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Sample records for automated blood sampling

  1. Manual versus automated blood sampling

    DEFF Research Database (Denmark)

    Teilmann, A C; Kalliokoski, Otto; Sørensen, Dorte B

    2014-01-01

    corticosterone metabolites, and expressed more anxious behavior than did the mice of the other groups. Plasma corticosterone levels of mice subjected to tail blood sampling were also elevated, although less significantly. Mice subjected to automated blood sampling were less affected with regard to the parameters......Facial vein (cheek blood) and caudal vein (tail blood) phlebotomy are two commonly used techniques for obtaining blood samples from laboratory mice, while automated blood sampling through a permanent catheter is a relatively new technique in mice. The present study compared physiological parameters......, glucocorticoid dynamics as well as the behavior of mice sampled repeatedly for 24 h by cheek blood, tail blood or automated blood sampling from the carotid artery. Mice subjected to cheek blood sampling lost significantly more body weight, had elevated levels of plasma corticosterone, excreted more fecal...

  2. Automated system for fractionation of blood samples

    Energy Technology Data Exchange (ETDEWEB)

    Lee, N. E.; Genung, R. K.; Johnson, W. F.; Mrochek, J. E.; Scott, C. D.

    1978-01-01

    A prototype system for preparing multiple fractions of blood components (plasma, washed red cells, and hemolysates) using automated techniques has been developed. The procedure is based on centrifugal separation and differential pressure-induced transfer in a rotor that has been designed to process numerous samples simultaneously. Red cells are sedimented against the outer walls of the sample chamber, and plasma is syphoned, by imposition of eithr a slight positive or negative pressure, into individual reservoirs in a collection ring. Washing of cells is performed in situ; samples of washed cells, either packed or in saline solution, can be recovered. Cellular hemolysates are prepared and automatically transferred to individual, commercially available collection vials ready for storage in liquid nitrogen or immediate analysis. The system has potential application in any biomedical area which requires high sample throughput and in which one or more of the blood fractions will be used. A separate unit has been designed and developed for the semiautomated cleaning of the blood processing vessel.

  3. Automated blood sampling systems for positron emission tomography

    International Nuclear Information System (INIS)

    Eriksson, L.; Holte, S.; Bohm, C.; Kesselberg, M.; Hovander, B.

    1988-01-01

    An automated blood sampling system has been constructed and evaluated. Two different detector units in the blood sampling system are compared. Results from studies of blood-brain barrier transfer of a C-11 labelled receptor antagonist will be discussed

  4. An automated blood sampling system used in positron emission tomography

    International Nuclear Information System (INIS)

    Eriksson, L.; Bohm, C.; Kesselberg, M.

    1988-01-01

    Fast dynamic function studies with positron emission tomography (PET), has the potential to give accurate information of physiological functions of the brain. This capability can be realised if the positron camera system accurately quantitates the tracer uptake in the brain with sufficiently high efficiency and in sufficiently short time intervals. However, in addition, the tracer concentration in blood, as a function of time, must be accurately determined. This paper describes and evaluates an automated blood sampling system. Two different detector units are compared. The use of the automated blood sampling system is demonstrated in studies of cerebral blood flow, in studies of the blood-brain barrier transfer of amino acids and of the cerebral oxygen consumption. 5 refs.; 7 figs

  5. Automated Blood Sample Preparation Unit (ABSPU) for Portable Microfluidic Flow Cytometry.

    Science.gov (United States)

    Chaturvedi, Akhil; Gorthi, Sai Siva

    2017-02-01

    Portable microfluidic diagnostic devices, including flow cytometers, are being developed for point-of-care settings, especially in conjunction with inexpensive imaging devices such as mobile phone cameras. However, two pervasive drawbacks of these have been the lack of automated sample preparation processes and cells settling out of sample suspensions, leading to inaccurate results. We report an automated blood sample preparation unit (ABSPU) to prevent blood samples from settling in a reservoir during loading of samples in flow cytometers. This apparatus automates the preanalytical steps of dilution and staining of blood cells prior to microfluidic loading. It employs an assembly with a miniature vibration motor to drive turbulence in a sample reservoir. To validate performance of this system, we present experimental evidence demonstrating prevention of blood cell settling, cell integrity, and staining of cells prior to flow cytometric analysis. This setup is further integrated with a microfluidic imaging flow cytometer to investigate cell count variability. With no need for prior sample preparation, a drop of whole blood can be directly introduced to the setup without premixing with buffers manually. Our results show that integration of this assembly with microfluidic analysis provides a competent automation tool for low-cost point-of-care blood-based diagnostics.

  6. Validation of a fully automated robotic setup for preparation of whole blood samples for LC-MS toxicology analysis

    DEFF Research Database (Denmark)

    Andersen, David Wederkinck; Rasmussen, Brian; Linnet, Kristian

    2012-01-01

    -mass spectrometry using several preparation techniques, including protein precipitation, solid-phase extraction and centrifugation, without any manual intervention. Pipetting of a known aliquot of whole blood was achieved by integrating a balance and performing gravimetric measurements. The system was able......A fully automated setup was developed for preparing whole blood samples using a Tecan Evo workstation. By integrating several add-ons to the robotic platform, the flexible setup was able to prepare samples from sample tubes to a 96-well sample plate ready for injection on liquid chromatography...... to handle 1,073 of 1,092 (98.3%) samples of whole blood from forensic material, including postmortem samples, without any need for repeating sample preparation. Only three samples required special treatment such as dilution. The addition of internal and calibration standards were validated by pipetting...

  7. Correction of an input function for errors introduced with automated blood sampling

    Energy Technology Data Exchange (ETDEWEB)

    Schlyer, D.J.; Dewey, S.L. [Brookhaven National Lab., Upton, NY (United States)

    1994-05-01

    Accurate kinetic modeling of PET data requires an precise arterial plasma input function. The use of automated blood sampling machines has greatly improved the accuracy but errors can be introduced by the dispersion of the radiotracer in the sampling tubing. This dispersion results from three effects. The first is the spreading of the radiotracer in the tube due to mass transfer. The second is due to the mechanical action of the peristaltic pump and can be determined experimentally from the width of a step function. The third is the adsorption of the radiotracer on the walls of the tubing during transport through the tube. This is a more insidious effect since the amount recovered from the end of the tube can be significantly different than that introduced into the tubing. We have measured the simple mass transport using [{sup 18}F]fluoride in water which we have shown to be quantitatively recovered with no interaction with the tubing walls. We have also carried out experiments with several radiotracers including [{sup 18}F]Haloperidol, [{sup 11}C]L-deprenyl, [{sup 18}]N-methylspiroperidol ([{sup 18}F]NMS) and [{sup 11}C]buprenorphine. In all cases there was some retention of the radiotracer by untreated silicone tubing. The amount retained in the tubing ranged from 6% for L-deprenyl to 30% for NMS. The retention of the radiotracer was essentially eliminated after pretreatment with the relevant unlabeled compound. For example less am 2% of the [{sup 18}F]NMS was retained in tubing treated with unlabelled NMS. Similar results were obtained with baboon plasma although the amount retained in the untreated tubing was less in all cases. From these results it is possible to apply a mathematical correction to the measured input function to account for mechanical dispersion and to apply a chemical passivation to the tubing to reduce the dispersion due to adsorption of the radiotracer on the tubing walls.

  8. [Automated serial diagnosis of donor blood samples. Ergonomic and economic organization structure].

    Science.gov (United States)

    Stoll, T; Fischer-Fröhlich, C L; Mayer, G; Hanfland, P

    1990-01-01

    A comprehensive computer-aided administration-system for blood-donors is presented. Ciphered informations of barcode-labels allow the automatic and nevertheless selective pipetting of samples by pipetting-robots. Self-acting analysis-results are transferred to a host-computer in order to actualize a donor data-base.

  9. Development of an Automated and Sensitive Microfluidic Device for Capturing and Characterizing Circulating Tumor Cells (CTCs from Clinical Blood Samples.

    Directory of Open Access Journals (Sweden)

    Priya Gogoi

    Full Text Available Current analysis of circulating tumor cells (CTCs is hindered by sub-optimal sensitivity and specificity of devices or assays as well as lack of capability of characterization of CTCs with clinical biomarkers. Here, we validate a novel technology to enrich and characterize CTCs from blood samples of patients with metastatic breast, prostate and colorectal cancers using a microfluidic chip which is processed by using an automated staining and scanning system from sample preparation to image processing. The Celsee system allowed for the detection of CTCs with apparent high sensitivity and specificity (94% sensitivity and 100% specificity. Moreover, the system facilitated rapid capture of CTCs from blood samples and also allowed for downstream characterization of the captured cells by immunohistochemistry, DNA and mRNA fluorescence in-situ hybridization (FISH. In a subset of patients with prostate cancer we compared the technology with a FDA-approved CTC device, CellSearch and found a higher degree of sensitivity with the Celsee instrument. In conclusion, the integrated Celsee system represents a promising CTC technology for enumeration and molecular characterization.

  10. Automated multi-dimensional liquid chromatography : sample preparation and identification of peptides from human blood filtrate

    NARCIS (Netherlands)

    Machtejevas, Egidijus; John, Harald; Wagner, Knut; Standker, Ludger; Marko-Varga, Gyorgy; Georg Forssmann, Wolf; Bischoff, Rainer; K. Unger, Klaus

    2004-01-01

    A comprehensive on-line sample clean-up with an integrated two-dimensional HPLC system was developed for the analysis of natural peptides. Samples comprised of endogenous peptides with molecular weights up to 20 kDa were generated from human hemofiltrate (HF) obtained from patients with chronic

  11. Automated extraction of DNA from blood and PCR setup using a Tecan Freedom EVO liquid handler for forensic genetic STR typing of reference samples

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Frøslev, Tobias G; Frank-Hansen, Rune

    2011-01-01

    17025 using the Qiagen MagAttract DNA Mini M48 kit (Qiagen GmbH, Hilden, Germany) from fresh whole blood and blood from deceased individuals. The workflow was simplified by returning the DNA extracts to the original tubes minimizing the risk of misplacing samples. The tubes that originally contained...... the samples were washed with MilliQ water before the return of the DNA extracts. The PCR was setup in 96-well microtiter plates. The methods were validated for the kits: AmpFlSTR Identifiler, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, WI......). The automated protocols allowed for extraction and addition of PCR master mix of 96 samples within 3.5h. In conclusion, we demonstrated that (1) DNA extraction with magnetic beads and (2) PCR setup for accredited, forensic genetic short tandem repeat typing can be implemented on a simple automated liquid...

  12. Manual versus automated streaking system in clinical microbiology laboratory: Performance evaluation of Previ Isola for blood culture and body fluid samples.

    Science.gov (United States)

    Choi, Qute; Kim, Hyun Jin; Kim, Jong Wan; Kwon, Gye Cheol; Koo, Sun Hoe

    2018-01-04

    The process of plate streaking has been automated to improve routine workflow of clinical microbiology laboratories. Although there were many evaluation reports about the inoculation of various body fluid samples, few evaluations have been reported for blood. In this study, we evaluated the performance of automated inoculating system, Previ Isola for various routine clinical samples including blood. Blood culture, body fluid, and urine samples were collected. All samples were inoculated on both sheep blood agar plate (BAP) and MacConkey agar plate (MCK) using Previ Isola and manual method. We compared two methods in aspect of quality and quantity of cultures, and sample processing time. To ensure objective colony counting, an enumeration reading reference was made through a preliminary experiment. A total of 377 nonduplicate samples (102 blood culture, 203 urine, 72 body fluid) were collected and inoculated. The concordance rate of quality was 100%, 97.0%, and 98.6% in blood, urine, and other body fluids, respectively. In quantitative aspect, it was 98.0%, 97.0%, and 95.8%, respectively. The Previ Isola took a little longer to inoculate the specimen than manual method, but the hands-on time decreased dramatically. The shortened hands-on time using Previ Isola was about 6 minutes per 10 samples. We demonstrated that the Previ Isola showed high concordance with the manual method in the inoculation of various body fluids, especially in blood culture sample. The use of Previ Isola in clinical microbiology laboratories is expected to save considerable time and human resources. © 2018 Wiley Periodicals, Inc.

  13. Designing an automated blood fractionation system.

    Science.gov (United States)

    McQuillan, Adrian C; Sales, Sean D

    2008-04-01

    UK Biobank will be collecting blood samples from a cohort of 500 000 volunteers and it is expected that the rate of collection will peak at approximately 3000 blood collection tubes per day. These samples need to be prepared for long-term storage. It is not considered practical to manually process this quantity of samples so an automated blood fractionation system is required. Principles of industrial automation were applied to the blood fractionation process leading to the requirement of developing a vision system to identify the blood fractions within the blood collection tube so that the fractions can be accurately aspirated and dispensed into micro-tubes. A prototype was manufactured and tested on a range of human blood samples collected in different tube types. A specially designed vision system was capable of accurately measuring the position of the plasma meniscus, plasma/buffy coat interface and the red cells/buffy coat interface within a vacutainer. A rack of 24 vacutainers could be processed in blood fractionation system offers a solution to the problem of processing human blood samples collected in vacutainers in a consistent manner and provides a means of ensuring data and sample integrity.

  14. Automated high-capacity on-line extraction and bioanalysis of dried blood spot samples using liquid chromatography/high-resolution accurate mass spectrometry.

    Science.gov (United States)

    Oliveira, Regina V; Henion, Jack; Wickremsinhe, Enaksha R

    2014-11-30

    Pharmacokinetic data to support clinical development of pharmaceuticals are routinely obtained from liquid plasma samples. The plasma samples require frozen shipment and storage and are extracted off-line from the liquid chromatography/tandem mass spectrometry (LC/MS/MS) systems. In contrast, the use of dried blood spot (DBS) sampling is an attractive alternative in part due to its benefits in microsampling as well as simpler sample storage and transport. However, from a practical aspect, sample extraction from DBS cards can be challenging as currently performed. The goal of this report was to integrate automated serial extraction of large numbers of DBS cards with on-line liquid chromatography/high-resolution accurate mass spectrometry (LC/HRAMS) bioanalysis. An automated system for direct DBS extraction coupled to a LC/HRAMS was employed for the quantification of midazolam (MDZ) and α-hydroxymidazolam (α-OHMDZ) in human blood. The target analytes were directly extracted from the DBS cards onto an on-line chromatographic guard column followed by HRAMS detection. No additional sample treatment was required. The automated DBS LC/HRAMS method was developed and validated, based on the measurement at the accurate mass-to-charge ratio of the target analytes to ensure specificity for the assay. The automated DBS LC/HRAMS method analyzed a DBS sample within 2 min without the need for punching or additional off-line sample treatment. The fully automated analytical method was shown to be sensitive and selective over the concentration range of 5 to 2000 ng/mL. Intra- and inter-day precision and accuracy was less than 15% (less than 20% at the LLOQ). The validated method was successfully applied to measure MDZ and α-OHMDZ in an incurred human sample after a single 7.5 mg dose of MDZ. The direct DBS LC/HRAMS method demonstrated successful implementation of automated DBS extraction and bioanalysis for MDZ and α-OHMDZ. This approach has the potential to promote workload

  15. Validation of an semi-automated multi component method using protein precipitation LC-MS-MS for the analysis of whole blood samples

    DEFF Research Database (Denmark)

    Slots, Tina

    BACKGROUND: Solid phase extraction (SPE) are one of many multi-component methods, but can be very time-consuming and labour-intensive. Protein precipitation is, on the other hand, a much simpler and faster sample pre-treatment than SPE, and protein precipitation also has the ability to cover...... a wider range of components. AIM: The aim was to develop a robust semi-automated analytical method for whole blood samples based on a protein precipitation method already used in the lab (Sørensen and Hasselstrøm, 2013). The setup should improve the speed, robustness, and reliability of anteand post...

  16. Automated counting of white blood cells in synovial fluid.

    NARCIS (Netherlands)

    R. de Jonge (Robert); R.W. Brouwer (Reinoud); M. Smit (Marij); M. de Frankrijker-Merkestijn; R.J. Dolhain; J.M.W. Hazes (Mieke); A.W. van Toorenenbergen (Albert); J. Lindemans (Jan)

    2004-01-01

    textabstractOBJECTIVES: To evaluate the performance of automated leucocyte (white blood cell; WBC) counting by comparison with manual counting. METHODS: The number of WBC was determined in heparinized synovial fluid samples by the use of (i) a standard urine cytometer (Kova) and a

  17. Novel automated blood separations validate whole cell biomarkers.

    Directory of Open Access Journals (Sweden)

    Douglas E Burger

    Full Text Available Progress in clinical trials in infectious disease, autoimmunity, and cancer is stymied by a dearth of successful whole cell biomarkers for peripheral blood lymphocytes (PBLs. Successful biomarkers could help to track drug effects at early time points in clinical trials to prevent costly trial failures late in development. One major obstacle is the inaccuracy of Ficoll density centrifugation, the decades-old method of separating PBLs from the abundant red blood cells (RBCs of fresh blood samples.To replace the Ficoll method, we developed and studied a novel blood-based magnetic separation method. The magnetic method strikingly surpassed Ficoll in viability, purity and yield of PBLs. To reduce labor, we developed an automated platform and compared two magnet configurations for cell separations. These more accurate and labor-saving magnet configurations allowed the lymphocytes to be tested in bioassays for rare antigen-specific T cells. The automated method succeeded at identifying 79% of patients with the rare PBLs of interest as compared with Ficoll's uniform failure. We validated improved upfront blood processing and show accurate detection of rare antigen-specific lymphocytes.Improving, automating and standardizing lymphocyte detections from whole blood may facilitate development of new cell-based biomarkers for human diseases. Improved upfront blood processes may lead to broad improvements in monitoring early trial outcome measurements in human clinical trials.

  18. Novel automated blood separations validate whole cell biomarkers.

    Science.gov (United States)

    Burger, Douglas E; Wang, Limei; Ban, Liqin; Okubo, Yoshiaki; Kühtreiber, Willem M; Leichliter, Ashley K; Faustman, Denise L

    2011-01-01

    Progress in clinical trials in infectious disease, autoimmunity, and cancer is stymied by a dearth of successful whole cell biomarkers for peripheral blood lymphocytes (PBLs). Successful biomarkers could help to track drug effects at early time points in clinical trials to prevent costly trial failures late in development. One major obstacle is the inaccuracy of Ficoll density centrifugation, the decades-old method of separating PBLs from the abundant red blood cells (RBCs) of fresh blood samples. To replace the Ficoll method, we developed and studied a novel blood-based magnetic separation method. The magnetic method strikingly surpassed Ficoll in viability, purity and yield of PBLs. To reduce labor, we developed an automated platform and compared two magnet configurations for cell separations. These more accurate and labor-saving magnet configurations allowed the lymphocytes to be tested in bioassays for rare antigen-specific T cells. The automated method succeeded at identifying 79% of patients with the rare PBLs of interest as compared with Ficoll's uniform failure. We validated improved upfront blood processing and show accurate detection of rare antigen-specific lymphocytes. Improving, automating and standardizing lymphocyte detections from whole blood may facilitate development of new cell-based biomarkers for human diseases. Improved upfront blood processes may lead to broad improvements in monitoring early trial outcome measurements in human clinical trials.

  19. Validation of a fully automated solid‐phase extraction and ultra‐high‐performance liquid chromatography–tandem mass spectrometry method for quantification of 30 pharmaceuticals and metabolites in post‐mortem blood and brain samples

    DEFF Research Database (Denmark)

    Nielsen, Marie Katrine Klose; Nedahl, Michael; Johansen, Sys Stybe

    2018-01-01

    In this study, we present the validation of an analytical method capable of quantifying 30 commonly encountered pharmaceuticals and metabolites in whole blood and brain tissue from forensic cases. Solid‐phase extraction was performed by a fully automated robotic system, thereby minimising manual...... labour and human error while increasing sample throughput, robustness, and traceability. The method was validated in blood in terms of selectivity, linear range, matrix effect, extraction recovery, process efficiency, carry‐over, stability, precision, and accuracy. Deuterated analogues of each analyte....../kg. Thus, the linear range covered both therapeutic and toxic levels. The method showed acceptable accuracy and precision, with accuracies ranging from 80 to 118% and precision below 19% for the majority of the analytes. Linear range, matrix effect, extraction recovery, process efficiency, precision...

  20. Correlation between glucose concentrations in serum, plasma, and whole blood measured by a point-of-care glucometer and serum glucose concentration measured by an automated biochemical analyzer for canine and feline blood samples.

    Science.gov (United States)

    Tauk, Barbara S; Drobatz, Kenneth J; Wallace, Koranda A; Hess, Rebecka S

    2015-06-15

    To investigate the correlation between glucose concentrations in serum, plasma, and whole blood measured by a point-of-care glucometer (POCG) and serum glucose concentration measured by a biochemical analyzer. Prospective clinical study. 96 blood samples from 80 dogs and 90 blood samples from 65 cats. Serum, plasma, and whole blood were obtained from each blood sample. The glucose concentrations in serum, plasma, and whole blood measured by a POCG were compared with the serum glucose concentration measured by a biochemical analyzer by use of the Lin concordance correlation coefficient (ρc) and Bland-Altman plots. For both canine and feline samples, glucose concentrations in serum and plasma measured by the POCG were more strongly correlated with the serum glucose concentration measured by the biochemical analyzer (ρc, 0.98 for both canine serum and plasma; ρc, 0.99 for both feline serum and plasma) than was that in whole blood (ρc, 0.62 for canine samples; ρc, 0.90 for feline samples). The mean difference between the glucose concentrations determined by the biochemical analyzer and the POCG in serum, plasma, and whole blood was 0.4, 0.3, and 31 mg/dL, respectively, for canine samples and 7, 6, and 32 mg/dL, respectively, for feline samples. Results indicated that use of a POCG to measure glucose concentrations in serum or plasma may increase the accuracy and reliability of diagnostic and treatment decisions associated with glucose homeostasis disorders in dogs and cats.

  1. Neonatal blood gas sampling methods

    African Journals Online (AJOL)

    adequate collateral circulation before radial arterial puncture may not be a reliable predictor of subsequent risk of vascular injury.75. Conclusion and recommendations. Indwelling arterial catheters remain a practical, reliable and accurate method of neonatal blood gas sampling, provided they are inserted and maintained ...

  2. Automated sampling and control of gaseous simulations

    KAUST Repository

    Huang, Ruoguan

    2013-05-04

    In this work, we describe a method that automates the sampling and control of gaseous fluid simulations. Several recent approaches have provided techniques for artists to generate high-resolution simulations based on a low-resolution simulation. However, often in applications the overall flow in the low-resolution simulation that an animator observes and intends to preserve is composed of even lower frequencies than the low resolution itself. In such cases, attempting to match the low-resolution simulation precisely is unnecessarily restrictive. We propose a new sampling technique to efficiently capture the overall flow of a fluid simulation, at the scale of user\\'s choice, in such a way that the sampled information is sufficient to represent what is virtually perceived and no more. Thus, by applying control based on the sampled data, we ensure that in the resulting high-resolution simulation, the overall flow is matched to the low-resolution simulation and the fine details on the high resolution are preserved. The samples we obtain have both spatial and temporal continuity that allows smooth keyframe matching and direct manipulation of visible elements such as smoke density through temporal blending of samples. We demonstrate that a user can easily configure a simulation with our system to achieve desired results. © 2013 Springer-Verlag Berlin Heidelberg.

  3. Percutaneous umbilical cord blood sampling - slideshow

    Science.gov (United States)

    ... page: //medlineplus.gov/ency/presentations/100196.htm Percutaneous umbilical cord blood sampling - series—Normal anatomy To use the ... or blood disorder, your doctor may recommend percutaneous umbilical cord blood sampling (PUBS), which is performed at 18 ...

  4. Is automated platelet counting still a problem in thrombocytopenic blood?

    Directory of Open Access Journals (Sweden)

    Raimundo Antônio Gomes Oliveira

    Full Text Available CONTEXT: Reliable platelet counting is crucial for indicating prophylactic platelet transfusion in thrombocytopenic patients. OBJECTIVE: To evaluate the precision and accuracy of platelet counting for thrombocytopenic patients, using four different automated counters in comparison with the Brecher & Cronkite reference method recommended by the International Committee for Standardization in Hematology (ICSH. TYPE OF STUDY: Automated platelet counting assessment in thrombocytopenic patients. SETTING: Hematology Laboratory, Hospital do Servidor Público Estadual de São Paulo, and the Hematology Division of Instituto Adolfo Lutz, São Paulo, SP, Brazil. MAIN MEASUREMENTS: Brecher & Cronkite reference method and four different automated platelet counters. PARTICIPANTS: 43 thrombocytopenic patients with platelet counts of less than 30,000/µl RESULTS: The ADVIA-120 (Bayer, Coulter STKS, H1 System (Technicom-Bayer and Coulter T-890 automatic instruments presented great precision and accuracy in relation to laboratory thrombocytopenic samples obtained by diluting blood from normal donors. However, when thrombocytopenic patients were investigated, all the counters except ADVIA (which is based on volume and refraction index showed low accuracy when compared to the Brecher & Cronkite reference method (ICSH. The ADVIA counter showed high correlation (r = 0.947. However, all counters showed flags in thrombocytopenic samples. CONCLUSION: The Brecher & Cronkite reference method should always be indicated in thrombocytopenic patients for platelet counts below 30,000 plt /µl obtained in one dimensional counters.

  5. High throughput sample processing and automated scoring

    Directory of Open Access Journals (Sweden)

    Gunnar eBrunborg

    2014-10-01

    Full Text Available The comet assay is a sensitive and versatile method for assessing DNA damage in cells. In the traditional version of the assay, there are many manual steps involved and few samples can be treated in one experiment. High throughput modifications have been developed during recent years, and they are reviewed and discussed. These modifications include accelerated scoring of comets; other important elements that have been studied and adapted to high throughput are cultivation and manipulation of cells or tissues before and after exposure, and freezing of treated samples until comet analysis and scoring. High throughput methods save time and money but they are useful also for other reasons: large-scale experiments may be performed which are otherwise not practicable (e.g., analysis of many organs from exposed animals, and human biomonitoring studies, and automation gives more uniform sample treatment and less dependence on operator performance. The high throughput modifications now available vary largely in their versatility, capacity, complexity and costs. The bottleneck for further increase of throughput appears to be the scoring.

  6. Automated extraction of lysergic acid diethylamide (LSD) and N-demethyl-LSD from blood, serum, plasma, and urine samples using the Zymark RapidTrace with LC/MS/MS confirmation.

    Science.gov (United States)

    de Kanel, J; Vickery, W E; Waldner, B; Monahan, R M; Diamond, F X

    1998-05-01

    A forensic procedure for the quantitative confirmation of lysergic acid diethylamide (LSD) and the qualitative confirmation of its metabolite, N-demethyl-LSD, in blood, serum, plasma, and urine samples is presented. The Zymark RapidTrace was used to perform fully automated solid-phase extractions of all specimen types. After extract evaporation, confirmations were performed using liquid chromatography (LC) followed by positive electrospray ionization (ESI+) mass spectrometry/mass spectrometry (MS/MS) without derivatization. Quantitation of LSD was accomplished using LSD-d3 as an internal standard. The limit of quantitation (LOQ) for LSD was 0.05 ng/mL. The limit of detection (LOD) for both LSD and N-demethyl-LSD was 0.025 ng/mL. The recovery of LSD was greater than 95% at levels of 0.1 ng/mL and 2.0 ng/mL. For LSD at 1.0 ng/mL, the within-run and between-run (different day) relative standard deviation (RSD) was 2.2% and 4.4%, respectively.

  7. Efeitos quantitativos da estocagem de sangue periférico nas determinações do hemograma automatizado Storage effects on peripheral blood samples as identified from automated hemograms

    Directory of Open Access Journals (Sweden)

    Michele Dalanhol

    2010-02-01

    Full Text Available O presente trabalho avaliou as possíveis alterações em vários parâmetros do hemograma (contagem de eritrócitos totais, hematócrito, concentração de hemoglobina, volume corpuscular médio (VCM, hemoglobina corpuscular média (HCM, concentração da hemoglobina corpuscular média (CHCM, contagem total de leucócitos e contagem de plaquetas, frente a diferentes tempos de armazenamento da amostra em ambiente refrigerado a 4ºC e temperatura ambiente. As determinações foram realizadas através do contador automatizado Sysmex® XT2000i. As amostras sanguíneas foram obtidas de indivíduos sem alterações hematológicas diagnosticadas, sem clínica de doença, sendo considerados indivíduos com valores de referências hematológicos normais. Os resultados foram avaliados através de análise estatística descritiva e comparação de médias através da análise de variância (ANOVA. Os resultados dos parâmetros CHCM e contagem de plaquetas mostraram diferenças estatisticamente significativas para ambas as temperaturas de estocagem. Na temperatura ambiente, os parâmetros que também apresentaram diferença estatística significante foram para o hematócrito, VCM e índice de variação do tamanho dos eritrócitos ( RDW. Conclui-se, portanto, que os resultados dos hemogramas liberados pelo aparelho analisado podem ser satisfatórios quando realizados entre 12 a 24 horas após a coleta da amostra para a maioria dos parâmetros avaliados.This study evaluated possible alterations in different hematologic parameters [total erythrocyte count, hematocrit, hemoglobin concentration, mean corpuscular volume (MCV, mean corpuscular hemoglobin (MCH, mean corpuscular hemoglobin concentration (MHCM, total leukocyte count and platelet count], of blood samples submitted to varied storage times at both 4ºC and at room temperature. The analyses were performed using a Sysmex XT2000i automated hematology analyzer. Written consent was obtained from donors

  8. High plasma corticosterone levels persist during frequent automatic blood sampling in rats

    DEFF Research Database (Denmark)

    Abelson, Klas S P; Adem, Bashir; Royo, Felix

    2005-01-01

    the importance of considering the frequency of blood withdrawal during automated blood sampling. This parameter may have an impact on the experimental results when using blood corticosterone levels as a stress marker, but also during any in vivo study where blood is collected, since high corticosterone levels...

  9. Transfusion management using a remote-controlled, automated blood storage.

    Science.gov (United States)

    Pagliaro, Pasqualepaolo; Turdo, Rosalia

    2008-04-01

    Generally, the safety of transfusion terapies for patients depends in part on the distribution of the blood products. The prevention of adverse events can be aided by technological means, which, besides improving the traceability of the process, make errors less likely. In this context, the latest frontier in automation and computerisation is the remote-controlled, automated refrigerator for blood storage. Computer cross-matching is an efficient and safe method for assigning blood components, based on Information Technology applied to typing and screening. This method can be extended to the management of an automated blood refrigerator, the programme of which is interfaced with the Transfusion Service's information system. The connection we made in our Service between EmoNet and Hemosafe enables real-time, remote-controlled management of the following aspects of blood component distribution: a) release of autologous and allogeneic units already allocated to a patient, b) release of available units, which can be allocated by remote-control to known patients, in the presence of a valid computer cross-match, c) release of O-negative units of blood for emergencies. Our system combines an information database, which enables computer cross-matching, with an automated refrigerator for blood storage with controlled access managed remotely by the Transfusion Service. The effectiveness and safety of the system were validated during the 4 months of its routine use in the Transfusion Service's outpatient department. The safety and efficiency of the distribution of blood products can and must be increased by the use of technological innovations. With the EmoNet/Hemosafe system, the responsibility for the remote-controlled distribution of red blood cell concentrates remains with the chief of the Transfusion Services, through the use of automated computer procedures and supported by continuous training of technicians and nursing staff.

  10. Automation of a dust sampling train | Akinola | Journal of Modeling ...

    African Journals Online (AJOL)

    The results obtained from this work show that the flue gas sampling process can be automated using microprocessor-based control system and a sampling train. Using the sampling train developed by the British Coal Utilization Research Association (BCURA) with both the internal and external flow-meter arrangements, the ...

  11. [Clinical application of automated digital image analysis for morphology review of peripheral blood leukocyte].

    Science.gov (United States)

    Xing, Ying; Yan, Xiaohua; Pu, Chengwei; Shang, Ke; Dong, Ning; Wang, Run; Wang, Jianzhong

    2016-03-01

    To explore the clinical application of automated digital image analysis in leukocyte morphology examination when review criteria of hematology analyzer are triggered. The reference range of leukocyte differentiation by automated digital image analysis was established by analyzing 304 healthy blood samples from Peking University First Hospital. Six hundred and ninty-seven blood samples from Peking University First Hospital were randomly collected from November 2013 to April 2014, complete blood cells were counted on hematology analyzer, blood smears were made and stained at the same time. Blood smears were detected by automated digital image analyzer and the results were checked (reclassification) by a staff with abundant morphology experience. The same smear was examined manually by microscope. The results by manual microscopic differentiation were used as"golden standard", and diagnostic efficiency of abnormal specimens by automated digital image analysis was calculated, including sensitivity, specificity and accuracy. The difference of abnormal leukocytes detected by two different methods was analyzed in 30 samples of hematological and infectious diseases. Specificity of identifying abnormalities of white blood cells by automated digital image analysis was more than 90% except monocyte. Sensitivity of neutrophil toxic abnormities (including Döhle body, toxic granulate and vacuolization) was 100%; sensitivity of blast cells, immature granulates and atypical lymphocytes were 91.7%, 60% to 81.5% and 61.5%, respectively. Sensitivity of leukocyte differential count was 91.8% for neutrophils, 88.5% for lymphocytes, 69.1% for monocytes, 78.9% for eosinophils and 36.3 for basophils. The positive rate of recognizing abnormal cells (blast, immature granulocyte and atypical lymphocyte) by manual microscopic method was 46.7%, 53.3% and 10%, respectively. The positive rate of automated digital image analysis was 43.3%, 60% and 10%, respectively. There was no statistic

  12. Blood sampling from adrenal gland vein

    International Nuclear Information System (INIS)

    Sun Yong; Ni Caifang

    2009-01-01

    Adrenal gland vein sampling is an interventional method to get the blood samples from the adrenal gland vein. The blood is obtained via a catheter which is selectively inserted in the adrenal gland vein. This technique is mainly used to be diagnostic for primary hyperaldosteronism. A full knowledge of the anatomy and variations of the adrenal gland vein, serious preoperative preparation and skilled catheterization manipulation are necessary for obtaining sufficient blood sample and for reducing the occurrence of complications. Providing the physicians with definite diagnostic evidence and being technically feasible, adrenal gland vein sampling should become one of the routine examinations for clarifying the cause of primary hyperaldosteronism. (authors)

  13. Automated measurement of retinal blood vessel tortuosity

    Science.gov (United States)

    Joshi, Vinayak; Reinhardt, Joseph M.; Abramoff, Michael D.

