositories of different scale (European Cancer Study, National Cancer Institute and School of Public Health Biorepository, University of California, Berkeley) are used to illustrate challenges faced by investigators and the ways to overcome them. New software and biorepository technologies are being developed by many companies that will help to bring biological banking to a new level required by molecular epidemiology of the 21st century.« less

  1. Trace element distribution in the water and sediments of certain storage lakes from the Jijia catchment, (Romania)

    NASA Astrophysics Data System (ADS)

    Dughila, A.; Iancu, O. G.; Romanescu, G. T.

    2012-04-01

    The present study aims at investigating the concentrations and distribution levels of a series of trace elements in water and sediment samples collected from six storage lakes located in the Jijia catchment - NE of Romania. The lakes are multi-purpose water reservoirs, three of them being mainly used as a source of municipal drinking water, or for fishing, irrigation for the farms in the area, protection against floods and the regulation of river flows. By contrast, agricultural wastes, fertilizers, raw sewage effluents and road runoff constitute the predominant anthropogenic sources, which supply the lakes in question with Cd, Cu, Pb and Zn. The present study was conducted on a series of 63 sediment samples and 18 water samples, collected from the same locations, in order to establish the distribution levels of certain trace elements from the water through sediments. Sediment cores were collected from two sections across each lake by means of a motor boat, using a system that consists of a graduated sampling tube (0.9 m in length and 72.5 mm in diameter) made of Plexiglas (Eijkelkamp sample tube guide). Prior to the analyses, the samples were air-dried, ground and homogenized using an agate mortar, oven-dried at 50 °C for 6 days and then sieved through 63 µm sieves. The sediment and water samples were subjected to a digestion technique with concentrated nitric acid using a microwave oven (Berghof type), and analyzed for the following elements: Pb, Zn, Cu, Cd, Cr and Ni. The total concentration of the elements was measured through atomic absorption spectrometry (AAS) with an RSD of < 10 % from solutions. The vertical distribution of most elements in the cores examined could be characterized as relatively uniform, with higher concentrations for those collected from the lakes which are more influenced by anthropogenic factors, compared to those situated in forested areas. The lake-water quality characteristics were below the recommended drinking water standards imposed by the current legislation (MMGA Ord. No. 161/16.02.2006 - Normative regarding the classification of surface waters in order to establish the ecological status of water bodies, which combines European and Romanian provisions), with the exception of copper (with very high concentrations in all the water samples), lead and cadmium. Keywords: AAS, Jijia catchment (Romania), lake water, sediment core, trace elements

  2. Ethical issues in the export, storage and reuse of human biological samples in biomedical research: perspectives of key stakeholders in Ghana and Kenya.

    PubMed

    Tindana, Paulina; Molyneux, Catherine S; Bull, Susan; Parker, Michael

    2014-10-18

    For many decades, access to human biological samples, such as cells, tissues, organs, blood, and sub-cellular materials such as DNA, for use in biomedical research, has been central in understanding the nature and transmission of diseases across the globe. However, the limitations of current ethical and regulatory frameworks in sub-Saharan Africa to govern the collection, export, storage and reuse of these samples have resulted in inconsistencies in practice and a number of ethical concerns for sample donors, researchers and research ethics committees. This paper examines stakeholders' perspectives of and responses to the ethical issues arising from these research practices. We employed a qualitative strategy of inquiry for this research including in-depth interviews and focus group discussions with key research stakeholders in Kenya (Nairobi and Kilifi), and Ghana (Accra and Navrongo). The stakeholders interviewed emphasised the compelling scientific importance of sample export, storage and reuse, and acknowledged the existence of some structures governing these research practices, but they also highlighted the pressing need for a number of practical ethical concerns to be addressed in order to ensure high standards of practice and to maintain public confidence in international research collaborations. These concerns relate to obtaining culturally appropriate consent for sample export and reuse, understanding cultural sensitivities around the use of blood samples, facilitating a degree of local control of samples and sustainable scientific capacity building. Drawing on these findings and existing literature, we argue that the ethical issues arising in practice need to be understood in the context of the interactions between host research institutions and local communities and between collaborating institutions. We propose a set of 'key points-to-consider' for research institutions, ethics committees and funding agencies to address these issues.

  3. Exhaled nitric oxide in mylar balloons: influence of storage time, humidity and temperature.

    PubMed Central

    Bodini, Alessandro; Pijnenburg, Mariëlle W H; Boner, Atillio L; de Jongste, Johan C

    2003-01-01

    BACKGROUND: Mylar balloons are used to collect exhaled air for analysis of fractional nitric oxide concentration (FENO). AIM: We studied the effect of storage conditions on the stability of nitric oxide (NO) in mylar balloons. METHODS: Exhaled air samples and calibration gases were stored in mylar balloons at 4, 21 and 37 degrees C, with or without silica gel. NO was measured after 0, 6, 9, 24 and 48 h. Scheffe F-tests were used to compare NO values. RESULTS: NO remained stable in balloons for 9 h at all temperatures, without silica gel. NO increased between 9 and 48 h, but only with low initial FENO. Silica gel increased variability. CONCLUSIONS: FENO in mylar balloons is stable for at least 9 h. The storage temperature is not critical, but silica gel increases variability. PMID:12745548

  4. Planning for hazardous campus waste collection.

    PubMed

    Liu, Kun-Hsing; Shih, Shao-Yang; Kao, Jehng-Jung

    2011-05-15

    This study examines a procedure developed for planning a nation-wide hazardous campus waste (HCW) collection system. Alternative HCW plans were designed for different collection frequencies, truckloads, storage limits, and also for establishing an additional transfer station. Two clustering methods were applied to group adjacent campuses into clusters based on their locations, HCW quantities, the type of vehicles used and collection frequencies. Transportation risk, storage risk, and collection cost are the major criteria used to evaluate the feasibility of each alternative. Transportation risk is determined based on the accident rates for each road type and collection distance, while storage risk is calculated by estimating the annual average HCW quantity stored on campus. Alternatives with large trucks can reduce both transportation risk and collection cost, but their storage risks would be significantly increased. Alternatives that collect neighboring campuses simultaneously can effectively reduce storage risks as well as collection cost if the minimum quantity to collect for each group of neighboring campuses can be properly set. The three transfer station alternatives evaluated for northern Taiwan are cost effective and involve significantly lower transportation risk. The procedure proposed is expected to facilitate decision making and to support analyses for formulating a proper nation-wide HCW collection plan. Copyright © 2011 Elsevier B.V. All rights reserved.

  5. Minimally invasive collection of adipose tissue facilitates the study of eco-physiology in small-bodied mammals

    Treesearch

    Jeff Clerc; Theodore J. Weller; Jeffrey B. Schineller; Joseph M. Szewczak; Diana Fisher

    2016-01-01

    Adipose tissue is the primary fuel storage for vertebrates and is an important component of energy budgets during periods of peak energetic demands. Investigating the composition of adipose tissue can provide information about energetics, migration, reproduction, and other life-history traits. Until now, most field methods for sampling the adipose tissue of...

  6. Study of the hadronic decays of [chi][sub c] states

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

    Bai, J.Z.; Bian, J.G.; Chen, G.P.

    1999-10-01

    Hadronic decays of the [ital P]-wave spin-triplet charmonium states [chi][sub cJ](J=0,1,2) are studied using a sample of [psi](2S) decays collected by the BES detector operating at the BEPC storage ring. Branching fractions for the decays [chi][sub c1][r arrow]K[sub S][sup 0]K[sup +][pi][sup [minus

  7. Recalcitrant Behavior of Cherrybark Oak Seed: An FT-IR Study of Desiccation Sensitivity in Quercus pagoda Raf. Acorns

    Treesearch

    Sharon Sowa; Kristina F. Connor

    2003-01-01

    The recalcitrant behavior of cherrybark oak (Quercus pagoda Raf.) acorns was examined in terms of effects of moisture content on seed storage longevity and (short term) seed germination. Seed samples collected over two consecutive years were fully hydrated, then subjected to drying under ambient conditions of temperature and relative humidity on the...

  8. Seminal plasma affects sperm sex sorting in boars.

    PubMed

    Alkmin, Diego V; Parrilla, Inmaculada; Tarantini, Tatiana; Del Olmo, David; Vazquez, Juan M; Martinez, Emilio A; Roca, Jordi

    2016-04-01

    Two experiments were conducted in boar semen samples to evaluate how both holding time (24h) and the presence of seminal plasma (SP) before sorting affect sperm sortability and the ability of sex-sorted spermatozoa to tolerate liquid storage. Whole ejaculate samples were divided into three aliquots immediately after collection: one was diluted (1:1, v/v) in Beltsville thawing solution (BTS; 50% SP); the SP of the other two aliquots was removed and the sperm pellets were diluted with BTS + 10% of their own SP (10% SP) or BTS alone (0% SP). The three aliquots of each ejaculate were divided into two portions, one that was processed immediately for sorting and a second that was sorted after 24h storage at 15-17°C. In the first experiment, the ability to exhibit well-defined X- and Y-chromosome-bearing sperm peaks (split) in the cytometry histogram and the subsequent sorting efficiency were assessed (20 ejaculates). In contrast with holding time, the SP proportion influenced the parameters examined, as evidenced by the higher number of ejaculates exhibiting split and better sorting efficiency (P<0.05) in semen samples with 0-10% SP compared with those with 50% SP. In a second experiment, the quality (viability, total and progressive motility) and functionality (plasma membrane fluidity and intracellular generation of reactive oxygen species) of sex-sorted spermatozoa were evaluated after 0, 72 and 120h storage at 15-17°C (10 ejaculates). Holding time and SP proportion did not influence the quality or functionality of stored sex-sorted spermatozoa. In conclusion, a holding time as long as 24h before sorting did not negatively affect sex sorting efficiency or the ability of sorted boar spermatozoa to tolerate long-term liquid storage. A high proportion of SP (50%) in the semen samples before sorting reduced the number of ejaculates to be sorted and negatively influenced the sorting efficiency, but did not affect the ability of sex-sorted spermatozoa to tolerate liquid storage.

  9. Major element concentrations in six Alaskan arctic rivers from melt to freeze-up

    NASA Astrophysics Data System (ADS)

    Douglas, T. A.; Barker, A.; Jacobson, A. D.; McClelland, J. W.; Khosh, M. S.; Lehn, G. O.

    2010-12-01

    It is increasingly evident that permafrost in the Arctic is responding to climate warming. An expected response to this warming is permafrost degradation and the downward migration of the seasonally thawed (active) layer into previously frozen material. This could allow weathering of previously frozen soils and influence surface water biogeochemistry in Arctic rivers. The weathering signal would most likely be evident in surface waters during summer and early fall base flow when the active layer is at its deepest extent. Studies collecting water samples from spring through late fall could capture these flows. Fieldwork in remote regions often requires long storage times for samples prior to analysis. One aspect of our study was to investigate whether waters collected for major element analyses should be preserved by acidification. We collected up to 60 surface water samples from each of six rivers between April and October, 2009. Two rivers were underlain by organic rich permafrost, two of the rivers drained mountainous bedrock, and two rivers were underlain by a combination of both bedrock and organic rich permafrost. We collected duplicate samples from each river. Samples were filtered in the field to less than 0.45 microns and collected into high density polyethylene bottles. Waters were stored for six months prior to analysis. One set of samples was analyzed without acidification while the second set was acidified with nitric acid to a pH of 2 after 6 months of storage. Concentrations of sodium, potassium, magnesium, calcium, ammonium, fluoride, chloride, nitrate, sulfate and phosphate were measured from all samples by ion chromatography. Sulfate, calcium, magnesium, sodium and potassium concentrations increase steadily through the summer from the end of spring melt into early fall. In all six rivers the potassium concentrations are unaffected by acidification. In the two streams underlain by organic rich permafrost and in one of the bedrock streams the calcium, magnesium, and sodium values are similar for both the acidified and unacidified samples. In the other rivers, however, the calcium, magnesium and sodium concentrations yield complex behavior. From spring melt into early summer the cation values are identical in the acidified and unacidified samples. However, in late summer and early fall calcium, magnesium, and sodium concentrations are roughly ten percent higher in the acidified samples than in the unacidified ones. Our results suggest an enhanced mineral weathering signal in the watersheds in the late summer which is evidenced by steadily increasing sulfate and cation concentrations throughout the summer. Potassium does not appear to be affected by acidification. Some cation species respond to acidification, especially in waters representing baseflow. However, the cause(s) for the higher values in some acidified samples is unknown.

  10. Sampling and pyrosequencing methods for characterizing bacterial communities in the human gut using 16S sequence tags.

    PubMed

    Wu, Gary D; Lewis, James D; Hoffmann, Christian; Chen, Ying-Yu; Knight, Rob; Bittinger, Kyle; Hwang, Jennifer; Chen, Jun; Berkowsky, Ronald; Nessel, Lisa; Li, Hongzhe; Bushman, Frederic D

    2010-07-30

    Intense interest centers on the role of the human gut microbiome in health and disease, but optimal methods for analysis are still under development. Here we present a study of methods for surveying bacterial communities in human feces using 454/Roche pyrosequencing of 16S rRNA gene tags. We analyzed fecal samples from 10 individuals and compared methods for storage, DNA purification and sequence acquisition. To assess reproducibility, we compared samples one cm apart on a single stool specimen for each individual. To analyze storage methods, we compared 1) immediate freezing at -80 degrees C, 2) storage on ice for 24 or 3) 48 hours. For DNA purification methods, we tested three commercial kits and bead beating in hot phenol. Variations due to the different methodologies were compared to variation among individuals using two approaches--one based on presence-absence information for bacterial taxa (unweighted UniFrac) and the other taking into account their relative abundance (weighted UniFrac). In the unweighted analysis relatively little variation was associated with the different analytical procedures, and variation between individuals predominated. In the weighted analysis considerable variation was associated with the purification methods. Particularly notable was improved recovery of Firmicutes sequences using the hot phenol method. We also carried out surveys of the effects of different 454 sequencing methods (FLX versus Titanium) and amplification of different 16S rRNA variable gene segments. Based on our findings we present recommendations for protocols to collect, process and sequence bacterial 16S rDNA from fecal samples--some major points are 1) if feasible, bead-beating in hot phenol or use of the PSP kit improves recovery; 2) storage methods can be adjusted based on experimental convenience; 3) unweighted (presence-absence) comparisons are less affected by lysis method.

  11. Effect of chilling temperature on the long-term survival of rabbit spermatozoa held either in a tris-based or a jellified extender.

    PubMed

    Rosato, Mp; Iaffaldano, N

    2011-04-01

    As the preservation of the fertilizing capacity of rabbit spermatozoa for several days after semen collection remains a major target for the artificial insemination programs of rabbit breeding, a study was conducted to compare the efficacy of 5 or 15°C as holding temperature in lengthening the preservability of rabbit semen quality during 192 h of storage both in a solid (Cunigel) and a liquid (Tris-Citric acid-Glucose; TCG) extender. Six pooled semen samples (two ejaculates/male; two-three males/pool) were taken and made four aliquots: two aliquots were tenfold diluted with the TCG extender, whereas the other two were tenfold diluted with the Cunigel extender. One aliquot per diluent was stored at 5°C and the second one at 15°C. Sperm motility (light microscope), viability (SyBr-PI staining), plasma membrane functional integrity (Hypo-osmotic swelling test) and acrosome integrity (PSA-FITC staining) were recorded at 0, 48, 120 and 192 h of storage. In liquid-stored spermatozoa, mass motility and viability were significantly higher (p ≤ 0.05) in samples stored at 5°C than at 15°C at all the storage times; at 5°C resulted also higher (p ≤ 0.05) the percentages of both forward motility at 48 h and sperm functional integrity at 120 and 192 h of storage, whereas chilling temperature did not affect acrosome integrity. With the Cunigel extender, all the semen qualitative parameters were significantly higher in sample stored at 5 than 15°C over storage time (p ≤ 0.05); only acrosome integrity at 192 h was not different according to the chilling temperatures. In conclusion, 5°C were better than 15°C for the long-term storage of rabbit semen both in the TCG and Cunigel extender. © 2010 Blackwell Verlag GmbH.

  12. The Astromaterials X-Ray Computed Tomography Laboratory at Johnson Space Center

    NASA Technical Reports Server (NTRS)

    Zeigler, R. A.; Coleff, D. M.; McCubbin, F. M.