    2010-03-01

    Abnormalities in the vascular pattern of the retina are associated with retinal diseases and are also risk factors for systemic diseases, especially cardiovascular diseases. The three-dimensional retinal vascular pattern is mostly formed congenitally, but is then modified over life, in response to aging, vessel wall dystrophies and long term changes in blood flow and pressure. A characteristic of the vascular pattern that is appreciated by clinicians is vascular tortuosity, i.e. how curved or kinked a blood vessel, either vein or artery, appears along its course. We developed a new quantitative metric for vascular tortuosity, based on the vessel's angle of curvature, length of the curved vessel over its chord length (arc to chord ratio), number of curvature sign changes, and combined these into a unidimensional metric, Tortuosity Index (TI). In comparison to other published methods this method can estimate appropriate TI for vessels with constant curvature sign and vessels with equal arc to chord ratios, as well. We applied this method to a dataset of 15 digital fundus images of 8 patients with Facioscapulohumeral muscular dystrophy (FSHD), and to the other publically available dataset of 60 fundus images of normal cases and patients with hypertensive retinopathy, of which the arterial and venous tortuosities have also been graded by masked experts (ophthalmologists). The method produced exactly the same rank-ordered list of vessel tortuosity (TI) values as obtained by averaging the tortuosity grading given by 3 ophthalmologists for FSHD dataset and a list of TI values with high ranking correlation with the ophthalmologist's grading for the other dataset. Our results show that TI has potential to detect and evaluate abnormal retinal vascular structure in early diagnosis and prognosis of retinopathies.

  14. Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies

    OpenAIRE

    Jo, Su Yeon; Lee, Ju Mi; Kim, Hye Lim; Sin, Kyeong Hwa; Lee, Hyeon Ji; Chang, Chulhun Ludgerus; Kim, Hyung-Hoi

    2016-01-01

    Background ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. Methods In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to...

  15. On-line Automated Sample Preparation-Capillary Gas Chromatography for the Analysis of Plasma Samples.

    NARCIS (Netherlands)

    Louter, A.J.H.; van der Wagt, R.A.C.A.; Brinkman, U.A.T.

    1995-01-01

    An automated sample preparation module, (the automated sample preparation with extraction columns, ASPEC), was interfaced with a capillary gas chromatograph (GC) by means of an on-column interface. The system was optimised for the determination of the antidepressant trazodone in plasma. The clean-up

  16. Preanalytical Blood Sampling Errors in Clinical Settings

    International Nuclear Information System (INIS)

    Zehra, N.; Malik, A. H.; Arshad, Q.; Sarwar, S.; Aslam, S.

    2016-01-01

    Background: Blood sampling is one of the common procedures done in every ward for disease diagnosis and prognosis. Daily hundreds of samples are collected from different wards but lack of appropriate knowledge of blood sampling by paramedical staff and accidental errors make the samples inappropriate for testing. Thus the need to avoid these errors for better results still remains. We carried out this research with an aim to determine the common errors during blood sampling; find factors responsible and propose ways to reduce these errors. Methods: A cross sectional descriptive study was carried out at the Military and Combined Military Hospital Rawalpindi during February and March 2014. A Venous Blood Sampling questionnaire (VBSQ) was filled by the staff on voluntary basis in front of the researchers. The staff was briefed on the purpose of the survey before filling the questionnaire. Sample size was 228. Results were analysed using SPSS-21. Results: When asked in the questionnaire, around 61.6 percent of the paramedical staff stated that they cleaned the vein by moving the alcohol swab from inward to outwards while 20.8 percent of the staff reported that they felt the vein after disinfection. On contrary to WHO guidelines, 89.6 percent identified that they had a habit of placing blood in the test tube by holding it in the other hand, which should actually be done after inserting it into the stand. Although 86 percent thought that they had ample knowledge regarding the blood sampling process but they did not practice it properly. Conclusion: Pre analytical blood sampling errors are common in our setup. Eighty six percent participants though thought that they had adequate knowledge regarding blood sampling, but most of them were not adhering to standard protocols. There is a need of continued education and refresher courses. (author)

  17. Fetal scalp blood sampling during labor

    DEFF Research Database (Denmark)

    Chandraharan, Edwin; Wiberg, Nana

    2014-01-01

    Fetal cardiotocography is characterized by low specificity; therefore, in an attempt to ensure fetal well-being, fetal scalp blood sampling has been recommended by most obstetric societies in the case of a non-reassuring cardiotocography. The scientific agreement on the evidence for using fetal...... scalp blood sampling to decrease the rate of operative delivery for fetal distress is ambiguous. Based on the same studies, a Cochrane review states that fetal scalp blood sampling increases the rate of instrumental delivery while decreasing neonatal acidosis, whereas the National Institute of Health...... and Clinical Excellence guideline considers that fetal scalp blood sampling decreases instrumental delivery without differences in other outcome variables. The fetal scalp is supplied by vessels outside the skull below the level of the cranial vault, which is likely to be compressed during contractions...

  18. An automated immunoradiometric assay of thyrotrophin (TSH) in dried blood filter paper spots

    International Nuclear Information System (INIS)

    John, R.; Woodhead, J.S.

    1982-01-01

    An immunoradiometric two-site assay for thyrotrophin (TSH) in dried blood filter paper spots is described. The assay is automated by means of the Kemtek 3000 automated immunoassay system. The technique uses a 6.0 mm disc punched from the dried blood samples collected as part of the screening programme for phenylketonuria. The method is sensitive and precise, and results correlate well with those obtained in TSH assays of serum samples. The procedure is rapid, results being available within 24 h of receipt of samples. Of 25204 specimens so far screened by this assay, 99.9% have TSH levels less than 15 mU/l. One false positive result has been obtained and six confirmed cases of neonatal hypothyroidism detected, giving a prevalence of 1 in 4200. (Auth.)

  19. Automated immunoradiometric assay of thyrotrophin (TSH) in dried blood filter paper spots

    Energy Technology Data Exchange (ETDEWEB)

    John, R.; Woodhead, J.S. (Welsh National School of Medicine, Cardiff (UK))

    1982-11-10

    An immunoradiometric two-site assay for thyrotrophin (TSH) in dried blood filter paper spots is described. The assay is automated by means of the Kemtek 3000 automated immunoassay system. The technique uses a 6.0 mm disc punched from the dried blood samples collected as part of the screening programme for phenylketonuria. The method is sensitive and precise, and results correlate well with those obtained in TSH assays of serum samples. The procedure is rapid, results being available within 24 h of receipt of samples. Of 25204 specimens so far screened by this assay, 99.9% have TSH levels less than 15 mU/l. One false positive result has been obtained and six confirmed cases of neonatal hypothyroidism detected, giving a prevalence of 1 in 4200.

  20. Blood Sample Transportation by Pneumatic Transportation Systems

    DEFF Research Database (Denmark)

    Nybo, Mads; Lund, Merete E; Titlestad, Kjell

    2018-01-01

    BACKGROUND: Pneumatic transportation systems (PTSs) are increasingly used for transportation of blood samples to the core laboratory. Many studies have investigated the impact of these systems on different types of analyses, but to elucidate whether PTSs in general are safe for transportation...... of blood samples, existing literature on the subject was systematically assessed. METHODS: A systematic literature review was conducted following the preferred reporting items for systematic reviews and metaanalyses (PRISMA) Statement guidelines to gather studies investigating the impact of PTS on analyses...... in blood samples. Studies were extracted from PubMed and Embase. The search period ended November 2016. RESULTS: A total of 39 studies were retrieved. Of these, only 12 studies were conducted on inpatients, mainly intensive care unit patients. Blood gases, hematology, and clinical chemistry were well...

  1. 21 CFR 864.9175 - Automated blood grouping and antibody test system.

    Science.gov (United States)

    2010-04-01

    ...) Identification. An automated blood grouping and antibody test system is a device used to group erythrocytes (red blood cells) and to detect antibodies to blood group antigens. (b) Classification. Class II (performance... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated blood grouping and antibody test system...

  2. [Comparison of arterial blood sample kits].

    Science.gov (United States)

    Calaf, N; Giner, J; Codina, E; Feixas, T; González, M; Casan, P

    2004-08-01

    Most inaccuracies in the analysis of gases and electrolytes in arterial blood samples are due to preanalytic factors, among which is the type of equipment used for blood collection. Our objective was to compare arterial blood gas sample kits used under clinical conditions and to evaluate the impact of delay in estimation on variability in results. In 2 types of study we compared 5 kits (Radiometer's Pico 70, Becton Dickinson's Preset, SIMS Portex's Pro-Vent, SIMS-Concord's Pulsator, and Marquest's Quick ABG). In the first study kitsyringe assignment was randomized for collecting arterial blood samples from 160 consecutive patients to evaluate practical aspects of using them and the presence of bubbles in the samples taken. The second study evaluated the effects of delays of 30 and 60 minutes in estimation and of the type of heparin used in 54 blood samples. The kits which produced the fewest bubbles, gave samples with the greatest stability, and had the least impact on ion concentration were Radiometer's Pico 70 and SIMS-Portex's Pro-vent.

  3. Non-Contact Conductivity Measurement for Automated Sample Processing Systems

    Science.gov (United States)

    Beegle, Luther W.; Kirby, James P.

    2012-01-01

    A new method has been developed for monitoring and control of automated sample processing and preparation especially focusing on desalting of samples before analytical analysis (described in more detail in Automated Desalting Apparatus, (NPO-45428), NASA Tech Briefs, Vol. 34, No. 8 (August 2010), page 44). The use of non-contact conductivity probes, one at the inlet and one at the outlet of the solid phase sample preparation media, allows monitoring of the process, and acts as a trigger for the start of the next step in the sequence (see figure). At each step of the muti-step process, the system is flushed with low-conductivity water, which sets the system back to an overall low-conductivity state. This measurement then triggers the next stage of sample processing protocols, and greatly minimizes use of consumables. In the case of amino acid sample preparation for desalting, the conductivity measurement will define three key conditions for the sample preparation process. First, when the system is neutralized (low conductivity, by washing with excess de-ionized water); second, when the system is acidified, by washing with a strong acid (high conductivity); and third, when the system is at a basic condition of high pH (high conductivity). Taken together, this non-contact conductivity measurement for monitoring sample preparation will not only facilitate automation of the sample preparation and processing, but will also act as a way to optimize the operational time and use of consumables

  4. System Design and Development of a Robotic Device for Automated Venipuncture and Diagnostic Blood Cell Analysis.

    Science.gov (United States)

    Balter, Max L; Chen, Alvin I; Fromholtz, Alex; Gorshkov, Alex; Maguire, Tim J; Yarmush, Martin L

    2016-10-01

    Diagnostic blood testing is the most prevalent medical procedure performed in the world and forms the cornerstone of modern health care delivery. Yet blood tests are still predominantly carried out in centralized labs using large-volume samples acquired by manual venipuncture, and no end-to-end solution from blood draw to sample analysis exists today. Our group is developing a platform device that merges robotic phlebotomy with automated diagnostics to rapidly deliver patient information at the site of the blood draw. The system couples an image-guided venipuncture robot, designed to address the challenges of routine venous access, with a centrifuge-based blood analyzer to obtain quantitative measurements of hematology. In this paper, we first present the system design and architecture of the integrated device. We then perform a series of in vitro experiments to evaluate the cannulation accuracy of the system on blood vessel phantoms. Next, we assess the effects of vessel diameter, needle gauge, flow rate, and viscosity on the rate of sample collection. Finally, we demonstrate proof-of-concept of a white cell assay on the blood analyzer using in vitro human samples spiked with fluorescently labeled microbeads.

  5. Improving blood sample logistics using simulation

    DEFF Research Database (Denmark)

    Jørgensen, Pelle Morten Thomas; Jacobsen, Peter

    2012-01-01

    Using simulation as an approach to display and improve internal logistics and handling at hospitals has great potential. This research will show how a simulation model can be used to evaluate changes made to two different cases of transportation of blood samples at a hospital, by evaluating...

  6. A bench-top automated workstation for nucleic acid isolation from clinical sample types.

    Science.gov (United States)

    Thakore, Nitu; Garber, Steve; Bueno, Arial; Qu, Peter; Norville, Ryan; Villanueva, Michael; Chandler, Darrell P; Holmberg, Rebecca; Cooney, Christopher G

    2018-04-18

    Systems that automate extraction of nucleic acid from cells or viruses in complex clinical matrices have tremendous value even in the absence of an integrated downstream detector. We describe our bench-top automated workstation that integrates our previously-reported extraction method - TruTip - with our newly-developed mechanical lysis method. This is the first report of this method for homogenizing viscous and heterogeneous samples and lysing difficult-to-disrupt cells using "MagVor": a rotating magnet that rotates a miniature stir disk amidst glass beads confined inside of a disposable tube. Using this system, we demonstrate automated nucleic acid extraction from methicillin-resistant Staphylococcus aureus (MRSA) in nasopharyngeal aspirate (NPA), influenza A in nasopharyngeal swabs (NPS), human genomic DNA from whole blood, and Mycobacterium tuberculosis in NPA. The automated workstation yields nucleic acid with comparable extraction efficiency to manual protocols, which include commercially-available Qiagen spin column kits, across each of these sample types. This work expands the scope of applications beyond previous reports of TruTip to include difficult-to-disrupt cell types and automates the process, including a method for removal of organics, inside a compact bench-top workstation. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. Optimizing centrifugation of coagulation samples in laboratory automation.

    Science.gov (United States)

    Suchsland, Juliane; Friedrich, Nele; Grotevendt, Anne; Kallner, Anders; Lüdemann, Jan; Nauck, Matthias; Petersmann, Astrid

    2014-08-01

    High acceleration centrifugation conditions are used in laboratory automation systems to reduce the turnaround time (TAT) of clinical chemistry samples, but not of coagulation samples. This often requires separate sample flows. The CLSI guideline and manufacturers recommendations for coagulation assays aim at reducing platelet counts. For measurement of prothrombin time (PT) and activated partial thromboplastin time (APTT) platelet counts (Plt) below 200×10(9)/L are recommended. Other coagulation assays may require even lower platelet counts, e.g., less than 10 × 10(9)/L. Unifying centrifugation conditions can facilitate the integration of coagulation samples in the overall workflow of a laboratory automation system. We evaluated centrifugation conditions of coagulation samples by using high acceleration centrifugation conditions (5 min; 3280×g) in a single and two consecutive runs. RESULTS of coagulation assays [PT, APTT, coagulation factor VIII (F. VIII) and protein S] and platelet counts were compared after the first and second centrifugation. Platelet counts below 200×10(9)/L were obtained in all samples after the first centrifugation and less than 10 × 10(9)/L was obtained in 73% of the samples after a second centrifugation. Passing-Bablok regression analyses showed an equal performance of PT, APTT and F. VIII after first and second centrifugation whereas protein S measurements require a second centrifugation. Coagulation samples can be integrated into the workflow of a laboratory automation system using high acceleration centrifugation. A single centrifugation was sufficient for PT, APTT and F. VIII whereas two successive centrifugations appear to be sufficient for protein S activity.

  8. Automating Groundwater Sampling At Hanford, The Next Step

    International Nuclear Information System (INIS)

    Connell, C.W.; Conley, S.F.; Hildebrand, R.D.; Cunningham, D.E.

    2010-01-01

    Historically, the groundwater monitoring activities at the Department of Energy's Hanford Site in southeastern Washington State have been very 'people intensive.' Approximately 1500 wells are sampled each year by field personnel or 'samplers.' These individuals have been issued pre-printed forms showing information about the well(s) for a particular sampling evolution. This information is taken from 2 official electronic databases: the Hanford Well information System (HWIS) and the Hanford Environmental Information System (HEIS). The samplers used these hardcopy forms to document the groundwater samples and well water-levels. After recording the entries in the field, the samplers turned the forms in at the end of the day and other personnel posted the collected information onto a spreadsheet that was then printed and included in a log book. The log book was then used to make manual entries of the new information into the software application(s) for the HEIS and HWIS databases. A pilot project for automating this extremely tedious process was lauched in 2008. Initially, the automation was focused on water-level measurements. Now, the effort is being extended to automate the meta-data associated with collecting groundwater samples. The project allowed electronic forms produced in the field by samplers to be used in a work flow process where the data is transferred to the database and electronic form is filed in managed records - thus eliminating manually completed forms. Elimating the manual forms and streamlining the data entry not only improved the accuracy of the information recorded, but also enhanced the efficiency and sampling capacity of field office personnel.

  9. 21 CFR 868.1100 - Arterial blood sampling kit.

    Science.gov (United States)

    2010-04-01

    ...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood samples... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Arterial blood sampling kit. 868.1100 Section 868...

  10. Automated nucleic acid amplification testing in blood banks: An additional layer of blood safety

    Directory of Open Access Journals (Sweden)

    Pragati Chigurupati

    2015-01-01

    Full Text Available Context: A total of 30 million blood components are transfused each year in India. Blood safety thus becomes a top priority, especially with a population of around 1.23 billion and a high prevalence rate of human immunodeficiency virus (HIV, hepatitis B virus (HBV and hepatitis C virus (HCV in general population. Nucleic acid amplification testing (NAT in blood donor screening has been implemented in many developed countries to reduce the risk of transfusion-transmitted viral infections (TTIs. NAT takes care of the dynamics of window period of viruses and offers the safest blood pack for donation. Aims: The aim of this study is to show the value of NAT in blood screening. Settings and Design: Dhanavantari Blood Bank, Rajahmundry, Andhra Pradesh, India. Subjects and Methods: Over a period of 1 year from January 2012 to December 2012, a total number of 15,000 blood donor samples were subjected to tests for HIV, HBV, and HCV by enzyme-linked immunosorbent assay (ELISA method and 8000 ELISA nonreactive samples were subjected for NAT using multiplex polymerase chain reaction technology. Results: Of the 15,000 donors tested, 525 were seroreactive. In 8000 ELISA negative blood samples subjected to NAT, 4 donor samples were reactive for HBV. The NAT yield was 1 in 2000. Conclusions: NAT could detect HIV, HBV, and HCV cases in blood donor samples those were undetected by serological tests. NAT could interdict 2500 infectious donations among our approximate 5 million annual blood donations.

  11. Comparison of a human portable blood glucose meter, veterinary portable blood glucose meter, and automated chemistry analyzer for measurement of blood glucose concentrations in dogs.

    Science.gov (United States)

    Johnson, Beth M; Fry, Michael M; Flatland, Bente; Kirk, Claudia A

    2009-12-01

    To compare blood glucose concentrations measured with 2 portable blood glucose meters (PBGMs) validated for use in dogs (PBGM-D) and humans (PBGM-H) and an automated chemistry analyzer. Validation study. 92 samples of fresh whole blood and plasma from 83 dogs with various diseases. Each PBGM was used to measure whole blood glucose concentration, and the automated analyzer was used to measure plasma glucose concentration. Passing-Bablok linear regression and Bland-Altman plots were used to determine correlations and bias between the PBGMs and the automated analyzer. Calculated acceptability limits based on combined inherent instrument imprecision were used with Bland-Altman plots to determine agreement. Clinical relevance was assessed via error grid analysis. Although correlation between results of both PBGMs and the standard analyzer was > 0.90, disagreement was greater than could be explained by instrument imprecision alone. Mean difference between PBGM-H and chemistry-analyzer values was -15.8 mg/dL. Mean difference between PBGM-D and chemistry-analyzer values was 2.4 mg/dL. Linear regression analysis revealed proportional bias of PBGM-H (greater disagreement at higher glucose concentrations); no proportional bias was detected for PBGM-D. No constant bias was detected for either PBGM. Error grid analysis revealed all measurements from both PBGMs were within zones without an anticipated effect on clinical outcome. Neither PBGM had exact agreement with the automated analyzer; however, the disagreement detected did not have serious clinical consequences. Our findings stressed the importance of using the same device for monitoring trends in dogs and using instrument-specific reference ranges.

  12. Small sample sorting of primary adherent cells by automated micropallet imaging and release.

    Science.gov (United States)

    Shah, Pavak K; Herrera-Loeza, Silvia Gabriela; Sims, Christopher E; Yeh, Jen Jen; Allbritton, Nancy L

    2014-07-01

    Primary patient samples are the gold standard for molecular investigations of tumor biology yet are difficult to acquire, heterogeneous in nature and variable in size. Patient-derived xenografts (PDXs) comprised of primary tumor tissue cultured in host organisms such as nude mice permit the propagation of human tumor samples in an in vivo environment and closely mimic the phenotype and gene expression profile of the primary tumor. Although PDX models reduce the cost and complexity of acquiring sample tissue and permit repeated sampling of the primary tumor, these samples are typically contaminated by immune, blood, and vascular tissues from the host organism while also being limited in size. For very small tissue samples (on the order of 10(3) cells) purification by fluorescence-activated cell sorting (FACS) is not feasible while magnetic activated cell sorting (MACS) of small samples results in very low purity, low yield, and poor viability. We developed a platform for imaging cytometry integrated with micropallet array technology to perform automated cell sorting on very small samples obtained from PDX models of pancreatic and colorectal cancer using antibody staining of EpCAM (CD326) as a selection criteria. These data demonstrate the ability to automate and efficiently separate samples with very low number of cells. © 2014 International Society for Advancement of Cytometry.

  13. One-step sample preparation of positive blood cultures for the direct detection of methicillin-sensitive and -resistant Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci within one hour using the automated GenomEra CDX™ PCR system.

    Science.gov (United States)

    Hirvonen, J J; von Lode, P; Nevalainen, M; Rantakokko-Jalava, K; Kaukoranta, S-S

    2012-10-01

    A method for the rapid detection of methicillin-sensitive and -resistant Staphylococcus aureus (MSSA and MRSA, respectively) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) with a straightforward sample preparation protocol of blood cultures using an automated homogeneous polymerase chain reaction (PCR) assay, the GenomEra™ MRSA/SA (Abacus Diagnostica Oy, Turku, Finland), is presented. In total, 316 BacT/Alert (bioMérieux, Marcy l'Etoile, France) and 433 BACTEC (Becton Dickinson, Sparks, MD, USA) blood culture bottles were analyzed, including 725 positive cultures containing Gram-positive cocci in clusters (n = 419) and other Gram stain forms (n = 361), as well as 24 signal- and growth-negative bottles. Detection sensitivities for MSSA, MRSA, and MRCoNS were 99.4 % (158/159), 100.0 % (9/9), and 99.3 % (132/133), respectively. One false-positive MRSA result was detected from a non-staphylococci-containing bottle, yielding a specificity of 99.8 %. The lowest detectable amount of viable cells in the blood culture sample was 4 × 10(4) CFU/mL. The results were available within one hour after microbial growth detection and the two-step, time-resolved fluorometric (TRF) measurement mode employed by the GenomEra CDX™ instrument showed no interference from blood, charcoal, or culture media. The method described lacks all sample purification steps and allows reliable and simplified pathogen detection also in clinical microbiology laboratory settings without specialized molecular microbiology competence.

  14. It's in your blood: spectral biomarker candidates for urinary bladder cancer from automated FTIR spectroscopy.

    Science.gov (United States)

    Ollesch, Julian; Heinze, Margot; Heise, H Michael; Behrens, Thomas; Brüning, Thomas; Gerwert, Klaus

    2014-04-01

    Blood samples of urinary bladder cancer (UBC) patients and patients with urinary tract infection were analysed with advanced automated high throughput Fourier transform infrared (HT-FTIR)-spectroscopy. Thin dried film samples were robotically prepared on multi-well titer plates (MTP) for absorbance measurements in transmission mode. Within the absorbance, 1st and 2nd derivative spectra of serum and two plasma preparations, discriminative patterns were identified and validated using bioinformatic tools. The optimal spectral resolution for data acquisition was determined. An accurate discrimination of the patient groups was achieved with three different independent spectral variable sets. The HT-FTIR blood test may support future clinical diagnostics. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Cardiovascular Risk in Hypertension in Relation to Achieved Blood Pressure Using Automated Office Blood Pressure Measurement.

    Science.gov (United States)

    Myers, Martin G; Kaczorowski, Janusz; Dolovich, Lisa; Tu, Karen; Paterson, J Michael

    2016-10-01

    The SPRINT (Systolic Blood Pressure Intervention Trial) reported that some older, higher risk patients might benefit from a target systolic blood pressure (BP) of <120 versus <140 mm Hg. However, it is not yet known how the BP target and measurement methods used in SPRINT relate to cardiovascular outcomes in real-world practice. SPRINT used the automated office BP technique, which requires the patient to be resting quietly and alone, with multiple readings being recorded automatically using an electronic oscillometric sphygmomanometer. We studied the relationship between achieved automated office BP at baseline and cardiovascular events in 6183 community-dwelling residents of Ontario aged ≥66 years who were receiving antihypertensive therapy and followed for a mean of 4.6 years. Adjusted hazard ratios (95% confidence intervals) were computed for 10 mm Hg increments in achieved automated office BP at baseline using Cox proportional hazards regression and the BP category with the lowest event rate as the reference category. Based on 904 fatal and nonfatal cardiovascular events, the nadir of cardiovascular events was at the systolic pressure category of 110 to 119 mm Hg, which was lower than the next highest category of 120 to 129 mm Hg (hazard ratio 1.30 [1.01, 1.66]). The hazard ratio for diastolic pressure was relatively unchanged above 60 mm Hg. Pulse pressure exhibited an increase in hazard ratio (1.33 [1.02, 1.72]) at ≥80 mm Hg. These results using automated office BP measurement in a usual treatment setting extend the finding in SPRINT of an optimum target systolic BP of <120 mm Hg to routine clinical practice. © 2016 American Heart Association, Inc.

  16. Automated high-resolution NMR with a sample changer

    International Nuclear Information System (INIS)

    Wade, C.G.; Johnson, R.D.; Philson, S.B.; Strouse, J.; McEnroe, F.J.

    1989-01-01

    Within the past two years, it has become possible to obtain high-resolution NMR spectra using automated commercial instrumentation. Software control of all spectrometer functions has reduced most of the tedious manual operations to typing a few computer commands or even making selections from a menu. Addition of an automatic sample changer is the next natural step in improving efficiency and sample throughput; it has a significant (and even unexpected) impact on how NMR laboratories are run and how it is taught. Such an instrument makes even sophisticated experiments routine, so that people with no previous exposure to NMR can run these experiments after a training session of an hour or less. This A/C Interface examines the impact of such instrumentation on both the academic and the industrial laboratory

  17. Identification and red blood cell automated counting from blood smear images using computer-aided system.

    Science.gov (United States)

    Acharya, Vasundhara; Kumar, Preetham

    2018-03-01

    Red blood cell count plays a vital role in identifying the overall health of the patient. Hospitals use the hemocytometer to count the blood cells. Conventional method of placing the smear under microscope and counting the cells manually lead to erroneous results, and medical laboratory technicians are put under stress. A computer-aided system will help to attain precise results in less amount of time. This research work proposes an image-processing technique for counting the number of red blood cells. It aims to examine and process the blood smear image, in order to support the counting of red blood cells and identify the number of normal and abnormal cells in the image automatically. K-medoids algorithm which is robust to external noise is used to extract the WBCs from the image. Granulometric analysis is used to separate the red blood cells from the white blood cells. The red blood cells obtained are counted using the labeling algorithm and circular Hough transform. The radius range for the circle-drawing algorithm is estimated by computing the distance of the pixels from the boundary which automates the entire algorithm. A comparison is done between the counts obtained using the labeling algorithm and circular Hough transform. Results of the work showed that circular Hough transform was more accurate in counting the red blood cells than the labeling algorithm as it was successful in identifying even the overlapping cells. The work also intends to compare the results of cell count done using the proposed methodology and manual approach. The work is designed to address all the drawbacks of the previous research work. The research work can be extended to extract various texture and shape features of abnormal cells identified so that diseases like anemia of inflammation and chronic disease can be detected at the earliest.

  18. Blood donors and factors impacting the blood donation decision: motives for donating blood in Turkish sample.

    Science.gov (United States)

    Karacan, Eda; Cengiz Seval, Guldane; Aktan, Zeynep; Ayli, Meltem; Palabiyikoglu, Refia

    2013-12-01

    Donations in Turkey are insufficient to cover the high transfusion needs arising from large numbers of thalassemia and sickle cell anemia patients and increasing demands for blood due to advanced surgery and cancer treatment. The most acceptable means to get blood is voluntary blood donation and the blood donor system in Turkey mostly depends on a combination of voluntary and involuntary donors. The main aim of this study is to explore the motivations of Turkish voluntary blood donors toward blood donation and to determine predictors of blood donation motivation. A cross-sectional sample survey of active blood donors in Ankara, Turkey was conducted. The sample consisted of 189 male volunteer blood donor adults. Donors filled in a self-administered questionnaire including the measures of demographic information, empathetic concern, altruism, social responsibility and blood donation motivation questionnaire during donation. Factor analysis of Blood Donation Motivation Measure with varimax rotation revealed a three-factor solution named as "values and moral duty", "positive feelings and esteem" and "self-benefit and external reasons". The results with regression analyses showed that only social responsibility had an significant effect independent of age, income, and education on blood donation motivation. These result reflects that blood donation motivation not only linked to a high degree of altruistic reasons, but also to a combination of some self-regarding motives. Additionally, feelings of empathy or altruism may be less strong at the time the decision to help, other factors may have a larger influence on helping decisions. Copyright © 2013 Elsevier Ltd. All rights reserved.

  19. Congener Production in Blood Samples During Preparation and Storage

    DEFF Research Database (Denmark)

    Felby, Søren; Nielsen, Erik

    1995-01-01

    Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone......Retsmedicin, congener production, preparation, head space GC, acetone, isobutanol, storage, blood samples, n-propanol, methanol, methylethylketone...

  20. Evaluation of the quality of blood components obtained after automated separation of whole blood by a new multiunit processor

    DEFF Research Database (Denmark)

    Lagerberg, Johan W; Salado-Jimena, Jose A; Löf, Helena

    2013-01-01

    The Reveos system (Terumo BCT) is a fully automated device able to process four whole blood (WB) units simultaneously into a plasma unit, a red blood cell (RBC) unit, and an interim platelet (PLT) unit (IPU). Multiple IPUs can be pooled to form a transfusable PLT product. The aim of our study was...

  1. Impact of blood sampling in very preterm infants

    DEFF Research Database (Denmark)

    Madsen, L P; Rasmussen, M K; Bjerregaard, L L

    2000-01-01

    ; the groups were then subdivided into critically ill or not. Diagnostic blood sampling and blood transfusion events were recorded. In total, 1905 blood samples (5,253 analysis) were performed, corresponding to 0.7 samples (1.9 analysis) per day per infant. The highest frequencies were found during the first....../kg. For the extremely preterm infants a significant correlation between sampled and transfused blood volume was found (mean 37.1 and 33.3 ml/kg, respectively, r = + 0.71, p = 0.0003). The most frequently requested analyses were glucose, sodium and potassium. Few blood gas analyses were requested (1.9/ infant). No blood...... losses attributable to excessive generous sampling were detected. The results show an acceptable low frequency of sampling and transfusion events for infants of GA 28-32 weeks. The study emphasizes the necessity of thorough reflection and monitoring of blood losses when ordering blood sampling...

  2. Current advances and strategies towards fully automated sample preparation for regulated LC-MS/MS bioanalysis.

    Science.gov (United States)

    Zheng, Naiyu; Jiang, Hao; Zeng, Jianing

    2014-09-01

    Robotic liquid handlers (RLHs) have been widely used in automated sample preparation for liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalysis. Automated sample preparation for regulated bioanalysis offers significantly higher assay efficiency, better data quality and potential bioanalytical cost-savings. For RLHs that are used for regulated bioanalysis, there are additional requirements, including 21 CFR Part 11 compliance, software validation, system qualification, calibration verification and proper maintenance. This article reviews recent advances in automated sample preparation for regulated bioanalysis in the last 5 years. Specifically, it covers the following aspects: regulated bioanalysis requirements, recent advances in automation hardware and software development, sample extraction workflow simplification, strategies towards fully automated sample extraction, and best practices in automated sample preparation for regulated bioanalysis.

  3. Enantioselective determination of methylphenidate and ritalinic acid in whole blood from forensic cases using automated solid-phase extraction and liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Thomsen, Ragnar; B. Rasmussen, Henrik; Linnet, Kristian

    2012-01-01

    A chiral liquid chromatography tandem mass spectrometry (LC–MS-MS) method was developed and validated for quantifying methylphenidate and its major metabolite ritalinic acid in blood from forensic cases. Blood samples were prepared in a fully automated system by protein precipitation followed...

  4. Automated Device for Asynchronous Extraction of RNA, DNA, or Protein Biomarkers from Surrogate Patient Samples.

    Science.gov (United States)

    Bitting, Anna L; Bordelon, Hali; Baglia, Mark L; Davis, Keersten M; Creecy, Amy E; Short, Philip A; Albert, Laura E; Karhade, Aditya V; Wright, David W; Haselton, Frederick R; Adams, Nicholas M

    2016-12-01

    Many biomarker-based diagnostic methods are inhibited by nontarget molecules in patient samples, necessitating biomarker extraction before detection. We have developed a simple device that purifies RNA, DNA, or protein biomarkers from complex biological samples without robotics or fluid pumping. The device design is based on functionalized magnetic beads, which capture biomarkers and remove background biomolecules by magnetically transferring the beads through processing solutions arrayed within small-diameter tubing. The process was automated by wrapping the tubing around a disc-like cassette and rotating it past a magnet using a programmable motor. This device recovered biomarkers at ~80% of the operator-dependent extraction method published previously. The device was validated by extracting biomarkers from a panel of surrogate patient samples containing clinically relevant concentrations of (1) influenza A RNA in nasal swabs, (2) Escherichia coli DNA in urine, (3) Mycobacterium tuberculosis DNA in sputum, and (4) Plasmodium falciparum protein and DNA in blood. The device successfully extracted each biomarker type from samples representing low levels of clinically relevant infectivity (i.e., 7.3 copies/µL of influenza A RNA, 405 copies/µL of E. coli DNA, 0.22 copies/µL of TB DNA, 167 copies/µL of malaria parasite DNA, and 2.7 pM of malaria parasite protein). © 2015 Society for Laboratory Automation and Screening.

  5. Multidrug resistant Salmonellae isolated from blood culture samples ...

    African Journals Online (AJOL)

    This study investigates the prevalence of R-plasmids in Salmonella sp. isolated from blood samples of suspected typhoid patients in Warri, Nigeria. A total of 136 blood samples were collected between May and December,2009 and screened for the presence of Salmonellae using standard blood culture techniques of which ...

  6. Evaluation of preanalytical stability of the sample for complete blood count

    Directory of Open Access Journals (Sweden)

    Raghavendra Karanth P

    2011-09-01

    Full Text Available Objective: To evaluate the effect of time of incubation on complete blood count (CBC by using HmX analyzer. Methods: A cross sectional study was conducted at Manipal Acunova Limited, Bangalore. Ten blood samples which were stored at room temperature for CBC and differential count by using HmX analyzer were analyzed within one hour of draw and on 24, 48 and 72 hours of draw. Results: Differential count measured in an automated instrument changed over time. Conclusions: The finding of this study shows that some of CBC parameters can be changed with the incubation, therefore it is better to perform on a fresh sample.

  7. Blood samples in the neutron beam

    International Nuclear Information System (INIS)

    Anon.

    2012-01-01

    Whether crocodile, platypus, man, or chicken: Always the hemoglobin in the red blood cells has the same task. It carries oxygen from the lungs throughout the body. In investigative manner and international team around Dr. Andreas Stadler from the Julic Research Center has deciphered in detail, how and why the hemoglobines of these creatures nevertheless differ. Their findings are among others interesting for the research on artificial blood.

  8. Combination syringe provides air-free blood samples

    Science.gov (United States)

    Pool, S. L.

    1970-01-01

    Standard syringe and spinal needle are combined in unique manner to secure air-free blood samples. Combination syringe obtains air free samples because air bubbles become insignificant when samples greater than 1 cc are drawn.

  9. Establishing a novel automated magnetic bead-based method for the extraction of DNA from a variety of forensic samples.

    Science.gov (United States)

    Witt, Sebastian; Neumann, Jan; Zierdt, Holger; Gébel, Gabriella; Röscheisen, Christiane

    2012-09-01

    Automated systems have been increasingly utilized for DNA extraction by many forensic laboratories to handle growing numbers of forensic casework samples while minimizing the risk of human errors and assuring high reproducibility. The step towards automation however is not easy: The automated extraction method has to be very versatile to reliably prepare high yields of pure genomic DNA from a broad variety of sample types on different carrier materials. To prevent possible cross-contamination of samples or the loss of DNA, the components of the kit have to be designed in a way that allows for the automated handling of the samples with no manual intervention necessary. DNA extraction using paramagnetic particles coated with a DNA-binding surface is predestined for an automated approach. For this study, we tested different DNA extraction kits using DNA-binding paramagnetic particles with regard to DNA yield and handling by a Freedom EVO(®)150 extraction robot (Tecan) equipped with a Te-MagS magnetic separator. Among others, the extraction kits tested were the ChargeSwitch(®)Forensic DNA Purification Kit (Invitrogen), the PrepFiler™Automated Forensic DNA Extraction Kit (Applied Biosystems) and NucleoMag™96 Trace (Macherey-Nagel). After an extensive test phase, we established a novel magnetic bead extraction method based upon the NucleoMag™ extraction kit (Macherey-Nagel). The new method is readily automatable and produces high yields of DNA from different sample types (blood, saliva, sperm, contact stains) on various substrates (filter paper, swabs, cigarette butts) with no evidence of a loss of magnetic beads or sample cross-contamination. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  10. Automated force volume image processing for biological samples.

    Directory of Open Access Journals (Sweden)

    Pavel Polyakov

    2011-04-01

    Full Text Available Atomic force microscopy (AFM has now become a powerful technique for investigating on a molecular level, surface forces, nanomechanical properties of deformable particles, biomolecular interactions, kinetics, and dynamic processes. This paper specifically focuses on the analysis of AFM force curves collected on biological systems, in particular, bacteria. The goal is to provide fully automated tools to achieve theoretical interpretation of force curves on the basis of adequate, available physical models. In this respect, we propose two algorithms, one for the processing of approach force curves and another for the quantitative analysis of retraction force curves. In the former, electrostatic interactions prior to contact between AFM probe and bacterium are accounted for and mechanical interactions operating after contact are described in terms of Hertz-Hooke formalism. Retraction force curves are analyzed on the basis of the Freely Jointed Chain model. For both algorithms, the quantitative reconstruction of force curves is based on the robust detection of critical points (jumps, changes of slope or changes of curvature which mark the transitions between the various relevant interactions taking place between the AFM tip and the studied sample during approach and retraction. Once the key regions of separation distance and indentation are detected, the physical parameters describing the relevant interactions operating in these regions are extracted making use of regression procedure for fitting experiments to theory. The flexibility, accuracy and strength of the algorithms are illustrated with the processing of two force-volume images, which collect a large set of approach and retraction curves measured on a single biological surface. For each force-volume image, several maps are generated, representing the spatial distribution of the searched physical parameters as estimated for each pixel of the force-volume image.

  11. Studies of U in the blood of two population samples

    International Nuclear Information System (INIS)

    Segovia, N.; Olguin, M.E.; Romero, M.

    1986-01-01

    The present work, attempts to establish the statistical distribution of blood uranium in a population of the same community, similar in age and in living patterns. U traces were evaluated by a fission track technique both in whole blood and plasma samples. Dried samples were compressed into pellets and irradiated in a nuclear reactor using the external detector method. For U quantification, standard U samples were used. A comparative sampling of U content in blood samples from a group of radiation exposed workers and another of leukemia patients was also carried out. Results from the sampling groups are reported and discussed. (author)

  12. Detection of drugs in 275 alcohol-positive blood samples of Korean drivers.

    Science.gov (United States)

    Kim, Eunmi; Choe, Sanggil; Lee, Juseon; Jang, Moonhee; Choi, Hyeyoung; Chung, Heesun

    2016-08-01

    Since driving under the influence of drugs (DUID) is as dangerous as drink-driving, many countries regulate DUID by law. However, laws against the use of drugs while driving are not yet established in Korea. In order to investigate the type and frequency of drugs used by drivers in Korea, we analyzed controlled and non-controlled drugs in alcohol-positive blood samples. Total 275 blood samples were taken from Korean drivers, which were positive in roadside alcohol testing. The following analyses were performed: blood alcohol concentrations by GC; screening for controlled drugs by immunoassay and confirmation for positive samples by GC-MS. For the detection of DUID related drugs in blood samples, a total of 49 drugs were selected and were examined by GC-MS. For a rapid detection of these drugs, an automated identification software called "DrugMan" was used. Concentrations of alcohol in 275 blood samples ranged from 0.011 to 0.249% (average 0.119%). Six specimens showed positive results by immunoassay: one methamphetamine and five benzodiazepines I. By GC-MS confirmation, only benzodiazepines in four cases were identified, while methamphetamine and benzodiazepine in two cases were not detected from the presumptive positive blood samples. Using DrugMan, four drugs were detected; chlorpheniramine (5)*, diazepam (4), dextromethorphan (1) and doxylamine (1). In addition, ibuprofen (1), lidocaine (1) and topiramate (1) were also detected as general drugs in blood samples ('*' indicates frequency). The frequency of drug abuse by Korean drivers was relatively low and a total 14 cases were positive in 275 blood samples with a ratio of 5%. However it is necessary to analyze more samples including alcohol negative blood, and to expand the range of drug lists to get the detailed information. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  13. Automated Aqueous Sample Concentration Methods for in situ Astrobiological Instrumentation

    Science.gov (United States)

    Aubrey, A. D.; Grunthaner, F. J.

    2009-12-01

    The era of wet chemical experiments for in situ planetary science investigations is upon us, as evidenced by recent results from the surface of Mars by Phoenix’s microscopy, electrochemistry, and conductivity analyzer, MECA [1]. Studies suggest that traditional thermal volatilization methods for planetary science in situ investigations induce organic degradation during sample processing [2], an effect that is enhanced in the presence of oxidants [3]. Recent developments have trended towards adaptation of non-destructive aqueous extraction and analytical methods for future astrobiological instrumentation. Wet chemical extraction techniques under investigation include subcritical water extraction, SCWE [4], aqueous microwave assisted extraction, MAE, and organic solvent extraction [5]. Similarly, development of miniaturized analytical space flight instruments that require aqueous extracts include microfluidic capillary electrophoresis chips, μCE [6], liquid-chromatography mass-spectrometrometers, LC-MS [7], and life marker chips, LMC [8]. If organics are present on the surface of Mars, they are expected to be present at extremely low concentrations (parts-per-billion), orders of magnitude below the sensitivities of most flight instrument technologies. Therefore, it becomes necessary to develop and integrate concentration mechanisms for in situ sample processing before delivery to analytical flight instrumentation. We present preliminary results of automated solid-phase-extraction (SPE) sample purification and concentration methods for the treatment of highly saline aqueous soil extracts. These methods take advantage of the affinity of low molecular weight organic compounds with natural and synthetic scavenger materials. These interactions allow for the separation of target organic analytes from unfavorable background species (i.e. salts) during inline treatment, and a clever method for selective desorption is utilized to obtain concentrated solutions on the order

  14. The pathology of facial vein blood sampling in mice

    DEFF Research Database (Denmark)

    Hansen, Ket; Harslund, Jakob le Fèvre; Bollen, Peter

    2014-01-01

    Introduction: The use of retro-orbital blood sampling is prohibited in Denmark. For this reason, alternative methods are used for obtaining larger blood samples of a good quality. The facial vein is generally recommended for this. However, we have experienced discomfort for mice subjected to facial...... vein blood sampling. Therefore, we investigated if this technique was associated with pathological changes of the jaw region. Methods: 43 NMRI mice were subjected to facial vein blood sampling by using the lancet method during 12 months, starting at the age of 8 weeks. The mice were restrained manually...... by the scruff and a lancet was placed 2-3 mm caudally to the freckle on the lower jaw, and the skin was punctured. After sampling, brief compression by a cotton swab was applied, if bleeding did not stop. Two days after the last blood sampling, the mice were euthanized by an overdose of pentobarbital...

  15. Automated sampling and data processing derived from biomimetic membranes

    DEFF Research Database (Denmark)

    Perry, Mark; Vissing, Thomas; Boesen, P.

    2009-01-01

    Recent advances in biomimetic membrane systems have resulted in an increase in membrane lifetimes from hours to days and months. Long-lived membrane systems demand the development of both new automated monitoring equipment capable of measuring electrophysiological membrane characteristics and new...

  16. Automated sampling and data processing derived from biomimetic membranes

    International Nuclear Information System (INIS)

    Perry, M; Vissing, T; Hansen, J S; Nielsen, C H; Boesen, T P; Emneus, J

    2009-01-01

    Recent advances in biomimetic membrane systems have resulted in an increase in membrane lifetimes from hours to days and months. Long-lived membrane systems demand the development of both new automated monitoring equipment capable of measuring electrophysiological membrane characteristics and new data processing software to analyze and organize the large amounts of data generated. In this work, we developed an automated instrumental voltage clamp solution based on a custom-designed software controller application (the WaveManager), which enables automated on-line voltage clamp data acquisition applicable to long-time series experiments. We designed another software program for off-line data processing. The automation of the on-line voltage clamp data acquisition and off-line processing was furthermore integrated with a searchable database (DiscoverySheet(TM)) for efficient data management. The combined solution provides a cost efficient and fast way to acquire, process and administrate large amounts of voltage clamp data that may be too laborious and time consuming to handle manually. (communication)

  17. Payload specialist Reinhard Furrer show evidence of previous blood sampling

    Science.gov (United States)

    1985-01-01

    Payload specialist Reinhard Furrer shows evidence of previous blood sampling while Wubbo J. Ockels, Dutch payload specialist (only partially visible), extends his right arm after a sample has been taken. Both men show bruises on their arms.

  18. Neonatal blood gas sampling methods | Goenka | South African ...

    African Journals Online (AJOL)

    Blood gas sampling is part of everyday practice in the care of babies admitted to the neonatal intensive care unit, particularly for those receiving respiratory support. There is little published guidance that systematically evaluates the different methods of neonatal blood gas sampling, where each method has its individual ...

  19. Blood oxygen saturation determined by transmission spectrophotometry of hemolyzed blood samples

    Science.gov (United States)

    Malik, W. M.

    1967-01-01

    Use of the Lambert-Beer Transmission Law determines blood oxygen saturation of hemolyzed blood samples. This simplified method is based on the difference in optical absorption properties of hemoglobin and oxyhemoglobin.

  20. Sample preparation automation for dosing plutonium in urine

    International Nuclear Information System (INIS)

    Jeanmaire, Lucien; Ballada, Jean; Ridelle Berger, Ariane

    1969-06-01

    After having indicated that dosing urinary plutonium by using the Henry technique can be divided into three stages (plutonium concentration by precipitation, passing the solution on an anionic resin column and plutonium elution, and eluate evaporation to obtain a source of which the radioactivity is measured), and recalled that the automation of the second stage has been reported in another document, this document describes the automation of the first stage, i.e. obtaining from urine a residue containing the plutonium, and sufficiently mineralized to be analyzed by means of ion exchanging resins. Two techniques are proposed, leading to slightly different devices. The different operations to be performed are indicated. The different components of the apparatus are described: beakers, hot plate stirrers, reagent circuits, a system for supernatant suction, and a control-command circuit. The operation and use are then described, and results are given

  1. Automated washing of FTA Card punches and PCR setup for reference samples using a LIMS-controlled Sias Xantus automated liquid handler

    DEFF Research Database (Denmark)

    Stangegaard, Michael; Olsen, Addie Nina; Frøslev, Tobias G.

    2009-01-01

    We have implemented and validated automated methods for washing FTA Card punches containing buccal samples and subsequent PCR setup using a Sias Xantus automated liquid handler. The automated methods were controlled by worklists generated by our LabWare Laboratory Information Management System...

  2. Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

    DEFF Research Database (Denmark)

    Skogstrand, K.; Thorsen, P.; Vogel, I.

    2008-01-01

    increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected...... and stored for other purposes, justifies the study hereof. Blood samples were stored for 0, 4, 24, and 48 h at 4 degrees C, room temperature (RT), and at 35 degrees C, respectively, before they were separated into serum or plasma and frozen. Dried blood spot samples (DBSS) were stored for 0, 1, 2, 3, 7...

  3. Fully automated drug screening of dried blood spots using online LC-MS/MS analysis

    Directory of Open Access Journals (Sweden)

    Stefan Gaugler

    2018-01-01

    Full Text Available A new and fully automated workflow for the cost effective drug screening of large populations based on the dried blood spot (DBS technology was introduced in this study. DBS were prepared by spotting 15 μL of whole blood, previously spiked with alprazolam, amphetamine, cocaine, codeine, diazepam, fentanyl, lysergic acid diethylamide (LSD, 3,4-methylenedioxymethamphet-amine (MDMA, methadone, methamphetamine, morphine and oxycodone onto filter paper cards. The dried spots were scanned, spiked with deuterated standards and directly extracted. The extract was transferred online to an analytical LC column and then to the electrospray ionization tandem mass spectrometry system. All drugs were quantified at their cut-off level and good precision and correlation within the calibration range was obtained. The method was finally applied to DBS samples from two patients with back pain and codeine and oxycodone could be identified and quantified accurately below the level of misuse of 89.6 ng/mL and 39.6 ng/mL respectively.

  4. Appearance of granulated cells in blood films stained by automated aqueous versus methanolic Romanowsky methods.

    Science.gov (United States)

    Allison, Robin W; Velguth, Karen E

    2010-03-01

    Romanowsky stains are used routinely by veterinary clinical pathology laboratories for cytologic and blood film evaluations. Automated stainers are available for both aqueous and methanolic Romanowsky stains. Mast cell granules and canine distemper virus inclusions are known to stain differently by these 2 methods, but we have noticed differences in the staining characteristics of other granulated cells. The aim of this study was to investigate and document the variable appearance of basophils and large granular lymphocytes in blood films stained using aqueous and methanolic Romanowsky methods. Cytologic preparations from 1 canine mast cell tumor and blood films from 8 dogs, 1 cat, 1 rabbit, and 1 ostrich were stained using an automated aqueous stain (Aerospray 7120, with and without a predip fixative) and an automated methanolic stain (Hematek). Staining quality and intensity of the cytoplasmic granules in mast cells, basophils, and large granular lymphocytes was evaluated subjectively. Cytoplasmic granules of mast cells, basophils, and large granular lymphocytes stained poorly or not at all with the automated aqueous stain but stained prominently and were readily identified with the automated methanolic stain. Use of the predip fixative with the Aerospray method improved the visibility of basophil granules but not mast cell granules, and had a variable affect on the visibility of granules in large granular lymphocytes. Clinical pathologists should be aware of the staining methodology used on the slides they evaluate to avoid incorrect interpretation of granulated cell populations.

  5. An Automated Sample Preparation System for Large-Scale DNA Sequencing

    Science.gov (United States)

    Marziali, Andre; Willis, Thomas D.; Federspiel, Nancy A.; Davis, Ronald W.

    1999-01-01

    Recent advances in DNA sequencing technologies, both in the form of high lane-density gels and automated capillary systems, will lead to an increased requirement for sample preparation systems that operate at low cost and high throughput. As part of the development of a fully automated sequencing system, we have developed an automated subsystem capable of producing 10,000 sequence-ready ssDNA templates per day from libraries of M13 plaques at a cost of $0.29 per sample. This Front End has been in high throughput operation since June, 1997 and has produced > 400,000 high-quality DNA templates. PMID:10330125

  6. The effect of increased centrifugation temperature on the quality of red-blood-cell concentrates of automated whole blood processing.

    Science.gov (United States)

    Weinigel, C; Rummler, S; Barz, D

    2013-10-01

    There are manual and automated methods to separate whole blood (WB) available. The Atreus whole blood processing system is an automated method, which combines centrifugation and expression of components into a single device. A major difference to conventional methods is that centrifugation temperature is not controlled at 22°C. The aim of this study was to examine the influence of increased centrifugation temperatures on the quality of red-blood-cell concentrates (RCC) after active cooling of WB prior to processing. A total of 28 WB were processed: 16 at centrifugation temperatures of up to 28°C (1st protocol) and 12 at 34°C (2nd protocol). RCC quality parameters were tested weekly for 42 days. Red-blood-cell concentrates (RCC) quality complied with the European and German guidelines. Haemolysis was not significantly different throughout storage. Significant statistical differences were detected between both protocols in potassium concentration at the end of storage and in ATP levels at the day of processing. Centrifugation temperatures of up to 34°C are well tolerated by the red blood cells with minimal interference with the RCC quality parameters. © 2013 International Society of Blood Transfusion.

  7. Microfluidic cartridges for automated, point-of-care blood cell counting

    CSIR Research Space (South Africa)

    Smith, Suzanne

    2016-11-01

    Full Text Available cell counting to be performed. The functional steps within the microfluidic cartridge as well as the surrounding instrumentation required to control and test the cartridges in an automated fashion are described. The results recorded from 10 white blood...

  8. Are They Bloody Guilty? Blood Doping with Simulated Samples

    Science.gov (United States)

    Stuart, Parker E.; Lees, Kelsey D.; Milanick, Mark A.

    2014-01-01

    In this practice-based lab, students are provided with four Olympic athlete profiles and simulated blood and urine samples to test for illegal substances and blood-doping practices. Throughout the course of the lab, students design and conduct a testing procedure and use their results to determine which athletes won their medals fairly. All of the…

  9. Hematocrit, Anemia, and Arm Preference for Blood Sample ...

    African Journals Online (AJOL)

    and pattern of anemia, as well the arm preferences for blood sample collection among pregnant women in Enugu, South East Nigeria. Subjects and Methods: HCT was determined using venous blood of 200 antenatal women at the University of Nigeria Teaching Hospital (UNTH). Enugu, Nigeria. Questionnaires were used ...

  10. Non-terminal blood sampling techniques in guinea pigs.

    Science.gov (United States)

    Birck, Malene M; Tveden-Nyborg, Pernille; Lindblad, Maiken M; Lykkesfeldt, Jens

    2014-10-11

    Guinea pigs possess several biological similarities to humans and are validated experimental animal models(1-3). However, the use of guinea pigs currently represents a relatively narrow area of research and descriptive data on specific methodology is correspondingly scarce. The anatomical features of guinea pigs are slightly different from other rodent models, hence modulation of sampling techniques to accommodate for species-specific differences, e.g., compared to mice and rats, are necessary to obtain sufficient and high quality samples. As both long and short term in vivo studies often require repeated blood sampling the choice of technique should be well considered in order to reduce stress and discomfort in the animals but also to ensure survival as well as compliance with requirements of sample size and accessibility. Venous blood samples can be obtained at a number of sites in guinea pigs e.g., the saphenous and jugular veins, each technique containing both advantages and disadvantages(4,5). Here, we present four different blood sampling techniques for either conscious or anaesthetized guinea pigs. The procedures are all non-terminal procedures provided that sample volumes and number of samples do not exceed guidelines for blood collection in laboratory animals(6). All the described methods have been thoroughly tested and applied for repeated in vivo blood sampling in studies within our research facility.

  11. Astronaut Joseph Kerwin takes blood sample from Astronaut Charles Conrad

    Science.gov (United States)

    1973-01-01

    Scientist-Astronaut Joseph P. Kerwin (right), Skylab 2 science pilot and a doctor of medicine, takes a blood sample from Astronaut Charles Conrad Jr., Sylab 2 commander, as seen in this reproduction taken from a color television transmission made by a TV camera aboard the Skylab 1 and 2 space station cluster in Earth orbit. The blood sampling was part of the Skylab Hematology and Immunology Experiment M110 series.

  12. Thresholds for Diagnosing Hypertension Based on Automated Office Blood Pressure Measurements and Cardiovascular Risk.

    Science.gov (United States)

    Myers, Martin G; Kaczorowski, Janusz; Paterson, J Michael; Dolovich, Lisa; Tu, Karen

    2015-09-01

    The risk of cardiovascular events in relation to blood pressure is largely based on readings taken with a mercury sphygmomanometer in populations which differ from those of today in terms of hypertension severity and drug therapy. Given replacement of the mercury sphygmomanometer with electronic devices, we sought to determine the blood pressure threshold for a significant increase in cardiovascular risk using a fully automated device, which takes multiple readings with the subject resting quietly alone. Participants were 3627 community-dwelling residents aged >65 years untreated for hypertension. Automated office blood pressure readings were obtained in a community pharmacy with subjects seated and undisturbed. This method for recording blood pressure produces similar readings in different settings, including a pharmacy and family doctor's office providing the above procedures are followed. Subjects were followed for a mean (SD) of 4.9 (1.0) years for fatal and nonfatal cardiovascular events. Adjusted hazard ratios (95% confidence intervals) were computed for 10 mm Hg increments in blood pressure (mm Hg) using Cox proportional hazards regression and the blood pressure category with the lowest event rate as the reference category. A total of 271 subjects experienced a cardiovascular event. There was a significant (P=0.02) increase in the hazard ratio of 1.66 (1.09, 2.54) at a systolic blood pressure of 135 to 144 and 1.72 (1.21, 2.45; P=0.003) at a diastolic blood pressure of 80 to 89. A significant (P=0.03) increase in hazard ratio of 1.73 (1.04, 2.86) occurred with a pulse pressure of 80 to 89. These findings are consistent with a threshold of 135/85 for diagnosing hypertension in older subjects using automated office blood pressure. © 2015 American Heart Association, Inc.

  13. Evaluation of Sample Stability and Automated DNA Extraction for Fetal Sex Determination Using Cell-Free Fetal DNA in Maternal Plasma

    Directory of Open Access Journals (Sweden)

    Elena Ordoñez

    2013-01-01

    Full Text Available Objective. The detection of paternally inherited sequences in maternal plasma, such as the SRY gene for fetal sexing or RHD for fetal blood group genotyping, is becoming part of daily routine in diagnostic laboratories. Due to the low percentage of fetal DNA, it is crucial to ensure sample stability and the efficiency of DNA extraction. We evaluated blood stability at 4°C for at least 24 hours and automated DNA extraction, for fetal sex determination in maternal plasma. Methods. A total of 158 blood samples were collected, using EDTA-K tubes, from women in their 1st trimester of pregnancy. Samples were kept at 4°C for at least 24 hours before processing. An automated DNA extraction was evaluated, and its efficiency was compared with a standard manual procedure. The SRY marker was used to quantify cfDNA by real-time PCR. Results. Although lower cfDNA amounts were obtained by automated DNA extraction (mean 107,35 GE/mL versus 259,43 GE/mL, the SRY sequence was successfully detected in all 108 samples from pregnancies with male fetuses. Conclusion. We successfully evaluated the suitability of standard blood tubes for the collection of maternal blood and assessed samples to be suitable for analysis at least 24 hours later. This would allow shipping to a central reference laboratory almost from anywhere in Europe.

  14. EVALUATION OF ZEBU NELLORE CATTLE BLOOD SAMPLES USING THE CELL-DYN 3500 HEMATOLOGY ANALYZER

    Directory of Open Access Journals (Sweden)

    Alexandre Secorun Borges

    2014-12-01

    Full Text Available The Cell-dyn 3500 is a multiparameter flow cytometer, which may analyze samples from several species performing several simultaneous analyses. It is able to perform white blood cells, red blood cells and platelet counts, besides differential leukocyte counts, packed cell volume and hemoglobin determination. Cell-Dyn 3500 performs total leukocyte count both optically and by impedance. The equipment may choose one or other method, based on the reliability of the results. Erythrocyte and platelet counts are determined by impedance. Leukocyte differentiation is based on an optical principle, using separation in multiangular polarized light. The objective of this study was to compare the results of complete blood count of Zebu Nellore heifers from Celldyn 3500, with those obtained from a semi-automated cell counter (Celm CC 510 and the manual technique. Blood samples were collected from the jugular vein in 5 mL EDTA vacuum tubes from 58 Nellore heifers, at 24 months of age. Samples were processed in parallel in the three different techniques. Results were analyzed using paired t test, Pearson’s correlation and the Bland-Altmann method. There was a strong correlation for all parameters analyzed by Cell-Dyn 3500, manual method and semiautomated cell counter, except for basophils and monocytes counts. These results confirm that this analyzer is reliable for blood samples analysis of zebu cattle.

  15. Extensive monitoring through multiple blood samples in professional soccer players

    DEFF Research Database (Denmark)

    Heisterberg, Mette F; Fahrenkrug, Jan; Krustrup, Peter

    2013-01-01

    that may be related to changes in training pattern, match exposure or length of the match-season. Especially the end of the preparation-season and at the end of the competitive season seem to be time points were the blood-derived values indicate that the players are under excessive physical strain......ABSTRACT: The aim of this study was to make a comprehensive gathering of consecutive detailed blood samples from professional soccer players, and to analyze different blood parameters in relation to seasonal changes in training and match exposure.Blood samples were collected five times during a six...... months period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, players were tested for body composition, VO2max and physical performance by the Yo-Yo intermittent endurance sub-max test (IE2).Multiple variations in blood parameters occurred during...

  16. [Automated management of the Center of Transfusion Blood Bank equipped with Autogrouper 16 C].

    Science.gov (United States)

    Girard, M; Connes, Y; Picot, P

    1981-11-01

    We describe in this paper a first processing of direct connection and data management from the Autogrouper 16 C. It bas been used since November 1980 in the Centre Départemental de Tranfusion Sanguine des Hauts-de-Seine. The automation of validation procedures, updating blood donors' file and labelling blood units allows the suppression of clerical errors that are the major cause of Trannsfusion accidents. The progressive development of programming lets a good adaptation of working methods and makes the setting of the system much easier in the Blood Bank.

  17. Blood sampling and hemolysis affect concentration of plasma metabolites

    DEFF Research Database (Denmark)

    Theil, Peter Kappel; Pedersen, Lene Juul; Jensen, Margit Bak

    2012-01-01

    design and blood was collected after restraint via vein puncture 1, 4, 11, and 23 h after morning feeding. Plasma samples were categorized as without or with minor or major hemolysis [clear (n = 218), yellow (n = 97), or red (n = 37)] upon centrifugation. Plasma NEFA (P ...Two experiments were carried out to reveal and quantify plasma metabolites that are sensitive to hemolysis and animal stress due to the blood sampling procedure (vein puncture vs. catheter). In Exp. 1, 48 sows were fed 4 diets either once (0800 h) or twice daily (0800 h and 1500 h) in a crossover......, a subset of samples from 24 sows fed twice daily in Exp. 1 was combined with data obtained from 30 sows sampled using jugular vein catheters. All sows in Exp. 2 were fed twice daily (0800 h and 1500 h) and blood samples collected repeatedly 1, 4, 11, and 23 h after morning feeding (other conditions were...

  18. Non-terminal blood sampling techniques in Guinea pigs

    DEFF Research Database (Denmark)

    Birck, Malene Muusfeldt; Tveden-Nyborg, Pernille; Lindblad, Maiken Marie

    2014-01-01

    Guinea pigs possess several biological similarities to humans and are validated experimental animal models(1-3). However, the use of guinea pigs currently represents a relatively narrow area of research and descriptive data on specific methodology is correspondingly scarce. The anatomical feature...... not exceed guidelines for blood collection in laboratory animals(6). All the described methods have been thoroughly tested and applied for repeated in vivo blood sampling in studies within our research facility....... repeated blood sampling the choice of technique should be well considered in order to reduce stress and discomfort in the animals but also to ensure survival as well as compliance with requirements of sample size and accessibility. Venous blood samples can be obtained at a number of sites in guinea pigs e.......g., the saphenous and jugular veins, each technique containing both advantages and disadvantages(4,5). Here, we present four different blood sampling techniques for either conscious or anaesthetized guinea pigs. The procedures are all non-terminal procedures provided that sample volumes and number of samples do...

  19. Dried blood spots of pooled samples for RHD gene screening in blood donors of mixed ancestry.

    Science.gov (United States)

    Silva-Malta, M C F; Araujo, N C Fidélis; Vieira, O V Neves; Schmidt, L Cayres; Gonçalves, P de Cassia; Martins, M Lobato

    2015-10-01

    In this study, we present a strategy for RHD gene screening based on real-time polymerase chain reaction (PCR) using dried blood spots of pooled samples. Molecular analysis of blood donors may be used to detect RHD variants among the presumed D-negative individuals. RHD genotyping using pooled samples is a strategy to test a large number of samples at a more reasonable cost. RHD gene detection based on real-time PCR using dried blood spots of pooled samples was standardised and used to evaluate 1550 Brazilian blood donors phenotyped as RhD-negative. Positive results were re-evaluated by retesting single samples using real-time PCR and conventional multiplex PCR to amplify five RHD-specific exons. PCR-sequence-specific primers was used to amplify RHDψ allele. We devised a strategy for RHD gene screening using dried blood spots of five pooled samples. Among 1550 serologically D-negative blood donors, 58 (3.74%) had the RHD gene. The non-functional RHDψ allele was detected in 47 samples (3.02%). The present method is a promising strategy to detect the RHD gene among presumed RhD-negative blood donors, particularly for populations with African ancestry. © 2015 British Blood Transfusion Society.

  20. Strategies for Developing Sensitive and Automated LC-MS/MS Assays of a Pharmaceutical Compound and Its Metabolite from Whole Blood Matrix

    Directory of Open Access Journals (Sweden)

    Raymond N. Xu

    2010-04-01

    Full Text Available When compared with biological samples in other matrices (plasma, urine, etc. that are typically seen in bioanalytical applications, whole blood samples present unique challenges in method development, because of the viscous nature of blood and complexity of its constituents. In this article, we have developed and validated a series of quantitative bioanalytical methods for the determination of a pharmaceutical compound, Compound A, and its phosphate metabolite from whole blood matrices using liquid chromatography tandem mass spectrometry. All methods employed a simple protein precipitation procedure that was automated in 96-well format. The methods were subjected to vigorous tests in precision, accuracy, matrix effect, reproducibility, and robustness. Monolithic chromatography was used to improve sample throughput in one of the methods. The results also demonstrated that proper sample preparation procedures, such as sample transfer and lysing of blood cells prior to the extraction, are key to reproducible results for pharmacokinetic parameter determination.

  1. Measurement and Comparison of Organic Compound Concentrations in Plasma, Whole Blood and Dried Blood Spot Samples

    Directory of Open Access Journals (Sweden)

    Stuart A Batterman

    2016-04-01

    Full Text Available The preferred sampling medium for measuring human exposures of persistent organic compounds (POPs is blood, and relevant sample types include whole blood, plasma, and dried blood spots (DBS. Because information regarding the performance and comparability of measurements across these sample types is limited, it is difficult to compare across studies. This study evaluates the performance of POP measurements in plasma, whole blood and DBS, and presents the distribution coefficients needed to convert concentrations among the three sample types. Blood samples were collected from adult volunteers, along with demographic and smoking information, and analyzed by GC/MS for organochlorine pesticides (OCPs, chlorinated hydrocarbons (CHCs, polychlorinated biphenyls (PCBs, and brominated diphenyl ethers (PBDEs. Regression models were used to evaluate the relationships between the sample types and possible effects of personal covariates. Distribution coefficients also were calculated using physically-based models.Across all compounds, concentrations in plasma were consistently the highest; concentrations in whole blood and DBS samples were comparable. Distribution coefficients for plasma to whole blood concentrations ranged from 1.74 to 2.26 for pesticides/CHCs, averaged 1.69 ± 0.06 for the PCBs, and averaged 1.65 ± 0.03 for the PBDEs. Regression models closely fit most chemicals (R2 > 0.80, and whole blood and DBS samples generally showed very good agreement. Distribution coefficients estimated using biologically-based models were near one and did not explain the observed distribution. Among the study population, median concentrations of several pesticides/CHCs and PBDEs exceeded levels reported in the 2007-2008 National Health and Nutrition Examination Survey, while levels of other OCPs and PBDEs were comparable or lower. Race and smoking status appeared to slightly affect plasma/blood concentration ratios for several POPs. The experimentally

  2. Detection of antileishmanial antibodies in blood sampled from blood bank donors in Istanbul.

    Science.gov (United States)

    Ates, Sezen Canim; Bagirova, Malahat; Allahverdiyev, Adil M; Baydar, Serap Yesilkir; Koc, Rabia Cakir; Elcicek, Serhat; Abamor, Emrah Sefik; Oztel, Olga Nehir

    2012-06-01

    According to the WHO, only 5-20% of the total cases of leishmaniasis are symptomatic leishmaniasis; the other cases are identified as asymptomatic leishmaniasis. In recent studies, it has been demonstrated that donor blood plays an important role in the epidemiology of asymptomatic leishmaniasis. However, the number of the studies on this subject is still insufficient. Additionally, donor blood samples obtained from Istanbul, which is the biggest metropolitan area in Turkey, have not been investigated with regard to Leishmania. Moreover, there is no information about the sensitivity of noninvasive serological methods that are used in the detection of leishmaniasis donor blood samples. Accordingly, this study aimed to investigate the presence of antileishmanial antibodies in blood samples obtained from blood bank donors in Istanbul, by using different serologic methods, and to determine the most sensitive detection method. Blood samples were taken from 188 healthy blood bank donors to the Capa Turkish Red Crescent Blood Bank (Istanbul, Turkey), and the presence of antileishmanial antibodies was measured by indirect immunofluorescent antibody test (IFAT), ELISA, immunochromatographic dipstick rapid test, and western blot (WB). Antileishmanial antibodies were determined in 12 out of 188 samples by IFAT (6.4%), and six out of these 12 donors were found to be positive at diagnostic titer 1:128 (3.2%). One hundred and eighty eight samples were investigated by ELISA and one (0.5%) of them gave a positive result. None of 188 samples provided a positive result by immunochromatographic test. WB applied to the 12 seroreactive donors showed that three out of 12 donors were positive. In this study, the presence of antileishmanial antibodies in blood samples of blood bank donors from Istanbul has been demonstrated by using feasible and low-cost serological methods. Additionally, in comparison with other simple and low-cost detection methods, WB was used for confirmation. IFAT

  3. SASSI: Subsystems for Automated Subsurface Sampling Instruments, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — Future robotic planetary exploration missions will benefit greatly from the ability to capture rock and/or regolith core samples that deliver the stratigraphy of the...