    2017-01-01

    The Astromaterials Acquisition and Curation Office at NASA's Johnson Space Center (hereafter JSC curation) is the past, present, and future home of all of NASA's astromaterials sample collections. JSC curation currently houses all or part of nine different sample collections: (1) Apollo samples (1969), (2) Lunar samples (1972), (3) Antarctic meteorites (1976), (4) Cosmic Dust particles (1981), (5) Microparticle Impact Collection (1985), (6) Genesis solar wind atoms (2004); (7) Stardust comet Wild-2 particles (2006), (8) Stardust interstellar particles (2006), and (9) Hayabusa asteroid Itokawa particles (2010). Each sample collection is housed in a dedicated clean room, or suite of clean rooms, that is tailored to the requirements of that sample collection. Our primary goals are to maintain the long-term integrity of the samples and ensure that the samples are distributed for scientific study in a fair, timely, and responsible manner, thus maximizing the return on each sample. Part of the curation process is planning for the future, and we also perform fundamental research in advanced curation initiatives. Advanced Curation is tasked with developing procedures, technology, and data sets necessary for curating new types of sample collections, or getting new results from existing sample collections [2]. We are (and have been) planning for future curation, including cold curation, extended curation of ices and volatiles, curation of samples with special chemical considerations such as perchlorate-rich samples, and curation of organically- and biologically-sensitive samples. As part of these advanced curation efforts we are augmenting our analytical facilities as well. A micro X-Ray computed tomography (micro-XCT) laboratory dedicated to the study of astromaterials will be coming online this spring within the JSC Curation office, and we plan to add additional facilities that will enable nondestructive (or minimally-destructive) analyses of astromaterials in the near future (micro-XRF, confocal imaging Raman Spectroscopy). These facilities will be available to: (1) develop sample handling and storage techniques for future sample return missions; (2) be utilized by PET for future sample return missions; (3) be used for retroactive PET (Positron Emission Tomography)-style analyses of our existing collections; and (4) for periodic assessments of the existing sample collections. Here we describe the new micro-XCT system, as well as some of the ongoing or anticipated applications of the instrument.

  13. Cryopreservation prevents iron-initiated highly unsaturated fatty acid loss during storage of human blood on chromatography paper at -20°C.

    PubMed

    Metherel, Adam H; Stark, Ken D

    2015-03-01

    Fingertip prick whole blood collection on chromatography paper is amenable to high-throughput fatty acid (FA) profiling for large clinical and field studies. However, sample storage is problematic because highly unsaturated FAs (HUFAs) in erythrocytes rapidly degrade in samples stored at -20°C. The aim of the current study was to determine the mechanism of HUFA degradation and to develop prevention protocols. Free fatty acid (FFA) standards and whole blood reference material from a single participant were used to examine sample storage at -20°C for up to 90 d in triplicate. Iron chelation with deferoxamine (0-5000 μg), antioxidant protection with butylated hydroxytoluene (50 μg), cryopreservation with glycerol, and blood drying were examined using whole blood on chromatography strips. Biological replicate blood samples from additional participants (n = 6) with a range of ω-3 (n-3) HUFA concentrations were similarly assessed. FFAs were relatively stable when stored on chromatography strips at -20°C. Glycerol treatment prevented HUFA degradation in whole blood reference material for 30 d (45 ± 0.4 to 46.8 ± 0.1, means ± SDs) compared to untreated saline controls (45.9 ± 1.0 to 6.8 ± 0.2). Pretreatment of paper for blood spots with deferoxamine and drying blood before storage slowed, but not entirely prevented, HUFA degradation over 30 d to 22% and 19% below baseline, respectively, compared to 86-92% in the controls. Protection against HUFA degradation with blood drying and glycerol treatment was confirmed in the biological replicate study and confirmed by prevention of cell lysis. HUFA degradation during storage at -20°C appears to be due to hemolysis and subsequent iron-initiated peroxidation. This degradation may be prevented by glycerol, iron chelation, and/or dried blood spotting. A more thorough understanding of methods to prevent degradation during storage is critical with increasing use of FA profiling in large clinical studies. © 2015 American Society for Nutrition.

  14. Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum.

    PubMed

    Rosypal, Alexa C; Pick, Leanne D; Hernandez, Jaime O Esquivel; Lindsay, David S

    2014-09-15

    Collection of blood samples from veterinary and wildlife patients is often challenging because the samples have to be collected on farm or in the wild under various environmental conditions. This poses many technical problems associated with venipuncture materials, their safe use and disposal, transportation and processing of collected samples. Dried blood spot (DBS) sample collection techniques offer a simple and practical alternative to traditional blood collection methods to obtain blood samples from animals for parasite antibody evaluation. The DBS collection devices are compact, simple to use, and are particularly useful for large number of samples. Additionally, DBS samples take up less space and they are easier to transport than traditional venipuncture-collected blood samples. Visceral leishmaniasis (VL) is a potentially fatal parasitic disease of dogs and humans and it is frequently diagnosed by antibody tests. Immunochromatographic tests (ICT) for antibodies to Leishmania infantum are commercially available for dogs and they produce qualitative results in minutes. Measurement of canine antibodies to L. infantum with the ICT using traditional venipuncture has been validated previously, but the use of DBS samples has not been evaluated using this method. The purpose of the present study was to determine the ability of DBS samples to detect antibodies to L. infantum in dogs using a commercial ICT assay. One hundred plasma samples from dogs experimentally infected with the LIVT-1 strain of L. infantum were collected by venipuncture and frozen. Individual samples were thawed, and then 80 μl plasma (2 drops) was aliquotted onto the 8-spoked disk pad on individual DBS sample collection devices (HemaSpot™, Spot-On Sciences, Austin, TX), dried, and stored in the dark at room temperature. After one month and six months, respectively, 2 spokes of the 8 spokes of the disk pad of each DBS sample were removed and eluted in 200 μl PBS. The eluate was used to test for antibodies in the ICT and compared to ICT results using thawed plasma (same initial source). Sensitivity and specificity of the ICT using DBS were determined by using ICT results from traditional blood collection samples for comparison. After 1 month, DBS samples showed 100% sensitivity and specificity when compared to ICT results on thawed plasma samples collected by traditional venipuncture. After six months storage at room temperature, DBS samples demonstrated 79% sensitivity and 100% specificity compared to traditional blood collection. Results from this study indicate that dried blood spot collection may be a useful tool for screening dogs for antibodies to L. infantum with the ICT assay. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Storage of cell samples for ToF-SIMS experiments-How to maintain sample integrity.

    PubMed

    Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Henss, Anja; Wenisch, Sabine; Janek, Jürgen

    2016-06-25

    In order to obtain comparable and reproducible results from time-of-flight secondary ion mass spectrometry (ToF-SIMS) analysis of biological cells, the influence of sample preparation and storage has to be carefully considered. It has been previously shown that the impact of the chosen preparation routine is crucial. In continuation of this work, the impact of storage needs to be addressed, as besides the fact that degradation will unavoidably take place, the effects of different storage procedures in combination with specific sample preparations remain largely unknown. Therefore, this work examines different wet (buffer, water, and alcohol) and dry (air-dried, freeze-dried, and critical-point-dried) storage procedures on human mesenchymal stem cell cultures. All cell samples were analyzed by ToF-SIMS immediately after preparation and after a storage period of 4 weeks. The obtained spectra were compared by principal component analysis with lipid- and amino acid-related signals known from the literature. In all dry storage procedures, notable degradation effects were observed, especially for lipid-, but also for amino acid-signal intensities. This leads to the conclusion that dried samples are to some extent easier to handle, yet the procedure is not the optimal storage solution. Degradation proceeds faster, which is possibly caused by oxidation reactions and cleaving enzymes that might still be active. Just as well, wet stored samples in alcohol struggle with decreased signal intensities from lipids and amino acids after storage. Compared to that, the wet stored samples in a buffered or pure aqueous environment revealed no degradation effects after 4 weeks. However, this storage bears a higher risk of fungi/bacterial contamination, as sterile conditions are typically not maintained. Thus, regular solution change is recommended for optimized storage conditions. Not directly exposing the samples to air, wet storage seems to minimize oxidation effects, and hence, buffer or water storage with regular renewal of the solution is recommended for short storage periods.

  16. Storage of cell samples for ToF-SIMS experiments—How to maintain sample integrity

    PubMed Central

    Schaepe, Kaija; Kokesch-Himmelreich, Julia; Rohnke, Marcus; Wagner, Alena-Svenja; Schaaf, Thimo; Henss, Anja; Wenisch, Sabine; Janek, Jürgen

    2016-01-01

    In order to obtain comparable and reproducible results from time-of-flight secondary ion mass spectrometry (ToF-SIMS) analysis of biological cells, the influence of sample preparation and storage has to be carefully considered. It has been previously shown that the impact of the chosen preparation routine is crucial. In continuation of this work, the impact of storage needs to be addressed, as besides the fact that degradation will unavoidably take place, the effects of different storage procedures in combination with specific sample preparations remain largely unknown. Therefore, this work examines different wet (buffer, water, and alcohol) and dry (air-dried, freeze-dried, and critical-point-dried) storage procedures on human mesenchymal stem cell cultures. All cell samples were analyzed by ToF-SIMS immediately after preparation and after a storage period of 4 weeks. The obtained spectra were compared by principal component analysis with lipid- and amino acid-related signals known from the literature. In all dry storage procedures, notable degradation effects were observed, especially for lipid-, but also for amino acid-signal intensities. This leads to the conclusion that dried samples are to some extent easier to handle, yet the procedure is not the optimal storage solution. Degradation proceeds faster, which is possibly caused by oxidation reactions and cleaving enzymes that might still be active. Just as well, wet stored samples in alcohol struggle with decreased signal intensities from lipids and amino acids after storage. Compared to that, the wet stored samples in a buffered or pure aqueous environment revealed no degradation effects after 4 weeks. However, this storage bears a higher risk of fungi/bacterial contamination, as sterile conditions are typically not maintained. Thus, regular solution change is recommended for optimized storage conditions. Not directly exposing the samples to air, wet storage seems to minimize oxidation effects, and hence, buffer or water storage with regular renewal of the solution is recommended for short storage periods. PMID:26810048

  17. The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol–chloroform–isoamyl alcohol DNA extraction

    PubMed Central

    Renshaw, Mark A; Olds, Brett P; Jerde, Christopher L; McVeigh, Margaret M; Lodge, David M

    2015-01-01

    Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult or where samples must be transported long distances for further processing and screening. This study demonstrates the successful preservation of eDNA at room temperature (20 °C) in two lysis buffers, CTAB and Longmire's, over a 2-week period of time. Moreover, the preserved eDNA samples were seamlessly integrated into a phenol–chloroform–isoamyl alcohol (PCI) DNA extraction protocol. The successful application of the eDNA extraction to multiple filter membrane types suggests the methods evaluated here may be broadly applied in future eDNA research. Our results also suggest that for many kinds of studies recently reported on macrobial eDNA, detection probabilities could have been increased, and at a lower cost, by utilizing the Longmire's preservation buffer with a PCI DNA extraction. PMID:24834966

  18. Banking of environmental samples for short-term biochemical and chemical monitoring of organic contamination in coastal marine environments: the GICBEM experience (1986-1990). Groupe Interface Chimie Biologie des Ecosystèmes, Marins.

    PubMed

    Garrigues, P; Narbonne, J F; Lafaurie, M; Ribera, D; Lemaire, P; Raoux, C; Michel, X; Salaun, J P; Monod, J L; Romeo, M

    1993-11-01

    The GICBEM (Groupe Interface Chimie Biologie des Ecosystèmes Marins) program consists of an evaluation of the ecosystem health status in the Mediterranean Sea mainly based on chemical and biochemical approaches. Specific chemical contaminants (polycyclic aromatic hydrocarbons (PAH), polychlorobiphenyls (PCB), heavy metals) in waters, sediments, and related biotransformation indicators in target organisms (mussels, fish) have been selected for a complete survey of the coastal waters. In order to provide an appropriate sampling program for standardization for each sampling cruise, various aspects have been studied: (a) parameters for the choice of the sample sites; (b) ways of collection the samples (waters, sediments, marine organisms); and (c) preparation of the samples for a short term storage on board ship and for further analyses in the ground laboratory. Methods of preparation and storage of the samples are described and could be used to initiate an environmental banking program including both possible retrospective analyses of chemical pollutants and biochemical indicators. Moreover, the correlation between chemicals (PAH) and biochemical (mixed function oxygenase activities) parameters has been studied and this demonstrates the capability of the enzyme activities as reliable pollution biomarkers.

  19. Autonomous water sampling for long-term monitoring of trace metals in remote environments.

    PubMed

    Kim, Hyojin; Bishop, James K B; Wood, Todd J; Fung, Inez Y

    2012-10-16

    A remotely controlled autonomous method for long-term high-frequency sampling of environmental waters in remote locations is described. The method which preserves sample integrity of dissolved trace metals and major ions for month-long periods employs a gravitational filtration system (GFS) that separates dissolved and particulate phases as samples are collected. The key elements of GFS are (1) a modified "air-outlet" filter holder to maximize filtration rate and thus minimize filtration artifacts; and (2) the direct delivery of filtrate to dedicated bottle sets for specific analytes. Depth and screen filter types were evaluated with depth filters showing best performance. GFS performance is validated using ground, stream, and estuary waters. Over 30 days of storage, samples with GFS treatment had average recoveries of 95 ± 19% and 105 ± 7% of Fe and Mn, respectively; without GFS treatment, average recoveries were only 16% and 18%. Dissolved major cations K, Mg, and Na were stable independent of collection methodology, whereas Ca in some groundwater samples decreased up to 42% without GFS due to CaCO(3) precipitation. In-field performance of GFS equipped autosamplers is demonstrated using ground and streamwater samples collected at the Angelo Coast Range Reserve, California from October 3 to November 4 2011.

  20. Effect of season on scrotal circumference, semen characteristics, seminal plasma composition and spermatozoa motility during liquid storage in INRA180 rams.

    PubMed

    Benmoula, Anass; Badi, Abdelmoughit; El Fadili, Moussa; El Khalil, Kaoutar; Allai, Larbi; El Hilali, Abderaouf; El Amiri, Bouchra

    2017-05-01

    The present study was undertaken to assess the effect of seasons on scrotal circumference, semen characteristics, seminal plasma composition, and sperm motility during liquid storage of INRA180 rams. The semen was collected from five mature INRA180 rams (2-3 years of age) during one year (from April 2014 to March 2015). Scrotal circumferences, semen characteristics, some biochemical parameters of seminal plasma were evaluated. Immediately after collection and evaluation, the semen was pooled and extended in skim milk (SM) at 15°C to reach 0.8×109 spermatozoa/ml. Thereafter, samples were evaluated at different storage times (0, 8, and 24h). The results showed that scrotal circumference, semen quality and the concentration of total protein in seminal plasma were relatively constant during the year (P>0.05). However, total lipid and cholesterol concentrations increased significantly (P<0.001) in winter and summer. The result showed also that progressive motility was higher in winter and summer after 24h of storage (P<0.01). In contrast, no difference was recorded regarding total motility (P>0.05). To conclude, the INRA180 rams have the ability to produce semen with high quality all over the year. The only parameters showing seasonal variations are cholesterol, total lipid, and progressive motility. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Antioxidant capacity of fresh and stored breast milk: is -80°C optimal temperature for freeze storage?

    PubMed

    Sari, Fatma Nur; Akdag, Arzu; Dizdar, Evrim Alyamac; Uras, Nurdan; Erdeve, Omer; Erel, Ozcan; Dilmen, Ugur

    2012-06-01

    To determine total antioxidant capacity and total oxidation status in fresh and freeze stored (at -80°C) breast milk during the stages of lactation. Samples of colostrum, transitional and mature milk were collected from 44 healthy women at 3, 8 and 30 days after birth. The total milk volume collected (6 ml) was divided in two aliquot parts: 3 ml for the fresh analysis which was done immediately after the extraction and 3 ml for storage under freezing conditions at -80°C for two months. The antioxidant status and oxidative stress of the fresh and stored breast milk were assessed via determination of total antioxidant capacity and total oxidation status. Antioxidant capacity of transitional and mature milk decreased (p = 0.0001, p = 0.028, respectively); however, antioxidant capacity of colostrum did not change by storage at -80°C (p > 0.05). Total antioxidant capacity of fresh and stored breast milk significantly decreased during the stages of lactation (p < 0.0001, p = 0.028, respectively). Total oxidation status showed no significant difference in fresh and stored breast milk during the stages of lactation (p > 0.05). Freeze storage of breast milk at -80°C for two months seems not to be the optimal condition to preserve the antioxidant capacity of breast milk.

  2. Evaluation of dried blood spot as an alternative sample collection method for hepatitis C virus RNA quantitation and genotyping using a commercial system.