  4. SASSI: Subsystems for Automated Subsurface Sampling Instruments, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — Autonomous surface sampling systems are necessary, near term, to construct a historical view of planetary significant events; as well as allow for the identification...

  5. An automated method for dynamic red blood cell aggregate detection in microfluidic flow.

    Science.gov (United States)

    Mehri, R; Niazi, E; Mavriplis, C; Fenech, M

    2018-01-31

    Red blood cell (RBC) aggregation is a unique phenomenon that occurs when red blood cells are subjected to low shear rates. Little is known about the sizes, shapes and behaviour of aggregates flowing in healthy humans. However, excessive aggregation has been shown to be an indication of pathological conditions. Therefore, characterizing RBC aggregates is important to medical research. The objective of this study was to develop a reliable technique based on image processing to assess and characterize human RBC aggregation subjected to controlled and measurable shear rates in a two-fluid flow microfluidic shearing system. Images of RBC suspensions at [Formula: see text], [Formula: see text] and [Formula: see text] entrained by a phosphate buffered saline solution in a PDMS microchannel were captured with a high speed camera. An algorithm for processing the RBC aggregate images is presented and validated (1) on a sample of known diameter hollow glass microspheres and (2) by comparing RBC aggregate size results with those of an ImageJ image processing technique and those obtained by manual detection by two independent researchers. The proposed image processing algorithm provides a very good agreement with the manufacturer data for the glass microspheres. It also performs well on the RBC suspension images, with errors of 2-4 [Formula: see text] with respect to the manual results. The proposed automated method for RBC aggregate detection is found to be reliable and fairly accurate and will serve researchers and, perhaps in the future, clinicians to assess healthy and pathological RBC aggregation under flowing conditions.

  6. Automated modal parameter estimation using correlation analysis and bootstrap sampling

    Science.gov (United States)

    Yaghoubi, Vahid; Vakilzadeh, Majid K.; Abrahamsson, Thomas J. S.

    2018-02-01

    The estimation of modal parameters from a set of noisy measured data is a highly judgmental task, with user expertise playing a significant role in distinguishing between estimated physical and noise modes of a test-piece. Various methods have been developed to automate this procedure. The common approach is to identify models with different orders and cluster similar modes together. However, most proposed methods based on this approach suffer from high-dimensional optimization problems in either the estimation or clustering step. To overcome this problem, this study presents an algorithm for autonomous modal parameter estimation in which the only required optimization is performed in a three-dimensional space. To this end, a subspace-based identification method is employed for the estimation and a non-iterative correlation-based method is used for the clustering. This clustering is at the heart of the paper. The keys to success are correlation metrics that are able to treat the problems of spatial eigenvector aliasing and nonunique eigenvectors of coalescent modes simultaneously. The algorithm commences by the identification of an excessively high-order model from frequency response function test data. The high number of modes of this model provides bases for two subspaces: one for likely physical modes of the tested system and one for its complement dubbed the subspace of noise modes. By employing the bootstrap resampling technique, several subsets are generated from the same basic dataset and for each of them a model is identified to form a set of models. Then, by correlation analysis with the two aforementioned subspaces, highly correlated modes of these models which appear repeatedly are clustered together and the noise modes are collected in a so-called Trashbox cluster. Stray noise modes attracted to the mode clusters are trimmed away in a second step by correlation analysis. The final step of the algorithm is a fuzzy c-means clustering procedure applied to

  7. A report of cerebral malaria treated with automated red blood cell exchange.

    Science.gov (United States)

    Anani, Waseem Q; Smith, Gerald P; Irani, Mehraboon; Puca, Kathleen E

    2017-04-01

    Adjunctive automated whole blood or red blood cell exchange (RBCEx) can rapidly decrease malarial hyperparasitemia. Several case reports and series suggest improvement in clinical symptomatology; however, recent Centers of Disease Control and Prevention (CDC) recommendations concluded that RBCEx has no efficacy as an adjunctive therapy. We present a case of mental status changes secondary to cerebral malaria treated with automated RBCEx resulting in rapid and dramatic neurologic improvement. An 84-year-old Somali woman presented with a 3-day history of altered mental status, spiking fevers, chills, bilateral leg pain and weakness, and intermittent diarrhea. Her travel history included a recent trip to Kenya for 1 month without antimalarial chemoprophylaxis. During the hospital stay, her health declined, and she became obtunded. Physical examination revealed fever, tachypnea, hypertension, hypoxia, and no response to verbal or physical stimuli. Her hemoglobin decreased from 12.6 to 6.5 g/dL with 12% intraerythrocytic parasitemia by thin smear. Intraerythrocytic trophozoites and banana-shaped gametocytes were present consistent with Plasmodium falciparum. An emergent 1.5-volume RBC mass automated RBCEx and quinidine infusion decreased her parasitemia to 2%. The patient's mental status improved throughout the procedure, and after the 2½-hour procedure, the patient was alert, oriented, and speaking coherently. The patient continued to receive quinidine and artesunate 1 day later from CDC. Automated RBCEx transfusion reduced the parasite burden and restored neurologic functioning in a patient with cerebral malaria while awaiting definitive treatment with artesunate. © 2017 AABB.

  8. Use of standard laboratory methods to obviate routine dithiothreitol treatment of blood samples with daratumumab interference.

    Science.gov (United States)

    Lintel, Nicholas J; Brown, Debra K; Schafer, Diane T; Tsimba-Chitsva, Farai M; Koepsell, Scott A; Shunkwiler, Sara M

    2017-01-01

    Daratumumab is an antibody currently used in the treatment of patients with refractory multiple myeloma. Blood samples from patients being treated with daratumumab may show panreactivity during pre-transfusion testing. To facilitate the provision of blood components for such patients, it is recommended that a baseline phenotype or genotype be established prior to starting treatment with daratumumab. If patient red blood cells (RBCs) require phenotyping after the start of daratumumab treatment, dithiothreitol (DTT) treatment of the patient's RBCs should be performed. The medical charts of four patients treated with daratumumab were reviewed. The individual number of doses ranged from 1 to 14; patient age ranged from 55 to 78 years; two men and two women were included in the review. Type and screen data were obtained from samples collected over 33 encounters with a range of 1 to 13 encounters per patient. All samples were tested initially by automated solid-phase testing. Any reactivity with solid phase led to tube testing with either low-ionic-strength saline, polyethylene glycol, or both. If incubation failed to eliminate the reactivity, the sample was sent to a reference laboratory for DTT treatment and phenotyping. Of the 33 samples tested, 23 (69.7%) samples had reactivity in solid-phase testing. In 8 of the 10 samples that did not react in solid-phase, testing was conducted more than four half-lives after the last dose of daratumumab. Of the 23 that had reactivity in solid-phase, 16 (69.6%) samples demonstrated loss of reactivity using common laboratory methods. For the seven patients whose sample reactivity was not initially eliminated, six were provided with phenotypically matched blood based on prior molecular testing. Only one sample was sent out for DTT treatment. These results suggest that daratumumab interference with pre-transfusion testing can be addressed using common laboratory methods. This finding could save time and money for laboratories that do

  9. Extensive monitoring through multiple blood samples in professional soccer players.

    Science.gov (United States)

    Heisterberg, Mette F; Fahrenkrug, Jan; Krustrup, Peter; Storskov, Anders; Kjær, Michael; Andersen, Jesper L

    2013-05-01

    The aim of this study was to make a comprehensive gathering of consecutive detailed blood samples from professional soccer players and to analyze different blood parameters in relation to seasonal changes in training and match exposure. Blood samples were collected 5 times during a 6-month period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, the players were tested for body composition, V[Combining Dot Above]O2max and physical performance by the Yo-Yo intermittent endurance submax test (IE2). Multiple variations in blood parameters occurred during the observation period, including a decrease in hemoglobin and an increase in hematocrit as the competitive season progressed. Iron and transferrin were stable, whereas ferritin showed a decrease at the end of the season. The immunoglobulin A (IgA) and IgM increased in the period with basal physical training and at the end of the season. Leucocytes decreased with increased physical training. Lymphocytes decreased at the end of the season. The V[Combining Dot Above]O2max decreased toward the end of the season, whereas no significant changes were observed in the IE2 test. The regular blood samples from elite soccer players reveal significant changes that may be related to changes in training pattern, match exposure, or length of the match season. Especially the end of the preparation season and at the end of the competitive season seem to be time points were the blood-derived values indicate that the players are under excessive physical strain and might be more subjected to a possible overreaching-overtraining conditions. We suggest that regular analyses of blood samples could be an important initiative to optimize training adaptation, training load, and game participation, but sampling has to be regular, and a database has to be built for each individual player.

  10. Determination of blood cell subtype concentrations from frozen whole blood samples using TruCount beads.

    Science.gov (United States)

    Langenskiöld, Cecilia; Mellgren, Karin; Abrahamsson, Jonas; Bemark, Mats

    2016-06-24

    In many studies it would be advantageous if blood samples could be collected and analyzed using flow cytometry at a later stage. Ideally, sample collection should involve little hands-on time, allow for long-term storage, and minimally influence the samples. Here we establish a flow cytometry antibody panel that can be used to determine granulocytes, monocytes, and lymphocyte subset concentrations in fresh and frozen whole blood using TruCount technology. The panel can be used on fresh whole-blood samples as well as whole-blood samples that have been frozen after mixing with 10% DMSO. Concentrations in frozen and fresh sample is highly correlated both when frozen within 4 h and the day after collection (r ≥ 0.98), and the estimated concentration in frozen samples was between 91 and 94% of that in fresh samples for all cell types. Using this method whole-blood samples can be frozen using a simple preparation method, and stored long-term before accurate determination of cell concentration. This allows for standardized analysis of the samples at a reference laboratory in multi-center studies. © 2016 International Clinical Cytometry Society. © 2016 International Clinical Cytometry Society.

  11. Development and Evaluation of a Blood Culture PCR Assay for Rapid Detection of Salmonella Paratyphi A in Clinical Samples

    Science.gov (United States)

    Zhou, Liqing; Jones, Claire; Gibani, Malick M.; Dobinson, Hazel; Thomaides-Brears, Helena; Shrestha, Sonu; Blohmke, Christoph J.; Darton, Thomas C.; Pollard, Andrew J.

    2016-01-01

    Background Enteric fever remains an important cause of morbidity in many low-income countries and Salmonella Paratyphi A has emerged as the aetiological agent in an increasing proportion of cases. Lack of adequate diagnostics hinders early diagnosis and prompt treatment of both typhoid and paratyphoid but development of assays to identify paratyphoid has been particularly neglected. Here we describe the development of a rapid and sensitive blood culture PCR method for detection of Salmonella Paratyphi A from blood, potentially allowing for appropriate diagnosis and antimicrobial treatment to be initiated on the same day. Methods Venous blood samples from volunteers experimentally challenged orally with Salmonella Paratyphi A, who subsequently developed paratyphoid, were taken on the day of diagnosis; 10 ml for quantitative blood culture and automated blood culture, and 5 ml for blood culture PCR. In the latter assay, bacteria were grown in tryptone soy broth containing 2.4% ox bile and micrococcal nuclease for 5 hours (37°C) before bacterial DNA was isolated for PCR detection targeting the fliC-a gene of Salmonella Paratyphi A. Results An optimized broth containing 2.4% ox bile and micrococcal nuclease, as well as a PCR test was developed for a blood culture PCR assay of Salmonella Paratyphi A. The volunteers diagnosed with paratyphoid had a median bacterial burden of 1 (range 0.1–6.9) CFU/ml blood. All the blood culture PCR positive cases where a positive bacterial growth was shown by quantitative blood culture had a bacterial burden of ≥ 0.3 CFU/ ml blood. The blood culture PCR assay identified an equal number of positive cases as automated blood culture at higher bacterial loads (≥0.3 CFU/ml blood), but utilized only half the volume of specimens. Conclusions The blood culture PCR method for detection of Salmonella Paratyphi A can be completed within 9 hours and offers the potential for same-day diagnosis of enteric fever. Using 5 ml blood, it exhibited a

  12. An automated blood vessel segmentation algorithm using histogram equalization and automatic threshold selection.

    Science.gov (United States)

    Saleh, Marwan D; Eswaran, C; Mueen, Ahmed

    2011-08-01

    This paper focuses on the detection of retinal blood vessels which play a vital role in reducing the proliferative diabetic retinopathy and for preventing the loss of visual capability. The proposed algorithm which takes advantage of the powerful preprocessing techniques such as the contrast enhancement and thresholding offers an automated segmentation procedure for retinal blood vessels. To evaluate the performance of the new algorithm, experiments are conducted on 40 images collected from DRIVE database. The results show that the proposed algorithm performs better than the other known algorithms in terms of accuracy. Furthermore, the proposed algorithm being simple and easy to implement, is best suited for fast processing applications.

  13. Single versus duplicate blood samples in ACTH stimulated adrenal vein sampling

    NARCIS (Netherlands)

    Dekkers, T.; Arntz, M.; Wilt, G.J. van der; Schultze Kool, L.J.; Sweep, F.C.; Hermus, A.R.M.M.; Lenders, J.W.M.; Deinum, J.

    2013-01-01

    BACKGROUND: Adrenal vein sampling (AVS) is the preferred test for subtyping primary aldosteronism. However, the procedure is technically demanding and costly. In AVS it is common practice to take duplicate blood samples at each location. In this paper we explore whether a single sample procedure

  14. Automated Sample Preparation for Radiogenic and Non-Traditional Metal Isotopes: Removing an Analytical Barrier for High Sample Throughput

    Science.gov (United States)

    Field, M. Paul; Romaniello, Stephen; Gordon, Gwyneth W.; Anbar, Ariel D.; Herrmann, Achim; Martinez-Boti, Miguel A.; Anagnostou, Eleni; Foster, Gavin L.

    2014-05-01

    MC-ICP-MS has dramatically improved the analytical throughput for high-precision radiogenic and non-traditional isotope ratio measurements, compared to TIMS. The generation of large data sets, however, remains hampered by tedious manual drip chromatography required for sample purification. A new, automated chromatography system reduces the laboratory bottle neck and expands the utility of high-precision isotope analyses in applications where large data sets are required: geochemistry, forensic anthropology, nuclear forensics, medical research and food authentication. We have developed protocols to automate ion exchange purification for several isotopic systems (B, Ca, Fe, Cu, Zn, Sr, Cd, Pb and U) using the new prepFAST-MC™ (ESI, Nebraska, Omaha). The system is not only inert (all-flouropolymer flow paths), but is also very flexible and can easily facilitate different resins, samples, and reagent types. When programmed, precise and accurate user defined volumes and flow rates are implemented to automatically load samples, wash the column, condition the column and elute fractions. Unattended, the automated, low-pressure ion exchange chromatography system can process up to 60 samples overnight. Excellent reproducibility, reliability, recovery, with low blank and carry over for samples in a variety of different matrices, have been demonstrated to give accurate and precise isotopic ratios within analytical error for several isotopic systems (B, Ca, Fe, Cu, Zn, Sr, Cd, Pb and U). This illustrates the potential of the new prepFAST-MC™ (ESI, Nebraska, Omaha) as a powerful tool in radiogenic and non-traditional isotope research.

  15. Quantification of multiple elements in dried blood spot samples

    DEFF Research Database (Denmark)

    Pedersen, Lise; Andersen-Ranberg, Karen; Hollergaard, Mads

    2017-01-01

    in venous blood. Samples with different hematocrit were spotted onto filter paper to assess hematocrit effect. RESULTS: The established method was precise and accurate for measurement of most elements in DBS. There was a significant but relatively weak correlation between measurement of the elements Mg, K......BACKGROUND: Dried blood spots (DBS) is a unique matrix that offers advantages compared to conventional blood collection making it increasingly popular in large population studies. We here describe development and validation of a method to determine multiple elements in DBS. METHODS: Elements were...... extracted from punches and analyzed using inductively coupled plasma-mass spectrometry (ICP-MS). The method was evaluated with quality controls with defined element concentration and blood spiked with elements to assess accuracy and imprecision. DBS element concentrations were compared with concentrations...

  16. Effects of blood sample handling procedures on measurable inflammatory markers in plasma, serum and dried blood spot samples

    DEFF Research Database (Denmark)

    Skogstrand, K.; Thorsen, P.; Vogel, I.

    2008-01-01

    , and 30 days at the same temperatures. 27 inflammatory markers in serum and plasma and 25 markers in DBSS were measured by a previously validated multiplex sandwich immunoassay using Luminex xMAP technology. The measurable concentrations of several cytokines in serum and plasma were significantly......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected...... increased when blood samples were stored for a period of time before the centrifugation, for certain cytokines more than 1000 fold compared to serum and plasma isolated and frozen immediately after venepuncture. The concentrations in serum generally increased more than in plasma. The measurable...

  17. [Evaluation of an automated streaking system of urine samples for urine cultures].

    Science.gov (United States)

    Bustamante, Verónica; Meza, Paulina; Román, Juan C; García, Patricia

    2014-12-01

    Automated systems have simplified laboratory workflow, improved standardization, traceability and diminished human errors and workload. Although microbiology laboratories have little automation, in recent years new tools for automating pre analytical steps have appeared. To assess the performance of an automated streaking machine for urine cultures and its agreement with the conventional manual plating method for semi quantitative colony counts. 495 urine samples for urinary culture were inoculated in CPS® agar using our standard protocol and the PREVI™ Isola. Rates of positivity, negativity, polymicrobial growth, bacterial species, colony counts and re-isolation requirements were compared. Agreement was achieved in 98.97% of the positive/negative results, in 99.39% of the polymicrobial growth, 99.76% of bacterial species isolated and in 98.56 % of colony counts. The need for re-isolation of colonies decreased from 12.1% to 1.1% using the automated system. PREVI™ Isola's performance was as expected, time saving and improving bacterial isolation. It represents a helpful tool for laboratory automation.

  18. Automated injection of a radioactive sample for preparative HPLC with feedback control

    International Nuclear Information System (INIS)

    Iwata, Ren; Yamazaki, Shigeki

    1990-01-01

    The injection of a radioactive reaction mixture into a preparative HPLC column has been automated with computer control for rapid purification of routinely prepared positron emitting radiopharmaceuticals. Using pneumatic valves, a motor-driven pump and a liquid level sensor, two intelligent injection methods for the automation were compared with regard to efficient and rapid sample loading into a 2 mL loop of the 6-way valve. One, a precise but rather slow method, was demonstrated to be suitable for purification of 18 F-radiopharmaceuticals, while the other, due to its rapid operation, was more suitable for 11 C-radiopharmaceuticals. A sample volume of approx 0.5 mL can be injected onto a preparative HPLC column with over 90% efficiency with the present automated system. (author)

  19. Prospective comparison of a PCR assay and a microbiological culture technique for identification of pathogens from blood and non-blood samples in septic patients.

    Science.gov (United States)

    Plettig, Runa; Nowak, Andreas; Balau, Veronika; Hahnenkamp, Klaus; Usichenko, Taras

    2015-01-01

    Molecular amplification techniques are suggested to be a useful adjunct in early detection of pathogens in septic patients. The aim was to study the feasibility of a polymerase chain reaction (PCR) assay compared to the standard microbiological culture (MC) technique in identification of pathogenic microorganisms from blood and non-blood samples in septic patients. Samples for pathogen identification were taken during febrile septic episodes (SE) in 54 patients with sepsis and analyzed using both MC and PCR. Semi-automated multiplex PCR, provided by Philips Medical Systems, was able to detect nine different pathogens. The accuracy of pathogen identification using PCR vs. MC as well as the time-saving effect of PCR on the potential decision-making process for antimicrobial therapy was evaluated. In a total of 258 samples taken during 87 SE, both methods yielded more pathogens from the non-blood than blood samples (87 % vs. 45 %; p = 0.002). PCR identified more pathogens than MC in the blood samples (98 vs. 21; p technique. In the non-blood samples, PCR was comparable to that of MC.

  20. Fetal scalp blood sampling in labor - a review

    DEFF Research Database (Denmark)

    Jørgensen, Jan Stener; Weber, Tom

    2014-01-01

    During the 1970s and 1980s, electronic fetal monitoring and fetal scalp blood sampling (FBS) were introduced without robust evidence. With a methodical review of the published literature, and using one randomized controlled trial, seven controlled studies, nine randomized studies of various...... in the interpretation of CTG patterns. This article is protected by copyright. All rights reserved....

  1. A simple method for repetitive blood sampling of cattle

    African Journals Online (AJOL)

    A simple practical technique is described for the collection of blood samples from at least six catile concurrenily at short time interuals for up to 48 h. The technique is advantageous in that it is carried out under minimum srress conditions. 'n Eenvoudige praktiese tegniek vir die gelyktydige trekking van bloedmonsters van ten ...

  2. Efficiency of Extraction of Trace metals from Blood samples using ...

    African Journals Online (AJOL)

    MICHAEL HORSFALL

    Efficiency of Extraction of Trace metals from Blood samples using Wet Digestion and. Microwave Digestion Techniques. *1M. I. YAHAYA; A. SHEHU; F.G. DABAI. Department of Applied Chemistry, Federal University Dutsin – Ma, Katsina State, Nigeria. Biology Department, Kebbi State College of Basic and Advanced Studies ...

  3. Improved sample capsule for determination of oxygen in hemolyzed blood

    Science.gov (United States)

    Malik, W. M.

    1967-01-01

    Sample capsule for determination of oxygen in hemolyzed blood consists of a measured section of polytetrafluoroethylene tubing equipped at each end with a connector and a stopcock valve. This method eliminates errors from air entrainment or from the use of mercury or syringe lubricant.

  4. Predicting blood transfusion using automated analysis of pulse oximetry signals and laboratory values.

    Science.gov (United States)

    Shackelford, Stacy; Yang, Shiming; Hu, Peter; Miller, Catriona; Anazodo, Amechi; Galvagno, Samuel; Wang, Yulei; Hartsky, Lauren; Fang, Raymond; Mackenzie, Colin

    2015-10-01

    Identification of hemorrhaging trauma patients and prediction of blood transfusion needs in near real time will expedite care of the critically injured. We hypothesized that automated analysis of pulse oximetry signals in combination with laboratory values and vital signs obtained at the time of triage would predict the need for blood transfusion with accuracy greater than that of triage vital signs or pulse oximetry analysis alone. Continuous pulse oximetry signals were recorded for directly admitted trauma patients with abnormal prehospital shock index (heart rate [HR] / systolic blood pressure) of 0.62 or greater. Predictions of blood transfusion within 24 hours were compared using Delong's method for area under the receiver operating characteristic (AUROC) curves to determine the optimal combination of triage vital signs (prehospital HR + systolic blood pressure), pulse oximetry features (40 waveform features, O2 saturation, HR), and laboratory values (hematocrit, electrolytes, bicarbonate, prothrombin time, international normalization ratio, lactate) in multivariate logistic regression models. We enrolled 1,191 patients; 339 were excluded because of incomplete data; 40 received blood within 3 hours; and 14 received massive transfusion. Triage vital signs predicted need for transfusion within 3 hours (AUROC, 0.59) and massive transfusion (AUROC, 0.70). Pulse oximetry for 15 minutes predicted transfusion more accurately than triage vital signs for both time frames (3-hour AUROC, 0.74; p = 0.004) (massive transfusion AUROC, 0.88; p transfusion prediction (3-hour AUROC, 0.84; p transfusion AUROC, 0.91; p blood transfusion during trauma resuscitation more accurately than triage vital signs or pulse oximetry analysis alone. Results suggest automated calculations from a noninvasive vital sign monitor interfaced with a point-of-care laboratory device may support clinical decisions by recognizing patients with hemorrhage sufficient to need transfusion. Epidemiologic

  5. Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies.

    Science.gov (United States)

    Jo, Su Yeon; Lee, Ju Mi; Kim, Hye Lim; Sin, Kyeong Hwa; Lee, Hyeon Ji; Chang, Chulhun Ludgerus; Kim, Hyung Hoi

    2017-03-01

    ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol. The AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods. ABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do.

  6. Whole blood solubilization and discoloration before LSC of yttrium samples

    International Nuclear Information System (INIS)

    Lima, Marina F.; Leonardo, Lucio

    2009-01-01

    Liquid scintillation counting of whole blood and animal tissues samples could be severely impaired owing to quenching by their compounds. The objective of this previous study is preparing one protocol of 90 Y measurement to apply in biodistribution and dosimetry studies of radiopharmaceuticals labeled with this isotope and other beta emitters in in vivo and ex vivo samples. The first parameters considered to choose a method were: the largest blood sample per collection (80μl-90μl), attending the collection limit of less than 7.5% of total circulating blood volume for in vivo samples. Other parameters were the use of EDTA and cyclohexydine as solubilization and lytic agents, HNO 3 and H 2 O 2 as mineralizing agents and NH 4 OH as neutralization agent. One samples batch was tested in a water bath under the lower temperature to prevent the volume lose of the ionic phase. Other samples batch was mineralized over a hot-plate at 120 deg C following the currently largest sample amounts processing procedure in our laboratory by using HNO 3 and H 2 O 2 . Results show the contribution of the blood fragments as quenching in the region A ( 3 and/or NH 4 OH in the hot-plate digestion. As expected, the measurements in the three spectral regions show the proteins and colored fragments were completely removed by the hot-plate digestion. The rate between efficiency and 90 Sr- 90 Y concentration had not significant differences in the range between 120 Bq and 1200 Bq. (author)

  7. A blood sampling microsystem for pharmacokinetic applications: design, fabrication, and initial results.

    Science.gov (United States)

    Li, Tao; Barnett, Adam; Rogers, Karen L; Gianchandani, Yogesh B

    2009-12-21

    This paper describes a microsystem for automated blood sampling from laboratory mice used in pharmacokinetic studies. Intended to be mounted as a "backpack" on a mouse, it uses a microneedle, reservoir, and an actuator to instantaneously prick the animal for a time-point sample, eliminating the need for a tethered catheter with large dead volume. The blood is collected by capillary effect through a 31-33 gauge microneedle (250-210 microm OD) into a approximately 1 microL micromachined steel reservoir. The voice coil actuator provides a peak force of approximately 300 mN, which amply exceeds the measured piercing force of mouse skin (i.e., 60-85 mN for a 31-gauge needle with 12 degrees bevel). The sampling system was tested in vitro using a mock vessel with adjustable pressure; the reservoir was filled in pressure. The system may also be used to sample interstitial fluid, but the absence of blood pressure makes it necessary to enhance the capillary effect of the needle. This is accomplished by either electropolishing the inner surface to make it more hydrophilic or using a polymer wire insert to increase the surface area. The steel surface of the reservoir is also coated with silicon oxynitride by plasma-enhanced chemical vapor deposition to improve its hydrophilicity. Blood from fresh bovine tissue was collected into the reservoir to simulate interstitial fluid sampling. In vivo tests on live, anesthetized mice resulted in successful collection of blood into the reservoir. The possible integration of the device in microanalytical systems and the device scalability for multisampling are discussed.

  8. Evaluation of portable blood glucose meters using canine and feline pooled blood samples.

    Science.gov (United States)

    Mori, A; Oda, H; Onozawa, E; Shono, S; Takahashi, T; Yamashita, S; Fujimoto, H; Sako, T

    2016-12-01

    This study evaluated the accuracy and reproducibility of a human portable blood glucose meter (PBGM) for canine and feline whole blood. Reference plasma glucose values (RPGV) were concurrently measured using glucose oxidation methods. Fifteen healthy dogs and 6 healthy cats were used for blood sampling. Blood glucose concentrations and hematocrits were adjusted using pooled blood samples for our targeted values. A positive correlation between the PBGM and RPGV was found for both dogs (y = 0.877, x = -24.38, r = 0.9982, n = 73) and cats (y = 1.048, x = -27.06, r = 0.9984, n = 69). Acceptable results were obtained in error grid analysis between PBGM and RPGV in both dogs and cats; 100% of these results were within zones A and B. Following ISO recommendations, a PBGM is considered accurate if 95% of the measurements are within ± 15 mg/dl of the RPGV when the glucose concentration is cats (39%, 27 of 69 cats). Blood samples with high hematocrits induced lower whole blood glucose values measured by the PBGM than RPGV under hypoglycemic, normoglycemic, and hyperglycemic conditions in both dogs and cats. Therefore, this device is not clinically useful in dogs and cats. New PBGMs which automatically compensate for the hematocrit should be developed in veterinary practice.

  9. Feasibility of surface sampling in automated inspection of concrete aggregates during bulk transport on a conveyor

    NARCIS (Netherlands)

    Bakker, M.C.M.; Di Maio, F.; Lotfi, S.; Bakker, M.; Hu, M.; Vahidi, A.

    2017-01-01

    Automated optic inspection of concrete aggregates for pollutants (e.g. wood, plastics, gypsum and brick) is required to establish the suitability for reuse in new concrete products. Inspection is more efficient when directly sampling the materials on the conveyor belt instead of feeding them in a

  10. Novel diffusion cell for in vitro transdermal permeation, compatible with automated dynamic sampling

    NARCIS (Netherlands)

    Bosman, I.J; Lawant, A.L; Avegaart, S.R.; Ensing, K; de Zeeuw, R.A

    The development of a new diffusion cell for in vitro transdermal permeation is described. The so-called Kelder cells were used in combination with the ASPEC system (Automatic Sample Preparation with Extraction Columns), which is designed for the automation of solid-phase extractions (SPE). Instead

  11. Thermophilic Campylobacter spp. in turkey samples: evaluation of two automated enzyme immunoassays and conventional microbiological techniques

    DEFF Research Database (Denmark)

    Borck, Birgitte; Stryhn, H.; Ersboll, A.K.

    2002-01-01

    Aims: To determine the sensitivity and specificity of two automated enzyme immunoassays (EIA), EiaFoss and Minividas, and a conventional microbiological culture technique for detecting thermophilic Campylobacter spp. in turkey samples. Methods and Results: A total of 286 samples (faecal, meat......, neckskin and environmental samples) were collected over a period of 4 months at a turkey slaughterhouse and meat-cutting plant in Denmark. Faecal and environmental samples were tested by the conventional culture method and by the two EIAs, whereas meat and neckskin samples were tested by the two EIAs only...

  12. Performance of Gram staining on blood cultures flagged negative by an automated blood culture system.

    Science.gov (United States)

    Peretz, A; Isakovich, N; Pastukh, N; Koifman, A; Glyatman, T; Brodsky, D

    2015-08-01

    Blood is one of the most important specimens sent to a microbiology laboratory for culture. Most blood cultures are incubated for 5-7 days, except in cases where there is a suspicion of infection caused by microorganisms that proliferate slowly, or infections expressed by a small number of bacteria in the bloodstream. Therefore, at the end of incubation, misidentification of positive cultures and false-negative results are a real possibility. The aim of this work was to perform a confirmation by Gram staining of the lack of any microorganisms in blood cultures that were identified as negative by the BACTEC™ FX system at the end of incubation. All bottles defined as negative by the BACTEC FX system were Gram-stained using an automatic device and inoculated on solid growth media. In our work, 15 cultures that were defined as negative by the BACTEC FX system at the end of the incubation were found to contain microorganisms when Gram-stained. The main characteristic of most bacteria and fungi growing in the culture bottles that were defined as negative was slow growth. This finding raises a problematic issue concerning the need to perform Gram staining of all blood cultures, which could overload the routine laboratory work, especially laboratories serving large medical centers and receiving a large number of blood cultures.

  13. Integrated Automation of High-Throughput Screening and Reverse Phase Protein Array Sample Preparation

    DEFF Research Database (Denmark)

    Pedersen, Marlene Lemvig; Block, Ines; List, Markus

    multiplexing readouts, but this has a natural limitation. High-content screening via image acquisition and analysis allows multiplexing of few parameters, but is connected to substantial time consumption and complex logistics. We report on integration of Reverse Phase Protein Arrays (RPPA)-based readouts...... into automated robotic high-throughput screens, which allows subsequent protein quantification. In this integrated solution, samples are directly forwarded to automated cell lysate preparation and preparation of dilution series, including reformatting to a protein spotter-compatible format after the high...

  14. ARAM: an automated image analysis software to determine rosetting parameters and parasitaemia in Plasmodium samples.

    Science.gov (United States)

    Kudella, Patrick Wolfgang; Moll, Kirsten; Wahlgren, Mats; Wixforth, Achim; Westerhausen, Christoph

    2016-04-18

    Rosetting is associated with severe malaria and a primary cause of death in Plasmodium falciparum infections. Detailed understanding of this adhesive phenomenon may enable the development of new therapies interfering with rosette formation. For this, it is crucial to determine parameters such as rosetting and parasitaemia of laboratory strains or patient isolates, a bottleneck in malaria research due to the time consuming and error prone manual analysis of specimens. Here, the automated, free, stand-alone analysis software automated rosetting analyzer for micrographs (ARAM) to determine rosetting rate, rosette size distribution as well as parasitaemia with a convenient graphical user interface is presented. Automated rosetting analyzer for micrographs is an executable with two operation modes for automated identification of objects on images. The default mode detects red blood cells and fluorescently labelled parasitized red blood cells by combining an intensity-gradient with a threshold filter. The second mode determines object location and size distribution from a single contrast method. The obtained results are compared with standardized manual analysis. Automated rosetting analyzer for micrographs calculates statistical confidence probabilities for rosetting rate and parasitaemia. Automated rosetting analyzer for micrographs analyses 25 cell objects per second reliably delivering identical results compared to manual analysis. For the first time rosette size distribution is determined in a precise and quantitative manner employing ARAM in combination with established inhibition tests. Additionally ARAM measures the essential observables parasitaemia, rosetting rate and size as well as location of all detected objects and provides confidence intervals for the determined observables. No other existing software solution offers this range of function. The second, non-malaria specific, analysis mode of ARAM offers the functionality to detect arbitrary objects

  15. On-chip acoustophoretic isolation of microflora including S. typhimurium from raw chicken, beef and blood samples.

    Science.gov (United States)

    Ngamsom, Bongkot; Lopez-Martinez, Maria J; Raymond, Jean-Claude; Broyer, Patrick; Patel, Pradip; Pamme, Nicole

    2016-04-01

    Pathogen analysis in food samples routinely involves lengthy growth-based pre-enrichment and selective enrichment of food matrices to increase the ratio of pathogen to background flora. Similarly, for blood culture analysis, pathogens must be isolated and enriched from a large excess of blood cells to allow further analysis. Conventional techniques of centrifugation and filtration are cumbersome, suffer from low sample throughput, are not readily amenable to automation and carry a risk of damaging biological samples. We report on-chip acoustophoresis as a pre-analytical technique for the resolution of total microbial flora from food and blood samples. The resulting 'clarified' sample is expected to increase the performance of downstream systems for the specific detection of the pathogens. A microfluidic chip with three inlets, a central separation channel and three outlets was utilized. Samples were introduced through the side inlets, and buffer solution through the central inlet. Upon ultrasound actuation, large debris particles (10-100 μm) from meat samples were continuously partitioned into the central buffer channel, leaving the 'clarified' outer sample streams containing both, the pathogenic cells and the background flora (ca. 1 μm) to be collected over a 30 min operation cycle before further analysis. The system was successfully tested with Salmonella typhimurium-spiked (ca. 10(3)CFU mL(-1)) samples of chicken and minced beef, demonstrating a high level of the pathogen recovery (60-90%). When applied to S. typhimurium contaminated blood samples (10(7)CFU mL(-1)), acoustophoresis resulted in a high depletion (99.8%) of the red blood cells (RBC) which partitioned in the buffer stream, whilst sufficient numbers of the viable S. typhimurium remained in the outer channels for further analysis. These results indicate that the technology may provide a generic approach for pre-analytical sample preparation prior to integrated and automated downstream detection of

  16. Feasibility of automated speech sample collection with stuttering children using interactive voice response (IVR) technology.

    Science.gov (United States)

    Vogel, Adam P; Block, Susan; Kefalianos, Elaina; Onslow, Mark; Eadie, Patricia; Barth, Ben; Conway, Laura; Mundt, James C; Reilly, Sheena

    2015-04-01

    To investigate the feasibility of adopting automated interactive voice response (IVR) technology for remotely capturing standardized speech samples from stuttering children. Participants were 10 6-year-old stuttering children. Their parents called a toll-free number from their homes and were prompted to elicit speech from their children using a standard protocol involving conversation, picture description and games. The automated IVR system was implemented using an off-the-shelf telephony software program and delivered by a standard desktop computer. The software infrastructure utilizes voice over internet protocol. Speech samples were automatically recorded during the calls. Video recordings were simultaneously acquired in the home at the time of the call to evaluate the fidelity of the telephone collected samples. Key outcome measures included syllables spoken, percentage of syllables stuttered and an overall rating of stuttering severity using a 10-point scale. Data revealed a high level of relative reliability in terms of intra-class correlation between the video and telephone acquired samples on all outcome measures during the conversation task. Findings were less consistent for speech samples during picture description and games. Results suggest that IVR technology can be used successfully to automate remote capture of child speech samples.

  17. Trends and applications of integrated automated ultra-trace sample handling and analysis (T9)

    International Nuclear Information System (INIS)

    Kingston, H.M.S.; Ye Han; Stewart, L.; Link, D.

    2002-01-01

    Full text: Automated analysis, sub-ppt detection limits, and the trend toward speciated analysis (rather than just elemental analysis) force the innovation of sophisticated and integrated sample preparation and analysis techniques. Traditionally, the ability to handle samples at ppt and sub-ppt levels has been limited to clean laboratories and special sample handling techniques and equipment. The world of sample handling has passed a threshold where older or 'old fashioned' traditional techniques no longer provide the ability to see the sample due to the influence of the analytical blank and the fragile nature of the analyte. When samples require decomposition, extraction, separation and manipulation, application of newer more sophisticated sample handling systems are emerging that enable ultra-trace analysis and species manipulation. In addition, new instrumentation has emerged which integrate sample preparation and analysis to enable on-line near real-time analysis. Examples of those newer sample-handling methods will be discussed and current examples provided as alternatives to traditional sample handling. Two new techniques applying ultra-trace microwave energy enhanced sample handling have been developed that permit sample separation and refinement while performing species manipulation during decomposition. A demonstration, that applies to semiconductor materials, will be presented. Next, a new approach to the old problem of sample evaporation without losses will be demonstrated that is capable of retaining all elements and species tested. Both of those methods require microwave energy manipulation in specialized systems and are not accessible through convection, conduction, or other traditional energy applications. A new automated integrated method for handling samples for ultra-trace analysis has been developed. An on-line near real-time measurement system will be described that enables many new automated sample handling and measurement capabilities. This

  18. Comparison of Proteins in Whole Blood and Dried Blood Spot Samples by LC/MS/MS

    Science.gov (United States)

    Chambers, Andrew G.; Percy, Andrew J.; Hardie, Darryl B.; Borchers, Christoph H.

    2013-09-01

    Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach.

  19. Integrated Automation of High-Throughput Screening and Reverse Phase Protein Array Sample Preparation

    DEFF Research Database (Denmark)

    Pedersen, Marlene Lemvig; Block, Ines; List, Markus

    into automated robotic high-throughput screens, which allows subsequent protein quantification. In this integrated solution, samples are directly forwarded to automated cell lysate preparation and preparation of dilution series, including reformatting to a protein spotter-compatible format after the high......High-throughput screening of genome wide siRNA- or compound libraries is currently applied for drug target and drug discovery. Commonly, these approaches deal with sample numbers ranging from 100,000 to several millions. Efforts to decrease costs and to increase information gained include......-throughput screening. Tracking of huge sample numbers and data analysis from a high-content screen to RPPAs is accomplished via MIRACLE, a custom made software suite developed by us. To this end, we demonstrate that the RPPAs generated in this manner deliver reliable protein readouts and that GAPDH and TFR levels can...

  20. Partial Red Blood Cell Exchange in Children and Young Patients with Sickle Cell Disease: Manual Versus Automated Procedure.

    Science.gov (United States)

    Escobar, Carlos; Moniz, Marta; Nunes, Pedro; Abadesso, Clara; Ferreira, Teresa; Barra, António; Lichtner, Anabela; Loureiro, Helena; Dias, Alexandra; Almeida, Helena

    2017-10-31

    The benefits of manual versus automated red blood cell exchange have rarely been documented and studies in young sickle cell disease patients are scarce. We aim to describe and compare our experience in these two procedures. Young patients (≤ 21 years old) who underwent manual- or automated-red blood cell exchange for prevention or treatment of sickle cell disease complications were included. Clinical, technical and hematological data were prospectively recorded and analyzed. Ninety-four red blood cell exchange sessions were performed over a period of 68 months, including 57 manual and 37 automated, 63 for chronic complications prevention, 30 for acute complications and one in the pre-operative setting. Mean decrease in sickle hemoglobin levels was higher in automated-red blood cell exchange (p sickle hemoglobin level decrease per volume removed (p cell exchange and access alarm on automated-red blood cell exchange. No major complication or alloimunization was recorded. Automated-red blood cell exchange decreased sickle hemoglobin levels more efficiently than manual procedure in the setting of acute and chronic complications of sickle cell disease, with minor technical concerns mainly due to vascular access. The threshold of sickle hemoglobin should be individualized for clinical and hematological goals. In our cohort of young patients, the need for an acceptable venous access was a limiting factor, but iron-overload was avoided. Automated red blood cell exchange is safe and well tolerated. It permits a higher sickle hemoglobin removal efficacy, better volume status control and iron-overload avoidance.

  1. Advancing haemostasis automation--successful implementation of robotic centrifugation and sample processing in a tertiary service hospital.

    Science.gov (United States)

    Sédille-Mostafaie, Nazanin; Engler, Hanna; Lutz, Susanne; Korte, Wolfgang

    2013-06-01

    Laboratories today face increasing pressure to automate operations due to increasing workloads and the need to reduce expenditure. Few studies to date have focussed on the laboratory automation of preanalytical coagulation specimen processing. In the present study, we examined whether a clinical chemistry automation protocol meets the preanalytical requirements for the analyses of coagulation. During the implementation of laboratory automation, we began to operate a pre- and postanalytical automation system. The preanalytical unit processes blood specimens for chemistry, immunology and coagulation by automated specimen processing. As the production of platelet-poor plasma is highly dependent on optimal centrifugation, we examined specimen handling under different centrifugation conditions in order to produce optimal platelet deficient plasma specimens. To this end, manually processed models centrifuged at 1500 g for 5 and 20 min were compared to an automated centrifugation model at 3000 g for 7 min. For analytical assays that are performed frequently enough to be targets for full automation, Passing-Bablok regression analysis showed close agreement between different centrifugation methods, with a correlation coefficient between 0.98 and 0.99 and a bias between -5% and +6%. For seldom performed assays that do not mandate full automation, the Passing-Bablok regression analysis showed acceptable to poor agreement between different centrifugation methods. A full automation solution is suitable and can be recommended for frequent haemostasis testing.

  2. On-chip sample preparation for complete blood count from raw blood.

    Science.gov (United States)

    Nguyen, John; Wei, Yuan; Zheng, Yi; Wang, Chen; Sun, Yu

    2015-03-21

    This paper describes a monolithic microfluidic device capable of on-chip sample preparation for both RBC and WBC measurements from whole blood. For the first time, on-chip sample processing (e.g. dilution, lysis, and filtration) and downstream single cell measurement were fully integrated to enable sample preparation and single cell analysis from whole blood on a single device. The device consists of two parallel sub-systems that perform sample processing and electrical measurements for measuring RBC and WBC parameters. The system provides a modular environment capable of handling solutions of various viscosities by adjusting the length of channels and precisely controlling mixing ratios, and features a new 'offset' filter configuration for increased duration of device operation. RBC concentration, mean corpuscular volume (MCV), cell distribution width, WBC concentration and differential are determined by electrical impedance measurement. Experimental characterization of over 100,000 cells from 10 patient blood samples validated the system's capability for performing on-chip raw blood processing and measurement.

  3. Design aspects of automation system for initial processing of fecal samples

    International Nuclear Information System (INIS)

    Sawant, Pramilla D.; Prabhu, Supreetha P.; Suja, A.; Wankhede, Sonal; Chaudhary, Seema; Rao, D.D.; Pradeepkumar, K.S.; Das, A.P.; Badodkar, B.D.

    2014-01-01

    The procedure for initial handling of the fecal samples at Bioassay Lab., Trombay is as follows: overnight fecal samples are collected from the worker in a kit consisting of a polythene bag placed in a wide mouth polythene container closed with an inner lid and a screw cap. Occupational worker collects the sample in the polythene bag. On receiving the sample, the polythene container along with the sample is weighed, polythene bag containing fecal sample is lifted out of the container using a pair of tongs placed inside a crucible and ashed inside a muffle furnace at 450℃. After complete ashing, the crucible containing white ash is taken-up for further radiochemical processing. This paper describes the various steps in developing a prototype automated system for initial handling of fecal samples. The proposed system for handling and processing of fecal samples is proposed to automate the above. The system once developed will help eliminate manual intervention till the ashing stage and reduce the biological hazard involved in handling such samples mentioned procedure

  4. [DNA quantification of blood samples pre-treated with pyramidon].

    Science.gov (United States)

    Zhu, Chuan-Hong; Zheng, Dao-Li; Ni, Rao-Zhi; Wang, Hai-Sheng; Ning, Ping; Fang, Hui; Liu, Yan

    2014-06-01

    To study DNA quantification and STR typing of samples pre-treated with pyramidon. The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

  5. Automatic sample changer control software for automation of neutron activation analysis process in Malaysian Nuclear Agency

    Science.gov (United States)

    Yussup, N.; Ibrahim, M. M.; Rahman, N. A. A.; Mokhtar, M.; Salim, N. A. A.; Soh@Shaari, S. C.; Azman, A.; Lombigit, L.; Azman, A.; Omar, S. A.

    2018-01-01

    Most of the procedures in neutron activation analysis (NAA) process that has been established in Malaysian Nuclear Agency (Nuclear Malaysia) since 1980s were performed manually. These manual procedures carried out by the NAA laboratory personnel are time consuming and inefficient especially for sample counting and measurement process. The sample needs to be changed and the measurement software needs to be setup for every one hour counting time. Both of these procedures are performed manually for every sample. Hence, an automatic sample changer system (ASC) that consists of hardware and software is developed to automate sample counting process for up to 30 samples consecutively. This paper describes the ASC control software for NAA process which is designed and developed to control the ASC hardware and call GammaVision software for sample measurement. The software is developed by using National Instrument LabVIEW development package.

  6. Portable Automation of Static Chamber Sample Collection for Quantifying Soil Gas Flux

    Energy Technology Data Exchange (ETDEWEB)

    Davis, Morgan P.; Groh, Tyler A.; Parkin, Timothy B.; Williams, Ryan J.; Isenhart, Thomas M.; Hofmockel, Kirsten S.

    2018-01-01

    Quantification of soil gas flux using the static chamber method is labor intensive. The number of chambers that can be sampled is limited by the spacing between chambers and the availability of trained research technicians. An automated system for collecting gas samples from chambers in the field would eliminate the need for personnel to return to the chamber during a flux measurement period and would allow a single technician to sample multiple chambers simultaneously. This study describes Chamber Automated Sampling Equipment (FluxCASE) to collect and store chamber headspace gas samples at assigned time points for the measurement of soil gas flux. The FluxCASE design and operation is described, and the accuracy and precision of the FluxCASE system is evaluated. In laboratory measurements of nitrous oxide (N2O), carbon dioxide (CO2), and methane (CH4) concentrations of a standardized gas mixture, coefficients of variation associated with automated and manual sample collection were comparable, indicating no loss of precision. In the field, soil gas fluxes measured from FluxCASEs were in agreement with manual sampling for both N2O and CO2. Slopes of regression equations were 1.01 for CO2 and 0.97 for N2O. The 95% confidence limits of the slopes of the regression lines included the value of one, indicating no bias. Additionally, an expense analysis found a cost recovery ranging from 0.6 to 2.2 yr. Implementing the FluxCASE system is an alternative to improve the efficiency of the static chamber method for measuring soil gas flux while maintaining the accuracy and precision of manual sampling.

  7. Highly Reproducible Automated Proteomics Sample Preparation Workflow for Quantitative Mass Spectrometry.

    Science.gov (United States)

    Fu, Qin; Kowalski, Michael P; Mastali, Mitra; Parker, Sarah J; Sobhani, Kimia; van den Broek, Irene; Hunter, Christie L; Van Eyk, Jennifer E

    2018-01-05

    Sample preparation for protein quantification by mass spectrometry requires multiple processing steps including denaturation, reduction, alkylation, protease digestion, and peptide cleanup. Scaling these procedures for the analysis of numerous complex biological samples can be tedious and time-consuming, as there are many liquid transfer steps and timed reactions where technical variations can be introduced and propagated. We established an automated sample preparation workflow with a total processing time for 96 samples of 5 h, including a 2 h incubation with trypsin. Peptide cleanup is accomplished by online diversion during the LC/MS/MS analysis. In a selected reaction monitoring (SRM) assay targeting 6 plasma biomarkers and spiked β-galactosidase, mean intraday and interday cyclic voltammograms (CVs) for 5 serum and 5 plasma samples over 5 days were samples repeated on 3 separate days had total CVs below 20%. Similar results were obtained when the workflow was transferred to a second site: 93% of peptides had CVs below 20%. An automated trypsin digestion workflow yields uniformly processed samples in less than 5 h. Reproducible quantification of peptides was observed across replicates, days, instruments, and laboratory sites, demonstrating the broad applicability of this approach.

  8. Home blood pressure measurement in elderly patients with cognitive impairment: comparison of agreement between relative-measured blood pressure and automated blood pressure measurement.

    Science.gov (United States)

    Plichart, Matthieu; Seux, Marie-Laure; Caillard, Laure; Chaussade, Edouard; Vidal, Jean-Sébastien; Boully, Clémence; Hanon, Olivier

    2013-08-01

    Home blood pressure measurement (HBPM) is recommended by guidelines for hypertension management. However, this method might be difficult to use in elderly individuals with cognitive disorders. Our aim was to assess the agreement and the feasibility of HBPM by a relative as compared with 24-h ambulatory blood pressure monitoring (ABPM) in elderly patients with dementia. Sixty outpatients with dementia aged 75 years and older with office hypertension (≥140/90 mmHg) were subjected successively to HBPM by a trained relative and 24-h ABPM. The order of the two methods was randomized. Current guidelines' thresholds for the diagnosis of hypertension were used. The mean (SD) age of the patients was 80.8 (6.1) years (55% women) and the mean (SD) mini-mental state examination score was 20.1 (6.9). The feasibility of relative-HBPM was very high, with a 97% success rate (defined by ≥12/18 measurements reported). The blood pressure measurements were highly correlated between the two methods (r=0.75 and 0.64 for systolic blood pressure and diastolic blood pressure, respectively; Pmethods for the diagnosis of sustained hypertension and white-coat hypertension was excellent (overall agreement, 92%; κ coefficient, 0.81; 95% CI, 0.61-0.93). Similar results were found for daytime-ABPM. In cognitively impaired elderly patients, HBPM by a relative using an automated device was a good alternative to 24-h ABPM.

  9. Sex identification of polar bears from blood and tissue samples

    Science.gov (United States)

    Amstrup, Steven C.; Garner, G.W.; Cronin, M.A.; Patton, J.C.

    1993-01-01

    Polar bears (Ursus maritimus) can be adversely affected by hunting and other human perturbations because of low population densities and low reproduction rates. The sustainable take of adult females may be as low as 1.5% of the population. Females and accompanying young are most vulnerable to hunting, and hunters have not consistently reported the sex composition of the harvest, therefore a method to confirm the sexes of polar bears harvested in Alaska is needed. Evidence of the sex of harvested animals is often not available, but blood or other tissue samples often are. We extracted DNA from tissue and blood samples, and amplified segments of zinc finger (ZFX and ZFY) genes from both X and Y chromosomes with the polymerase chain reaction. Digestion of amplified portions of the X chromosome with the restriction enzyme HaeIII resulted in subdivision of the original amplified segment into four smaller fragments. Digestion with HaeIII did not subdivide the original segment amplified from the Y chromosome. The differing fragment sizes produced patterns in gel electrophoresis that distinguished samples from male and female bears 100% of the time. This technique is applicable to the investigation of many wildlife management and research questions.

  10. Oak ridge national laboratory automated clean chemistry for bulk analysis of environmental swipe samples

    Energy Technology Data Exchange (ETDEWEB)

    Bostick, Debra A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Hexel, Cole R. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Ticknor, Brian W. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Tevepaugh, Kayron N. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Metzger, Shalina C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2016-11-01

    To shorten the lengthy and costly manual chemical purification procedures, sample preparation methods for mass spectrometry are being automated using commercial-off-the-shelf (COTS) equipment. This addresses a serious need in the nuclear safeguards community to debottleneck the separation of U and Pu in environmental samples—currently performed by overburdened chemists—with a method that allows unattended, overnight operation. In collaboration with Elemental Scientific Inc., the prepFAST-MC2 was designed based on current COTS equipment that was modified for U/Pu separations utilizing Eichrom™ TEVA and UTEVA resins. Initial verification of individual columns yielded small elution volumes with consistent elution profiles and good recovery. Combined column calibration demonstrated ample separation without crosscontamination of the eluent. Automated packing and unpacking of the built-in columns initially showed >15% deviation in resin loading by weight, which can lead to inconsistent separations. Optimization of the packing and unpacking methods led to a reduction in the variability of the packed resin to less than 5% daily. The reproducibility of the automated system was tested with samples containing 30 ng U and 15 pg Pu, which were separated in a series with alternating reagent blanks. These experiments showed very good washout of both the resin and the sample from the columns as evidenced by low blank values. Analysis of the major and minor isotope ratios for U and Pu provided values well within data quality limits for the International Atomic Energy Agency. Additionally, system process blanks spiked with 233U and 244Pu tracers were separated using the automated system after it was moved outside of a clean room and yielded levels equivalent to clean room blanks, confirming that the system can produce high quality results without the need for expensive clean room infrastructure. Comparison of the amount of personnel time necessary for successful manual vs

  11. Severe interference between retinal angiography and automated four-color flow cytometry analysis of blood mononuclear cells.

    Science.gov (United States)

    Bürgisser, Philippe; Vaudaux, Jean; Bart, Pierre-Alexandre

    2007-08-01

    Retinal angiography has become a widely used diagnostic tool. It requires the intravenous administration of the fluorescent dyes fluorescein and indocyanin green. We recently received blood taken 8 h after retinal angiography, without our knowing it. We describe the failure of an automated flow cytometry system in the enumeration of lymphocyte subpopulations in this sample. Cell enumeration was achieved by the use of the lyse-no wash MultiTEST procedure (Becton-Dickinson) together with the FACSCalibur cytometer. Absolute cell counts were obtained using TruCount beads. Data were analyzed automatically by the MultiSET and manually with the CellQuest softwares. The dot plots obtained with this sample looked quite abnormal. All monuclear cells stained brightly in the FITC channel irrespective of anti-CD3-FITC conjugate binding. This resulted in a major undercompensation for the increased spillover of the fluorescein emission into the PE-channel. PE-labeled cell and TruCount bead events coalesced. The MultiSET software failed to draw proper gatings and proved useless. Alternative manual gatings could partially rescue the analysis. Clinicians and cytometrists should be aware that, because of dye entry or binding, blood mononuclear cells collected shortly after retinal angiography are not suitable even for common cytometry applications. Copyright 2007 International Society for Analytical Cytology.

  12. In the aftermath of SPRINT: further comparison of unattended automated office blood pressure measurement and 24-hour blood pressure monitoring.

    Science.gov (United States)

    Seidlerová, Jitka; Gelžinský, Julius; Mateřánková, Markéta; Ceral, Jiří; König, Petr; Filipovský, Jan

    2018-03-22

    Several papers reported that unattended automated office blood pressure (uAutoOBP) is closely related to daytime ambulatory blood pressure monitoring (ABPM). In the present study, we aim to study uAutoOBP and its relation to 24-hour ABPM and ABPM variability. Stable treated hypertensive subjects were examined in two Czech academic hypertension centres. uAutoOBP was measured with the BP Tru device; attended BP three times with auscultatory method (AuscOBP) by the physician. ABPM was performed within one week from the clinical visit. Data on 98 subjects aged 67.7 ± 9.3 years with 24-hour ABPM 120.3 ± 10.6/72.7 ± 7.9 mm Hg are reported. uAutoOBP was lower than 24-hour (by -5.2 ± 11.3/-0.5 ± 6.9 mm Hg) and daytime (by -6.7 ± 12.82.4 ± 8.0 mm Hg) ABPM and the individual variability of the difference was very large (up to 30 mm Hg). The correlation coefficients between ABPM and uAutoOBP were similar compared to AuscOBP (p ≥ .17). Variability of uAutoOBP, but not AuscOBP, readings during one clinical visit was related to short-term blood pressure variability of ABPM. The difference between AuscOBP and uAutoOBP was larger in patients with white-coat effect compared to other blood pressure control groups (25.1 ± 7.0 vs. 2.2 ± 10.3 mm Hg; p = .0036). Our study shows that uAutoOBP is not good predictor of ambulatory blood pressure monitoring, not even of the daytime values. It might, however, indicate short-term blood pressure variability and, when compared with AuscOBP, also detect patients with white-coat effect.

  13. Automated Prediction of Catalytic Mechanism and Rate Law Using Graph-Based Reaction Path Sampling.

    Science.gov (United States)

    Habershon, Scott

    2016-04-12

    In a recent article [ J. Chem. Phys. 2015 , 143 , 094106 ], we introduced a novel graph-based sampling scheme which can be used to generate chemical reaction paths in many-atom systems in an efficient and highly automated manner. The main goal of this work is to demonstrate how this approach, when combined with direct kinetic modeling, can be used to determine the mechanism and phenomenological rate law of a complex catalytic cycle, namely cobalt-catalyzed hydroformylation of ethene. Our graph-based sampling scheme generates 31 unique chemical products and 32 unique chemical reaction pathways; these sampled structures and reaction paths enable automated construction of a kinetic network model of the catalytic system when combined with density functional theory (DFT) calculations of free energies and resultant transition-state theory rate constants. Direct simulations of this kinetic network across a range of initial reactant concentrations enables determination of both the reaction mechanism and the associated rate law in an automated fashion, without the need for either presupposing a mechanism or making steady-state approximations in kinetic analysis. Most importantly, we find that the reaction mechanism which emerges from these simulations is exactly that originally proposed by Heck and Breslow; furthermore, the simulated rate law is also consistent with previous experimental and computational studies, exhibiting a complex dependence on carbon monoxide pressure. While the inherent errors of using DFT simulations to model chemical reactivity limit the quantitative accuracy of our calculated rates, this work confirms that our automated simulation strategy enables direct analysis of catalytic mechanisms from first principles.

  14. RoboDiff: combining a sample changer and goniometer for highly automated macromolecular crystallography experiments.

    Science.gov (United States)

    Nurizzo, Didier; Bowler, Matthew W; Caserotto, Hugo; Dobias, Fabien; Giraud, Thierry; Surr, John; Guichard, Nicolas; Papp, Gergely; Guijarro, Matias; Mueller-Dieckmann, Christoph; Flot, David; McSweeney, Sean; Cipriani, Florent; Theveneau, Pascal; Leonard, Gordon A

    2016-08-01

    Automation of the mounting of cryocooled samples is now a feature of the majority of beamlines dedicated to macromolecular crystallography (MX). Robotic sample changers have been developed over many years, with the latest designs increasing capacity, reliability and speed. Here, the development of a new sample changer deployed at the ESRF beamline MASSIF-1 (ID30A-1), based on an industrial six-axis robot, is described. The device, named RoboDiff, includes a high-capacity dewar, acts as both a sample changer and a high-accuracy goniometer, and has been designed for completely unattended sample mounting and diffraction data collection. This aim has been achieved using a high level of diagnostics at all steps of the process from mounting and characterization to data collection. The RoboDiff has been in service on the fully automated endstation MASSIF-1 at the ESRF since September 2014 and, at the time of writing, has processed more than 20 000 samples completely automatically.

  15. How You Can Help Medical Research: Donating Your Blood, Tissue, and Other Samples

    Science.gov (United States)

    ... HUMAN SERVICES National Institutes of Health Donating Your Blood, Tissue, and Other Samples You have the choice to donate samples, such as blood and tissue, for medical research. Medical researchers use samples to ...

  16. Automated combustion accelerator mass spectrometry for the analysis of biomedical samples in the low attomole range.

    Science.gov (United States)

    van Duijn, Esther; Sandman, Hugo; Grossouw, Dimitri; Mocking, Johannes A J; Coulier, Leon; Vaes, Wouter H J

    2014-08-05

    The increasing role of accelerator mass spectrometry (AMS) in biomedical research necessitates modernization of the traditional sample handling process. AMS was originally developed and used for carbon dating, therefore focusing on a very high precision but with a comparably low sample throughput. Here, we describe the combination of automated sample combustion with an elemental analyzer (EA) online coupled to an AMS via a dedicated interface. This setup allows direct radiocarbon measurements for over 70 samples daily by AMS. No sample processing is required apart from the pipetting of the sample into a tin foil cup, which is placed in the carousel of the EA. In our system, up to 200 AMS analyses are performed automatically without the need for manual interventions. We present results on the direct total (14)C count measurements in <2 μL human plasma samples. The method shows linearity over a range of 0.65-821 mBq/mL, with a lower limit of quantification of 0.65 mBq/mL (corresponding to 0.67 amol for acetaminophen). At these extremely low levels of activity, it becomes important to quantify plasma specific carbon percentages. This carbon percentage is automatically generated upon combustion of a sample on the EA. Apparent advantages of the present approach include complete omission of sample preparation (reduced hands-on time) and fully automated sample analysis. These improvements clearly stimulate the standard incorporation of microtracer research in the drug development process. In combination with the particularly low sample volumes required and extreme sensitivity, AMS strongly improves its position as a bioanalysis method.

  17. Automated low energy photon absorption equipment for measuring internal moisture and density distributions of wood samples

    International Nuclear Information System (INIS)

    Tiitta, M.; Olkkonen, H.; Lappalainen, T.; Kanko, T.

    1993-01-01

    Automated equipment for measuring the moisture and density distributions of wood samples was developed. Using a narrow beam of gamma rays, the equipment scans the wood samples, which are placed on the moving belt. The moisture measurement is based on the 241 Am photon absorption technique (59.5 keV), where the difference of the linear absorption coefficients of the moist and dry wood is measured. The method requires no knowledge of the thickness of the specimen. The density estimation method is based on the measurement of the linear attenuation coefficient of wood. Comprehensive software including image processing was developed for treatment of the numerical values of the measurements. (author)

  18. Human mixed lymphocyte cultures. Evaluation of microculture technique utilizing the multiple automated sample harvester (MASH)

    Science.gov (United States)

    Thurman, G. B.; Strong, D. M.; Ahmed, A.; Green, S. S.; Sell, K. W.; Hartzman, R. J.; Bach, F. H.

    1973-01-01

    Use of lymphocyte cultures for in vitro studies such as pretransplant histocompatibility testing has established the need for standardization of this technique. A microculture technique has been developed that has facilitated the culturing of lymphocytes and increased the quantity of cultures feasible, while lowering the variation between replicate samples. Cultures were prepared for determination of tritiated thymidine incorporation using a Multiple Automated Sample Harvester (MASH). Using this system, the parameters that influence the in vitro responsiveness of human lymphocytes to allogeneic lymphocytes have been investigated. PMID:4271568

  19. On the improvement of blood sample collection at clinical laboratories.

    Science.gov (United States)

    Grasas, Alex; Ramalhinho, Helena; Pessoa, Luciana S; Resende, Mauricio G C; Caballé, Imma; Barba, Nuria

    2014-01-09

    Blood samples are usually collected daily from different collection points, such hospitals and health centers, and transported to a core laboratory for testing. This paper presents a project to improve the collection routes of two of the largest clinical laboratories in Spain. These routes must be designed in a cost-efficient manner while satisfying two important constraints: (i) two-hour time windows between collection and delivery, and (ii) vehicle capacity. A heuristic method based on a genetic algorithm has been designed to solve the problem of blood sample collection. The user enters the following information for each collection point: postal address, average collecting time, and average demand (in thermal containers). After implementing the algorithm using C programming, this is run and, in few seconds, it obtains optimal (or near-optimal) collection routes that specify the collection sequence for each vehicle. Different scenarios using various types of vehicles have been considered. Unless new collection points are added or problem parameters are changed substantially, routes need to be designed only once. The two laboratories in this study previously planned routes manually for 43 and 74 collection points, respectively. These routes were covered by an external carrier company. With the implementation of this algorithm, the number of routes could be reduced from ten to seven in one laboratory and from twelve to nine in the other, which represents significant annual savings in transportation costs. The algorithm presented can be easily implemented in other laboratories that face this type of problem, and it is particularly interesting and useful as the number of collection points increases. The method designs blood collection routes with reduced costs that meet the time and capacity constraints of the problem.

  20. Fully automated gamma spectrometry gauge observing possible radioactive contamination of melting-shop samples

    International Nuclear Information System (INIS)

    Kroos, J.; Westkaemper, G.; Stein, J.

    1999-01-01

    At Salzgitter AG, several monitoring systems have been installed to check the scrap transport by rail and by car. At the moment, the scrap transport by ship is reloaded onto wagons for monitoring afterwards. In the future, a detection system will be mounted onto a crane for a direct check on scrap upon the departure of ship. Furthermore, at Salzgitter AG Central Chemical Laboratory, a fully automated gamma spectrometry gauge is installed in order to observe a possible radioactive contamination of the products. The gamma spectrometer is integrated into the automated OE spectrometry line for testing melting shop samples after performing the OE spectrometry. With this technique the specific activity of selected nuclides and dose rate will be determined. The activity observation is part of the release procedure. The corresponding measurement data are stored in a database for quality management reasons. (author)

  1. Ambulatory blood pressure and blood lipids in a multiethnic sample of healthy adults.

    Science.gov (United States)

    James, Gary D; Van Berge-Landry, Helene M; Morrison, Lynn A; Reza, Angela M; Nicolaisen, Nicola M; Bindon, James R; Brown, Daniel E

    2013-01-01

    Elevated blood pressure (BP), elevated serum cholesterol, and aberrant lipoprotein fractions (low levels of high-density lipoprotein (HDL) and high levels of low-density lipoprotein fractions and triglycerides) have all been used as measures that assess the "metabolic syndrome" and more recently in indexes of allostatic load, which are designed to assess the degree of integrated metabolic pathology. While there are ample data regarding the interrelationships of these measures in various pathophysiological settings, there are limited data regarding the interrelationship of ambulatory BP (ABP) and blood lipids in healthy subjects. The present study evaluates ABP-blood lipid relationships in a multiethnic sample of healthy adults. The subjects were 37 men (age = 40.9 ± 10.7 years) and 42 women (age = 35.8 ± 10.4 years) who were employed as hotel workers in Hawaii. Each wore an ABP monitor for one midweek workday and had pressures averaged in three daily microenvironments (work, home, and during sleep). They also had fasting blood samples taken for lipid profiling. Multivariate analysis of covariance shows that there was a strong inverse relationship between HDL and both systolic (P act as a group in healthy adults but that higher HDL is associated with lower BP. This latter finding is consistent with research that shows that HDL promotes vasodilation via its effect on endothelial nitric oxide synthase. Copyright © 2013 Wiley Periodicals, Inc.

  2. Automated, Ultra-Sterile Solid Sample Handling and Analysis on a Chip

    Science.gov (United States)

    Mora, Maria F.; Stockton, Amanda M.; Willis, Peter A.

    2013-01-01

    There are no existing ultra-sterile lab-on-a-chip systems that can accept solid samples and perform complete chemical analyses without human intervention. The proposed solution is to demonstrate completely automated lab-on-a-chip manipulation of powdered solid samples, followed by on-chip liquid extraction and chemical analysis. This technology utilizes a newly invented glass micro-device for solid manipulation, which mates with existing lab-on-a-chip instrumentation. Devices are fabricated in a Class 10 cleanroom at the JPL MicroDevices Lab, and are plasma-cleaned before and after assembly. Solid samples enter the device through a drilled hole in the top. Existing micro-pumping technology is used to transfer milligrams of powdered sample into an extraction chamber where it is mixed with liquids to extract organic material. Subsequent chemical analysis is performed using portable microchip capillary electrophoresis systems (CE). These instruments have been used for ultra-highly sensitive (parts-per-trillion, pptr) analysis of organic compounds including amines, amino acids, aldehydes, ketones, carboxylic acids, and thiols. Fully autonomous amino acid analyses in liquids were demonstrated; however, to date there have been no reports of completely automated analysis of solid samples on chip. This approach utilizes an existing portable instrument that houses optics, high-voltage power supplies, and solenoids for fully autonomous microfluidic sample processing and CE analysis with laser-induced fluorescence (LIF) detection. Furthermore, the entire system can be sterilized and placed in a cleanroom environment for analyzing samples returned from extraterrestrial targets, if desired. This is an entirely new capability never demonstrated before. The ability to manipulate solid samples, coupled with lab-on-a-chip analysis technology, will enable ultraclean and ultrasensitive end-to-end analysis of samples that is orders of magnitude more sensitive than the ppb goal given

  3. Characteristics of a new fully programmable blood sampling device for monitoring blood radioactivity during PET

    International Nuclear Information System (INIS)

    Boellaard, R.; Lingen, A. van; Balen, S.C.M. van; Hoving, B.G.; Lammertsma, A.A.