    PubMed

    Mahajan, Supriya; Choudhary, Manish Chandra; Kumar, Guresh; Gupta, Ekta

    2018-06-01

    Dried blood spot (DBS) is a minimally invasive sampling method suitable for sample collection, storage and transportation in resource limited areas. Aim of this study was to compare the diagnostic utility of DBS with plasma sample for HCV RNA quantitation and genotyping using commercial systems. Plasma and DBS card spotted samples were collected from 95 HCV seropositive patients. Both types of samples were subjected to HCV RNA by real-time PCR (Abbott m2000rt, USA). Genotyping was performed using Abbott HCV genotype II kit (Abbott diagnostics, USA) in samples with viral load > 3 log 10  IU/ml. In both plasma and DBS, 14 (14.7%) samples were negative and 81 (85.3%) were positive for HCV RNA. Median viral load in plasma (3.78; range 0-7.43) log 10  IU/ml was comparable to DBS (3.93; range 0-7.24) log 10  IU/ml. DBS demonstrated sensitivity and specificity of 97.5 and 85.7% respectively, with positive predictive value (PPV) of 97.5% and negative predictive value (NPV) of 85.7%. DBS showed good correlation (r 2  = 0.866) and agreement (93.5%) with plasma. Genotyping in 20 patients showed 100% concordance between DBS and plasma samples. DBS showed good sensitivity and specificity as a sampling method for HCV RNA quantitation and genotyping.

  3. Analysis of ECs and related compounds in plasma: artifactual isomerization and ex vivo enzymatic generation of 2-MGs.

    PubMed

    Pastor, Antoni; Farré, Magí; Fitó, Montserrat; Fernandez-Aranda, Fernando; de la Torre, Rafael

    2014-05-01

    The analysis of peripheral endocannabinoids (ECs) is a good biomarker of the EC system. Their concentrations, from clinical studies, strongly depend on sample collection and time processing conditions taking place in clinical and laboratory settings. The analysis of 2-monoacylglycerols (MGs) (i.e., 2-arachidonoylglycerol or 2-oleoylglycerol) is a particularly challenging issue because of their ex vivo formation and chemical isomerization that occur after blood sample collection. We provide evidence that their ex vivo formation can be minimized by adding Orlistat, an enzymatic lipase inhibitor, to plasma. Taking into consideration the low cost of Orlistat, we recommend its addition to plasma collecting tubes while maintaining sample cold chain until storage. We have validated a method for the determination of the EC profile of a range of MGs and N-acylethanolamides in plasma that preserves the original isomer ratio of MGs. Nevertheless, the chemical isomerization of 2-MGs can only be avoided by an immediate processing and analysis of samples due to their instability during conservation. We believe that this new methodology can aid in the harmonization of the measurement of ECs and related compounds in clinical samples.

  4. Evaluation of the BD Vacutainer Plus Urine C&S Preservative Tubes compared with nonpreservative urine samples stored at 4°C and room temperature.

    PubMed

    Eisinger, Stephen W; Schwartz, Matthew; Dam, Lisa; Riedel, Stefan

    2013-09-01

    The stability of urine specimens submitted for culture remains a challenge for many laboratories because of delays in specimen transport. We evaluated the usefulness of BD Vacutainer Plus Urine C&S Preservative Tube in ensuring specimen stability. Clinical urine specimens collected in sterile collection cups (n = 110) were plated onto sheep blood and MacConkey agar following standard laboratory procedures guidelines. Thereafter, specimens were divided into 3 storage conditions: nonpreservative, refrigerated; nonpreservative, room temperature (RT); BD Vacutainer Plus Urine C&S Preservative Tube, RT. For each sample type, additional cultures were set up at 2, 4, 24, and 48 hours. Initially, 18 specimens had no growth, 32 showed mixed skin flora, and 60 yielded at least 1 uropathogen. Increased colony counts of uropathogens were observed for nonpreserved urine samples stored at RT; these changes were statistically significant. Minor differences between refrigerated urine samples and BD Vacutainer Plus Urine C&S Preservative Tube samples were seen but were not statistically significant. The use of preservative-containing collection tubes is desirable to ensure specimen stability when prompt processing or refrigeration is not feasible.

  5. Stability of Bovine viral diarrhea virus 1 nucleic acid in fetal bovine samples stored under different conditions.

    PubMed

    Ridpath, Julia F; Neill, John D; Chiang, Yu-Wei; Waldbillig, Jill

    2014-01-01

    Infection of pregnant cattle with both species of Bovine viral diarrhea virus (BVDV) can result in reproductive disease that includes fetal reabsorption, mummification, abortion, stillbirths, congenital defects affecting structural, neural, reproductive, and immune systems, and the birth of calves persistently infected with BVDV. Accurate diagnosis of BVDV-associated reproductive disease is important to control BVDV at the production unit level and assessment of the cost of BVDV infections in support of BVDV control programs. The purpose of the current study was to examine the stability of viral nucleic acid in fetal tissues exposed to different conditions, as measured by detection by polymerase chain reaction. Five different types of fetal tissue, including brain, skin and muscle, ear, and 2 different pooled organ samples, were subjected to conditions that mimicked those that might exist for samples collected after abortions in production settings or possible storage conditions after collection and prior to testing. In addition, tissues were archived for 36 months at -20°C and then retested, to mimic conditions that might occur in the case of retrospective surveillance studies. Brain tissue showed the highest stability under the conditions tested. The impact of fecal contamination was increased following archiving in all tissue types suggesting that, for long-term storage, effort should be made to reduce environmental contaminants before archiving.

  6. Measuring telomere length and telomere dynamics in evolutionary biology and ecology

    PubMed Central

    Nussey, Daniel H; Baird, Duncan; Barrett, Emma; Boner, Winnie; Fairlie, Jennifer; Gemmell, Neil; Hartmann, Nils; Horn, Thorsten; Haussmann, Mark; Olsson, Mats; Turbill, Chris; Verhulst, Simon; Zahn, Sandrine; Monaghan, Pat

    2014-01-01

    Telomeres play a fundamental role in the protection of chromosomal DNA and in the regulation of cellular senescence. Recent work in human epidemiology and evolutionary ecology suggests adult telomere length (TL) may reflect past physiological stress and predict subsequent morbidity and mortality, independent of chronological age. Several different methods have been developed to measure TL, each offering its own technical challenges. The aim of this review is to provide an overview of the advantages and drawbacks of each method for researchers, with a particular focus on issues that are likely to face ecologists and evolutionary biologists collecting samples in the field or in organisms that may never have been studied in this context before. We discuss the key issues to consider and wherever possible try to provide current consensus view regarding best practice with regard to sample collection and storage, DNA extraction and storage, and the five main methods currently available to measure TL. Decisions regarding which tissues to sample, how to store them, how to extract DNA, and which TL measurement method to use cannot be prescribed, and are dependent on the biological question addressed and the constraints imposed by the study system. What is essential for future studies of telomere dynamics in evolution and ecology is that researchers publish full details of their methods and the quality control thresholds they employ. PMID:25834722

  7. Exploring No-SQL alternatives for ALMA monitoring system

    NASA Astrophysics Data System (ADS)

    Shen, Tzu-Chiang; Soto, Ruben; Merino, Patricio; Peña, Leonel; Bartsch, Marcelo; Aguirre, Alvaro; Ibsen, Jorge

    2014-07-01

    The Atacama Large Millimeter /submillimeter Array (ALMA) will be a unique research instrument composed of at least 66 reconfigurable high-precision antennas, located at the Chajnantor plain in the Chilean Andes at an elevation of 5000 m. This paper describes the experience gained after several years working with the monitoring system, which has a strong requirement of collecting and storing up to 150K variables with a highest sampling rate of 20.8 kHz. The original design was built on top of a cluster of relational database server and network attached storage with fiber channel interface. As the number of monitoring points increases with the number of antennas included in the array, the current monitoring system demonstrated to be able to handle the increased data rate in the collection and storage area (only one month of data), but the data query interface showed serious performance degradation. A solution based on no-SQL platform was explored as an alternative to the current long-term storage system. Among several alternatives, mongoDB has been selected. In the data flow, intermediate cache servers based on Redis were introduced to allow faster streaming of the most recently acquired data to web based charts and applications for online data analysis.

  8. Filter paper collection of Plasmodium falciparum mRNA for detecting low-density gametocytes

    PubMed Central

    2012-01-01

    Background Accurate sampling of sub-microscopic gametocytes is necessary for epidemiological studies to identify the infectious reservoir of Plasmodium falciparum. Detection of gametocyte mRNA achieves sensitive detection, but requires careful handling of samples. Filter papers can be used for collecting RNA samples, but rigorous testing of their capacity to withstand adverse storage conditions has not been fully explored. Methods Three gametocyte dilutions: 10/μL, 1.0/μL and 0.1/μL were spotted onto Whatman™ 903 Protein Saver Cards, FTA Classic Cards and 3MM filter papers that were stored under frozen, cold chain or tropical conditions for up to 13 weeks . RNA was extracted, then detected by quantitative nucleic acid sequence-based amplification (QT-NASBA) and reverse-transcriptase PCR (RT-PCR). Results Successful gametocyte detection was more frequently observed from the Whatman 903 Protein Saver Card compared to the Whatman FTA Classic Card, by both techniques (p < 0.0001). When papers were stored at higher temperatures, a loss in sensitivity was experienced for the FTA Classic Card but not the 903 Protein Saver Card or Whatman 3MM filter paper. The sensitivity of gametocyte detection was decreased when papers were stored at high humidity. Conclusions This study indicates the Whatman 903 Protein Saver Card is better for Pfs25 mRNA sampling compared to the Whatman FTA Classic Card, and that the Whatman 3MM filter paper may prove to be a satisfactory cheaper option for Pfs25 mRNA sampling. When appropriately dried, filter papers provide a useful approach to Pfs25 mRNA sampling, especially in settings where storage in RNA-protecting buffer is not possible. PMID:22873569

  9. Filter paper collection of Plasmodium falciparum mRNA for detecting low-density gametocytes.

    PubMed

    Jones, Sophie; Sutherland, Colin J; Hermsen, Cornelus; Arens, Theo; Teelen, Karina; Hallett, Rachel; Corran, Patrick; van der Vegte-Bolmer, Marga; Sauerwein, Robert; Drakeley, Chris J; Bousema, Teun

    2012-08-08

    Accurate sampling of sub-microscopic gametocytes is necessary for epidemiological studies to identify the infectious reservoir of Plasmodium falciparum. Detection of gametocyte mRNA achieves sensitive detection, but requires careful handling of samples. Filter papers can be used for collecting RNA samples, but rigorous testing of their capacity to withstand adverse storage conditions has not been fully explored. Three gametocyte dilutions: 10/μL, 1.0/μL and 0.1/μL were spotted onto Whatman™ 903 Protein Saver Cards, FTA Classic Cards and 3MM filter papers that were stored under frozen, cold chain or tropical conditions for up to 13 weeks . RNA was extracted, then detected by quantitative nucleic acid sequence-based amplification (QT-NASBA) and reverse-transcriptase PCR (RT-PCR). Successful gametocyte detection was more frequently observed from the Whatman 903 Protein Saver Card compared to the Whatman FTA Classic Card, by both techniques (p<0.0001). When papers were stored at higher temperatures, a loss in sensitivity was experienced for the FTA Classic Card but not the 903 Protein Saver Card or Whatman 3MM filter paper. The sensitivity of gametocyte detection was decreased when papers were stored at high humidity. This study indicates the Whatman 903 Protein Saver Card is better for Pfs25 mRNA sampling compared to the Whatman FTA Classic Card, and that the Whatman 3MM filter paper may prove to be a satisfactory cheaper option for Pfs25 mRNA sampling. When appropriately dried, filter papers provide a useful approach to Pfs25 mRNA sampling, especially in settings where storage in RNA-protecting buffer is not possible.

  10. Optimization of analytical and pre-analytical conditions for MALDI-TOF-MS human urine protein profiles.

    PubMed

    Calvano, C D; Aresta, A; Iacovone, M; De Benedetto, G E; Zambonin, C G; Battaglia, M; Ditonno, P; Rutigliano, M; Bettocchi, C

    2010-03-11

    Protein analysis in biological fluids, such as urine, by means of mass spectrometry (MS) still suffers for insufficient standardization in protocols for sample collection, storage and preparation. In this work, the influence of these variables on healthy donors human urine protein profiling performed by matrix assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was studied. A screening of various urine sample pre-treatment procedures and different sample deposition approaches on the MALDI target was performed. The influence of urine samples storage time and temperature on spectral profiles was evaluated by means of principal component analysis (PCA). The whole optimized procedure was eventually applied to the MALDI-TOF-MS analysis of human urine samples taken from prostate cancer patients. The best results in terms of detected ions number and abundance in the MS spectra were obtained by using home-made microcolumns packed with hydrophilic-lipophilic balance (HLB) resin as sample pre-treatment method; this procedure was also less expensive and suitable for high throughput analyses. Afterwards, the spin coating approach for sample deposition on the MALDI target plate was optimized, obtaining homogenous and reproducible spots. Then, PCA indicated that low storage temperatures of acidified and centrifuged samples, together with short handling time, allowed to obtain reproducible profiles without artifacts contribution due to experimental conditions. Finally, interesting differences were found by comparing the MALDI-TOF-MS protein profiles of pooled urine samples of healthy donors and prostate cancer patients. The results showed that analytical and pre-analytical variables are crucial for the success of urine analysis, to obtain meaningful and reproducible data, even if the intra-patient variability is very difficult to avoid. It has been proven how pooled urine samples can be an interesting way to make easier the comparison between healthy and pathological samples and to individuate possible differences in the protein expression between the two sets of samples. Copyright 2009 Elsevier B.V. All rights reserved.

  11. Introduction to Field Water-Quality Methods for the Collection of Metals - 2007 Project Summary

    USGS Publications Warehouse

    Allen, Monica L.

    2008-01-01

    The U.S. Geological Survey (USGS), Region VI of the U.S. Environmental Protection Agency (USEPA), and the Osage Nation presented three 3-day workshops, in June-August 2007, entitled ?Introduction to Field Water-Quality Methods for the Collection of Metals.? The purpose of the workshops was to provide instruction to tribes within USEPA Region VI on various USGS surface-water measurement methods and water-quality sampling protocols for the collection of surface-water samples for metals analysis. Workshop attendees included members from over 22 tribes and pueblos. USGS instructors came from Oklahoma, New Mexico, and Georgia. Workshops were held in eastern and south-central Oklahoma and New Mexico and covered many topics including presampling preparation, water-quality monitors, and sampling for metals in surface water. Attendees spent one full classroom day learning the field methods used by the USGS Water Resources Discipline and learning about the complexity of obtaining valid water-quality and quality-assurance data. Lectures included (1) a description of metal contamination sources in surface water; (2) introduction on how to select field sites, equipment, and laboratories for sample analysis; (3) collection of sediment in surface water; and (4) utilization of proper protocol and methodology for sampling metals in surface water. Attendees also were provided USGS sampling equipment for use during the field portion of the class so they had actual ?hands-on? experience to take back to their own organizations. The final 2 days of the workshop consisted of field demonstrations of current USGS water-quality sample-collection methods. The hands-on training ensured that attendees were exposed to and experienced proper sampling procedures. Attendees learned integrated-flow techniques during sample collection, field-property documentation, and discharge measurements and calculations. They also used enclosed chambers for sample processing and collected quality-assurance samples to verify their techniques. Benefits of integrated water-quality sample-collection methods are varied. Tribal environmental programs now have the ability to collect data that are comparable across watersheds. The use of consistent sample collection, manipulation, and storage techniques will provide consistent quality data that will enhance the understanding of local water resources. The improved data quality also will help the USEPA better document the condition of the region?s water. Ultimately, these workshops equipped tribes to use uniform sampling methods and to provide consistent quality data that are comparable across the region.

  12. High Burnup Dry Storage Cask Research and Development Project, Final Test Plan

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

    None

    2014-02-27

    EPRI is leading a project team to develop and implement the first five years of a Test Plan to collect data from a SNF dry storage system containing high burnup fuel.12 The Test Plan defined in this document outlines the data to be collected, and the storage system design, procedures, and licensing necessary to implement the Test Plan.13 The main goals of the proposed test are to provide confirmatory data14 for models, future SNF dry storage cask design, and to support license renewals and new licenses for ISFSIs. To provide data that is most relevant to high burnup fuel inmore » dry storage, the design of the test storage system must mimic real conditions that high burnup SNF experiences during all stages of dry storage: loading, cask drying, inert gas backfilling, and transfer to the ISFSI for multi-year storage.15 Along with other optional modeling, SETs, and SSTs, the data collected in this Test Plan can be used to evaluate the integrity of dry storage systems and the high burnup fuel contained therein over many decades. It should be noted that the Test Plan described in this document discusses essential activities that go beyond the first five years of Test Plan implementation.16 The first five years of the Test Plan include activities up through loading the cask, initiating the data collection, and beginning the long-term storage period at the ISFSI. The Test Plan encompasses the overall project that includes activities that may not be completed until 15 or more years from now, including continued data collection, shipment of the Research Project Cask to a Fuel Examination Facility, opening the cask at the Fuel Examination Facility, and examining the high burnup fuel after the initial storage period.« less

  13. Enrichment of free-living amoebae in biofilms developed at upper water levels in drinking water storage towers: An inter- and intra-seasonal study.