    2001-01-01

    The first performance tests of a new fully programmable blood sampling device for monitoring blood radioactivity during positron emission tomography (PET) are described. Blood is withdrawn through 1-mm internal diameter tubing using an infusion pump which can be operated at rates varying from 0 to 600 ml/h. Activity in blood is measured by a 6-cm-thick bismuth germanate crystal connected to a photomultiplier tube and multichannel analyser (MCA) which are positioned within 6 cm lead shielding. Positioning of the tubing is an exact and simple procedure. The minimal readout time of the MCA is 1 s. Two independent energy windows can be set. Operation of the pump and MCA is fully controlled by a PC, i.e. sampling time, interval time and pump rate can be varied at any time during the PET scan by user-defined scripts. A number of characteristics of the new system were studied, such as sensitivity, dead time, linearity, effect of background radiation and pump rate as a function of input pressure. In addition, dispersion was measured as a function of pump rate. Finally, first clinical results were compared with manual samples. The sensitivity equalled 0.7 and 0.2 cps/Bq for 511- and 1022-keV 30% energy windows, respectively, and the system dead time was 500 ns. The system remained linear within 2% with activity concentrations up to 2.5 MBq/cc. Short-term reproducibility was better than 3% for a 1-h period. Long-term reproducibility was about 5% (1SD), which was mainly caused by variation in the diameter of the tubing. If the device was positioned in such a way that maximum shielding was directed towards the patient, the effects of background radiation from the patient on the measured activity concentration for clinically relevant conditions was minimal ( -1 were observed for pump rates higher than 300 ml/h, indicating that the system dispersion is small. Clinical data showed an excellent agreement to within 3% (1SD) between the results obtained with the new system and

  4. Retinal blood vessel segmentation in high resolution fundus photographs using automated feature parameter estimation

    Science.gov (United States)

    Orlando, José Ignacio; Fracchia, Marcos; del Río, Valeria; del Fresno, Mariana

    2017-11-01

    Several ophthalmological and systemic diseases are manifested through pathological changes in the properties and the distribution of the retinal blood vessels. The characterization of such alterations requires the segmentation of the vasculature, which is a tedious and time-consuming task that is infeasible to be performed manually. Numerous attempts have been made to propose automated methods for segmenting the retinal vasculature from fundus photographs, although their application in real clinical scenarios is usually limited by their ability to deal with images taken at different resolutions. This is likely due to the large number of parameters that have to be properly calibrated according to each image scale. In this paper we propose to apply a novel strategy for automated feature parameter estimation, combined with a vessel segmentation method based on fully connected conditional random fields. The estimation model is learned by linear regression from structural properties of the images and known optimal configurations, that were previously obtained for low resolution data sets. Our experiments in high resolution images show that this approach is able to estimate appropriate configurations that are suitable for performing the segmentation task without requiring to re-engineer parameters. Furthermore, our combined approach reported state of the art performance on the benchmark data set HRF, as measured in terms of the F1-score and the Matthews correlation coefficient.

  5. Construction and calibration of a low cost and fully automated vibrating sample magnetometer

    Energy Technology Data Exchange (ETDEWEB)

    El-Alaily, T.M., E-mail: toson_alaily@yahoo.com [Physics Department, Faculty of Science, Tanta University, Tanta (Egypt); El-Nimr, M.K.; Saafan, S.A.; Kamel, M.M.; Meaz, T.M. [Physics Department, Faculty of Science, Tanta University, Tanta (Egypt); Assar, S.T. [Engineering Physics and Mathematics Department, Faculty of Engineering, Tanta University, Tanta (Egypt)

    2015-07-15

    A low cost vibrating sample magnetometer (VSM) has been constructed by using an electromagnet and an audio loud speaker; where both are controlled by a data acquisition device. The constructed VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. The apparatus has been calibrated and tested by using magnetic hysteresis data of some ferrite samples measured by two scientifically calibrated magnetometers; model (Lake Shore 7410) and model (LDJ Electronics Inc. Troy, MI). Our VSM lab-built new design proved success and reliability. - Highlights: • A low cost automated vibrating sample magnetometer VSM has been constructed. • The VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. • The VSM has been calibrated and tested by using some measured ferrite samples. • Our VSM lab-built new design proved success and reliability.

  6. An Automated Method to Quantify Radiation Damage in Human Blood Cells

    Energy Technology Data Exchange (ETDEWEB)

    Gordon K. Livingston, Mark S. Jenkins and Akio A. Awa

    2006-07-10

    Cytogenetic analysis of blood lymphocytes is a well established method to assess the absorbed dose in persons exposed to ionizing radiation. Because mature lymphocytes circulate throughout the body, the dose to these cells is believed to represent the average whole body exposure. Cytogenetic methods measure the incidence of structural aberrations in chromosomes as a means to quantify DNA damage which occurs when ionizing radiation interacts with human tissue. Methods to quantify DNA damage at the chromosomal level vary in complexity and tend to be laborious and time consuming. In a mass casualty scenario involving radiological/nuclear materials, the ability to rapidly triage individuals according to radiation dose is critically important. For high-throughput screening for dicentric chromosomes, many of the data collection steps can be optimized with motorized microscopes coupled to automated slide scanning platforms.

  7. Whole blood samples for adrenocorticotrophic hormone measurement can be stored at room temperature for 4 hours

    DEFF Research Database (Denmark)

    Christensen, Mette; Madsen, Rikke Fogt; Møller, Line Rosengreen

    2016-01-01

    INTRODUCTION: The aim of this study was to investigate and compare the stability of adrenocorticotrophic hormone (ACTH) in whole blood stored on ice and at room temperature for up to 48 hours. This study differs from previous studies by a larger data material. MATERIALS AND METHODS: EDTA-blood...... samples from 30 patients were collected, aliquoted and stored on ice or at room temperature for 0, 2, 4, 24, or 48 h before centrifugation, and the plasma was stored frozen until analysis. All samples were analyzed using an automated electrochemiluminescence immunoassay on cobas 6000 e601. The change...... in ACTH concentration was illustrated as ACTH recovery compared to standard conditions defined as samples stored immediately on ice, centrifuged and plasma frozen within 1 h. A change in ACTH concentration of more than 10% was considered to be of clinical relevance. RESULTS: The results showed...

  8. Comparison of blood RNA isolation methods from samples stabilized in Tempus tubes and stored at a large human biobank.

    Science.gov (United States)

    Aarem, Jeanette; Brunborg, Gunnar; Aas, Kaja K; Harbak, Kari; Taipale, Miia M; Magnus, Per; Knudsen, Gun Peggy; Duale, Nur

    2016-09-01

    More than 50,000 adult and cord blood samples were collected in Tempus tubes and stored at the Norwegian Institute of Public Health Biobank for future use. In this study, we systematically evaluated and compared five blood-RNA isolation protocols: three blood-RNA isolation protocols optimized for simultaneous isolation of all blood-RNA species (MagMAX RNA Isolation Kit, both manual and semi-automated protocols; and Norgen Preserved Blood RNA kit I); and two protocols optimized for large RNAs only (Tempus Spin RNA, and Tempus 6-port isolation kit). We estimated the following parameters: RNA quality, RNA yield, processing time, cost per sample, and RNA transcript stability of six selected mRNAs and 13 miRNAs using real-time qPCR. Whole blood samples from adults (n = 59 tubes) and umbilical cord blood (n = 18 tubes) samples collected in Tempus tubes were analyzed. High-quality blood-RNAs with average RIN-values above seven were extracted using all five RNA isolation protocols. The transcript levels of the six selected genes showed minimal variation between the five protocols. Unexplained differences within the transcript levels of the 13 miRNA were observed; however, the 13 miRNAs had similar expression direction and they were within the same order of magnitude. Some differences in the RNA processing time and cost were noted. Sufficient amounts of high-quality RNA were obtained using all five protocols, and the Tempus blood RNA system therefore seems not to be dependent on one specific RNA isolation method.

  9. Autoverification of the automated blood cell counter (CBC in a reference laboratory in Bogota, Colombia

    Directory of Open Access Journals (Sweden)

    Oscar Martinez-Nieto

    2015-12-01

    Full Text Available ABSTRACT Introduction: The clinical laboratory is part of the group of actors in health systems that are under increasing pressure by users and administrators to increase their productivity in order to respond efficiently to the increased volume of patients, optimizing costs and professional time. This pressure forced laboratories to perform a full review of their procedures and develop technical, logistical and computational tools to enable excellent response times. Objective: This study aimed to evaluate the implementation of the automated blood cell counter autoverification process and its impact on the safety of patients. Methods: Verification rules were designed in the connectivity software, based on manual validation criteria for laboratory professionals, according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI Guideline Auto10-A and the International Consensus Group for Hematology Review (ISLH. The autoverification percentage was established, and non-conforming product (NCP percentages were estimated before and after the procedure. Pilot tests were also performed in different days so as to adjust the process. Results: 53.4% of automated blood cell counters autoverification were achieved, and, subsequently in the audit of 18 months, 60% was reached due to verification adjustments in the delta programmed filter. The NCPs rose from 0.065% to 0.0036% from the beginning to the end of the process. Conclusion: The autoverification process enabled to reduce the variability associated with human intervention, therefore the professional is able to focus on the pathological report analysis, reducing the risk of errors and advocating greater importance on patient safety.

  10. Automated sample exchange and tracking system for neutron research at cryogenic temperatures

    Science.gov (United States)

    Rix, J. E.; Weber, J. K. R.; Santodonato, L. J.; Hill, B.; Walker, L. M.; McPherson, R.; Wenzel, J.; Hammons, S. E.; Hodges, J.; Rennich, M.; Volin, K. J.

    2007-01-01

    An automated system for sample exchange and tracking in a cryogenic environment and under remote computer control was developed. Up to 24 sample "cans" per cycle can be inserted and retrieved in a programed sequence. A video camera acquires a unique identification marked on the sample can to provide a record of the sequence. All operations are coordinated via a LABVIEW™ program that can be operated locally or over a network. The samples are contained in vanadium cans of 6-10mm in diameter and equipped with a hermetically sealed lid that interfaces with the sample handler. The system uses a closed-cycle refrigerator (CCR) for cooling. The sample was delivered to a precooling location that was at a temperature of ˜25K, after several minutes, it was moved onto a "landing pad" at ˜10K that locates the sample in the probe beam. After the sample was released onto the landing pad, the sample handler was retracted. Reading the sample identification and the exchange operation takes approximately 2min. The time to cool the sample from ambient temperature to ˜10K was approximately 7min including precooling time. The cooling time increases to approximately 12min if precooling is not used. Small differences in cooling rate were observed between sample materials and for different sample can sizes. Filling the sample well and the sample can with low pressure helium is essential to provide heat transfer and to achieve useful cooling rates. A resistive heating coil can be used to offset the refrigeration so that temperatures up to ˜350K can be accessed and controlled using a proportional-integral-derivative control loop. The time for the landing pad to cool to ˜10K after it has been heated to ˜240K was approximately 20min.

  11. Quantification of 31 illicit and medicinal drugs and metabolites in whole blood by fully automated solid-phase extraction and ultra-performance liquid chromatography-tandem mass spectrometry

    DEFF Research Database (Denmark)

    Bjørk, Marie Kjærgaard; Simonsen, Kirsten Wiese; Andersen, David Wederkinck

    2013-01-01

    An efficient method for analyzing illegal and medicinal drugs in whole blood using fully automated sample preparation and short ultra-high-performance liquid chromatography–tandem mass spectrometry (MS/MS) run time is presented. A selection of 31 drugs, including amphetamines, cocaine, opioids...

  12. Rapid and Automated Determination of Plutonium and Neptunium in Environmental Samples

    DEFF Research Database (Denmark)

    Qiao, Jixin

    This thesis presents improved analytical methods for rapid and automated determination of plutonium and neptunium in environmental samples using sequential injection (SI) based chromatography and inductively coupled plasma mass spectrometry (ICP-MS). The progress of methodology development in thi...... environmental risk monitoring and assessment, emergency preparedness and surveillance of contaminated areas....... for rapid and simultaneous determination of plutonium and neptunium within an SI system (Paper VI). The results demonstrate that the developed methods in this study are reliable and efficient for accurate assays of trace levels of plutonium and neptunium as demanded in different situations including...

  13. A continuous flow from sample collection to data acceptability determination using an automated system

    International Nuclear Information System (INIS)

    Fisk, J.F.; Leasure, C.; Sauter, A.D.

    1993-01-01

    In its role as regulator, EPA is the recipient of enormous reams of analytical data, especially within the Superfund Program. In order to better manage the volume of paper that comes in daily, Superfund has required its laboratories to provide data that is contained on reporting forms to be delivered also on a diskette for uploading into data bases for various purposes, such as checking for contractual compliance, tracking quality assurance parameters, and, ultimately, for reviewing the data by computer. This last area, automated review of the data, has generated programs that are not necessarily appropriate for use by clients other than Superfund. Such is the case with Los Alamos National Laboratory's Environmental Chemistry Group and its emerging subcontractor community, designed to meet the needs of the remedial action program at LANL. LANL is in the process of implementing an automated system that will be used from the planning stage of sample collection to the production of a project-specific report on analytical data quality. Included are electronic scheduling and tracking of samples, data entry, checking and transmission, data assessment and qualification for use, and report generation that will tie the analytical data quality back to the performance criteria defined prior to sample collection. Industry standard products will be used (e.g., ORACLE, Microsoft Excel) to ensure support for users, prevent dependence on proprietary software, and to protect LANL's investment for the future

  14. Fluidic automation of nitrate and nitrite bioassays in whole blood by dissolvable-film based centrifugo-pneumatic actuation

    DEFF Research Database (Denmark)

    Nwankire, Charles E.; Chan, Di-Sien S.; Gaughran, Jennifer

    2013-01-01

    This paper demonstrates the full centrifugal microfluidic integration and automation of all liquid handling steps of a 7-step fluorescence-linked immunosorbent assay (FLISA) for quantifying nitrate and nitrite levels in whole blood within about 15 min. The assay protocol encompasses the extraction...

  15. Evaluation of an automated protocol for efficient and reliable DNA extraction of dietary samples.

    Science.gov (United States)

    Wallinger, Corinna; Staudacher, Karin; Sint, Daniela; Thalinger, Bettina; Oehm, Johannes; Juen, Anita; Traugott, Michael

    2017-08-01

    Molecular techniques have become an important tool to empirically assess feeding interactions. The increased usage of next-generation sequencing approaches has stressed the need of fast DNA extraction that does not compromise DNA quality. Dietary samples here pose a particular challenge, as these demand high-quality DNA extraction procedures for obtaining the minute quantities of short-fragmented food DNA. Automatic high-throughput procedures significantly decrease time and costs and allow for standardization of extracting total DNA. However, these approaches have not yet been evaluated for dietary samples. We tested the efficiency of an automatic DNA extraction platform and a traditional CTAB protocol, employing a variety of dietary samples including invertebrate whole-body extracts as well as invertebrate and vertebrate gut content samples and feces. Extraction efficacy was quantified using the proportions of successful PCR amplifications of both total and prey DNA, and cost was estimated in terms of time and material expense. For extraction of total DNA, the automated platform performed better for both invertebrate and vertebrate samples. This was also true for prey detection in vertebrate samples. For the dietary analysis in invertebrates, there is still room for improvement when using the high-throughput system for optimal DNA yields. Overall, the automated DNA extraction system turned out as a promising alternative to labor-intensive, low-throughput manual extraction methods such as CTAB. It is opening up the opportunity for an extensive use of this cost-efficient and innovative methodology at low contamination risk also in trophic ecology.

  16. Measurement of thyroxine and cortisol in canine and feline blood samples using two immunoassay analysers.

    Science.gov (United States)

    Higgs, P; Costa, M; Freke, A; Papasouliotis, K

    2014-03-01

    The AIA-360 (Tosoh Corporation) is an automated immunoassay analyser. The aims of this study were to estimate the precision of thyroxine and cortisol AIA-360 immunoassays in canine and feline samples and to compare the results produced with those obtained by a chemiluminescence analyser (Immulite® 1000, Siemens). Blood samples from 240 clinical cases (60 dogs and 60 cats for both thyroxine and cortisol) were analysed using both instruments. Deming regression calculations showed excellent correlation (thyroxine, canine rs  = 0 · 94, feline rs  = 0 · 97; cortisol, canine rs  = 0 · 97, feline rs  = 0 · 97). Agreement between the two instruments was examined by Bland-Altman difference plots, which identified wide confidence intervals and outliers for thyroxine (canine n = 6, feline n = 4) and cortisol (canine n = 3, feline n = 4) results. Inter/intra-run precision of the AIA-360 was excellent for both cortisol and thyroxine (coefficients of variation cortisol and thyroxine in canine and feline samples demonstrating that the AIA-360 can be used in clinical practice. The agreement studies suggest that the results from the AIA-360 cannot be used interchangeably with those generated by the Immulite 1000 and should be interpreted using reference intervals that have been established specific to the AIA-360. © 2014 British Small Animal Veterinary Association.

  17. Automatic collection of bovine blood samples | Hale | South African ...

    African Journals Online (AJOL)

    A technique is described which allows automatic collection of jugular venous blood from tethered cows. In this system, blood is pumped continuously from an intravenous cannula which has a double lumen while an anticoagulant is pumped through the second opening. Diluted blood is collected in a fraction collector which ...

  18. An Energy Efficient Adaptive Sampling Algorithm in a Sensor Network for Automated Water Quality Monitoring.

    Science.gov (United States)

    Shu, Tongxin; Xia, Min; Chen, Jiahong; Silva, Clarence de

    2017-11-05

    Power management is crucial in the monitoring of a remote environment, especially when long-term monitoring is needed. Renewable energy sources such as solar and wind may be harvested to sustain a monitoring system. However, without proper power management, equipment within the monitoring system may become nonfunctional and, as a consequence, the data or events captured during the monitoring process will become inaccurate as well. This paper develops and applies a novel adaptive sampling algorithm for power management in the automated monitoring of the quality of water in an extensive and remote aquatic environment. Based on the data collected on line using sensor nodes, a data-driven adaptive sampling algorithm (DDASA) is developed for improving the power efficiency while ensuring the accuracy of sampled data. The developed algorithm is evaluated using two distinct key parameters, which are dissolved oxygen (DO) and turbidity. It is found that by dynamically changing the sampling frequency, the battery lifetime can be effectively prolonged while maintaining a required level of sampling accuracy. According to the simulation results, compared to a fixed sampling rate, approximately 30.66% of the battery energy can be saved for three months of continuous water quality monitoring. Using the same dataset to compare with a traditional adaptive sampling algorithm (ASA), while achieving around the same Normalized Mean Error (NME), DDASA is superior in saving 5.31% more battery energy.

  19. An Energy Efficient Adaptive Sampling Algorithm in a Sensor Network for Automated Water Quality Monitoring

    Directory of Open Access Journals (Sweden)

    Tongxin Shu

    2017-11-01

    Full Text Available Power management is crucial in the monitoring of a remote environment, especially when long-term monitoring is needed. Renewable energy sources such as solar and wind may be harvested to sustain a monitoring system. However, without proper power management, equipment within the monitoring system may become nonfunctional and, as a consequence, the data or events captured during the monitoring process will become inaccurate as well. This paper develops and applies a novel adaptive sampling algorithm for power management in the automated monitoring of the quality of water in an extensive and remote aquatic environment. Based on the data collected on line using sensor nodes, a data-driven adaptive sampling algorithm (DDASA is developed for improving the power efficiency while ensuring the accuracy of sampled data. The developed algorithm is evaluated using two distinct key parameters, which are dissolved oxygen (DO and turbidity. It is found that by dynamically changing the sampling frequency, the battery lifetime can be effectively prolonged while maintaining a required level of sampling accuracy. According to the simulation results, compared to a fixed sampling rate, approximately 30.66% of the battery energy can be saved for three months of continuous water quality monitoring. Using the same dataset to compare with a traditional adaptive sampling algorithm (ASA, while achieving around the same Normalized Mean Error (NME, DDASA is superior in saving 5.31% more battery energy.

  20. Effect of order of draw of blood samples during phlebotomy on routine biochemistry results.

    Science.gov (United States)

    Sulaiman, Raashda A; Cornes, Michael P; Whitehead, Simon J; Othonos, Nadia; Ford, Clare; Gama, Rousseau

    2011-11-01

    To investigate whether incorrect order of draw of blood samples during phlebotomy causes in vitro potassium ethylenediaminetetraacetic acid EDTA (kEDTA) contamination of blood samples. Serum kEDTA, potassium, calcium, magnesium, alkaline phosphatase, zinc and iron concentrations were measured in blood samples drawn before and after collecting blood into kEDTA containing sample tubes by an experienced phlebotomist using the Sarstedt Safety Monovette system. EDTA was undetectable in all samples. The concentrations of other analytes were similar in blood samples drawn before and after collection of the EDTA blood sample. Order of draw of blood samples using the Sarstedt Safety Monovette system has no effect on serum biochemistry results, when samples are taken by an experienced phlebotomist.

  1. Comparison of the Omron HEM-713C automated blood pressure monitor with a standard ausculatory method using a mercury manometer.

    Science.gov (United States)

    Mufunda, J; Sparks, B; Chifamba, J; Dakwa, C; Matenga, J A; Adams, J M; Sparks, H V

    1996-08-01

    To compare the Omron HEM-713C automated blood pressure machine with the standard ausculatory method using a mercury manometer. Blood pressures of randomly selected subjects were measured using both the Omron HEM-713C and the mercury manometer. Dombotombo surburb in Marondera, Zimbabwe. One hundred and sixteen subjects 25 years and above (47 males and 69 females) randomly selected in Marondera. Systolic blood pressure and diastolic blood pressure. The Omron HEM-713C passed with a grade B for both systolic and diastolic blood pressures when using the British Hypertension Society protocol. It also passed both systolic and diastolic criteria for Association of the Advancement of Medical Instrumentation. The Omron HEM-713C compares well with the standard mercury manometer, we therefore recommend its use in both research and clinical applications which require blood pressure measurements.

  2. Automated Clean Chemistry for Bulk Analysis of Environmental Swipe Samples - FY17 Year End Report

    Energy Technology Data Exchange (ETDEWEB)

    Ticknor, Brian W. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Metzger, Shalina C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); McBay, Eddy H. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Hexel, Cole R. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Tevepaugh, Kayron N. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Bostick, Debra A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)

    2017-11-30

    Sample preparation methods for mass spectrometry are being automated using commercial-off-the-shelf (COTS) equipment to shorten lengthy and costly manual chemical purification procedures. This development addresses a serious need in the International Atomic Energy Agency’s Network of Analytical Laboratories (IAEA NWAL) to increase efficiency in the Bulk Analysis of Environmental Samples for Safeguards program with a method that allows unattended, overnight operation. In collaboration with Elemental Scientific Inc., the prepFAST-MC2 was designed based on COTS equipment. It was modified for uranium/plutonium separations using renewable columns packed with Eichrom TEVA and UTEVA resins, with a chemical separation method based on the Oak Ridge National Laboratory (ORNL) NWAL chemical procedure. The newly designed prepFAST-SR has had several upgrades compared with the original prepFAST-MC2. Both systems are currently installed in the Ultra-Trace Forensics Science Center at ORNL.

  3. Automated high-volume aerosol sampling station for environmental radiation monitoring

    Energy Technology Data Exchange (ETDEWEB)

    Toivonen, H.; Honkamaa, T.; Ilander, T.; Leppaenen, A.; Nikkinen, M.; Poellaenen, R.; Ylaetalo, S

    1998-07-01

    An automated high-volume aerosol sampling station, known as CINDERELLA.STUK, for environmental radiation monitoring has been developed by the Radiation and Nuclear Safety Authority (STUK), Finland. The sample is collected on a glass fibre filter (attached into a cassette), the airflow through the filter is 800 m{sup 3}/h at maximum. During the sampling, the filter is continuously monitored with Na(I) scintillation detectors. After the sampling, the large filter is automatically cut into 15 pieces that form a small sample and after ageing, the pile of filter pieces is moved onto an HPGe detector. These actions are performed automatically by a robot. The system is operated at a duty cycle of 1 d sampling, 1 d decay and 1 d counting. Minimum detectable concentrations of radionuclides in air are typically 1Ae10 x 10{sup -6} Bq/m{sup 3}. The station is equipped with various sensors to reveal unauthorized admittance. These sensors can be monitored remotely in real time via Internet or telephone lines. The processes and operation of the station are monitored and partly controlled by computer. The present approach fulfils the requirements of CTBTO for aerosol monitoring. The concept suits well for nuclear material safeguards, too 10 refs.

  4. Comparison of performance of two Treponema pallidum automated chemiluminescent immunoassays in blood donors.

    Science.gov (United States)

    Sommese, Linda; Sabia, Chiara; Esposito, Antonella; Iannone, Carmela; Montesano, Maria Lourdes; Napoli, Claudio

    2016-01-01

    The recrudescence of syphilis is leading to the development of new serological tests. The goal of this study was to compare the performance of the more recent Elecsys Syphilis assay, the Electro Chemiluminescence Immunoassay (ECLIA), with the former Architect Syphilis TP assay, the Chemiluminescent Microparticle Immunoassay (CMIA), for the detection of antibodies against Treponema pallidum in blood donors. Serum samples of 5543 voluntary blood donors were screened in parallel with two tests. All repeatedly reactive (RR) samples by one or both assays were further analysed for confirmation by immmunoblot INNO-LIA and TPHA. Of 32 RR samples by CMIA, 21 were confirmed positive; of 21 RR samples by ECLIA, 20 were confirmed positive. The sensitivities of CMIA and ECLIA were 100% and 95.24% (95% CI = 85.71-100), respectively, not significant (p > 0.05). The specificity and predictive positive value (PPV) of CMIA were 99.86% (95% CI = 99.74-99.94) and 72.41%, respectively, while the specificity and PPV of ECLIA were both 100%, being statistically significant (p = 0.01 for both). The overall agreement was 99.80% and the Cohen's kappa coefficients was 0.79. In conclusion, the recent Elecsys Syphilis assay could represent another reliable assay for blood donor screening.

  5. Qualitative and quantitative cell recovery in umbilical cord blood processed by two automated devices in routine cord blood banking: a comparative study.

    Science.gov (United States)

    Solves, Pilar; Planelles, Dolores; Mirabet, Vicente; Blanquer, Amando; Carbonell-Uberos, Francisco

    2013-07-01

    Volume reduction is a widely used procedure in umbilical cord blood banking. It concentrates progenitor cells by reducing plasma and red blood cells, thereby optimising the use of storage space. Sepax and AXP are automated systems specifically developed for umbilical cord blood processing. These systems basically consist of a bag processing set into which cord blood is transferred and a device that automatically separates the different components during centrifugation. The aim of this study was to analyse and compare cell recovery of umbilical cord blood units processed with Sepax and AXP at Valencia Cord Blood Bank. Cell counts were performed before and after volume reduction with AXP and Sepax. When analysing all the data (n =1,000 for AXP and n= 670 for Sepax), the percentages of total nucleated cell recovery and red blood cell depletion were 76.76 ± 7.51% and 88.28 ± 5.62%, respectively, for AXP and 78.81 ± 7.25% and 88.32 ± 7.94%, respectively, for Sepax (P recovery and viability in umbilical cord blood units were similar with both devices. Mononuclear cell recovery was significantly higher when the Sepax system was used. Both the Sepax and AXP automated systems achieve acceptable total nucleated cell recovery and good CD34(+) cell recovery after volume reduction of umbilical cord blood units and maintain cell viability. It should be noted that total nucleated cell recovery is significantly better with the Sepax system. Both systems deplete red blood cells efficiently, especially AXP which works without hydroxyethyl starch.

  6. A filter paper-based microdevice for low-cost, rapid, and automated DNA extraction and amplification from diverse sample types.

    Science.gov (United States)

    Gan, Wupeng; Zhuang, Bin; Zhang, Pengfei; Han, Junping; Li, Cai-Xia; Liu, Peng

    2014-10-07

    A plastic microfluidic device that integrates a filter disc as a DNA capture phase was successfully developed for low-cost, rapid and automated DNA extraction and PCR amplification from various raw samples. The microdevice was constructed by sandwiching a piece of Fusion 5 filter, as well as a PDMS (polydimethylsiloxane) membrane, between two PMMA (poly(methyl methacrylate)) layers. An automated DNA extraction from 1 μL of human whole blood can be finished on the chip in 7 minutes by sequentially aspirating NaOH, HCl, and water through the filter. The filter disc containing extracted DNA was then taken out directly for PCR. On-chip DNA purification from 0.25-1 μL of human whole blood yielded 8.1-21.8 ng of DNA, higher than those obtained using QIAamp® DNA Micro kits. To realize DNA extraction from raw samples, an additional sample loading chamber containing a filter net with an 80 μm mesh size was designed in front of the extraction chamber to accommodate sample materials. Real-world samples, including whole blood, dried blood stains on Whatman® 903 paper, dried blood stains on FTA™ cards, buccal swabs, saliva, and cigarette butts, can all be processed in the system in 8 minutes. In addition, multiplex amplification of 15 STR (short tandem repeat) loci and Sanger-based DNA sequencing of the 520 bp GJB2 gene were accomplished from the filters that contained extracted DNA from blood. To further prove the feasibility of integrating this extraction method with downstream analyses, "in situ" PCR amplifications were successfully performed in the DNA extraction chamber following DNA purification from blood and blood stains without DNA elution. Using a modified protocol to bond the PDMS and PMMA, our plastic PDMS devices withstood the PCR process without any leakage. This study represents a significant step towards the practical application of on-chip DNA extraction methods, as well as the development of fully integrated genetic analytical systems.

  7. Fully Automated Laser Ablation Liquid Capture Sample Analysis using NanoElectrospray Ionization Mass Spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Lorenz, Matthias [ORNL; Ovchinnikova, Olga S [ORNL; Van Berkel, Gary J [ORNL

    2014-01-01

    RATIONALE: Laser ablation provides for the possibility of sampling a large variety of surfaces with high spatial resolution. This type of sampling when employed in conjunction with liquid capture followed by nanoelectrospray ionization provides the opportunity for sensitive and prolonged interrogation of samples by mass spectrometry as well as the ability to analyze surfaces not amenable to direct liquid extraction. METHODS: A fully automated, reflection geometry, laser ablation liquid capture spot sampling system was achieved by incorporating appropriate laser fiber optics and a focusing lens into a commercially available, liquid extraction surface analysis (LESA ) ready Advion TriVersa NanoMate system. RESULTS: Under optimized conditions about 10% of laser ablated material could be captured in a droplet positioned vertically over the ablation region using the NanoMate robot controlled pipette. The sampling spot size area with this laser ablation liquid capture surface analysis (LA/LCSA) mode of operation (typically about 120 m x 160 m) was approximately 50 times smaller than that achievable by direct liquid extraction using LESA (ca. 1 mm diameter liquid extraction spot). The set-up was successfully applied for the analysis of ink on glass and paper as well as the endogenous components in Alstroemeria Yellow King flower petals. In a second mode of operation with a comparable sampling spot size, termed laser ablation/LESA , the laser system was used to drill through, penetrate, or otherwise expose material beneath a solvent resistant surface. Once drilled, LESA was effective in sampling soluble material exposed at that location on the surface. CONCLUSIONS: Incorporating the capability for different laser ablation liquid capture spot sampling modes of operation into a LESA ready Advion TriVersa NanoMate enhanced the spot sampling spatial resolution of this device and broadened the surface types amenable to analysis to include absorbent and solvent resistant

  8. Artificial Neural Network for Total Laboratory Automation to Improve the Management of Sample Dilution.

    Science.gov (United States)

    Ialongo, Cristiano; Pieri, Massimo; Bernardini, Sergio

    2017-02-01

    Diluting a sample to obtain a measure within the analytical range is a common task in clinical laboratories. However, for urgent samples, it can cause delays in test reporting, which can put patients' safety at risk. The aim of this work is to show a simple artificial neural network that can be used to make it unnecessary to predilute a sample using the information available through the laboratory information system. Particularly, the Multilayer Perceptron neural network built on a data set of 16,106 cardiac troponin I test records produced a correct inference rate of 100% for samples not requiring predilution and 86.2% for those requiring predilution. With respect to the inference reliability, the most relevant inputs were the presence of a cardiac event or surgery and the result of the previous assay. Therefore, such an artificial neural network can be easily implemented into a total automation framework to sensibly reduce the turnaround time of critical orders delayed by the operation required to retrieve, dilute, and retest the sample.

  9. Automated MALDI Matrix Coating System for Multiple Tissue Samples for Imaging Mass Spectrometry

    Science.gov (United States)

    Mounfield, William P.; Garrett, Timothy J.

    2012-03-01

    Uniform matrix deposition on tissue samples for matrix-assisted laser desorption/ionization (MALDI) is key for reproducible analyte ion signals. Current methods often result in nonhomogenous matrix deposition, and take time and effort to produce acceptable ion signals. Here we describe a fully-automated method for matrix deposition using an enclosed spray chamber and spray nozzle for matrix solution delivery. A commercial air-atomizing spray nozzle was modified and combined with solenoid controlled valves and a Programmable Logic Controller (PLC) to control and deliver the matrix solution. A spray chamber was employed to contain the nozzle, sample, and atomized matrix solution stream, and to prevent any interference from outside conditions as well as allow complete control of the sample environment. A gravity cup was filled with MALDI matrix solutions, including DHB in chloroform/methanol (50:50) at concentrations up to 60 mg/mL. Various samples (including rat brain tissue sections) were prepared using two deposition methods (spray chamber, inkjet). A linear ion trap equipped with an intermediate-pressure MALDI source was used for analyses. Optical microscopic examination showed a uniform coating of matrix crystals across the sample. Overall, the mass spectral images gathered from tissues coated using the spray chamber system were of better quality and more reproducible than from tissue specimens prepared by the inkjet deposition method.

  10. Design of an automated rapid vapor concentrator and its application in nitroaromatic vapor sampling

    Science.gov (United States)

    Gehrke, Mark; Kapila, Shubhender; Hambacker, Kurt L.; Flanigan, Virgil I.

    2000-08-01

    An automated, rapid-cycling vapor concentrator and sample introduction device was designed and evaluated. The device consists of an inert deactivated fused silica capillary sampling loop. The temperature of the loop was manipulated through contact with a cold plate or a hot plate, maintained at pre-selected temperatures with a thermoelectric cooler and heating cartridge, respectively. The position of the loop was controlled with a stepper motor under microprocessor control. The low mass of the loop permit its rapid cooling and heating. This permits efficient trapping of adsorptive vapors such as the nitroaromatics from the air stream and also allows rapid and quantitative transfer of the trapped analytes to the detection system. The use of at thermoelectric cooler permits variable trapping temperatures and increased sampling selectivity without the use of cumbersome cryogenic fluids. Chemically inert sampling train surfaces prevent analyte loss due to irreversible adsorption and cross contamination between samples. The device was evaluated for rapid analysis of nitroaromatic and chlorinated aromatic vapors from air stream at trace concentrations with a selective electron capture detection system. Trapping efficiencies of > 95 percent can be readily obtained with the device for nitroaromatics at ppb and sub ppb concentrations.