    PubMed

    Taravaud, Alexandre; Ali, Myriam; Lafosse, Bernard; Nicolas, Valérie; Féliers, Cédric; Thibert, Sylvie; Lévi, Yves; Loiseau, Philippe M; Pomel, Sébastien

    2018-08-15

    Free-living amoebae (FLA) are ubiquitous organisms present in various natural and artificial environments, such as drinking water storage towers (DWST). Some FLA, such as Acanthamoeba sp., Naegleria fowleri, and Balamuthia mandrillaris, can cause severe infections at ocular or cerebral level in addition to being potential reservoirs of other pathogens. In this work, the abundance and diversity of FLA was evaluated in two sampling campaigns: one performed over five seasons in three DWST at three different levels (surface, middle and bottom) in water and biofilm using microscopy and PCR, and one based on the kinetics analysis in phase contrast and confocal microscopy of biofilm samples collected every two weeks during a 3-month period at the surface and at the bottom of a DWST. In the seasonal study, the FLA were detected in each DWST water in densities of ~20 to 25amoebaeL -1 . A seasonal variation of amoeba distribution was observed in water samples, with maximal densities in summer at ~30amoebaeL -1 and minimal densities in winter at ~16amoebaeL -1 . The FLA belonging to the genus Acanthamoeba were detected in two spring sampling campaigns, suggesting a possible seasonal appearance of this potentially pathogenic amoeba. Interestingly, a 1 log increase of amoebae density was observed in biofilm samples collected at the surface of all DWST compared to the middle and the bottom where FLA were at 0.1-0.2amoebae/cm 2 . In the kinetics study, an increase of amoebae density, total cell density, and biofilm thickness was observed as a function of time at the surface of the DWST, but not at the bottom. To our knowledge, this study describes for the first time a marked higher FLA density in biofilms collected at upper water levels in DWST, constituting a potential source of pathogenic micro-organisms. Copyright © 2018 Elsevier B.V. All rights reserved.

  14. Evaluation of human papillomavirus detection by Abbott m2000 system on samples collected by FTA Elute™ Card in a Chinese HIV-1 positive population.

    PubMed

    Qin, Yu; Zhang, Hongyun; Marlowe, Natalia; Fei, Mandong; Yu, Judy; Lei, Xiaoqin; Yu, Lulu; Zhang, Jia; Cao, Di; Ma, Li; Chen, Wen

    2016-12-01

    HIV+/AIDS women have an increased risk of developing into CIN and cervical cancer compared to the general population. Limited medical resource and the lack of AIDS relevant knowledge impair the coverage and efficiency of cervical cancer screening. To compare the clinical performance of self-collected dry storage medium (FTA Elute card) and physician-collected PreservCyt medium in detection of high risk human papillomavirus (HR HPV) among HIV-1 positive population. Three hundred HIV-1 positive women (aged 25-65) were recruited from Yunnan infectious hospital. Two cervicovaginal samples were collected from each participant: one was collected by the women themselves and applied on a FTA Elute card; the other one was collected by a physician and stored in PreservCyt solution. All the samples were tested for 14 HR HPV using Abbott RealTime High Risk HPV assay. Biopsies were taken for histological diagnosis if any abnormal impression was noticed under colposcopy. 291 (97.0%) of participants were eligible for this study. 101 (34.70%) participants were found HR HPV positive in both FTA card and PreservCyt samples, and 19 (6.53%) women were diagnosed as CIN2+. The HR HPV positive rate on samples collected by FTA Elute card and PreservCyt solution was 42.61% and 39.86%, respectively. The overall agreement was 87% (kappa=0.731) between FTA card and PreservCyt. The clinical sensitivity and specificity of FTA card and PreservCyt were 100%, 61.39% and 100%, 64.33%, respectively. In this study, FTA Elute card demonstrated a good performance on self-collected sample for HR HPV detection in HIV-1 positive population. For the women from low-resource area with HIV-1 infection, FTA Elute card could be an attractive sample collection method for cervical cancer screening. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Optimized Standard Operating Procedures for the Analysis of Cerebrospinal Fluid Aβ42 and the Ratios of Aβ Isoforms Using Low Protein Binding Tubes.

    PubMed

    Vanderstichele, Hugo Marcel Johan; Janelidze, Shorena; Demeyer, Leentje; Coart, Els; Stoops, Erik; Herbst, Victor; Mauroo, Kimberley; Brix, Britta; Hansson, Oskar

    2016-05-31

    Reduced cerebrospinal fluid (CSF) concentration of amyloid-β1-42 (Aβ1-42) reflects the presence of amyloidopathy in brains of subjects with Alzheimer's disease (AD). To qualify the use of Aβ1-42/Aβ1-40 for improvement of standard operating procedures (SOP) for measurement of CSF Aβ with a focus on CSF collection, storage, and analysis. Euroimmun ELISAs for CSF Aβ isoforms were used to set up a SOP with respect to recipient properties (low binding, polypropylene), volume of tubes, freeze/thaw cycles, addition of detergents (Triton X-100, Tween-20) in collection or storage tubes or during CSF analysis. Data were analyzed with linear repeated measures and mixed effects models. Optimization of CSF analysis included a pre-wash of recipients (e.g., tubes, 96-well plates) before sample analysis. Using the Aβ1-42/Aβ1-40 ratio, in contrast to Aβ1-42, eliminated effects of tube type, additional freeze/thaw cycles, or effect of CSF volumes for polypropylene storage tubes. 'Low binding' tubes reduced the loss of Aβ when aliquoting CSF or in function of additional freeze/thaw cycles. Addition of detergent in CSF collection tubes resulted in an almost complete absence of variation in function of collection procedures, but affected the concentration of Aβ isoforms in the immunoassay. The ratio of Aβ1-42/Aβ1-40 is a more robust biomarker than Aβ1-42 toward (pre-) analytical interfering factors. Further, 'low binding' recipients and addition of detergent in collection tubes are able to remove effects of SOP-related confounding factors. Integration of the Aβ1-42/Aβ1-40 ratio and 'low-binding tubes' into guidance criteria may speed up worldwide standardization of CSF biomarker analysis.

  16. Tracer techniques for urine volume determination and urine collection and sampling back-up system

    NASA Technical Reports Server (NTRS)

    Ramirez, R. V.

    1971-01-01

    The feasibility, functionality, and overall accuracy of the use of lithium were investigated as a chemical tracer in urine for providing a means of indirect determination of total urine volume by the atomic absorption spectrophotometry method. Experiments were conducted to investigate the parameters of instrumentation, tracer concentration, mixing times, and methods for incorporating the tracer material in the urine collection bag, and to refine and optimize the urine tracer technique to comply with the Skylab scheme and operational parameters of + or - 2% of volume error and + or - 1% accuracy of amount of tracer added to each container. In addition, a back-up method for urine collection and sampling system was developed and evaluated. This back-up method incorporates the tracer technique for volume determination in event of failure of the primary urine collection and preservation system. One chemical preservative was selected and evaluated as a contingency chemical preservative for the storage of urine in event of failure of the urine cooling system.

  17. Understanding biological mechanisms underlying adverse birth outcomes in developing countries: protocol for a prospective cohort (AMANHI bio-banking) study.

    PubMed

    Baqui, Abdullah H; Khanam, Rasheda; Rahman, Mohammad Sayedur; Ahmed, Aziz; Rahman, Hasna Hena; Moin, Mamun Ibne; Ahmed, Salahuddin; Jehan, Fyezah; Nisar, Imran; Hussain, Atiya; Ilyas, Muhammad; Hotwani, Aneeta; Sajid, Muhammad; Qureshi, Shahida; Zaidi, Anita; Sazawal, Sunil; Ali, Said M; Deb, Saikat; Juma, Mohammed Hamad; Dhingra, Usha; Dutta, Arup; Ame, Shaali Makame; Hayward, Caroline; Rudan, Igor; Zangenberg, Mike; Russell, Donna; Yoshida, Sachiyo; Polašek, Ozren; Manu, Alexander; Bahl, Rajiv

    2017-12-01

    The AMANHI study aims to seek for biomarkers as predictors of important pregnancy-related outcomes, and establish a biobank in developing countries for future research as new methods and technologies become available. AMANHI is using harmonised protocols to enrol 3000 women in early pregnancies (8-19 weeks of gestation) for population-based follow-up in pregnancy up to 42 days postpartum in Bangladesh, Pakistan and Tanzania, with collection taking place between August 2014 and June 2016. Urine pregnancy tests will be used to confirm reported or suspected pregnancies for screening ultrasound by trained sonographers to accurately date the pregnancy. Trained study field workers will collect very detailed phenotypic and epidemiological data from the pregnant woman and her family at scheduled home visits during pregnancy (enrolment, 24-28 weeks, 32-36 weeks & 38+ weeks) and postpartum (days 0-6 or 42-60). Trained phlebotomists will collect maternal and umbilical blood samples, centrifuge and obtain aliquots of serum, plasma and the buffy coat for storage. They will also measure HbA1C and collect a dried spot sample of whole blood. Maternal urine samples will also be collected and stored, alongside placenta, umbilical cord tissue and membrane samples, which will both be frozen and prepared for histology examination. Maternal and newborn stool (for microbiota) as well as paternal and newborn saliva samples (for DNA extraction) will also be collected. All samples will be stored at -80°C in the biobank in each of the three sites. These samples will be linked to numerous epidemiological and phenotypic data with unique study identification numbers. AMANHI biobank proves that biobanking is feasible to implement in LMICs, but recognises that biobank creation is only the first step in addressing current global challenges.

  18. Influence of storage environment, maturity stage and pre-storage disinfection treatments on tomato fruit quality during winter in KwaZulu-Natal, South Africa.

    PubMed

    Tolesa, G N; Workneh, T S

    2017-09-01

    The aim of this study was to explore influence of evaporative cooling (EC), pre-storage disinfection treatments and maturity stage at harvest on postharvest quality of tomato fruit. The tomato samples ( Lycopersicon esculentum Mill. cv. Nemonetta) were harvested, stored for 28 days and data were collected every seven days. The pH, total titratable acidity (TTA), total soluble solids (TSS), firmness, colour, weight loss (PWL) and marketability percentage were analysed. The temperature difference between ambient storage and EC at the fan varied between 4 and 7 °C, the relative humidity (RH) varied between 31 and 86%, while at different locations inside the EC it varied between 2-3 °C and 5-8%, respectively. Maturity had significant influence on the overall quality of tomatoes. The pH value of green, pink and red tomato was 4.86 and 5.03. The TTA content, the TSS content significantly affected over the 14 days of storage. TSS:TA was found to be in the range of 7.8-33.9. The EC storage shows a higher firmness and hue angle, when compared to the ambient conditions stored tomatoes. Compared to ambient storage, EC storage reduced the PWL by 7-10% over 30 days, while ambient storage took 15 days. EC storage and pre-storage treatments improved the shelf-life and marketability of tomatoes. However, variation in temperature and RH inside EC could affect the storability of the produce.

  19. Changes in bacteria recoverable from subsurface volcanic rock samples during storage at 4{degrees}C

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

    Haldeman, D.L.; Amy, P.S.; White, D.C.

    1994-08-01

    The abundance of viable microorganisms recovered from deep subsurface volcanic rock samples increased after rock perturbation and storage for 1 week at 4{degrees}C, while the diversity and evenness of recoverable heterotrophic bacterial communities generally decreased. One sample of each morphologically distinct colony type, recovered both before and after storage of U12n rock samples, was purified and characterized by fatty acid methyl ester (MIDI) and API rapid NFT strips. As determined by MIDI cluster analysis, the composition of the recoverable microbial communities changed with storage of rock samples; some groups of organisms were recovered only before, only after, or at bothmore » sample times. In general, the isolates recovered only after storage of rock samples had a greater ability to utilize the carbohydrates included in API test strips and had faster generation times than isolates recovered only on initial plating. The nutritional versatility and faster growth rates of organisms recovered in higher proportions after sample storage provide evidence that some microbial community changes may be due to the proliferation of a few bacterial types. However, because some new genera are recovered only after storage, the possibility also exists that dormant bacterial types are resuscitated during sample perturbation and storage. 30 refs., 1 fig., 5 tabs.« less

  20. Uranium Enrichment Safeguards

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

    Demuth, Scott F.; Trahan, Alexis Chanel

    2017-06-26

    DIV of facility layout, material flows, and other information provided in the DIQ. Material accountancy through an annual PIV and a number of interim inventory verifications, including UF6 cylinder identification and counting, NDA of cylinders, and DA on a sample collection of UF6. Application of C/S technologies utilizing seals and tamper-indicating devices (TIDs) on cylinders, containers, storage rooms, and IAEA instrumentation to provide continuity of knowledge between inspection. Verification of the absence of undeclared material and operations, especially HEU production, through SNRIs, LFUA of cascade halls, and environmental swipe sampling

  1. The nail and hair in forensic science.

    PubMed

    Daniel, C Ralph; Piraccini, Bianca Maria; Tosti, Antonella

    2004-02-01

    Drugs, chemicals, and biological substances accumulate and are stored in hair and nails where they can be detected and measured. Advantages of analyzing hair and nail samples also include their easy and non-invasive collection, the small sample size required for analysis, and their easy storage at room temperature. We report 3 examples of heavy metal poisoning diagnosed because of the hair or nail symptoms. Drugs and toxins that can be detected in hair and nails are reviewed and the application of hair/nail analysis in general and in forensic medicine is discussed.

  2. Human Milk Adrenomedullin Is Unstable During Cold Storage at 4°C.

    PubMed

    Peila, Chiara; Coscia, Alessandra; Bertino, Enrico; Li Volti, Giovanni; Galvano, Fabio; Barbagallo, Ignazio; Gazzolo, Diego

    2017-11-01

    Under some circumstances human milk (HM) extraction and refrigerated storage may be necessary. Depending on the length and on the type of cold storage, milk may lose some important properties, but current advices on safe HM storage are discordant. Moreover until now no data in literature were present on the effect of prolonged cold storage on biologically active components of the HM such as adrenomedullin (AM). This important peptide is involved in response to hypoxia and inflammation, associated with neovascularization, in several tissues. The aim is to evaluate: (a) the presence of AM in preterm and term HM and (b) the concentration of AM in refrigerated milk at 4°C at 24-hour intervals, up to 96 hours of storage. The experiment was repeated four times. Immediately after collection, each HM sample deriving from each mother was divided into two parts as follows: "Pool" line and "Single Mother" line. One part (Pool line) was pooled and then divided into five aliquots. The other part (Single Mother line) was divided into five aliquots. From each line, one aliquot was analyzed within 3 hours, while the others were stored in the refrigerator for 24, 48, 72, and 96 hours, respectively, and then analyzed. AM levels were determined using a specific ELISA test. AM was detectable in all samples. Its concentration was significantly higher in preterm milk with respect to term milk (p < 0.05). Significant differences were observed during the cold storage: the AM levels decreased steadily during the storage and the remaining concentration at 96 hours is ∼2%. This study provides evidences regarding the presence of AM in HM, regardless of the gestational age. In particular, the refrigeration of fresh HM in controlled conditions significantly affected its bioactivity and nutritional quality related with AM, already at 24 hours.

  3. Agreement for HPV genotyping detection between self-collected specimens on a FTA cartridge and clinician-collected specimens

    PubMed Central

    Guan, YaoYao; Gravitt, Patti E.; Howard, Roslyn; Eby, Yolanda J.; Wang, Shaoming; Li, Belinda; Feng, Changyan; Qiao, You-Lin; Castle, Philip E.

    2016-01-01

    The current method of transporting self-collected cervicovaginal specimen for HPV DNA testing relies on liquid based medium, which is challenging and expensive to transport. A novel, dry storage and transportation device, Whatman indicating FTA™ Elute Cartridge, avoids some of the pitfalls of liquid-based medium. This method has been shown to be comparable to liquid-based collection medium, but relative performance of self-collected (SC) and clinician-collected (CC) samples onto FTA cards has not been reported. The objective of this study is to compare the analytic performance of self- and clinician-collected samples onto FTA cartridges for the detection of carcinogenic HPV using Linear Array. There was a 91% agreement, 69% positive agreement, and kappa of 0.75 between the clinician-collected and self-collected specimens for detection of any carcinogenic HPV genotype. When the HPV results were categorized hierarchically according to cervical cancer risk, there was no difference in the distribution of the HPV results for the clinician- and self-collected specimens (p = 0.7). This study concludes that FTA elute cartridge is a promising method of specimen transport for cervical cancer screening programs considering using self-collected specimen and HPV testing. Larger studies with clinical endpoints are now needed to assess the clinical performance. PMID:23370404

  4. Metabolomics of AS-1 RBC storage

    PubMed Central

    Roback, John D.; Josephson, Cassandra D.; Waller, Edmund K.; Newman, James L.; Karatela, Sulaiman; Uppal, Karan; Jones, Dean; Zimring, James C.; Dumont, Larry J.

    2014-01-01

    Background Population based investigations suggest that red blood cells (RBCs) are therapeutically effective when collected, processed and stored for up to 42 days under validated conditions prior to transfusion. However, some retrospective clinical studies have shown worse patient outcomes when transfused RBCs have been stored for the longest times. Furthermore, studies of RBC persistence in the circulation after transfusion have suggested that considerable donor-to-donor variability exists, and may affect transfusion efficacy. To understand the limitations of current blood storage technologies and to develop approaches to improve RBC storage and transfusion efficacy, we investigated the global metabolic alterations that occur when RBCs are stored in AS-1 (AS1-RBC). Methods Leukoreduced AS1-RBC units prepared from 9 volunteer research donors (12 total donated units) were serially sampled for metabolomics analysis over 42 days of refrigerated storage. Samples were tested by GC/MS and LC/MS/MS, and specific biochemical compounds were identified by comparison to a library of purified standards. Results Over three experiments, 185–264 defined metabolites were quantified in stored RBC samples. Kinetic changes in these biochemicals confirmed known alterations in glycolysis and other pathways previously identified in RBCs stored in SAGM (SAGM-RBC). Furthermore, we identified additional alterations not previously seen in SAGM-RBCs (e.g., stable pentose phosphate pathway flux, progressive decreases in oxidized glutathione), and we delineated changes occurring in other metabolic pathways not previously studied (e.g., S-adenosyl methionine cycle). These data are presented in the context of a detailed comparison with previous studies of SAGM-RBCs from human donors and murine AS1-RBCs. Conclusion Global metabolic profiling of AS1-RBCs revealed a number of biochemical alterations in stored blood that may affect RBC viability during storage as well as therapeutic effectiveness of stored RBCs in transfusion recipients. Significance These results provide future opportunities to more clearly pinpoint the metabolic defects during RBC storage, to identify biomarkers for donor screening and prerelease RBC testing, and to develop improved RBC storage solutions and methodologies. PMID:24636780

  5. Customized electric power storage device for inclusion in a collective microgrid

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

    Robinett, III, Rush D.; Wilson, David G.; Goldsmith, Steven Y.

    An electric power storage device is described herein, wherein the electric power storage device is included in a microgrid. The electric power storage device has at least one of a charge rate, a discharge rate, or a power retention capacity that has been customized for a collective microgrid. The collective microgrid includes at least two connected microgrids. The at least one of the charge rate, the discharge rate, or the power retention capacity of the electric power storage device is computed based at least in part upon specified power source parameters in the at least two connected microgrids and specifiedmore » load parameters in the at least two connected microgrids.« less

  6. 28 CFR 115.289 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Data Collection and Review § 115.289 Data storage, publication, and destruction. (a) The agency shall ensure that data collected...

  7. 28 CFR 115.289 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Data Collection and Review § 115.289 Data storage, publication, and destruction. (a) The agency shall ensure that data collected...

  8. 28 CFR 115.289 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Community Confinement Facilities Data Collection and Review § 115.289 Data storage, publication, and destruction. (a) The agency shall ensure that data collected...

  9. Perceptions of firearms and suicide: The role of misinformation in storage practices and openness to means safety measures.

    PubMed

    Anestis, Michael D; Butterworth, Sarah E; Houtsma, Claire

    2018-02-01

    Firearm ownership and unsafe storage increase risk for suicide. Little is known regarding factors that influence storage practices and willingness to engage in means safety. Utilizing Amazon's Mechanical Turk program, we recruited an online sample of 300 adults living in the US who own at least one firearm. Firearm storage practices and openness to means safety measures were assessed using items designed for this study. Data were collected and analyzed in 2017. Firearms stored in non-secure locations and without a locking device were associated with lower beliefs in the relationship between firearm storage and suicide risk. Fearlessness about death moderated the association between current secure versus non-secure storage and beliefs regarding firearm storage and suicide risk, in that storage practices and beliefs were more strongly related at higher levels of fearlessness about death. For both secure and locked storage of a firearm, there was a significant indirect effect of current storage practices on willingness to engage in means safety in the future through current beliefs regarding the relationship between firearm storage and suicide risk. Unsafe storage practices were largely associated with an unwillingness to store firearms more safely or to allow a trusted peer to temporarily store the firearm outside the home in order to prevent their own or someone else's suicide. Self-report and cross-sectional data were used. Results may not generalize to non-firearm owners. Firearm owners are prone to inaccurate beliefs about the relationship between firearms and suicide. These beliefs may influence both current firearm storage practices and the willingness to engage in means safety. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. Quality control of human tissues--experience from the Indiana University Cancer Center-Lilly Research Labs human tissue bank.

    PubMed

    Sandusky, George E; Teheny, Katie Heinz; Esterman, Mike; Hanson, Jeff; Williams, Stephen D

    2007-01-01

    The success of molecular research and its applications in both the clinical and basic research arenas is strongly dependent on the collection, handling, storage, and quality control of fresh human tissue samples. This tissue bank was set up to bank fresh surgically obtained human tissue using a Clinical Annotated Tissue Database (CATD) in order to capture the associated patient clinical data and demographics using a one way patient encryption scheme to protect patient identification. In this study, we determined that high quality of tissue samples is imperative for both genomic and proteomic molecular research. This paper also contains a brief compilation of the literature involved in the patient ethics, patient informed consent, patient de-identification, tissue collection, processing, and storage as well as basic molecular research generated from the tissue bank using good clinical practices. The current applicable rules, regulations, and guidelines for handling human tissues are briefly discussed. More than 6,610 cancer patients have been consented (97% of those that were contacted by the consenter) and 16,800 tissue specimens have been banked from these patients in 9 years. All samples collected in the bank were QC'd by a pathologist. Approximately 1,550 tissue samples have been requested for use in basic, clinical, and/or biomarker cancer research studies. Each tissue aliquot removed from the bank for a research study were evaluated by a second H&E, if the samples passed the QC, they were submitted for genomic and proteomic molecular analysis/study. Approximately 75% of samples evaluated were of high histologic quality and used for research studies. Since 2003, we changed the patient informed consent to allow the tissue bank to gather more patient clinical follow-up information. Ninety two percent of the patients (1,865 patients) signed the new informed consent form and agreed to be re-contacted for follow-up information on their disease state. In addition, eighty five percent of patients (1,584) agreed to be re-contacted to provide a biological fluid sample to be used for biomarker research.

  11. Selection of Russian Plutonium Beryllium Sources for Inclusion in the Nuclear Mateirals Information Program Archive

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

    Narlesky, Joshua E; Padilla, Dennis D; Watts, Joe

    2009-01-01

    Throughout the 1960s and 1970s, the former Soviet Union produced and exported Plutonium-Beryllium (PuBe) neutron sources to various Eastern European countries. The Russian sources consist of an intermetallic compound of plutonium and beryllium encapsulated in an inner welded, sealed capsule and consisting of a body and one or more covers. The amount of plutonium in the sources ranges from 0.002 g up to 15 g. A portion of the sources was originally exported to East Germany. A portion of these sources were acquired by Los Alamos National Laboratory (LANL) in the late 1990s for destruction in the Offsite Source Recoverymore » Program. When the OSRP was canceled, the remaining 88 PuBe neutron sources were packaged and stored in a 55-gal drum at T A-55. This storage configuration is no longer acceptable for PuBe sources, and the sources must either be repackaged or disposed of. Repackaging would place the sources into Hagan container, and depending on the dose rates, some sources may be packaged individually increasing the footprint and cost of storage. In addition, each source will be subject to leak-checking every six months. Leaks have already been detected in some of the sources, and due to the age of these sources, it is likely that additional leaks may be detected over time, which will increase the overall complexity of handling and storage. Therefore, it was decided that the sources would be disposed of at the Waste Isolation Pilot Plant (WIPP) due to the cost and labor associated with continued storage at TA-55. However, the plutonium in the sources is of Russian origin and needs to be preserved for research purposes. Therefore, it is important that a representative sample of the sources retained and archived for future studies. This report describes the criteria used to obtain a representative sample of the sources. Nine Russian PuBe neutron sources have been selected out of a collection of 77 sources for inclusion in the NMIP archive. Selection criteria were developed so that the largest sources that are representative of the collection are included. One representative source was chosen for every 20 sources in the collection, and effort was made to preserve sources unique to the collection. In total, four representative sources and five unique sources were selected for the archive. The archive samples contain 40 grams of plutonium with an isotopic composition similar to that of weapon grade material and three grams of plutonium with an isotopic composition similar to that of reactor grade plutonium.« less

  12. Design and calibration of a vacuum compatible scanning tunneling microscope

    NASA Technical Reports Server (NTRS)

    Abel, Phillip B.

    1990-01-01

    A vacuum compatible scanning tunneling microscope was designed and built, capable of imaging solid surfaces with atomic resolution. The single piezoelectric tube design is compact, and makes use of sample mounting stubs standard to a commercially available surface analysis system. Image collection and display is computer controlled, allowing storage of images for further analysis. Calibration results from atomic scale images are presented.

  13. Effects of Storage on Sediment Toxicity, Bioaccumulation Potential, and Chemistry

    DTIC Science & Technology

    1991-01-01

    and tested with organisms used by the US Army Engineer District, New York. Test sedi- ments were collected from Westchester Creek (WC), Gowanus Bay ...Ms. Carole Brown, ERSD, obtained the sediment samples. Dr. Eric Crecelius, Battelle Pacific Northwest Laboratories, Sequim , WA, coordinated chemical...other sites, suspected of containing contam- inated sediment, were Westchester Creek (WC), Gowanus Bay (GB), and Arthur Kill (AK), all located in

  14. Viscous-sludge sample collector

    DOEpatents

    Not Available

    1979-01-01

    A vertical core sample collection system for viscous sludge is disclosed. A sample tube's upper end has a flange and is attached to a piston. The tube and piston are located in the upper end of a bore in a housing. The bore's lower end leads outside the housing and has an inwardly extending rim. Compressed gas, from a storage cylinder, is quickly introduced into the bore's upper end to rapidly accelerate the piston and tube down the bore. The lower end of the tube has a high sludge entering velocity to obtain a full-length sludge sample without disturbing strata detail. The tube's downward motion is stopped when its upper end flange impacts against the bore's lower end inwardly extending rim.

  15. Viscous sludge sample collector

    DOEpatents

    Beitel, George A [Richland, WA

    1983-01-01

    A vertical core sample collection system for viscous sludge. A sample tube's upper end has a flange and is attached to a piston. The tube and piston are located in the upper end of a bore in a housing. The bore's lower end leads outside the housing and has an inwardly extending rim. Compressed gas, from a storage cylinder, is quickly introduced into the bore's upper end to rapidly accelerate the piston and tube down the bore. The lower end of the tube has a high sludge entering velocity to obtain a full-length sludge sample without disturbing strata detail. The tube's downward motion is stopped when its upper end flange impacts against the bore's lower end inwardly extending rim.

  16. Characterization of Qatar's surface carbonates for CO2 capture and thermochemical energy storage

    NASA Astrophysics Data System (ADS)

    Kakosimos, Konstantinos E.; Al-Haddad, Ghadeer; Sakellariou, Kyriaki G.; Pagkoura, Chrysa; Konstandopoulos, Athanasios G.

    2017-06-01

    Samples of surface carbonates were collected from three different areas of the Qatar peninsula. We employed material characterization techniques to examine the morphology and composition of the samples, while their CO2 capture capacity was assessed via multiple successive calcination-carbonation cycles. Our samples were mainly calcite and dolomite based. Calcite samples showed higher initial capacity of around 11 mmol CO2 g-1 which decayed rapidly to less than 2 mmol CO2 g-1. On the other hand, dolomite samples showed an excellent stability (˜15 cycles) with a capacity of 6 mmol CO2 g-1. The performance of the dolomite samples is better compared to other similar natural samples, from literature. A promising result for future studies towards improving their performance by physical and chemical modification.

  17. Gas analyses from the Pu'u O'o eruption in 1985, Kilauea volcano, Hawaii

    USGS Publications Warehouse

    Greenland, L.P.

    1986-01-01

    Volcanic gas samples were collected from July to November 1985 from a lava pond in the main eruptive conduit of Pu'u O'o from a 2-week-long fissure eruption and from a minor flank eruption of Pu'u O'o. The molecular composition of these gases is consistent with thermodynamic equilibrium at a temperature slightly less than measured lava temperatures. Comparison of these samples with previous gas samples shows that the composition of volatiles in the magma has remained constant over the 3-year course of this episodic east rift eruption of Kilauea volcano. The uniformly carbon depleted nature of these gases is consistent with previous suggestions that all east rift eruptive magmas degas during prior storage in the shallow summit reservoir of Kilauea. Minor compositional variations within these gas collections are attributed to the kinetics of the magma degassing process. ?? 1986 Springer-Verlag.

  18. Sarcomere length influences postmortem proteolysis of excised bovine semitendinosus muscle.

    PubMed

    Weaver, A D; Bowker, B C; Gerrard, D E

    2008-08-01

    The interaction between sarcomere length and postmortem proteolysis as related to meat tenderness is not clear. The extent of thick and thin filament overlap alters actomyosin binding and may alter substrate availability during aging-induced tenderization. The objective of this study was to determine the influence of sarcomere length on proteolytic degradation in beef. Strips from bovine semitendinosus were either stretched 40% and restrained or allowed to shorten unrestrained in an ice bath. After rigor completion, 0.6-cm cross sections were fabricated and were randomly assigned to 2, 4, 7, or 10 d of aging treatments. Myofibrils were isolated for sarcomere length determination. Samples were collected and frozen for shear force analysis, and muscle proteins were extracted for SDS-PAGE and Western blotting analyses to determine troponin T (TnT) proteolysis. Sarcomere length was greater (P < 0.01) in stretched muscle samples compared with shortened samples (2.57 vs. 1.43 microm, respectively). Correspondingly, shear force values were greater (P < 0.05) in shortened samples than stretched samples. Western blots revealed the presence of 3 major intact TnT bands that diminished with time postmortem and 4 bands (TnT degradation products) that accumulated during postmortem storage. Quantification of intact TnT showed increased (P < 0.05) proteolysis at 4 and 7 d postmortem in samples with long sarcomeres. By 10 d, only traces of the greatest molecular weight intact TnT band were evident in both shortened and stretched samples, suggesting this TnT band may be more susceptible to proteolysis than other intact TnT bands. Degradation products of TnT appeared earlier postmortem in samples with long sarcomeres. The 30-kDa TnT fragment appeared after 7 d of postmortem storage in samples with long sarcomeres but not until 10 d in muscle containing short sarcomeres. Collectively, these data show that postmortem TnT proteolysis is sarcomere length-dependent and suggest that thick and thin filament overlap may influence the postmortem aging process in beef.

  19. The Japanese Society of Pathology Guidelines on the handling of pathological tissue samples for genomic research: Standard operating procedures based on empirical analyses.

    PubMed

    Kanai, Yae; Nishihara, Hiroshi; Miyagi, Yohei; Tsuruyama, Tatsuhiro; Taguchi, Kenichi; Katoh, Hiroto; Takeuchi, Tomoyo; Gotoh, Masahiro; Kuramoto, Junko; Arai, Eri; Ojima, Hidenori; Shibuya, Ayako; Yoshida, Teruhiko; Akahane, Toshiaki; Kasajima, Rika; Morita, Kei-Ichi; Inazawa, Johji; Sasaki, Takeshi; Fukayama, Masashi; Oda, Yoshinao

    2018-02-01

    Genome research using appropriately collected pathological tissue samples is expected to yield breakthroughs in the development of biomarkers and identification of therapeutic targets for diseases such as cancers. In this connection, the Japanese Society of Pathology (JSP) has developed "The JSP Guidelines on the Handling of Pathological Tissue Samples for Genomic Research" based on an abundance of data from empirical analyses of tissue samples collected and stored under various conditions. Tissue samples should be collected from appropriate sites within surgically resected specimens, without disturbing the features on which pathological diagnosis is based, while avoiding bleeding or necrotic foci. They should be collected as soon as possible after resection: at the latest within about 3 h of storage at 4°C. Preferably, snap-frozen samples should be stored in liquid nitrogen (about -180°C) until use. When intending to use genomic DNA extracted from formalin-fixed paraffin-embedded tissue, 10% neutral buffered formalin should be used. Insufficient fixation and overfixation must both be avoided. We hope that pathologists, clinicians, clinical laboratory technicians and biobank operators will come to master the handling of pathological tissue samples based on the standard operating procedures in these Guidelines to yield results that will assist in the realization of genomic medicine. © 2018 The Authors. Pathology International published by Japanese Society of Pathology and John Wiley & Sons Australia, Ltd.