  11. Screening for neonatal hypothyroidism by thyroxine and thyrotrophin radioimmunoassays using dried blood samples on filter paper

    International Nuclear Information System (INIS)

    Beckers, C.; Cornette, C.; Francois, B.; Bouckaert, A.

    1979-01-01

    A routine and automated methodology for thyroxine (T4) and thyrotrophin (TSH) radioimmunoassay (RIA) using dried blood samples on filter paper is described. T4-RIA was performed on one single dot (5 mm diameter equivalent to 4 μl of serum) while two dots were necessary for TSH-RIA. Reference filter papers were introduced in each assay for quality control. In a preliminary study on 4,155 neonates, samples generally obtained between the 5th-7th day gave a mean 'dot-T4' of 97.95 +- 36.04 nmol/l and a mean 'dot-TSH' of 10.19 mU/l +- 8.25, corresponding to 2.47 mU/l of serum. Within an 18-month period (November 1976 - April 1978), a total of 16,522 neonates have been screened allowing detection of three cases of congenital hypothyroidism (incidence 1:5507), two cases of congenitally low TBG and thirty-three cases of transient hypothyroidism. (author)

  12. 21 CFR 864.5200 - Automated cell counter.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Automated cell counter. 864.5200 Section 864.5200 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED...-automated device used to count red blood cells, white blood cells, or blood platelets using a sample of the...

  13. Bacterial Isolates from Blood Samples of Patients in University of ...

    African Journals Online (AJOL)

    The thioglychollate broth was sub cultured onto blood agar plate for anaerobic incubation, while the brain heart infusion broth was sub cultured onto chocolate, blood agar and McConkey agar for aerobic incubation. The anaerobic incubation was done by putting the culture plates into an anaerobic jar and filling it with ...

  14. Development of an animal-borne blood sample collection device and its deployment for the determination of cardiovascular and stress hormones in phocid seals.

    Science.gov (United States)

    Takei, Yoshio; Suzuki, Ippei; Wong, Marty K S; Milne, Ryan; Moss, Simon; Sato, Katsufumi; Hall, Ailsa

    2016-10-01

    An animal-borne blood sampler with data-logging functions was developed for phocid seals, which collected two blood samples for the comparison of endocrinological/biochemical parameters under two different conditions. The sampler can be triggered by preset hydrostatic pressure, acceleration (descending or ascending), temperature, and time, and also manually by light. The sampling was reliable with 39/50 (78%) successful attempts to collect blood samples. Contamination of fluids in the tubing to the next blood sample was seals ( Phoca vitulina ), the automated blood-sampling method was less stressful than direct blood withdrawal, as evidenced by lower levels of stress hormones ( P seal blood. Using the sampler, plasma levels of cardiovascular hormones, atrial natriuretic peptide (ANP), AVP, and ANG II were compared in grey seals ( Halichoerus grypus ), between samples collected when the animals were on land and in the water. HPLC analyses determined that [Met 12 ] ANP (1-28) and various forms of angiotensins (ANG II, III, and IV) were circulating in seal blood. Although water immersion profoundly changes the plasma levels of cardiovascular hormones in terrestrial mammals, there were only tendencies toward an increase in ANP ( P = 0.069) and a decrease in AVP ( P = 0.074) in the seals. These results suggest that cardiovascular regulation in phocid seals may have undergone adaptation during evolution of the carnivore to a semiaquatic lifestyle. Copyright © 2016 the American Physiological Society.

  15. Performance of an app measuring spot quality in dried blood spot sampling

    NARCIS (Netherlands)

    Veenhof, Herman

    2016-01-01

    Introduction: The Dried Blood Spot sampling (DBS) method gives patients and health care workers the opportunity for remote sampling using a drop of blood from a fingerprick on a sampling card which can be send to the laboratory by mail. Laboratory analysts frequently reject DBS samples because of

  16. Diagnosis of Carrion's disease by direct blood PCR in thin blood smear negative samples.

    Directory of Open Access Journals (Sweden)

    Juana del Valle Mendoza

    Full Text Available Bartonella bacilliformis is the etiologic agent of Carrion's disease. This disease has two well established phases, the most relevant being the so called Oroya Fever, in which B. bacilliformis infect the erythrocytes resulting in severe anemia and transient immunosuppression, with a high lethality in the absence of adequate antibiotic treatment. The presence of B. bacilliformis was studied in 113 blood samples suspected of Carrion's disease based on clinical criteria, despite the absence of a positive thin blood smear, by two different PCR techniques (using Bartonella-specific and universal 16S rRNA gene primers, and by bacterial culture. The specific 16S rRNA gene primers revealed the presence of 21 B. bacilliformis and 1 Bartonella elizabethae, while universal primers showed both the presence of 3 coinfections in which a concomitant pathogen was detected plus Bartonella, in addition to the presence of infections by other microorganisms such as Agrobacterium or Bacillus firmus. These data support the need to implement molecular tools to diagnose Carrion's disease.

  17. Rapid and automated determination of plutonium and neptunium in environmental samples

    Energy Technology Data Exchange (ETDEWEB)

    Qiao, J.

    2011-03-15

    This thesis presents improved analytical methods for rapid and automated determination of plutonium and neptunium in environmental samples using sequential injection (SI) based chromatography and inductively coupled plasma mass spectrometry (ICP-MS). The progress of methodology development in this work consists of 5 subjects stated as follows: 1) Development and optimization of an SI-anion exchange chromatographic method for rapid determination of plutonium in environmental samples in combination of inductively coupled plasma mass spectrometry detection (Paper II); (2) Methodology development and optimization for rapid determination of plutonium in environmental samples using SI-extraction chromatography prior to inductively coupled plasma mass spectrometry (Paper III); (3) Development of an SI-chromatographic method for simultaneous determination of plutonium and neptunium in environmental samples (Paper IV); (4) Investigation of the suitability and applicability of 242Pu as a tracer for rapid neptunium determination using anion exchange chromatography in an SI-network coupled with inductively coupled plasma mass spectrometry (Paper V); (5) Exploration of macro-porous anion exchange chromatography for rapid and simultaneous determination of plutonium and neptunium within an SI system (Paper VI). The results demonstrate that the developed methods in this study are reliable and efficient for accurate assays of trace levels of plutonium and neptunium as demanded in different situations including environmental risk monitoring and assessment, emergency preparedness and surveillance of contaminated areas. (Author)

  18. Automated Three-Dimensional Microbial Sensing and Recognition Using Digital Holography and Statistical Sampling

    Directory of Open Access Journals (Sweden)

    Inkyu Moon

    2010-09-01

    Full Text Available We overview an approach to providing automated three-dimensional (3D sensing and recognition of biological micro/nanoorganisms integrating Gabor digital holographic microscopy and statistical sampling methods. For 3D data acquisition of biological specimens, a coherent beam propagates through the specimen and its transversely and longitudinally magnified diffraction pattern observed by the microscope objective is optically recorded with an image sensor array interfaced with a computer. 3D visualization of the biological specimen from the magnified diffraction pattern is accomplished by using the computational Fresnel propagation algorithm. For 3D recognition of the biological specimen, a watershed image segmentation algorithm is applied to automatically remove the unnecessary background parts in the reconstructed holographic image. Statistical estimation and inference algorithms are developed to the automatically segmented holographic image. Overviews of preliminary experimental results illustrate how the holographic image reconstructed from the Gabor digital hologram of biological specimen contains important information for microbial recognition.

  19. Are Flow Injection-based Approaches Suitable for Automated Handling of Solid Samples?

    DEFF Research Database (Denmark)

    Miró, Manuel; Hansen, Elo Harald; Cerdà, Victor

    Flow-based approaches were originally conceived for liquid-phase analysis, implying that constituents in solid samples generally had to be transferred into the liquid state, via appropriate batch pretreatment procedures, prior to analysis. Yet, in recent years, much effort has been focused...... electrolytic or aqueous leaching, on-line dialysis/microdialysis, in-line filtration, and pervaporation-based procedures have been successfully implemented in continuous flow/flow injection systems. In this communication, the new generation of flow analysis, including sequential injection, multicommutated flow...... with the potential hyphenation with modern analytical instrumentation for automated monitoring of the content of targeted species in the on-line generated extracts [3,4]. [1] Z.-L. Zhi, A. Ríos, M. Valcárcel, Crit. Rev. Anal. Chem., 26 (1996) 239. [2] M. Miró, E.H. Hansen, R. Chomchoei, W. Frenzel, TRAC-Trends Anal...

  20. Effect of Sample Storage Temperature and Time Delay on Blood Gases, Bicarbonate and pH in Human Arterial Blood Samples.

    Science.gov (United States)

    Mohammadhoseini, Elham; Safavi, Enayat; Seifi, Sepideh; Seifirad, Soroush; Firoozbakhsh, Shahram; Peiman, Soheil

    2015-03-01

    Results of arterial blood gas analysis can be biased by pre-analytical factors, such as time interval before analysis, temperature during storage and syringe type. To investigate the effects of samples storage temperature and time delay on blood gases, bicarbonate and PH results in human arterial blood samples. 2.5 mL arterial blood samples were drawn from 45 patients via an indwelling Intraarterial catheter. Each sample was divided into five equal samples and stored in multipurpose tuberculin plastic syringes. Blood gas analysis was performed on one of five samples as soon as possible. Four other samples were divided into two groups stored at 22°C and 0°C. Blood gas analyses were repeated at 30 and 60 minutes after sampling. PaO2 of the samples stored at 0°C was increased significantly after 60 minutes (P = 0.007). The PaCO2 of the samples kept for 30 and 60 minutes at 22°C was significantly higher than primary result (P = 0.04, P samples stored at 22°C, pH decreased significantly after 30 and 60 minutes (P = 0.017, P = 0.001). There were no significant differences in other results of samples stored at 0°C or 22°C after 30 or 60 minutes. In samples stored in plastic syringes, overestimation of PaO2 levels should be noted if samples cooled before analysis. In samples stored in plastic syringes, it is not necessary to store samples in iced water when analysis delayed up to one hour.

  1. The quantitative regional cerebral blood flow measurement with autoradiography method using 123I-IMP SPECT. Evaluation of arterialized venous blood sampling as a substitute for arterial blood sampling

    International Nuclear Information System (INIS)

    Ohnishi, Takashi; Yano, Takao; Nakano, Shinichi; Jinnouchi, Seishi; Nagamachi, Shigeki; Flores, L. II; Nakahara, Hiroshi; Watanabe, Katsushi.

    1996-01-01

    The purpose of this study is validation of calibrating a standard input function in autoradiography (ARG) method by one point venous blood sampling as a substitute for that by one point arterial blood sampling. Ten and 20 minutes after intravenous constant infusion of 123 I-IMP, arterialized venous blood sampling from a dorsal vein were performed on 15 patients having ischemic cerebrovascular disease. And arterial blood sampling from radial artery was performed 10 min after 123 I-IMP infusion. The mean difference rates of integrated input function between calibrated standard input function by arterial blood sampling at 10 min and that by venous blood sampling were 4.1±3% and 9.3±5.4% at 10 and 20 min after 123 I-IMP infusion, respectively. The ratio of venous blood radioactivity to arterial blood radioactivity at 10 min after 123 I-IMP infusion was 0.96±0.02. There was an excellent correlation between ARG method CBF values obtained by arterial blood sampling at 10 min and those obtained by arterialized venous blood sampling at 10 min. In conclusion, a substitution by arterialized venous blood sampling from dorsal hand vein for artery can be possible. The optimized time for arterialized venous blood sampling was 10 min after 123 I-IMP infusion. (author)

  2. An Automated Algorithm to Screen Massive Training Samples for a Global Impervious Surface Classification

    Science.gov (United States)

    Tan, Bin; Brown de Colstoun, Eric; Wolfe, Robert E.; Tilton, James C.; Huang, Chengquan; Smith, Sarah E.

    2012-01-01

    An algorithm is developed to automatically screen the outliers from massive training samples for Global Land Survey - Imperviousness Mapping Project (GLS-IMP). GLS-IMP is to produce a global 30 m spatial resolution impervious cover data set for years 2000 and 2010 based on the Landsat Global Land Survey (GLS) data set. This unprecedented high resolution impervious cover data set is not only significant to the urbanization studies but also desired by the global carbon, hydrology, and energy balance researches. A supervised classification method, regression tree, is applied in this project. A set of accurate training samples is the key to the supervised classifications. Here we developed the global scale training samples from 1 m or so resolution fine resolution satellite data (Quickbird and Worldview2), and then aggregate the fine resolution impervious cover map to 30 m resolution. In order to improve the classification accuracy, the training samples should be screened before used to train the regression tree. It is impossible to manually screen 30 m resolution training samples collected globally. For example, in Europe only, there are 174 training sites. The size of the sites ranges from 4.5 km by 4.5 km to 8.1 km by 3.6 km. The amount training samples are over six millions. Therefore, we develop this automated statistic based algorithm to screen the training samples in two levels: site and scene level. At the site level, all the training samples are divided to 10 groups according to the percentage of the impervious surface within a sample pixel. The samples following in each 10% forms one group. For each group, both univariate and multivariate outliers are detected and removed. Then the screen process escalates to the scene level. A similar screen process but with a looser threshold is applied on the scene level considering the possible variance due to the site difference. We do not perform the screen process across the scenes because the scenes might vary due to

  3. Blood samples in the neutron beam; Blutproben im Neutronenstrahl

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    2012-07-01

    Whether crocodile, platypus, man, or chicken: Always the hemoglobin in the red blood cells has the same task. It carries oxygen from the lungs throughout the body. In investigative manner and international team around Dr. Andreas Stadler from the Julic Research Center has deciphered in detail, how and why the hemoglobines of these creatures nevertheless differ. Their findings are among others interesting for the research on artificial blood.

  4. Comparison of QIAsymphony automated and QIAamp manual DNA extraction systems for measuring Epstein-Barr virus DNA load in whole blood using real-time PCR.

    Science.gov (United States)

    Laus, Stella; Kingsley, Lawrence A; Green, Michael; Wadowsky, Robert M

    2011-11-01

    Automated and manual extraction systems have been used with real-time PCR for quantification of Epstein-Barr virus [human herpesvirus 4 (HHV-4)] DNA in whole blood, but few studies have evaluated relative performances. In the present study, the automated QIAsymphony and manual QIAamp extraction systems (Qiagen, Valencia, CA) were assessed using paired aliquots derived from clinical whole-blood specimens and an in-house, real-time PCR assay. The detection limits using the QIAsymphony and QIAamp systems were similar (270 and 560 copies/mL, respectively). For samples estimated as having ≥10,000 copies/mL, the intrarun and interrun variations were significantly lower using QIAsymphony (10.0% and 6.8%, respectively), compared with QIAamp (18.6% and 15.2%, respectively); for samples having ≤1000 copies/mL, the two variations ranged from 27.9% to 43.9% and were not significantly different between the two systems. Among 68 paired clinical samples, 48 pairs yielded viral loads ≥1000 copies/mL under both extraction systems. Although the logarithmic linear correlation from these positive samples was high (r(2) = 0.957), the values obtained using QIAsymphony were on average 0.2 log copies/mL higher than those obtained using QIAamp. Thus, the QIAsymphony and QIAamp systems provide similar EBV DNA load values in whole blood. Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.

  5. The impact of different blood sampling methods on laboratory rats under different types of anaesthesia

    DEFF Research Database (Denmark)

    Toft, Martin Fitzner; Petersen, Mikke Haxø; Dragsted, Nils

    2006-01-01

    Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO?), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were...... showed lower increases in blood pressure after, and fewer fluctuations in body temperature during sampling, and the post-anaesthetic effects of isoflurane, if any, seemed to disappear immediately after sampling. It is, therefore, concluded that blood sampling in rats by jugular puncture seems...

  6. A simple and automated sample preparation system for subsequent halogens determination: Combustion followed by pyrohydrolysis.

    Science.gov (United States)

    Pereira, L S F; Pedrotti, M F; Vecchia, P Dalla; Pereira, J S F; Flores, E M M

    2018-06-20

    A simple and automated system based on combustion followed by a pyrohydrolysis reaction was proposed for further halogens determination. This system was applied for digestion of soils containing high (90%) and also low (10%) organic matter content for further halogens determination. The following parameters were evaluated: sample mass, use of microcrystalline cellulose and heating time. For analytes absorption, a diluted alkaline solution (6 mL of 25 mmol L -1  NH 4 OH) was used in all experiments. Up to 400 mg of soil with high organic matter content and 100 mg of soil with low organic matter content (mixed with 400 mg of cellulose) could be completely digested using the proposed system. Quantitative results for all halogens were obtained using less than 12 min of sample preparation step (about 1.8 min for sample combustion and 10 min for pyrohydrolysis). The accuracy was evaluated using a certified reference material of coal and spiked samples. No statistical difference was observed between the certified values and results obtained by the proposed method. Additionally, the recoveries obtained using spiked samples were in the range of 98-103% with relative standard deviation values lower than 5%. The limits of quantification obtained for F, Cl, Br and I for soil with high (400 mg of soil) and low (100 mg of soil) organic matter were in the range of 0.01-2 μg g -1 and 0.07-59 μg g -1 , respectively. The proposed system was considered as a simple and suitable alternative for soils digestion for further halogens determination by ion chromatography and inductively coupled plasma mass spectrometry techniques. Copyright © 2018 Elsevier B.V. All rights reserved.

  7. An artificial pancreas for automated blood glucose control in patients with Type 1 diabetes

    DEFF Research Database (Denmark)

    Schmidt, Signe; Boiroux, Dimitri; Ranjan, Ajenthen

    2015-01-01

    Automated glucose control in patients with Type 1 diabetes is much-coveted by patients, relatives and healthcare professionals. It is the expectation that a system for automated control, also know as an artificial pancreas, will improve glucose control, reduce the risk of diabetes complications...

  8. Air bubbles and hemolysis of blood samples during transport by pneumatic tube systems.

    Science.gov (United States)

    Mullins, Garrett R; Bruns, David E

    2017-10-01

    Transport of blood samples through pneumatic tube systems (PTSs) generates air bubbles in transported blood samples and, with increasing duration of transport, the appearance of hemolysis. We investigated the role of air-bubble formation in PTS-induced hemolysis. Air was introduced into blood samples for 0, 1, 3 or 5min to form air bubbles. Hemolysis in the blood was assessed by (H)-index, lactate dehydrogenase (LD) and potassium in plasma. In an effort to prevent PTS-induced hemolysis, blood sample tubes were completely filled, to prevent air bubble formation, and compared with partially filled samples after PTS transport. We also compared hemolysis in anticoagulated vs clotted blood subjected to PTS transport. As with transport through PTSs, the duration of air bubble formation in blood by a gentle stream of air predicted the extent of hemolysis as measured by H-index (pblood sample prevented bubble formation and fully protected the blood from PTS-induced hemolysis (pblood developed less foaming during PTS transport and was partially protected from hemolysis vs anticoagulated blood as indicated by lower LD (psample transport. Prevention of air bubble formation in blood samples during PTS transport protects samples from hemolysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. ABO and D typing and alloantibody screening in marrow samples: relevance to intraosseous blood transfusion.

    Science.gov (United States)

    Bäckman, Sari; Ångerman-Haasmaa, Susanne; Jousi, Milla; Siitonen, Sanna; Salmela, Katja

    2018-03-01

    Blood transfusion through the intraosseous route is gaining popularity in emergency medicine. Pretransfusion peripheral blood (PB) samples are usually not available in these patients, leading to discrepancies in blood group typing and a possible delay in transferring to group-specific blood products. The aim of this study was to assess the feasibility of ABO and D typing and red blood cell alloantibody screening in marrow (BM) samples. Direct and reverse ABO typing, D typing, and a two-cell alloantibody screen were performed in EDTA-anticoagulated BM samples with standard manual column agglutination techniques. EDTA-anticoagulated PB samples were used as controls. The mean age of the study subjects (n = 71) was 47 years (range, 1-82 years). All ABO groups and both D+ and D- types were represented. In all subjects, concordant results were observed for all analyses in BM and PB samples. In 15 (21%) of the samples, a discrepancy of one reaction strength step (1+) was observed in at least one of the analyses (Cohen's weighted κ = 0.993); this did not affect interpretation of the results. Blood group typing and alloantibody screening are feasible in BM samples, providing proof-of-concept that intraosseous samples for blood group serologic analyses can be collected from emergency patients before intraosseous blood transfusion. This will enable a timely transfer to group-specific blood products and enable conservation of the valuable universal-donor blood products. © 2018 AABB.

  10. A technique for extracting blood samples from mice in fire toxicity tests

    Science.gov (United States)

    Bucci, T. J.; Hilado, C. J.; Lopez, M. T.

    1976-01-01

    The extraction of adequate blood samples from moribund and dead mice has been a problem because of the small quantity of blood in each animal and the short time available between the animals' death and coagulation of the blood. These difficulties are particularly critical in fire toxicity tests because removal of the test animals while observing proper safety precautions for personnel is time-consuming. Techniques for extracting blood samples from mice were evaluated, and a technique was developed to obtain up to 0.8 ml of blood from a single mouse after death. The technique involves rapid exposure and cutting of the posterior vena cava and accumulation of blood in the peritoneal space. Blood samples of 0.5 ml or more from individual mice have been consistently obtained as much as 16 minutes after apparent death. Results of carboxyhemoglobin analyses of blood appeared reproducible and consistent with carbon monoxide concentrations in the exposure chamber.

  11. Invention and validation of an automated camera system that uses optical character recognition to identify patient name mislabeled samples.

    Science.gov (United States)

    Hawker, Charles D; McCarthy, William; Cleveland, David; Messinger, Bonnie L

    2014-03-01

    Mislabeled samples are a serious problem in most clinical laboratories. Published error rates range from 0.39/1000 to as high as 1.12%. Standardization of bar codes and label formats has not yet achieved the needed improvement. The mislabel rate in our laboratory, although low compared with published rates, prompted us to seek a solution to achieve zero errors. To reduce or eliminate our mislabeled samples, we invented an automated device using 4 cameras to photograph the outside of a sample tube. The system uses optical character recognition (OCR) to look for discrepancies between the patient name in our laboratory information system (LIS) vs the patient name on the customer label. All discrepancies detected by the system's software then require human inspection. The system was installed on our automated track and validated with production samples. We obtained 1 009 830 images during the validation period, and every image was reviewed. OCR passed approximately 75% of the samples, and no mislabeled samples were passed. The 25% failed by the system included 121 samples actually mislabeled by patient name and 148 samples with spelling discrepancies between the patient name on the customer label and the patient name in our LIS. Only 71 of the 121 mislabeled samples detected by OCR were found through our normal quality assurance process. We have invented an automated camera system that uses OCR technology to identify potential mislabeled samples. We have validated this system using samples transported on our automated track. Full implementation of this technology offers the possibility of zero mislabeled samples in the preanalytic stage.

  12. Cerebral blood flow SPET in transient global amnesia with automated ROI analysis by 3DSRT

    Energy Technology Data Exchange (ETDEWEB)

    Takeuchi, Ryo [Division of Nuclear Medicine, Nishi-Kobe Medical Center, Kohjidai 5-7-1, 651-2273, Nishi-ku, Kobe-City, Hyogo (Japan); Matsuda, Hiroshi [Department of Radiology, National Center Hospital for Mental, Nervous and Muscular Disorders, National Center of Neurology and Psychiatry, Tokyo (Japan); Yoshioka, Katsunori [Daiichi Radioisotope Laboratories, Ltd., Tokyo (Japan); Yonekura, Yoshiharu [Biomedical Imaging Research Center, University of Fukui, Fukui (Japan)

    2004-04-01

    The aim of this study was to determine the areas involved in episodes of transient global amnesia (TGA) by calculation of cerebral blood flow (CBF) using 3DSRT, fully automated ROI analysis software which we recently developed. Technetium-99m l,l-ethyl cysteinate dimer single-photon emission tomography ({sup 99m}Tc-ECD SPET) was performed during and after TGA attacks on eight patients (four men and four women; mean study interval, 34 days). The SPET images were anatomically standardized using SPM99 followed by quantification of 318 constant ROIs, grouped into 12 segments (callosomarginal, precentral, central, parietal, angular, temporal, posterior cerebral, pericallosal, lenticular nucleus, thalamus, hippocampus and cerebellum), in each hemisphere to calculate segmental CBF (sCBF) as the area-weighted mean value for each of the respective 12 segments based on the regional CBF in each ROI. Correlation of the intra- and post-episodic sCBF of each of the 12 segments of the eight patients was estimated by scatter-plot graphical analysis and Pearson's correlation test with Fisher's Z-transformation. For the control, {sup 99m}Tc-ECD SPET was performed on eight subjects (three men and five women) and repeated within 1 month; the correlation between the first and second sCBF values of each of the 12 segments was evaluated in the same way as for patients with TGA. Excellent reproducibility between the two sCBF values was found in all 12 segments of the control subjects. However, a significant correlation between intra- and post-episodic sCBF was not shown in the thalamus or angular segments of TGA patients. The present study was preliminary, but at least suggested that thalamus and angular regions are closely involved in the symptoms of TGA. (orig.)

  13. Cerebral blood flow SPET in transient global amnesia with automated ROI analysis by 3DSRT

    International Nuclear Information System (INIS)

    Takeuchi, Ryo; Matsuda, Hiroshi; Yoshioka, Katsunori; Yonekura, Yoshiharu

    2004-01-01

    The aim of this study was to determine the areas involved in episodes of transient global amnesia (TGA) by calculation of cerebral blood flow (CBF) using 3DSRT, fully automated ROI analysis software which we recently developed. Technetium-99m l,l-ethyl cysteinate dimer single-photon emission tomography ( 99m Tc-ECD SPET) was performed during and after TGA attacks on eight patients (four men and four women; mean study interval, 34 days). The SPET images were anatomically standardized using SPM99 followed by quantification of 318 constant ROIs, grouped into 12 segments (callosomarginal, precentral, central, parietal, angular, temporal, posterior cerebral, pericallosal, lenticular nucleus, thalamus, hippocampus and cerebellum), in each hemisphere to calculate segmental CBF (sCBF) as the area-weighted mean value for each of the respective 12 segments based on the regional CBF in each ROI. Correlation of the intra- and post-episodic sCBF of each of the 12 segments of the eight patients was estimated by scatter-plot graphical analysis and Pearson's correlation test with Fisher's Z-transformation. For the control, 99m Tc-ECD SPET was performed on eight subjects (three men and five women) and repeated within 1 month; the correlation between the first and second sCBF values of each of the 12 segments was evaluated in the same way as for patients with TGA. Excellent reproducibility between the two sCBF values was found in all 12 segments of the control subjects. However, a significant correlation between intra- and post-episodic sCBF was not shown in the thalamus or angular segments of TGA patients. The present study was preliminary, but at least suggested that thalamus and angular regions are closely involved in the symptoms of TGA. (orig.)

  14. Microassay for interferon, using [3H]uridine, microculture plates, and a multiple automated sample harvester.

    Science.gov (United States)

    Richmond, J Y; Polatnick, J; Knudsen, R C

    1980-01-01

    A microassay for interferon is described which uses target cells grown in microculture wells, [3H]uridine to measure vesicular stomatitis virus replication in target cells, and a multiple automated sample harvester to collect the radioactively labeled viral ribonucleic acid onto glass fiber filter disks. The disks were placed in minivials, and radioactivity was counted in a liquid scintillation spectrophotometer. Interferon activity was calculated as the reciprocal of the highest titer which inhibited the incorporation of [3H]uridine into viral ribonucleic acid by 50%. Interferon titers determined by the microassay were similar to the plaque reduction assay when 100 plaque-forming units of challenge vesicular stomatitis virus was used. However, it was found that the interferon titers decreased approximately 2-fold for each 10-fold increase in the concentration of challenge vesicular stomatitis virus when tested in the range of 10(2) to 10(5) plaque-forming units. Interferon titers determined by the microassay show a high degree of repeatability, and the assay can be used to measure small and large numbers of interferon samples. PMID:6155105

  15. Automation and integration of multiplexed on-line sample preparation with capillary electrophoresis for DNA sequencing

    Energy Technology Data Exchange (ETDEWEB)

    Tan, H.

    1999-03-31

    The purpose of this research is to develop a multiplexed sample processing system in conjunction with multiplexed capillary electrophoresis for high-throughput DNA sequencing. The concept from DNA template to called bases was first demonstrated with a manually operated single capillary system. Later, an automated microfluidic system with 8 channels based on the same principle was successfully constructed. The instrument automatically processes 8 templates through reaction, purification, denaturation, pre-concentration, injection, separation and detection in a parallel fashion. A multiplexed freeze/thaw switching principle and a distribution network were implemented to manage flow direction and sample transportation. Dye-labeled terminator cycle-sequencing reactions are performed in an 8-capillary array in a hot air thermal cycler. Subsequently, the sequencing ladders are directly loaded into a corresponding size-exclusion chromatographic column operated at {approximately} 60 C for purification. On-line denaturation and stacking injection for capillary electrophoresis is simultaneously accomplished at a cross assembly set at {approximately} 70 C. Not only the separation capillary array but also the reaction capillary array and purification columns can be regenerated after every run. DNA sequencing data from this system allow base calling up to 460 bases with accuracy of 98%.

  16. Effects of Blood Sample Collection Pre- and Post- Slaughter, Edta ...

    African Journals Online (AJOL)

    The samples were immediately subjected to Wet mount (WM), Haemotocrit centrifugation test (HCT) and Thin smear (TS) tests. The results revealed that, of the 100 samples examined, 19 (19%) were positive for the presence of Microfilaria spp while 6(6%) yielded Trypanosome spp. Of the 19 samples detected having ...

  17. Forensic Identification of Human Blood: comparison of two one-step presumptive tests for blood screening of crime scene samples.

    Directory of Open Access Journals (Sweden)

    Ana Flávia Belchior Andrade

    2014-08-01

    Full Text Available Blood is the most common body fluid found at crime scenes. One-step presumptive tests have been designed as a rapid immunological test for the qualitative detection of human hemoglobin in stool samples (faecal occult blood their usefulness for forensic purposes has been demonstrated before. In this study we compare Hexagon OBTI kit and FOB One-step Bioeasy kit sensitivity in the analysis of diluted blood samples. With Hexagon OBTI, positive test results are achieved in whole blood dilutions up to 1:1.000. Sensitivity decreased with aged samples, if samples were not stored under low temperatures regardless of which presumptive test is used. Whole blood tests must take into consideration that “hook” effect may interfere. Comparing both tests, OBTI Hexagon Kit is more sensible to detect diluted blood, showing a wider detection window in all conditions. This is interesting when analyzing forensic samples as forensic analysts usually do not know about the history of the analyzed sample before its collection.

  18. Validation of curve-fitting method for blood retention of 99mTc-GSA. Comparison with blood sampling method

    International Nuclear Information System (INIS)

    Ha-Kawa, Sang Kil; Suga, Yutaka; Kouda, Katsuyasu; Ikeda, Koshi; Tanaka, Yoshimasa

    1997-01-01

    We investigated a curve-fitting method for the rate of blood retention of 99m Tc-galactosyl serum albumin (GSA) as a substitute for the blood sampling method. Seven healthy volunteers and 27 patients with liver disease underwent 99m Tc-GSA scanning. After normalization of the y-intercept as 100 percent, a biexponential regression curve for the precordial time-activity curve provided the percent injected dose (%ID) of 99m Tc-GSA in the blood without blood sampling. The discrepancy between %ID obtained by the curve-fitting method and that by the multiple blood samples was minimal in normal volunteers 3.1±2.1% (mean±standard deviation, n=77 sampling). Slightly greater discrepancy was observed in patients with liver disease (7.5±6.1%, n=135 sampling). The %ID at 15 min after injection obtained from the fitted curve was significantly greater in patients with liver cirrhosis than in the controls (53.2±11.6%, n=13; vs. 31.9±2.8%, n=7, p 99m Tc-GSA and the plasma retention rate for indocyanine green (r=-0.869, p 99m Tc-GSA and could be a substitute for the blood sampling method. (author)

  19. Comparison of QIAsymphony Automated and QIAamp Manual DNA Extraction Systems for Measuring Epstein-Barr Virus DNA Load in Whole Blood Using Real-Time PCR

    OpenAIRE

    Laus, Stella; Kingsley, Lawrence A.; Green, Michael; Wadowsky, Robert M.

    2011-01-01

    Automated and manual extraction systems have been used with real-time PCR for quantification of Epstein-Barr virus [human herpesvirus 4 (HHV-4)] DNA in whole blood, but few studies have evaluated relative performances. In the present study, the automated QIAsymphony and manual QIAamp extraction systems (Qiagen, Valencia, CA) were assessed using paired aliquots derived from clinical whole-blood specimens and an in-house, real-time PCR assay. The detection limits using the QIAsymphony and QIAam...

  20. Continuous quality control of the blood sampling procedure using a structured observation scheme

    DEFF Research Database (Denmark)

    Seemann, T. L.; Nybo, M.

    2015-01-01

    Background: An important preanalytical factor is the blood sampling procedure and its adherence to the guidelines, i.e. CLSI and ISO 15189, in order to ensure a consistent quality of the blood collection. Therefore, it is critically important to introduce quality control on this part of the process....... As suggested by the EFLM working group on the preanalytical phase we introduced continuous quality control of the blood sampling procedure using a structured observation scheme to monitor the quality of blood sampling performed on an everyday basis. Materials and methods: Based on our own routines the EFLM....... Conclusion: It is possible to establish a continuous quality control on blood sampling. It has been well accepted by the staff and we have already been able to identify critical areas in the sampling process. We find that continuous auditing increase focus on the quality of blood collection which ensures...

  1. The impact of different blood sampling methods on laboratory rats under different types of anaesthesia

    DEFF Research Database (Denmark)

    Toft, Martin Fitzner; Petersen, Mikke Haxø; Dragsted, Nils

    2006-01-01

    Rats with implanted telemetry transponders were blood sampled by jugular puncture, periorbital puncture or tail vein puncture, or sampled by jugular puncture in carbon dioxide (CO?), isoflurane or without anaesthesia in a crossover design. Heart rate, blood pressure and body temperature were...... for rats sampled from the tail vein, which showed fluctuations in body temperature in excess of 30 h after sampling. Increases in heart rate and blood pressure within the first hours after sampling indicated that periorbital puncture was the method that had the largest acute impact on the rats...... registered for three days after sampling. Initially blood pressure increased, but shortly after sampling it decreased, which led to increased heart rate. Sampling induced rapid fluctuations in body temperature, and an increase in body temperature. Generally, rats recovered from sampling within 2-3 h, except...

  2. Validation of a less invasive blood sampling technique in rabies serology using reduviid bugs (Triatominae, Hemiptera).