  20. Impact of Nisin-Activated Packaging on Microbiota of Beef Burgers during Storage.

    PubMed

    Ferrocino, Ilario; Greppi, Anna; La Storia, Antonietta; Rantsiou, Kalliopi; Ercolini, Danilo; Cocolin, Luca

    2016-01-15

    Beef burgers were stored at 4°C in a vacuum in nisin-activated antimicrobial packaging. Microbial ecology analyses were performed on samples collected between days 0 and 21 of storage to discover the population diversity. Two batches were analyzed using RNA-based denaturing gradient gel electrophoresis (DGGE) and pyrosequencing. The active packaging retarded the growth of the total viable bacteria and lactic acid bacteria. Culture-independent analysis by pyrosequencing of RNA extracted directly from meat showed that Photobacterium phosphoreum, Lactococcus piscium, Lactobacillus sakei, and Leuconostoc carnosum were the major operational taxonomic units (OTUs) shared between control and treated samples. Beta diversity analysis of the 16S rRNA sequence data and RNA-DGGE showed a clear separation between two batches based on the microbiota. Control samples from batch B showed a significant high abundance of some taxa sensitive to nisin, such as Kocuria rhizophila, Staphylococcus xylosus, Leuconostoc carnosum, and Carnobacterium divergens, compared to control samples from batch A. However, only from batch B was it possible to find a significant difference between controls and treated samples during storage due to the active packaging. Predicted metagenomes confirmed differences between the two batches and indicated that the use of nisin-based antimicrobial packaging can determine a reduction in the abundance of specific metabolic pathways related to spoilage. The present study aimed to assess the viable bacterial communities in beef burgers stored in nisin-based antimicrobial packaging, and it highlights the efficacy of this strategy to prolong beef burger shelf life. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  1. 77 FR 13151 - Agency Information Collection Activities: Proposed Collection; Comments Requested; Records and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-03-05

    ... Data: Daily Summaries, Records of Production, Storage, and Disposition, and Supporting Data by Licensed... approved collection. (2) Title of the Form/Collection: Records and Supporting Data: Daily Summaries, Records of Production, Storage and Disposition and Supporting Data by Explosives Manufacturers. (3) Agency...

  2. Self-irradiation damage to the local structure of plutonium and plutonium intermetallics

    NASA Astrophysics Data System (ADS)

    Booth, C. H.; Jiang, Yu; Medling, S. A.; Wang, D. L.; Costello, A. L.; Schwartz, D. S.; Mitchell, J. N.; Tobash, P. H.; Bauer, E. D.; McCall, S. K.; Wall, M. A.; Allen, P. G.

    2013-03-01

    The effect of self-irradiation damage on the local structure of δ-Pu, PuAl2, PuGa3, and other Pu intermetallics has been determined for samples stored at room temperature using the extended x-ray absorption fine-structure (EXAFS) technique. These measurements indicate that the intermetallic samples damage at a similar rate as indicated in previous studies of PuCoGa5. In contrast, δ-Pu data indicate a much slower damage accumulation rate. To explore the effect of storage temperature and possible room temperature annealing effects, we also collected EXAFS data on a δ-Pu sample that was held at less than 32 K for a two month period. This sample damaged much more quickly. In addition, the measurable damage was annealed out at above only 135 K. Data from samples of δ-Pu with different Ga concentrations and results on all samples collected from different absorption edges are also reported. These results are discussed in terms of the vibrational properties of the materials and the role of Ga in δ-Pu as a network former.

  3. Comparison of quality attributes of buffalo meat curry at different storage temperature.

    PubMed

    Kandeepan, Gurunathan; Anjaneyulu, Anne Seet Ram; Kondaiah, Napa; Mendiratta, Sanjod Kumar

    2011-01-01

    The product quality of curry is determined by the food animal source, raw materials and the method of processing. Moreover the scientific information on processing and quality of traditional buffalo meat curry from different groups of buffaloes is not available. This study was undertaken to develop processed curry from different buffalo groups and to compare its quality during storage at ambient and refrigeration temperature. The meat samples were collected from the longissimus dorsi muscle of the carcasses from each group of buffaloes slaughtered according to the traditional halal method. Buffalo meat curry was prepared in a pressure cooker with the standardized formulation. This final product was subjected to evaluation of quality and shelf life. To evaluate the effect of different groups of meat samples on the quality of curry, product yield, pH, proximate composition, water activity (aw), thiobarbituric acid reactive substances (TBARS), calorific value, sensory attributes and microbiological assay were determined The energy of meat curry from young buffaloes was significantly lower than the meat curry from spent animal groups. The overall acceptability of curry decreased significantly during 3 days ambient storage compared to refrigeration storage. Scientific processing by adopting good manufacturing practices and suitable packaging helped greatly to improve the shelf life of the ambient temperature stored buffalo meat curry. Buffalo meat curry from young male group showed better product characteristics and overall acceptability scores than spent buffalo group.

  4. Use of FTA cards for the storage of breast carcinoma nucleic acid on fine-needle aspiration samples.

    PubMed

    Peluso, Anna Lucia; Cascone, Anna Maria; Lucchese, Lucrezia; Cozzolino, Immacolata; Ieni, Antonio; Mignogna, Chiara; Pepe, Stefano; Zeppa, Pio

    2015-10-01

    The preservation and storage of nucleic acids is important for DNA molecular techniques. The material obtained by fine-needle aspiration (FNA) is often scanty and can not be wasted. FTA cards are filter papers that immobilize and stabilize nucleic acids and can be stored at room temperature. The current study evaluated whether nucleic acids of breast carcinoma cells, obtained by FNA in a clinical setting, may be collected, stored, and preserved on FTA cards. Thirty breast carcinoma, 5 non-Hodgkin lymphoma (NHL), and 5 benign reactive lymph node (RLN) cell samples obtained by FNA were stored at -80 °C and on FTA cards. DNA extraction and polymerase chain reaction were performed on cells at -80 °C and on 2 punched disks of FTA cards. Fifty nanograms of extracted DNA from both sample types were used to amplify the Janus Kinase 2 (JAK2) gene. The mean value of DNA extracted from breast carcinoma cells was 28.19 ng/µL for that stored at -80 °C and 3.28 ng/µL for that stored on FTA cards. Agarose gel analysis demonstrated expected bands of DNA in 29 cases (97%) with both methods. The mean value of DNA extracted from NHL and RLN samples was 37.54 ng/µL and 4.28 ng/µL, respectively, and agarose gel analysis demonstrated bands of high molecular weight DNA in both methods. Significant differences in DNA yield were found between storage at -80 °C and FTA cards (P<.0001), but no differences were detected between 260/280 nm ratios in breast carcinoma and NHL/RLN samples. FTA cards can be conveniently used for the storage of breast carcinoma cells obtained by FNA, thus providing a reliable alternative to traditional methods. © 2015 American Cancer Society.

  5. Variability of alpha-tocopherol values associated with procurement, storage, and freezing of equine serum and plasma samples.

    PubMed

    Craig, A M; Blythe, L L; Rowe, K E; Lassen, E D; Barrington, R; Walker, K C

    1992-12-01

    Recent evidence concerning the pathogenesis of equine degenerative myeloencephalopathy indicated that low blood alpha-tocopherol values are a factor in the disease process. Variables that could be introduced by a veterinarian procuring, transporting, or storing samples were evaluated for effects on alpha-tocopherol concentration in equine blood. These variables included temperature; light; exposure to the rubber stopper of the evacuated blood collection tube; hemolysis; duration of freezing time, with and without nitrogen blanketing; and repeated freeze/thaw cycles. It was found that hemolysis caused the greatest change in high-performance liquid chromatography-measured serum alpha-tocopherol values, with mean decrease of 33% (P < 0.001). Lesser, but significant (P < 0.01) changes in serum alpha-tocopherol values were an approximate 10% decrease when refrigerated blood was left in contact with the red rubber stopper of the blood collection tube for 72 hours and an approximate 5% increase when blood was stored at 20 to 25 C (room temperature) for 72 hours. Repeated freeze/thaw cycles resulted in a significant (P < 0.05) 3% decrease in alpha-tocopherol values in heparinized plasma by the third thawing cycle. Freezer storage for a 3-month period without nitrogen blanketing resulted in slight (2%) decrease in mean serum alpha-tocopherol values, whereas values in serum stored for an identical period under nitrogen blanketing did not change. A significant (P < 0.001) mean decrease (10.3%) in alpha-tocopherol values was associated with freezer (-16 C) storage of nitrogen blanketed serum for 6 months.(ABSTRACT TRUNCATED AT 250 WORDS)

  6. Independent Verification Survey of the Clean Coral Storage Pile at the Johnston Atoll Plutonium Contaminated Soil Remediation Project

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

    Wilson-Nichols, M.J.; Egidi, P.V.; Roemer, E.K.

    2000-09-01

    f I The Oak Ridge National Laboratory (ORNL) Environmental Technology Section conducted an independent verification (IV) survey of the clean storage pile at the Johnston Atoll Plutonium Contaminated Soil Remediation Project (JAPCSRP) from January 18-25, 1999. The goal of the JAPCSRP is to restore a 24-acre area that was contaminated with plutonium oxide particles during nuclear testing in the 1960s. The selected remedy was a soil sorting operation that combined radiological measurements and mining processes to identify and sequester plutonium-contaminated soil. The soil sorter operated from about 1990 to 1998. The remaining clean soil is stored on-site for planned beneficialmore » use on Johnston Island. The clean storage pile currently consists of approximately 120,000 m3 of coral. ORNL conducted the survey according to a Sampling and Analysis Plan, which proposed to provide an IV of the clean pile by collecting a minimum number (99) of samples. The goal was to ascertain wi th 95% confidence whether 97% of the processed soil is less than or equal to the accepted guideline (500-Bq/kg or 13.5-pCi/g) total transuranic (TRU) activity.« less

  7. Field Degassing as a New Sampling Method for 14C Analyses in Old Groundwater

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

    Yokochi, Reika; Bernier, Ryan; Purtschert, Roland

    Radiocarbon ( 14C) activity in groundwater can be used to determine subsurface residence time up to ~40 kyr, providing crucial information on dynamic properties of groundwater and on paleoclimate. However, commonly applied sampling methods for dissolved inorganic carbon (DIC- 14C) are prone to low level of modern atmospheric contamination, resulting in underestimation of groundwater ages that cluster around 30–40 kyr. We extract CO 2 gas from groundwater using a device originally developed for studies of noble gas radionuclides. Carbon is collected in the gas phase, eliminating the possibility of fostering microbial activities and aqueous chemical reactions during sample storage. Thismore » method collects CO 2- 14C and radiokrypton ( 81Kr and 85Kr) samples simultaneously. The presence of any shorter-lived 85Kr is used to evaluate the degree of atmospheric contamination during sampling or mixing of young groundwater. Most groundwater samples showed lower CO 2- 14C activities than those of DIC- 14C, presumably due to the absence of atmospheric contamination. Samples with 81Kr age exceeding 150 kyr have no detectable CO 2- 14C except where mixing sources of young groundwater is suspected. Furthermore these field data serve as confirmations for the reliability of the newly presented sample collection and CO 2- 14C method, and for the outstanding roles of radiokrypton isotopes in characterizing old groundwater.« less

  8. Field Degassing as a New Sampling Method for 14C Analyses in Old Groundwater

    DOE PAGES

    Yokochi, Reika; Bernier, Ryan; Purtschert, Roland; ...

    2017-09-07

    Radiocarbon ( 14C) activity in groundwater can be used to determine subsurface residence time up to ~40 kyr, providing crucial information on dynamic properties of groundwater and on paleoclimate. However, commonly applied sampling methods for dissolved inorganic carbon (DIC- 14C) are prone to low level of modern atmospheric contamination, resulting in underestimation of groundwater ages that cluster around 30–40 kyr. We extract CO 2 gas from groundwater using a device originally developed for studies of noble gas radionuclides. Carbon is collected in the gas phase, eliminating the possibility of fostering microbial activities and aqueous chemical reactions during sample storage. Thismore » method collects CO 2- 14C and radiokrypton ( 81Kr and 85Kr) samples simultaneously. The presence of any shorter-lived 85Kr is used to evaluate the degree of atmospheric contamination during sampling or mixing of young groundwater. Most groundwater samples showed lower CO 2- 14C activities than those of DIC- 14C, presumably due to the absence of atmospheric contamination. Samples with 81Kr age exceeding 150 kyr have no detectable CO 2- 14C except where mixing sources of young groundwater is suspected. Furthermore these field data serve as confirmations for the reliability of the newly presented sample collection and CO 2- 14C method, and for the outstanding roles of radiokrypton isotopes in characterizing old groundwater.« less

  9. Evaluation of the Flinders Technology Associates Cards for Storage and Temperature Challenges in Field Conditions for Foot-and-Mouth Disease Virus Surveillance.

    PubMed

    Madhanmohan, M; Yuvaraj, S; Manikumar, K; Kumar, R; Nagendrakumar, S B; Rana, S K; Srinivasan, V A

    2016-12-01

    Foot-and-mouth disease virus (FMDV) samples transported to the laboratory from far and inaccessible areas for diagnosis and identification of FMDV pose a major problem in a tropical country like India, where wide fluctuation of temperature over a large geographical area is common. Inadequate storage methods lead to spoilage of FMDV samples collected from clinically positive animals in the field. Such samples are declared as non-typeable by the typing laboratories with the consequent loss of valuable epidemiological data. In this study, an attempt was made to evaluate the robustness of Flinders Technology Associates (FTA) cards for storage and transportation of FMDV samples in different climatic conditions which will be useful for FMDV surveillance. Simulation transport studies were conducted using FTA impregnated FMDV samples during post-monsoon (September-October 2010) and summer season (May-June 2012). FMDV genome or serotype could be identified from the FTA cards after the simulation transport studies with varying temperature (22-45°C) and relative humidity (20-100%). The stability of the viral RNA, the absence of infectivity and ease of processing the sample for molecular methods make the FTA cards an useful option for transport of FMDV genome for identification and type determination. The method can be used routinely for FMDV research as it is economical and the cards can be transported easily in envelopes by regular courier/postal systems. The absence of live virus in FTA card can be viewed as an advantage as it restricts the risk of transmission of live virus. © 2015 Blackwell Verlag GmbH.

  10. Evaluation of Nobuto filter paper strips for the detection of avian influenza virus antibody in waterfowl

    USGS Publications Warehouse

    Dusek, Robert J.; Hall, Jeffrey S.; Nashold, Sean W.; Teslaa, Joshua L.; Ip, Hon S.

    2011-01-01

    The utility of using Nobuto paper strips for the detection of avian influenza antibodies was examined in mallards (Anas platyrhynchos) experimentally infected with low pathogenic avian influenza viruses. Blood was collected 2 wk after infection and was preserved either as serum or whole blood absorbed onto Nobuto strips. Analysis of samples using a commercially available blocking enzyme-linked immunosorbent assay revealed comparable results (???96% sensitivity for all methods) between sera stored at -30 C and the Nobuto strip preservation method even when the Nobuto strips were stored up to 3 mo at room temperature (RT). Significant differences were detected in the ratio of sample absorbance to negative control absorbance for Nobuto strips stored at RT compared with sera stored at -30 C, although these differences did not affect the ability of the test to reliably detect positive and negative samples. Nobuto strips are a convenient and sensitive alternative to the collection of serum samples when maintaining appropriate storage temperatures is difficult. ?? 2011 American Association of Avian Pathologists.

  11. Silk-based blood stabilization for diagnostics.

    PubMed

    Kluge, Jonathan A; Li, Adrian B; Kahn, Brooke T; Michaud, Dominique S; Omenetto, Fiorenzo G; Kaplan, David L

    2016-05-24

    Advanced personalized medical diagnostics depend on the availability of high-quality biological samples. These are typically biofluids, such as blood, saliva, or urine; and their collection and storage is critical to obtain reliable results. Without proper temperature regulation, protein biomarkers in particular can degrade rapidly in blood samples, an effect that ultimately compromises the quality and reliability of laboratory tests. Here, we present the use of silk fibroin as a solid matrix to encapsulate blood analytes, protecting them from thermally induced damage that could be encountered during nonrefrigerated transportation or freeze-thaw cycles. Blood samples are recovered by simple dissolution of the silk matrix in water. This process is demonstrated to be compatible with a number of immunoassays and provides enhanced sample preservation in comparison with traditional air-drying paper approaches. Additional processing can remediate interactions with conformational structures of the silk protein to further enhance blood stabilization and recovery. This approach can provide expanded utility for remote collection of blood and other biospecimens empowering new modalities of temperature-independent remote diagnostics.

  12. 28 CFR 115.389 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Juvenile Facilities Data Collection and Review § 115.389 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115...

  13. 28 CFR 115.189 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Lockups Data Collection and Review § 115.189 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115.187 are...

  14. 28 CFR 115.389 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Juvenile Facilities Data Collection and Review § 115.389 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115...

  15. 28 CFR 115.189 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Lockups Data Collection and Review § 115.189 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115.187 are...

  16. 28 CFR 115.89 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 28 Judicial Administration 2 2014-07-01 2014-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Adult Prisons and Jails Data Collection and Review § 115.89 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to...

  17. 28 CFR 115.89 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 28 Judicial Administration 2 2012-07-01 2012-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Adult Prisons and Jails Data Collection and Review § 115.89 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to...

  18. 28 CFR 115.89 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Adult Prisons and Jails Data Collection and Review § 115.89 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to...

  19. 28 CFR 115.189 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Lockups Data Collection and Review § 115.189 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115.187 are...

  20. 28 CFR 115.389 - Data storage, publication, and destruction.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 28 Judicial Administration 2 2013-07-01 2013-07-01 false Data storage, publication, and... ELIMINATION ACT NATIONAL STANDARDS Standards for Juvenile Facilities Data Collection and Review § 115.389 Data storage, publication, and destruction. (a) The agency shall ensure that data collected pursuant to § 115...

  1. Challenges for conducting blood collection and biochemical analysis in a large multicenter school-based study with adolescents: lessons from ERICA in Brazil.

    PubMed

    Cureau, Felipe Vogt; Bloch, Katia Vergetti; Henz, Aline; Schaan, Camila W; Klein, Carlos Henrique; Oliveira, Cecília Lacroix de; Giannini, Denise Tavares; Leon, Elisa Brosina de; Abreu, Gabriela de Azevedo; Telo, Gabriela Heiden; Dias, Glauber Monteiro; Carvalho, Kênia Mara Baiocchi de; Barufaldi, Laura Augusta; Kuschnir, Maria Cristina Caetano; Szklo, Moyses; Montenegro, Renan; Silva, Thiago Luiz Nogueira da; Ekelund, Ulf; Schaan, Beatriz D

    2017-05-18

    The Study of Cardiovascular Risk in Adolescents (ERICA) is a pioneering study that aimed to assess the prevalence of cardiovascular risk factors, including metabolic syndrome components in Brazilian adolescents. This study aims to describe the methodological aspects related to blood collection as well as to report pertaining results of the preparation, transport, storage, and exams in ERICA. Exams in ERICA were performed in a single laboratory and blood samples were collected in schools in a standardized manner. Logistics involved air transportation of samples to the reference laboratory with controlled temperature since sample collection. The serum was stored in local biorepositories in four centers to be used in future analyses. During the study, 284,247 exams were performed and rate of participation in exams was 56.2%, thus involving 40,732 adolescents. From the total, 92.6% of the samples reached the reference laboratory maintaining the temperature between 0-10°C. No clinical significant changes in results due to temperature changes were identified. External quality control recorded satisfactory results in 98.7% of the evaluations. Four biorepositories with samples of 7,785 adolescents were created. Thus, we can consider that the logistics adopted in ERICA was fairly successful and description of this as well as the difficulties experienced in Brazil can inform and facilitate the planning of future studies, especially in developing countries.

  2. Geomechanical characterization of the Upper Carboniferous under thermal stress for the evaluation of a High Temperature - Mine Thermal Energy Storage (HT-MTES)

    NASA Astrophysics Data System (ADS)

    Hahn, Florian; Brüggemann, Nina; Bracke, Rolf; Alber, Michael

    2017-04-01

    The goal of this R&D project is to create a technically and economically feasible conceptual model for a High Temperature - Mine Thermal Energy Storage (HT-MTES) for the energetic reuse of a mine on the example of the Prosper-Haniel coal mine in Bottrop, Germany. This project is funded by the "Initiative Energy Storage" program of the German Federal Ministries BMWi, BMU and BMBF. At the end of 2018, the last operative coal mine in North Rhine-Westphalia, Germany (Prosper-Haniel), is going to be closed down, plugged and abandoned. Large amounts of subsurface infrastructures, resembled mainly by open parts of former galleries and mining faces are going to be flooded, after the mine is closed down and therefore have the potential to become an enormous geothermal reservoir for a seasonal heat storage. During the summer non-used (waste) heat from solar thermal power plants, garbage incineration, combined heat and power plants (CHP) or industrial production processes can be stored within dedicated drifts of the mine. During the winter season, this surplus heat can be extracted and directly utilized in commercial and/or residential areas. For the evaluation of such a HT-MTES within a former coal mine, the corresponding geomechanical parameters of the Upper Carboniferous under thermal stress needs to be evaluated. Therefore the main rock types of the Upper Carboniferous (claystone, siltstone and sandstone) are subject to a geomechanical characterization before and after thermal cyclic loadings of temperatures up to 200 °C. The samples have been collected directly from the coal mine Prosper-Haniel within a depth range of 1000 - 1200 m. Unconfined compressive and tensile strengths, as well as triaxial tests were performed at room temperature. Furthermore, a range of petrophysical properties like density, thin-section analysis and P-wave velocities were determined. First results show an indication that the overall strength properties of the samples are not effected by thermal cyclic loadings with temperatures of up to 200 °C. However, a reduction in the Young's modulus was observed in all samples, after thermal cyclic loads were induced. This effect is mainly correlated to a relaxation of the grain bonds and a pore space expansion. Currently, the experimental focus was set on the evaluation of the collected siltstone samples. Therefore further experiments are needed to undermine these results also for the claystone and sandstone samples.

  3. Maintenance of host DNA integrity in field-preserved mosquito (Diptera: Culicidae) blood meals for identification by DNA barcoding.

    PubMed

    Reeves, Lawrence E; Holderman, Chris J; Gillett-Kaufman, Jennifer L; Kawahara, Akito Y; Kaufman, Phillip E

    2016-09-15

    Determination of the interactions between hematophagous arthropods and their hosts is a necessary component to understanding the transmission dynamics of arthropod-vectored pathogens. Current molecular methods to identify hosts of blood-fed arthropods require the preservation of host DNA to serve as an amplification template. During transportation to the laboratory and storage prior to molecular analysis, genetic samples need to be protected from nucleases, and the degradation effects of hydrolysis, oxidation and radiation. Preservation of host DNA contained in field-collected blood-fed specimens has an additional caveat: suspension of the degradative effects of arthropod digestion on host DNA. Unless effective preservation methods are implemented promptly after blood-fed specimens are collected, host DNA will continue to degrade. Preservation methods vary in their efficacy, and need to be selected based on the logistical constraints of the research program. We compared four preservation methods (cold storage at -20 °C, desiccation, ethanol storage of intact mosquito specimens and crushed specimens on filter paper) for field storage of host DNA from blood-fed mosquitoes across a range of storage and post-feeding time periods. The efficacy of these techniques in maintaining host DNA integrity was evaluated using a polymerase chain reaction (PCR) to detect the presence of a sufficient concentration of intact host DNA templates for blood meal analysis. We applied a logistic regression model to assess the effects of preservation method, storage time and post-feeding time on the binomial response variable, amplification success. Preservation method, storage time and post-feeding time all significantly impacted PCR amplification success. Filter papers and, to a lesser extent, 95 % ethanol, were the most effective methods for the maintenance of host DNA templates. Amplification success of host DNA preserved in cold storage at -20 °C and desiccation was poor. Our data suggest that, of the methods tested, host DNA template integrity was most stable when blood meals were preserved using filter papers. Filter paper preservation is effective over short- and long-term storage, while ethanol preservation is only suitable for short-term storage. Cold storage at -20 °C, and desiccation of blood meal specimens, even for short time periods, should be avoided.

  4. Adsorbing/dissolving Lyoprotectant Matrix Technology for Non-cryogenic Storage of Archival Human Sera

    NASA Astrophysics Data System (ADS)

    Solivio, Morwena J.; Less, Rebekah; Rynes, Mathew L.; Kramer, Marcus; Aksan, Alptekin

    2016-04-01

    Despite abundant research conducted on cancer biomarker discovery and validation, to date, less than two-dozen biomarkers have been approved by the FDA for clinical use. One main reason is attributed to inadvertent use of low quality biospecimens in biomarker research. Most proteinaceous biomarkers are extremely susceptible to pre-analytical factors such as collection, processing, and storage. For example, cryogenic storage imposes very harsh chemical, physical, and mechanical stresses on biospecimens, significantly compromising sample quality. In this communication, we report the development of an electrospun lyoprotectant matrix and isothermal vitrification methodology for non-cryogenic stabilization and storage of liquid biospecimens. The lyoprotectant matrix was mainly composed of trehalose and dextran (and various low concentration excipients targeting different mechanisms of damage), and it was engineered to minimize heterogeneity during vitrification. The technology was validated using five biomarkers; LDH, CRP, PSA, MMP-7, and C3a. Complete recovery of LDH, CRP, and PSA levels was achieved post-rehydration while more than 90% recovery was accomplished for MMP-7 and C3a, showing promise for isothermal vitrification as a safe, efficient, and low-cost alternative to cryogenic storage.

  5. Changes in the free amino acid contents of honeys during storage at ambient temperature.

    PubMed

    Iglesias, M Teresa; Martín-Alvarez, Pedro J; Polo, M Carmen; de Lorenzo, Cristina; Gonzalez, Montserrat; Pueyo, Encarnación

    2006-11-29

    This study was carried out to establish the changes in the free amino acid contents of floral honeys, honeydew honeys, and blend honeys during storage at room temperature and to test the capacity of the amino acids to distinguish the origin of the honeys after storage. For this purpose, 54 artisanal honeys (39 floral, 5 honeydew, and 10 blend) were studied. Samples were taken from recently collected honeys and at 3, 6, 9, 12, 16, 20, and 24 months after harvesting. The contents of most of the free amino acids were found to decrease with storage time, with the greatest reduction observed in the first 9 months. The contents of the amino acids aspartic acid, beta-alanine, and proline increased in the first few months after storage, reaching maximum values at 6 months, suggesting the possible existence of enzymatic activities. The application of stepwise discriminant analysis to the free amino acid content data demonstrated that the contents of the amino acids valine, beta-alanine, gamma-aminobutyric acid, serine, isoleucine, alpha-alanine, ornithine, and glutamine correctly assigned 87% of honeys to their group of origin: floral, honeydew, or blend.

  6. Evaluation of the Universal Viral Transport system for long-term storage of virus specimens for microbial forensics.

    PubMed

    Hosokawa-Muto, Junji; Fujinami, Yoshihito; Mizuno, Natsuko

    2015-08-01

    Forensic microbial specimens, including bacteria and viruses, are collected at biocrime and bioterrorism scenes. Although it is preferable that the pathogens in these samples are alive and kept in a steady state, the samples may be stored for prolonged periods before analysis. Therefore, it is important to understand the effects of storage conditions on the pathogens contained within such samples. To evaluate the capacity to preserve viable virus and the viral genome, influenza virus was added to the transport medium of the Universal Viral Transport system and stored for over 3 months at various temperatures, after which virus titrations and quantitative analysis of the influenza hemagglutinin gene were performed. Although viable viruses became undetectable 29 days after the medium was stored at room temperature, viruses in the medium stored at 4°C were viable even after 99 days. A quantitative PCR analysis indicated that the hemagglutinin gene was maintained for 99 days at both 4°C and room temperature. Therefore, long-term storage at 4°C has little effect on viable virus and viral genes, so the Universal Viral Transport system can be useful for microbial forensics. This study provides important information for the handling of forensic virus specimens. Copyright © 2015 Elsevier Ltd and Faculty of Forensic and Legal Medicine. All rights reserved.

  7. Effect of red blood cell storage time on markers of hemolysis and inflammation in transfused very low birth weight infants.

    PubMed

    Kalhan, Tamara G; Bateman, David A; Bowker, Rakhee M; Hod, Eldad A; Kashyap, Sudha

    2017-12-01

    BackgroundProlonged storage of transfused red blood cells (RBCs) is associated with hemolysis in healthy adults and inflammation in animal models. We aimed to determine whether storage duration affects markers of hemolysis (e.g., serum bilirubin, iron, and non-transferrin-bound iron (NTBI)) and inflammation (e.g., interleukin (IL)-8 and monocyte chemoattractant protein (MCP)-1) in transfused very low birth weight (VLBW) infants.MethodsBlood samples from 23 independent transfusion events were collected by heel stick before and 2-6 h after transfusion.ResultsSerum iron, total bilirubin, NTBI, and MCP-1 levels were significantly increased after transfusion of RBCs (P<0.05 for each comparison). The storage age of transfused RBCs positively correlated with increases in NTBI following transfusion (P<0.001; R 2 =0.44). No associations between storage duration and changes in the other analytes were observed.ConclusionTransfusion of RBCs into VLBW infants is associated with increased markers of hemolysis and the inflammatory chemokine MCP-1. RBC-storage duration only correlated with increases in NTBI levels following transfusion. NTBI was only observed in healthy adults following 35 days of storage; however, this study suggests that VLBW infants are potentially more susceptible to produce this pathological form of iron, with increased levels observed after transfusion of only 20-day-old RBCs.

  8. Assessing the microbiological performance and potential cost of boiling drinking water in urban Zambia.

    PubMed

    Psutka, Rebecca; Peletz, Rachel; Michelo, Sandford; Kelly, Paul; Clasen, Thomas

    2011-07-15

    Boiling is the most common method of disinfecting water in the home and the benchmark against which other point-of-use water treatment is measured. In a six-week study in peri-urban Zambia, we assessed the microbiological effectiveness and potential cost of boiling among 49 households without a water connection who reported "always" or "almost always" boiling their water before drinking it. Source and household drinking water samples were compared weekly for thermotolerant coliforms (TTC), an indicator of fecal contamination. Demographics, costs, and other information were collected through surveys and structured observations. Drinking water samples taken at the household (geometric mean 7.2 TTC/100 mL, 95% CI, 5.4-9.7) were actually worse in microbiological quality than source water (geometric mean 4.0 TTC/100 mL, 95% CI, 3.1-5.1) (p < 0.001), although both are relatively low levels of contamination. Only 60% of drinking water samples were reported to have actually been boiled at the time of collection from the home, suggesting over-reporting and inconsistent compliance. However, these samples were of no higher microbiological quality. Evidence suggests that water quality deteriorated after boiling due to lack of residual protection and unsafe storage and handling. The potential cost of fuel or electricity for boiling was estimated at 5% and 7% of income, respectively. In this setting where microbiological water quality was relatively good at the source, safe-storage practices that minimize recontamination may be more effective in managing the risk of disease from drinking water at a fraction of the cost of boiling.

  9. Stability of Phosphatidylethanol in Dry Blood Spot Cards.

    PubMed

    Bakhireva, Ludmila N; Shrestha, Shikhar; Gutierrez, Hilda L; Berry, Mike; Schmitt, Cheryl; Sarangarm, Dusadee

    2016-05-01

    The analysis of phosphatidylethanol, a promising direct ethanol metabolite, in dry blood spots (PEth-DBS) is advantageous due to ease of storage, transportation and minimal invasiveness of capillary blood collection. One potential application of PEth-DBS is to confirm prenatal alcohol exposure in newborns suspected of FASD; however, stability of PEth-DBS is largely unknown. Phlebotomized samples from 31 adults with a history of alcoholism, admitted to the University of New Mexico Emergency Department, were analyzed for blood alcohol content and pipetted onto DBS cards (13 spots per patient). The first spot was analyzed within 2 weeks of collection for a baseline PEth; the remaining 12 spots were allocated into three temperature conditions (room temperature, 4°C, -80°C) for the repeated measures analysis. In addition, 5 newborn DBS samples with a baseline PEth>LOD were obtained from a prospective cohort at UNM and re-analyzed at 4 months after storage at -80°C. A mixed linear model was fitted to examine the effects of temperature, time and temperature-time interaction on PEth degradation over the first 9 months. The baseline PEth levels were 592.8 ± 86.7 ng/ml and 18.3 ± 4.8 ng/ml in adult and newborn samples, respectively. All DBS samples remained positive in successive samples in all temperature conditions. Results of mixed linear model demonstrated a significant effect of temperature (P < 0.001) on PEth degradation over 9 months. PEth-DBS appears to be relatively stable, especially when stored at lower temperatures. These initial results are encouraging and highlight the PEth-DBS potential in retrospective assessment of alcohol exposure. © The Author 2015. Medical Council on Alcohol and Oxford University Press. All rights reserved.

  10. Stability of Phosphatidylethanol in Dry Blood Spot Cards

    PubMed Central

    Bakhireva, Ludmila N.; Shrestha, Shikhar; Gutierrez, Hilda L.; Berry, Mike; Schmitt, Cheryl; Sarangarm, Dusadee

    2016-01-01

    Background The analysis of phosphatidylethanol, a promising direct ethanol metabolite, in dry blood spots (PEth-DBS) is advantageous due to ease of storage, transportation and minimal invasiveness of capillary blood collection. One potential application of PEth-DBS is to confirm prenatal alcohol exposure in newborns suspected of FASD; however, stability of PEth-DBS is largely unknown. Methods Phlebotomized samples from 31 adults with a history of alcoholism, admitted to the University of New Mexico Emergency Department, were analyzed for blood alcohol content and pipetted onto DBS cards (13 spots per patient). The first spot was analyzed within 2 weeks of collection for a baseline PEth; the remaining 12 spots were allocated into three temperature conditions (room temperature, 4°C, −80°C) for the repeated measures analysis. In addition, 5 newborn DBS samples with a baseline PEth>LOD were obtained from a prospective cohort at UNM and re-analyzed at 4 months after storage at −80°C. A mixed linear model was fitted to examine the effects of temperature, time and temperature–time interaction on PEth degradation over the first 9 months. Results The baseline PEth levels were 592.8 ± 86.7 ng/ml and 18.3 ± 4.8 ng/ml in adult and newborn samples, respectively. All DBS samples remained positive in successive samples in all temperature conditions. Results of mixed linear model demonstrated a significant effect of temperature (P < 0.001) on PEth degradation over 9 months. Conclusions PEth-DBS appears to be relatively stable, especially when stored at lower temperatures. These initial results are encouraging and highlight the PEth-DBS potential in retrospective assessment of alcohol exposure. PMID:26519350

  11. Characterization of the treefrog null allele, 1991

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

    Guttman, S.I.

    1992-04-01

    Spring peeper (Hyla crucifer) tadpoles collected from the waste storage area during the Biological and Ecological Site Characterization of the Feed Materials Production Center (FEMP) in 1986 and 1987 appeared to be unique. A null (inactive) allele was found at the glucose phosphate isomerase enzyme locus in significant frequencies (approximately 20%) each year; this allele did not appear to occur in the offsite sample collected approximately 15km from the FEMP. Null alleles at this locus have not been reported in other amphibian populations; when they have been found in other organisms they have invariably been lethal in the homozygous condition.

  12. Reconnaissance investigations of potential ground-water and sediment contamination at three former underground storage tank locations, Fort Jackson, South Carolina, 1994

    USGS Publications Warehouse

    Robertson, J.F.; Nagle, Douglas D.; Rhodes, Liesl C.

    1994-01-01

    Investigations to provide initial qualitative delineation of petroleum hydrocarbon contamination at three former underground storage tank locations at Fort Jackson, South Carolina, were made during March 1994. Ground-water and sediment samples were collected using direct-push technology and analyzed on-site with a gas chromatograph, which provided real-time, semi-quantitative data. In addition, ground-water and sediment samples were collected at selected sites for laboratory analyses to provide a confirmation of the on-site data. These analyses provided qualitative data on the lateral distri- bution of petroleum hydrocarbons. Petroleum hydrocarbons were detected by on-site analysis in ground-water samples from nine locations at Site 1062, suggesting the presence of a contaminant plume. Concentrations ranged from less than the minimum detection limit to 4,511 mg/L (micrograms per liter) for benzene, 15,594 mg/L for toluene, 16,501 mg/L for ethylbenzene, and 19,391 mg/L for total xylenes. Concentrations of Total Petroleum Hydrocarbons-Gasoline Range Organics ranged from 323 mg/L to 3,364 mg/L; Total Petroleum Hydrocarbons-Diesel Range Organics were not detected. Three samples from this site were analyzed for benzene, toluene, ethylbenzene, and total xylenes at a laboratory and results showed concentrations ranging from less than the minimum detection limit to 1,070 mg/L for benzene, 7,930 mg/L for toluene, 6,890 mg/L for ethylbenzene, and 1,524 mg/L for total xylenes. Petroleum hydro- carbons were detected by on-site analysis in only one sample at Site 2438. A concentration of 131,000 micrograms per kilogram Total Petroleum Hydrocarbons-Diesel Range Organics was detected in sample number GP-2-4-13.5. Petroleum hydrocarbons were detected by on-site analysis in only one ground-water sample from Site 2444. A concentration of 3,145 mg/L Total Petroleum Hydrocarbons-Gasoline Range Organics was detected at sampling location GP-3-2.

  13. Comparison of DNA preservation methods for environmental bacterial community samples

    USGS Publications Warehouse

    Gray, Michael A.; Pratte, Zoe A.; Kellogg, Christina A.

    2013-01-01

    Field collections of environmental samples, for example corals, for molecular microbial analyses present distinct challenges. The lack of laboratory facilities in remote locations is common, and preservation of microbial community DNA for later study is critical. A particular challenge is keeping samples frozen in transit. Five nucleic acid preservation methods that do not require cold storage were compared for effectiveness over time and ease of use. Mixed microbial communities of known composition were created and preserved by DNAgard™, RNAlater®, DMSO–EDTA–salt (DESS), FTA® cards, and FTA Elute® cards. Automated ribosomal intergenic spacer analysis and clone libraries were used to detect specific changes in the faux communities over weeks and months of storage. A previously known bias in FTA® cards that results in lower recovery of pure cultures of Gram-positive bacteria was also detected in mixed community samples. There appears to be a uniform bias across all five preservation methods against microorganisms with high G + C DNA. Overall, the liquid-based preservatives (DNAgard™, RNAlater®, and DESS) outperformed the card-based methods. No single liquid method clearly outperformed the others, leaving method choice to be based on experimental design, field facilities, shipping constraints, and allowable cost.

  14. Ensuring the Safety and Security of Frozen Lung Cancer Tissue Collections through the Encapsulation of Dried DNA.

    PubMed

    Washetine, Kevin; Kara-Borni, Mehdi; Heeke, Simon; Bonnetaud, Christelle; Félix, Jean-Marc; Ribeyre, Lydia; Bence, Coraline; Ilié, Marius; Bordone, Olivier; Pedro, Marine; Maitre, Priscilla; Tanga, Virginie; Gormally, Emmanuelle; Mossuz, Pascal; Lorimier, Philippe; Marquette, Charles Hugo; Mouroux, Jérôme; Cohen, Charlotte; Lassalle, Sandra; Long-Mira, Elodie; Clément, Bruno; Dagher, Georges; Hofman, Véronique; Hofman, Paul

    2018-06-11

    Collected specimens for research purposes may or may not be made available depending on their scarcity and/or on the project needs. Their protection against degradation or in the event of an incident is pivotal. Duplication and storage on a different site is the best way to assure their sustainability. The conservation of samples at room temperature (RT) by duplication can facilitate their protection. We describe a security system for the collection of non-small cell lung cancers (NSCLC) stored in the biobank of the Nice Hospital Center, France, by duplication and conservation of lyophilized (dried), encapsulated DNA kept at RT. Therefore, three frozen tissue collections from non-smoking, early stage and sarcomatoid carcinoma NSCLC patients were selected for this study. DNA was extracted, lyophilized and encapsulated at RT under anoxic conditions using the DNAshell technology. In total, 1974 samples from 987 patients were encapsulated. Six and two capsules from each sample were stored in the biobanks of the Nice and Grenoble (France) Hospitals, respectively. In conclusion, DNA maintained at RT allows for the conservation, duplication and durability of collections of interest stored in biobanks. This is a low-cost and safe technology that requires a limited amount of space and has a low environmental impact.

  15. Extracting metalworking fluid aerosol samples in cassettes by provisional ASTM and NIOSH methods.

    PubMed

    Harper, Martin

    2002-01-01

    Recent provisional methods for the determination of metalworking fluid aerosol in workplace air involve a solvent extraction procedure to separate the nonvolatile fraction of the fluid from insoluble material such as metal turnings and dirt. The procedure calls for preweighing a filter (W1) and assembling it into a cassette and taking a sample. In the laboratory the filter is removed from the cassette, desiccated to remove any collected water or other volatile substances, and weighed again (W2). The filter is then extracted in an organic solvent blend, allowed to dry, and weighed a final time (W3). The total weight collected by the filter is given by (W2-W1), and the weight of (nonvolatile) metalworking fluid collected is given by (W2-W3). The extraction step can take place within a cassette housing if it is relatively inert to the solvent blend used. The extraction of four metalworking fluids (straight oil, soluble oil, synthetic and semisynthetic) within disposable polypropylene cassettes was investigated using the same protocol used to evaluate the original method. For all fluids the extraction efficiency was greater than 95% with a precision better than 5%. The mean blank contribution to the extraction step was 16 micrograms. Blanks were also evaluated after storage, and after transport and storage. A small additional blank contribution could be removed by desiccation. The limits of detection and quantitation of the extraction step were calculated to be 28 and 94 micrograms, respectively.

  16. Viability and resistance of lactobacilli isolated from cocoa fermentation to simulated gastrointestinal digestive steps in soy yogurt.

    PubMed

    Saito, V S T; Dos Santos, T F; Vinderola, C G; Romano, C; Nicoli, J R; Araújo, L S; Costa, M M; Andrioli, J L; Uetanabaro, A P T

    2014-02-01

    To study the potential probiotic characteristics such as decrease of pH, microbial viability, and tolerance to simulated digestive steps of fermented soy beverage ("soy yogurt") produced with lactobacilli isolated from cocoa fermentation (Lactobacillus fermentum TcUESC01 and Lactobacillus plantarum TcUESC02) during fermentation and refrigerated storage. The sensory acceptance of the yogurts was also tested. Samples of soy yogurt produced with L. fermentum TcUESC01 or L. plantarum TcUESC02 were collected during fermentation (0, 4, 8, and 12 h) and refrigerated storage (1, 9, 18, and 27 d), and submitted to pH and bacterial viability determinations. Tolerance to simulated digestion steps was done with refrigerated storage samples at 9 °C. Simulated digestion was performed in 3 successive steps: exposure to pepsin-HCl solution, bile shock, and simulated small intestinal juice. During storage, a decrease in pH and lactobacillus viability was observed. L. fermentum TcUESC01 showed to be more resistant than L. plantarum TcUESC02 to simulated gastrointestinal digestion. All soy yogurts showed acceptable hedonic scores (greater than 5 in a 9-point hedonic scale ranging from "like extremely" to "dislike extremely") in sensory evaluation for flavor, aroma, color, consistency, and overall impression. L. plantarum TcUESC02 and, especially, L. fermentum TcUESC01 showed potential probiotic characteristics when considering pH, cell viability, and tolerance to simulated digestive steps and did not affect the sensory characteristics when supplemented to soy yogurt during storage. © 2014 Institute of Food Technologists®

  17. Alterations in hematologic indices during long-duration spaceflight.

    PubMed

    Kunz, Hawley; Quiriarte, Heather; Simpson, Richard J; Ploutz-Snyder, Robert; McMonigal, Kathleen; Sams, Clarence; Crucian, Brian

    2017-01-01

    Although a state of anemia is perceived to be associated with spaceflight, to date a peripheral blood hematologic assessment of red blood cell (RBC) indices has not been performed during long-duration space missions. This investigation collected whole blood samples from astronauts participating in up to 6-months orbital spaceflight, and returned those samples (ambient storage) to Earth for analysis. As samples were always collected near undock of a returning vehicle, the delay from collection to analysis never exceeded 48 h. As a subset of a larger immunologic investigation, a complete blood count was performed. A parallel stability study of the effect of a 48 h delay on these parameters assisted interpretation of the in-flight data. We report that the RBC and hemoglobin were significantly elevated during flight, both parameters deemed stable through the delay of sample return. Although the stability data showed hematocrit to be mildly elevated at +48 h, there was an in-flight increase in hematocrit that was ~3-fold higher in magnitude than the anticipated increase due to the delay in processing. While susceptible to the possible influence of dehydration or plasma volume alterations, these results suggest astronauts do not develop persistent anemia during spaceflight.

  18. The effect of sample storage on the performance and reproducibility of the galactomannan EIA test.

    PubMed

    Kimpton, George; White, P Lewis; Barnes, Rosemary A

    2014-08-01

    Galactomannan enzyme immune assay (GM EIA) is a nonculture test for detecting invasive aspergillosis (IA) forming a key part of diagnosis and management. Recent reports have questioned the reproducibility of indices after sample storage. To investigate this, 198 serum samples (72 from cases and 126 from controls) and 61 plasma samples (24 from cases and 37 from controls), initially tested between 2010 and 2013, were retested to determine any change in index. Data were also collected on circulatory protein levels for false-positive serum samples. Serum indices significantly declined on retesting (median: initial, 0.50, retest, 0.23; P < 0.0001). This was shown to be diagnosis dependent as the decline was apparent on retesting of control samples (median: initial 0.50, retest 0.12; P < 0.0001), but was not evident with case samples (median: initial, 0.80, retest, 0.80; P = 0.724). Plasma samples showed little change on reanalysis after long-term storage at 4°C. Retesting after freezing showed a decrease in index values for controls (median: initial 0.40, retest 0.26; P = 0.0505), but no significant change in cases. Circulatory proteins showed a correlation between serum albumin concentration and difference in index value on retesting. Overall, this study suggests that a lack of reproducibility in GM EIA positivity is only significant when disease is absent. Retesting after freezing helps to differentiate false-positive GM EIA results and, with consecutive positivity, could help to improve accuracy in predicting disease status. The freezing of samples prior to testing could potentially reduce false-positivity rates and the need to retest. © The Author 2014. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  19. Organic carbon storage and benthic consumption in sediments of northern fjords (60-80°N)

    NASA Astrophysics Data System (ADS)

    Włodarska-Kowalczuk, Maria; Zaborska, Agata; Jankowska, Emilia; Mazurkiewicz, Mikołaj

    2017-04-01

    Fjords have been recently recognized as hotspots of organic carbon storage, with organic carbon burial rates one hundred times larger than the global ocean average, accounting for 11% of global annual marine carbon burial (Smith et al. (2015) Nature Geoscience 8: 450-453). The organic carbon production and processing in coastal waters and sediments are controlled by environmental settings that are likely to be reshaped in the course of the global warming. The fastest and strongest changes are to occur in polar regions. In the present study we compare organic carbon stocks, accumulation and burial in temperate (Raunefjorden, Ullsfjorden, Balsfjorden) and Arctic (Hornsund, Kongsfjorden, Rijpfjorden) fjords located along the latitudinal/thermal gradient from the southern Norway (60 °N) to North of Svalbard (80 °N). The sediment cores were collected at 3 to 5 stations within the central basin at 150-300 m in each fjord during r/v Helmer Hansen and r/v Oceania cruises in 2014 and 2015. Vertical patterns of grain size and organic matter content and sources (Corg concentration, stable isotope δ13C signature, photosynthetic pigments concentration) have been analyzed. Sediment accumulation rates have been estimated with use of 210Pb dating method. Fresh carbon accumulation rate was estimated based on organic carbon concentration is surface sediments and mass sediment accumulation rate. The variability in metazoan productivity and carbon consumption (calculated based on the macro- and meiobenthic species biomass spectra in samples collected at the same stations) was also assessed to explore the patterns of biological controls of carbon storage in sediments. Carbon burial was estimated by multiplying organic carbon concentration in deepest sampled sediments and mass sediment accumulation rate. The effects of contrasting hydrological regimes and biological activity on the carbon storage in the studied fjords are discussed from the perspective of possible effects of climate warming driven changes on the Arctic fjordic sedimentary systems.

  20. Integrating clinical and biological information in a shanghai biobank: an introduction to the sample repository and information sharing platform project.

    PubMed

    Cui, Wenbin; Zheng, Peiyong; Yang, Jiahong; Zhao, Rong; Gao, Jiechun; Yu, Guangjun

    2015-02-01

    Biobanks are important resources and central tools for translational medicine, which brings scientific research outcomes to clinical practice. The key purpose of biobanking in translational medicine and other medical research is to provide biological samples that are integrated with clinical information. In 2008, the Shanghai Municipal Government launched the "Shanghai Tissue Bank" in an effort to promote research in translational medicine. Now a sharing service platform has been constructed to integrate clinical practice and biological information that can be used in diverse medical and pharmaceutical research studies. The platform collects two kinds of data: sample data and clinical data. The sample data are obtained from the hospital biobank management system, and mainly include the donors' age, gender, marital status, sample source, sample type, collection time, deposit time, and storage method. The clinical data are collected from the "Hospital-Link" system (a medical information sharing system that connects 23 tertiary hospitals in Shanghai). The main contents include donors' corresponding medication information, test reports, inspection reports, and hospital information. As of the end of September 2014, the project has a collection of 16,020 donors and 148,282 samples, which were obtained from 12 medical institutions, and automatically acquired donors' corresponding clinical data from the "Hospital-Link" system for 6830 occurrences. This project will contribute to scientific research at medical institutions in Shanghai, and will also support the development of the biopharmaceutical industry. In this article, we will describe the significance, the construction phases, the application prospects, and benefits of the sample repository and information sharing service platform.

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