    Science.gov (United States)

    Vos, Ad C; Müller, Thomas; Neubert, Larissa; Voigt, Christian C

    2010-03-01

    During serologic rabies surveys, bleeding is often difficult or almost impossible in small or endangered mammals such as bats. Therefore, the usefulness of an alternative, less invasive technique--that is, the use of blood-sucking reduviid bugs (Dipetalogaster maximus and Rhodnius prolixus)--was investigated. Bugs were used in combination with a conventional method (retro-orbitale bleeding) to obtain blood samples from the same individual NMRI-mice (Mus musculus) vaccinated against rabies. Rabies virus-neutralizing antibody (VNA) titers between paired blood samples obtained from the same mice were compared. The accuracy (between-method comparison), precision (repeatability of results), and robustness (influence of digestion on blood parameter) of the bug method was evaluated. VNA titers in the blood sample obtained from the bugs' crops were equivalent to those from samples collected directly from the mice. No differences between samples taken from different bugs that had fed on the same mouse were noted. In addition, there were no changes in VNA titers in blood samples collected from the triatomine bugs for up to 4 hr after completion of the blood meal. This study demonstrates that the application of blood-sucking bugs offers a validated alternative for obtaining blood samples to determine rabies virus-neutralizing antibody titers and is highly suitable for animals with limited or no accessibility of veins by conventional sampling techniques.

  3. Detection of Salmonella typhi by nested polymerase chain reaction in blood, urine, and stool samples

    NARCIS (Netherlands)

    Hatta, Mochammad; Smits, Henk L.

    2007-01-01

    A nested polymerase chain reaction (PCR) specific for Salmonella enterica serovar Typhi was used for the detection of the pathogen in blood, urine, and stool samples from 131 patients with clinical suspicion of typhoid fever. The sensitivity of blood culture, the PCRs with blood, urine, and feces,

  4. Automated analysis of carbon in powdered geological and environmental samples by Raman spectroscopy.

    Science.gov (United States)

    Sparkes, Robert; Hovius, Niels; Galy, Albert; Kumar, R Vasant; Liu, James T

    2013-07-01

    Raman spectroscopy can be used to assess the structure of naturally occurring carbonaceous materials (CM), which exist in a wide range of crystal structures. The sources of these geological and environmental materials include rocks, soils, river sediments, and marine sediment cores, all of which can contain carbonaceous material ranging from highly crystalline graphite to amorphous-like organic compounds. In order to fully characterize a geological sample and its intrinsic heterogeneity, several spectra must be collected and analyzed in a precise and repeatable manner. Here, we describe a suitable processing and analysis technique. We show that short-period ball-mill grinding does not introduce structural changes to semi-graphitized material and allows for easy collection of Raman spectra from the resulting powder. Two automated peak-fitting procedures are defined that allow for rapid processing of large datasets. For very disordered CM, Lorentzian profiles are fitted to five characteristic peaks, for highly graphitized material, three Voigt profiles are fitted. Peak area ratios and peak width measurements are used to classify each spectrum and allow easy comparison between samples. By applying this technique to samples collected in Taiwan after Typhoon Morakot, sources of carbon to offshore sediments have been identified. Carbon eroded from different areas of Taiwan can be seen mixed and deposited in the offshore flood sediments, and both graphite and amorphous-like carbon have been recycled from terrestrial to marine deposits. The practicality of this application illustrates the potential for this technique to be deployed to sediment-sourcing problems in a wide range of geological settings.

  5. A femoral arteriovenous shunt facilitates arterial whole blood sampling in animals

    International Nuclear Information System (INIS)

    Weber, Bruno; Burger, Cyrill; Buck, Alfred; Biro, Peter

    2002-01-01

    In this study we evaluated on-line continuous blood sampling in a femoral arteriovenous (a-v) shunt for use in quantitative tracer studies using gamma-emitting radionuclides in animals. The shunt consisted of 40 cm polyethylene tubing (PE-50) guided through a coincidence probe. Two three-way valves allowed blood pressure measurements and tracer injection. Blood flow in the shunt and the impulse response function (IRF) were assessed using heparinized human blood mixed with fluorine-18 fluorodeoxyglucose (FDG). In vivo experiments were performed in eight male rats (300-350 g) anaesthetized with halothane. In three rats, manual blood sampling was performed in parallel with on-line sampling. In another five animals, the arterial whole blood activity was recorded on-line for 40 min. For the experiments 150-180 MBq FDG was injected over 35 s. Blood flow in the shunt was 23.6, 29.2 and 42.8 ml/h at 100, 120 and 160 mmHg, respectively. The IRF was characterized by minimal dispersion (1-2 s FWHM). Deconvolution of the measured arterial input curves with the IRF changed the measured curve only minimally. Whole blood radioactivity concentration derived from manual and on-line sampling were in excellent agreement. The curves derived from on-line sampling were of high statistical quality. In conclusion, a femoral a-v shunt allows multiple manipulations such as measurement of the arterial whole blood activity, continuous blood pressure monitoring, injection of the tracer and collection of blood samples if necessary. It is not associated with blood loss if the collection of blood samples is not required. It is more convenient to use than manual sampling, the peak of the input curve is never missed and the input curves are of high statistical quality. (orig.)

  6. Automated Diagnosis of Myocardial Infarction ECG Signals Using Sample Entropy in Flexible Analytic Wavelet Transform Framework

    Directory of Open Access Journals (Sweden)

    Mohit Kumar

    2017-09-01

    Full Text Available Myocardial infarction (MI is a silent condition that irreversibly damages the heart muscles. It expands rapidly and, if not treated timely, continues to damage the heart muscles. An electrocardiogram (ECG is generally used by the clinicians to diagnose the MI patients. Manual identification of the changes introduced by MI is a time-consuming and tedious task, and there is also a possibility of misinterpretation of the changes in the ECG. Therefore, a method for automatic diagnosis of MI using ECG beat with flexible analytic wavelet transform (FAWT method is proposed in this work. First, the segmentation of ECG signals into beats is performed. Then, FAWT is applied to each ECG beat, which decomposes them into subband signals. Sample entropy (SEnt is computed from these subband signals and fed to the random forest (RF, J48 decision tree, back propagation neural network (BPNN, and least-squares support vector machine (LS-SVM classifiers to choose the highest performing one. We have achieved highest classification accuracy of 99.31% using LS-SVM classifier. We have also incorporated Wilcoxon and Bhattacharya ranking methods and observed no improvement in the performance. The proposed automated method can be installed in the intensive care units (ICUs of hospitals to aid the clinicians in confirming their diagnosis.

  7. Novel blood sampling method of an artificial endocrine pancreas via the cardiopulmonary bypass circuit.

    Science.gov (United States)

    Kawahito, Shinji; Higuchi, Seiichi; Mita, Naoji; Kitagawa, Tetsuya; Kitahata, Hiroshi

    2013-12-01

    We tried to perform continuous blood glucose monitoring during cardiovascular surgery involving cardiopulmonary bypass using an artificial endocrine pancreas (STG-22 or -55; Nikkiso, Tokyo, Japan); however, we often encountered problems during these procedures because insufficient blood was obtained for monitoring. Thus, we started performing the blood sampling via the venous side of the cardiopulmonary bypass circuit. As a result, continuous blood glucose monitoring using an artificial endocrine pancreas was proven to be stable and reliable during cardiovascular surgery involving cardiopulmonary bypass.

  8. A novel flow injection chemiluminescence method for automated and miniaturized determination of phenols in smoked food samples.

    Science.gov (United States)

    Vakh, Christina; Evdokimova, Ekaterina; Pochivalov, Aleksei; Moskvin, Leonid; Bulatov, Andrey

    2017-12-15

    An easily performed fully automated and miniaturized flow injection chemiluminescence (CL) method for determination of phenols in smoked food samples has been proposed. This method includes the ultrasound assisted solid-liquid extraction coupled with gas-diffusion separation of phenols from smoked food sample and analytes absorption into a NaOH solution in a specially designed gas-diffusion cell. The flow system was designed to focus on automation and miniaturization with minimal sample and reagent consumption by inexpensive instrumentation. The luminol - N-bromosuccinimide system in an alkaline medium was used for the CL determination of phenols. The limit of detection of the proposed procedure was 3·10 -8 ·molL -1 (0.01mgkg -1 ) in terms of phenol. The presented method demonstrated to be a good tool for easy, rapid and cost-effective point-of-need screening phenols in smoked food samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Total reflection x-ray analysis of metals in blood samples

    International Nuclear Information System (INIS)

    Nakamura, Takuya; Matsui, Hiroshi; Kawamata, Masaya

    2009-01-01

    The sample preparation for TXRF (total reflection X-ray fluorescence) quantitative analysis of trace elements in human blood samples was investigated. In the TXRF analysis, a solution sample is dropped and dried on a flat substrate, and then the dried residue is measured. In this case, the dried residue should be flat not to disturb X-ray total reflection on the substrate. In addition, it is required to simply measure the whole blood sample by TXRF method, although a serum is analyzed in many cases. Thus, we studied the optimum conditions of the sample preparation of the whole blood by adding the pure water to apply Hemolysis phenomenon, where blood cells are destroyed due to different of the osmotic pressure, leading to flat residue. It was found that the best S/B ratio was obtained when the whole blood was diluted 8 times with pure water. Moreover, it was investigated the influence of the surface chemical condition of the glass substrate on the shape of the dried reside of the blood sample. When the surface of the glass substrate was hydrophilic, the shape of the dried residues was not uniform, as a result, the quantitative data of TXRF analysis gave a large deviation. On the other hand, when the surface of the glass was hydrophobic, the shape of the residue was almost uniform, as a result, a good reproducibility was obtained. Another problem was an outer ring of the dried residue of the blood. This uneven ring absorbs the primary X-rays, caused to low determined quantitative data. Thus, we tried the heating way of the dropped blood sample at a high temperature of 200 degrees. In this case, the blood sample was dried immediately, and a flat homogeneous dried residue was obtained without the outer ring. Using the optimized conditions for sample preparation, human blood sample was quantitatively measured by TXRF and ICP-AES. A good agreement was obtained in TXRF and ICP-AES determinations; however, the measurement of Cl and Br will be an advantage of TXRF, because

  10. Rapid DNA analysis for automated processing and interpretation of low DNA content samples.

    Science.gov (United States)

    Turingan, Rosemary S; Vasantgadkar, Sameer; Palombo, Luke; Hogan, Catherine; Jiang, Hua; Tan, Eugene; Selden, Richard F

    2016-01-01

    Short tandem repeat (STR) analysis of casework samples with low DNA content include those resulting from the transfer of epithelial cells from the skin to an object (e.g., cells on a water bottle, or brim of a cap), blood spatter stains, and small bone and tissue fragments. Low DNA content (LDC) samples are important in a wide range of settings, including disaster response teams to assist in victim identification and family reunification, military operations to identify friend or foe, criminal forensics to identify suspects and exonerate the innocent, and medical examiner and coroner offices to identify missing persons. Processing LDC samples requires experienced laboratory personnel, isolated workstations, and sophisticated equipment, requires transport time, and involves complex procedures. We present a rapid DNA analysis system designed specifically to generate STR profiles from LDC samples in field-forward settings by non-technical operators. By performing STR in the field, close to the site of collection, rapid DNA analysis has the potential to increase throughput and to provide actionable information in real time. A Low DNA Content BioChipSet (LDC BCS) was developed and manufactured by injection molding. It was designed to function in the fully integrated Accelerated Nuclear DNA Equipment (ANDE) instrument previously designed for analysis of buccal swab and other high DNA content samples (Investigative Genet. 4(1):1-15, 2013). The LDC BCS performs efficient DNA purification followed by microfluidic ultrafiltration of the purified DNA, maximizing the quantity of DNA available for subsequent amplification and electrophoretic separation and detection of amplified fragments. The system demonstrates accuracy, precision, resolution, signal strength, and peak height ratios appropriate for casework analysis. The LDC rapid DNA analysis system is effective for the generation of STR profiles from a wide range of sample types. The technology broadens the range of sample

  11. Eosinophilia detected by automated blood cell counting in ambulatory North American outpatients. Incidence and clinical significance.

    Science.gov (United States)

    Brigden, M; Graydon, C

    1997-09-01

    To audit a cohort of ambulatory outpatients with eosinophilia detected on automated blood cell counting. Specific objectives included the determination of whether the eosinophilia had been anticipated, the etiology of the eosinophilia, the clinical follow-up and investigations performed on patients with eosinophilia, and the effect of the detection of eosinophilia on patient management and ultimate clinical outcome. A year-long retrospective review of all patients with an absolute eosinophil count of greater than 0.7 x 10(9)/L. A large outpatient laboratory system. The patient population was managed by family physicians and specialists. Data collection included the results of the hematology profile, the absolute eosinophil count, the clinical situation responsible for the hematologic profile determination, and the probable cause of eosinophilia. Individual physicians were surveyed to determine if discovery of the eosinophilia had changed patient management plan or clinical outcome. Out of 195,300 patients who had a hematology profile performed, 225 were found to have an absolute eosinophilia count higher than 0.7 x 10(9)/L. The overall incidence of eosinophilia in the study population was 0.1%. The eosinophilia was not anticipated in 85% of patients. No obvious cause was detected for the eosinophilia in 36% of patients. Various allergic diseases were responsible for the eosinophilia in the majority of the remaining patients. Fewer than 9% of individuals manifested a serious systemic illness or parasitemia. Further clinical follow-up had been performed in 69% of patients. Additional laboratory tests had been ordered in 59% of patients. The laboratory tests most frequently ordered were a repeat hematology profile or stool examinations for ova and parasites. In only two instances did the discovery of the eosinophilia appear to result in a significant change in patient management or ultimate clinical income. The vast majority of eosinophilias detected in ambulatory

  12. Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia.

    Science.gov (United States)

    Krleza, Jasna Lenicek

    2014-01-01

    Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about the laboratory's parent institution, patient population, types and frequencies of laboratory tests based on capillary blood samples, choice of reference intervals, and policies and procedures specifically related to capillary sampling. Sampling practices were compared with guidelines from the Clinical and Laboratory Standards Institute (CLSI) and the World Health Organization (WHO). Of the 204 laboratories surveyed, 174 (85%) responded with complete questionnaires. Among the 174 respondents, 155 (89%) reported that they routinely perform capillary sampling, which is carried out by laboratory staff in 118 laboratories (76%). Nearly half of respondent laboratories (48%) do not have a written protocol including order of draw for multiple sampling. A single puncture site is used to provide capillary blood for up to two samples at 43% of laboratories that occasionally or regularly perform such sampling. Most respondents (88%) never perform arterialisation prior to capillary blood sampling. Capillary blood sampling is highly prevalent in Croatia across different types of clinical facilities and patient populations. Capillary sampling procedures are not standardised in the country, and the rate of laboratory compliance with CLSI and WHO guidelines is low.

  13. Single injection 51Cr EDTA plasma clearance determination in children using capillary blood samples

    International Nuclear Information System (INIS)

    Broechner-Mortensen, J.; Christoffersen, J.

    1977-01-01

    The reliability of a determination of the total 51 Cr EDTA plasma clearance (e) (and with it the glomerular filtration rate), by a simplified single injection method (injected dose: 4.5 μCi per kg b.w.) using capillary blood samples (0.2 ml), was investigated in twenty children. Clearance values determined from capillary blood samples did not differ significantly from those measured simultaneously from venous blood samples, the mean ratio+-SD being 1.02+-0.06(n = 10). The reproducibility (total day-to-day variation) of E determined from capillary blood samples was 6.7% in children with decreased renal function (n = 3) and 6.9% in children with normal renal function (n = 7). The present data indicate that the use of capillary blood samples is an accurate and very precise approach for determination of E in children. (Auth.)

  14. Relative Abundance of Proteins in Blood Plasma Samples from Patients with Chronic Cerebral Ischemia.

    Science.gov (United States)

    Kaysheva, Anna L; Kopylov, Artur T; Ponomarenko, Elena A; Kiseleva, Olga I; Teryaeva, Nadezhda B; Potapov, Alexander A; Izotov, Alexander А; Morozov, Sergei G; Kudryavtseva, Valeria Yu; Archakov, Alexander I

    2018-03-01

    A comparative protein profile analysis of 17 blood plasma samples from patients with ischemia and 20 samples from healthy volunteers was carried out using ultra-high resolution mass spectrometry. The analysis of measurements was performed using the proteomics search engine OMSSA. Normalized spectrum abundance factor (NSAF) in the biological samples was assessed using SearchGUI. The findings of mass spectrometry analysis of the protein composition of blood plasma samples demonstrate that the depleted samples are quite similar in protein composition and relative abundance of proteins. By comparing them with the control samples, we have found a small group of 44 proteins characteristic of the blood plasma samples from patients with chronic cerebral ischemia. These proteins contribute to the processes of homeostasis maintenance, including innate immune response unfolding, the response of a body to stress, and contribution to the blood clotting cascade.

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

    Science.gov (United States)

    Elliott, Paul; Peakman, Tim C

    2008-04-01

    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.

  16. Effect of red blood cell aggregation and sedimentation on optical coherence tomography signals from blood samples

    International Nuclear Information System (INIS)

    Kirillin, M Yu; Priezzhev, A V; Tuchin, V V; Wang, R K; Myllylae, R

    2005-01-01

    In this work, Monte Carlo simulation is used to obtain model optical coherence tomography (OCT) signals from a horizontally orientated blood layer at different stages of red blood cell (RBC) aggregation and sedimentation processes. The parameters for aggregating and sedimenting blood cells were chosen based on the data available from the literature and our earlier experimental studies. We consider two different cases: a suspension of washed RBCs in physiological solution (where aggregation does not take place) and RBCs in blood plasma (which provides necessary conditions for aggregation). Good agreement of the simulation results with the available experimental data shows that the chosen optical parameters are reasonable. The dependence of the numbers of photons contributing to the OCT signal on the number of experienced scattering events was analysed for each simulated signal. It was shown that the maxima of these dependences correspond to the peaks in the OCT signals related to the interfaces between the layers of blood plasma and blood cells. Their positions can be calculated from the optical thicknesses of the layers, and the absorption and scattering coefficients of the media

  17. Influence of SPRINT Study Type Automated Office Blood Pressure Measurements on Hypertension Diagnosis in Kidney Transplant Patients

    Directory of Open Access Journals (Sweden)

    Akanksha Bhatnagar

    2018-03-01

    Full Text Available Background/Aims: We compare conventional office blood pressure measurements with automated SPRINT-study type readings in kidney transplant recipients in order to determine the impact of the white coat effect in a prospective observational study. Methods: Adult patients with a functional renal transplant not dependent on dialysis were eligible. Readings were taken in the office in presence of the physician with an oscillometric method. Afterwards, readings were repeated with the patients resting alone in a quiet examination room with an automated blood pressure monitor. After 5 minutes of rest, 3 readings were taken at 1 minute intervals, with an average of these 3 readings calculated by the monitor. Results: 120 patients with an average age of 58.5±12.2 years were included. Mean time since transplantation was 7.95±6.48 years. Mean eGFR (CKD-EPI was 48.5±18.3 ml/min. SPRINT-study type readings were significantly lower than office readings (139.01±18.45 vs. 149.00±21.02 mmHg systolic, p<0.001; 80.88±11.63 mmHg vs. 84.35±12.41 mmHg diastolic, p <0.001. Correlation analysis for many potentially influencing factors (diabetes mellitus, transplant vintage, proteinuria, age, immunosuppression, donor type was not significant but obese women were significantly more prone to white coat hypertension. Conclusion: Automated office blood pressure measurements should be considered the method of choice in kidney transplant recipients.

  18. Assessment of blood sample stability for complete blood count using the Sysmex XN-9000 and Mindray BC-6800 analyzers

    Directory of Open Access Journals (Sweden)

    Sabrina Buoro

    Full Text Available ABSTRACT BACKGROUND: Different hematological analyzers have different analytical performances that are often reflected in the criteria for sample stability of the complete blood count. This study aimed to assess the stability of several hematological parameters using the XN-9000 Sysmex and BC-6800 Mindray analyzers. METHODS: The impact of storage at room temperature and 4 °C was evaluated after 2, 4, 6, 8, 24, 36 and 48 h using ten normal and 40 abnormal blood samples. The variation from the baseline measurement was evaluated by the Steel-Dwass-Critchlow-Fligner test and by Bland-Altman plots, using quality specifications and critical difference as the total allowable variation. RESULTS: Red blood cells and reticulocyte parameters (i.e. hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration, red blood cell distribution width, immature reticulocyte fractions, low-fluorescence reticulocytes, middle-fluorescence reticulocytes, high fluorescence mononuclear cells showed less stability compared to leukocyte and platelet parameters (except for monocyte count and mean platelet volume. The bias for hematocrit, mean corpuscular volume, mean corpuscular hemoglobin concentration and red blood cell distribution width coefficient of variation was higher than the critical difference after 8 h using both analyzers. CONCLUSION: Blood samples measured with both analyzers do not show analytically significant changes in up to 2 h of storage at room temperature and 4 °C. However, the maximum time for analysis can be extended for up to 8 h when the bias is compared to the critical difference.

  19. Investigations into the environmental conditions experienced during ambient sample transport: impact to dried blood spot sample shipments.

    Science.gov (United States)

    Bowen, Chester L; Dopson, Wesley; Kemp, Daniel C; Lewis, Mark; Lad, Rakesh; Overvold, Carol

    2011-07-01

    Prior to bioanalysis, sample transport and storage are critical considerations in any pharmacokinetic or toxicokinetic study design. Care must be taken to ensure the shipment is properly packaged and tracked to make certain it arrives at the desired, final destination in the appropriate timeframe, and that the integrity of the sample is not compromised. When dealing with biological specimens, environmental conditions may have a deleterious effect on the stability and conditions of the sample. Currently, frozen plasma or blood samples are the matrix of choice within the pharmaceutical industry for analysis within both preclinical and clinical trials. Liquid samples are shipped and received frozen and, therefore, the assumption is made that the frozen conditions are maintained throughout the entire transit process. Dried blood spot and dried matrix spot samples are becoming popular alternatives to plasma sampling in many small- and even large-molecule applications. With the implementation of dried blood spot and dried matrix spot samples, shipping and storage occurs under ambient conditions. In this article we discuss various shipping containers for these samples, illustrate the environmental extremes encountered during the shipping process, demonstrate a cost-effective method of monitoring both temperature and humidity, and discuss validation steps that may be implemented to minimize the impact of these variables on your study design.

  20. [Detection of Candida DNA in simulated blood samples by polymerase chain reaction].

    Science.gov (United States)

    Ozer, Sinem; Yücesoy, Mine

    2007-07-01

    Although blood culture method is accepted as gold standard in the laboratory diagnosis of invasive candidiasis seen in immunocompromised patients, cultivation of blood is considered as not a reliable and rapid method for the diagnosis of candidemia, since it may be negative in approximately half of the patients, slow growth rate of Candida in routine culture media and requirement of large amounts of blood for the isolation. The aim of this study was to detect Candida DNA in simulated blood samples by using polymerase chain reaction (PCR). Simulated samples were prepared by using blood samples of healthy volunteers. These samples were inoculated into tubes with EDTA and BACTEC 9240 blood culture bottles in which no growth was detected and with standard strains of C. albicans, C. tropicalis, C. parapsilosis, C. krusei, Escherichia coli and Staphylococcus aureus together with the clinical isolates of Aspergillus fumigatus, C. kefyr, C. glabrata, C. lusitaniae, C. guilliermondii and Rhodotorula sp. Additionally, blood culture samples of 23 cases whose blood culture bottles signaled as positive and revealed growth of Candida in agar plates were examined. DNA extraction of all samples were performed according to the standard procedure proposed by the MN Nucleospin Tissue Kit (Macherey-Nagel, Germany) for tissue samples; following the pre-treatment with erythrocyte, leukocyte and fungus cell wall lysis buffers. DNAs were amplified with PCR, using primers specific for the 5S rDNA region (PCon 1 and PCon 2 primers) and PCR products were obtained by electrophoresis in 2% agarose gel. Presence of a 105 base pair (bp) product was considered as positive. The lowest detection limit of PCR has been determined as 10(2)-10(3) cfu/ml Candida for our simulated samples. The presence of a 105 bp band has been observed in samples prepared with all Candida strains included in the study. Blood samples spiked with E. coli, S. aureus, A. fumigatus and Rhodotorula sp. and negative blood

  1. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    Energy Technology Data Exchange (ETDEWEB)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E. (Vyzkumny Ustav Veterinarniho Lekarstvi, Brno-Medlanky (Czechoslovakia))

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples using ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC.

  2. Microcapillary blood sampling for serological examinations by radioimmunoassay (RIA) and enzyme immunoassay (ELISA)

    International Nuclear Information System (INIS)

    Rodak, L.; Smid, B.; Valicek, L.; Jurak, E.

    1984-01-01

    Methods were tested of sampling blood and blood serum for serological examinations on filtration paper and into heparinized glass capillaries with transfer into the dilution solution of the given composition. Samples were also examined for ACH virus antibodies. The suitability of the sampling was verified by an examination of samples usiOg ELISA and RIA methods. The results showed the suitability of sampling using microcapillaries. The titres of virus antibodies found using the ELISA and RIA methods were identical and the sensitivity of antibody detection was not reduced even after the sample had been stored for 60 days at a temperature of 20 degC. (B.S.)

  3. An artificial pancreas for automated blood glucose control in patients with Type 1 diabetes.

    Science.gov (United States)

    Schmidt, Signe; Boiroux, Dimitri; Ranjan, Ajenthen; Jørgensen, John Bagterp; Madsen, Henrik; Nørgaard, Kirsten

    2015-01-01

    Automated glucose control in patients with Type 1 diabetes is much-coveted by patients, relatives and healthcare professionals. It is the expectation that a system for automated control, also know as an artificial pancreas, will improve glucose control, reduce the risk of diabetes complications and markedly improve patient quality of life. An artificial pancreas consists of portable devices for glucose sensing and insulin delivery which are controlled by an algorithm residing on a computer. The technology is still under development and currently no artificial pancreas is commercially available. This review gives an introduction to recent progress, challenges and future prospects within the field of artificial pancreas research.

  4. ASPIRE: An automated sample positioning and irradiation system for radiation biology experiments at Inter University Accelerator Centre, New Delhi

    International Nuclear Information System (INIS)

    Kothari, Ashok; Barua, P.; Archunan, M.; Rani, Kusum; Subramanian, E.T.; Pujari, Geetanjali; Kaur, Harminder; Satyanarayanan, V.V.V.; Sarma, Asitikantha; Avasthi, D.K.

    2015-01-01

    An automated irradiation setup for biology samples has been built at Inter University Accelerator Centre (IUAC), New Delhi, India. It can automatically load and unload 20 biology samples in a run of experiment. It takes about 20 min [2% of the cell doubling time] to irradiate all the 20 samples. Cell doubling time is the time taken by the cells (kept in the medium) to grow double in numbers. The cells in the samples keep growing during entire of the experiment. The fluence irradiated to the samples is measured with two silicon surface barrier detectors. Tests show that the uniformity of fluence and dose of heavy ions reaches to 2% at the sample area in diameter of 40 mm. The accuracy of mean fluence at the center of the target area is within 1%. The irradiation setup can be used to the studies of radiation therapy, radiation dosimetry and molecular biology at the heavy ion accelerator. - Highlights: • Automated positioning and irradiation setup for biology samples at IUAC is built. • Loading and unloading of 20 biology samples can be automatically carried out. • Biologicals cells keep growing during entire experiment. • Fluence and dose of heavy ions are measured by two silicon barrier detectors. • Uniformity of fluence and dose of heavy ions at sample position reaches to 2%

  5. Use of filter paper blood samples for rabies antibody detection in foxes and raccoon dogs.

    Science.gov (United States)

    Wasniewski, Marine; Barrat, Jacques; Combes, Benoit; Guiot, Anne Laure; Cliquet, Florence

    2014-08-01

    The effectiveness of oral rabies vaccination in wildlife is usually evaluated by the detection of rabies antibodies. However, the assessment of rabies antibodies has several technical difficulties in the field, such as the collection, storage, transport and titration of blood samples, often of poor quality. The objective of this study was to assess the feasibility of collecting blood on a filter paper (FP) coupled with enzyme-linked immunosorbent assay (ELISA) titration of rabies antibodies in raccoon dogs and red foxes. The FP blood sampling method was found highly specific and repeatable in both species. Overall, results obtained with the FP sampling method were highly concordant with the conventional (venipuncture) sampling methods. Blood eluates from FP samples from foxes and raccoon dogs tested using ELISA showed concordance values of 92% and 95%, respectively, with serum samples tested using the seroneutralisation test and values of 95% and 91%, respectively, when the ELISA was used on both types of sample. The use of FP blood sampling coupled with the titration of rabies antibodies by ELISA provides a reliable alternative to conventional blood sampling and serum testing by seroneutralisation. This simple procedure is particularly attractive and cost-effective for assessing the effectiveness of oral rabies vaccination in field conditions. Copyright © 2014. Published by Elsevier B.V.

  6. A method to quantitate cerebral blood flow using a rotating gamma camera and iodine-123 iodoamphetamine with one blood sampling

    International Nuclear Information System (INIS)

    Iida, Hidehiro; Itoh, Hiroshi; Bloomfield, P.M.; Munaka, Masahiro; Higano, Shuichi; Murakami, Matsutaro; Inugami, Atsushi; Eberl, S.; Aizawa, Yasuo; Kanno, Iwao; Uemura, Kazuo

    1994-01-01

    A method has been developed to quantitate regional cerebral blood blow (rCBF) using iodine-123-labelled N-isopropyl-p-iodoamphetamine (IMP). This technique requires only two single-photon emission tomography (SPET) scans and one blood sample. Based on a two-compartment model, radioactivity concentrations in the brain for each scan time are calculated. A standard input function has been generated by combining the input functions from 12 independent studies prior to this work to avoid frequent arterial blood sampling, and one blood sample is taken at 10 min following IMP administration for calibration of the standard arterial input function. This calibration time was determined such that the integration of the first 40 min of the calibrated, combined input function agreed best with those from 12 individual input functions (the difference was 5.3% on average). This method was applied to eight subjects (two normals and six patients with cerebral infarction), and yielded rCBF values which agreed well with those obtained by a positron emission tomography H 2 15 O autoradiography method. This method was also found to provide rCBF values that were consistent with those obtained by the non-linear least squares fitting technique and those obtained by conventional microsphere model analysis. The optimum SPET scan times were found to be 40 and 180 min for the early and delayed scans, respectively. These scan times allow the use of a conventional rotating gamma camera for clinical purposes. V d values ranged between 10 and 40 ml/g depending on the pathological condition, thereby suggesting the importance of measuring V d for each ROI. In conclusion, optimization of the blood sampling time and the scanning time enabled quantitative measurement of rCBF with two SPET scans and one blood sample. (orig.)

  7. [Evaluation of the test strip Reflotron ALP (alkaline phosphatase) for blood samples of dogs and cats].

    Science.gov (United States)

    Hirschberger, J; Dietz, J; Baumeister, C; Kraft, M

    1998-09-01

    The test strip Reflotron ALP (alcaline phosphatase) was evaluated for use in canine and feline heparinized blood samples. The within-run and the day-to-day precision of blood and plasma samples was excellent. A haematocrit up to 60% of canine blood and up to 50% of feline blood had no influence on the measurements. Reflotron ALP was compared with ALP on Hitachi 717 (Boehringer Mannheim, Mannheim, Germany). The correlation of both methods was good for canine and feline blood samples. Reflotron ALP was higher than Hitachi ALP in both species. Despite an enormous deviation between Reflotron ALP and the reference method of canine and feline blood samples within the reference range, Reflotron ALP is a suited test for the detection of elevated ALP activity in canine and feline blood samples. Differences of the ALP activity might be caused by ALP isoenzymes. The activity of ALP isoenzymes depends on the method. The buffer systems of Reflotron ALP and Hitachi ALP are different. Significant exceeding of the reference range was reliably detected. In this investigation the study results for heparinized whole blood, heparinized plasma and serum are approximately the same.

  8. Detection of Salmonella spp. with the BACTEC 9240 Automated Blood Culture System in 2008 - 2014 in Southern Iran (Shiraz): Biogrouping, MIC, and Antimicrobial Susceptibility Profiles of Isolates

    Science.gov (United States)

    Anvarinejad, Mojtaba; Pouladfar, Gholam Reza; Pourabbas, Bahman; Amin Shahidi, Maneli; Rafaatpour, Noroddin; Dehyadegari, Mohammad Ali; Abbasi, Pejman; Mardaneh, Jalal

    2016-01-01

    Background Human salmonellosis continues to be a major international problem, in terms of both morbidity and economic losses. The antibiotic resistance of Salmonella is an increasing public health emergency, since infections from resistant bacteria are more difficult and costly to treat. Objectives The aims of the present study were to investigate the isolation of Salmonella spp. with the BACTEC automated system from blood samples during 2008 - 2014 in southern Iran (Shiraz). Detection of subspecies, biogrouping, and antimicrobial susceptibility testing by the disc diffusion and agar dilution methods were performed. Patients and Methods A total of 19 Salmonella spp. were consecutively isolated using BACTEC from blood samples of patients between 2008 and 2014 in Shiraz, Iran. The isolates were identified as Salmonella, based on biochemical tests embedded in the API-20E system. In order to characterize the biogroups and subspecies, biochemical testing was performed. Susceptibility testing (disc diffusion and agar dilution) and extended-spectrum β-lactamase (ESBL) detection were performed according to the clinical and laboratory standards institute (CLSI) guidelines. Results Of the total 19 Salmonella spp. isolates recovered by the BACTEC automated system, all belonged to the Salmonella enterica subsp. houtenae. Five isolates (26.5%) were resistant to azithromycin. Six (31.5%) isolates with the disc diffusion method and five (26.3%) with the agar dilution method displayed resistance to nalidixic acid (minimum inhibitory concentration [MIC] > 32 μg/mL). All nalidixic acid-resistant isolates were also ciprofloxacin-sensitive. All isolates were ESBL-negative. Twenty-one percent of isolates were found to be resistant to chloramphenicol (MIC ≥ 32 μg/mL), and 16% were resistant to ampicillin (MIC ≥ 32 μg/mL). Conclusions The results indicate that multidrug-resistant (MDR) strains of Salmonella are increasing in number, and fewer antibiotics may be useful for

  9. Human blood RNA stabilization in samples collected and transported for a large biobank

    Directory of Open Access Journals (Sweden)

    Duale Nur

    2012-09-01

    Full Text Available Abstract Background The Norwegian Mother and Child Cohort Study (MoBa is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems – the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes. Findings High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression.

  10. Human blood RNA stabilization in samples collected and transported for a large biobank

    Science.gov (United States)

    2012-01-01

    Background The Norwegian Mother and Child Cohort Study (MoBa) is a nation-wide population-based pregnancy cohort initiated in 1999, comprising more than 108.000 pregnancies recruited between 1999 and 2008. In this study we evaluated the feasibility of integrating RNA analyses into existing MoBa protocols. We compared two different blood RNA collection tube systems – the PAXgene™ Blood RNA system and the Tempus™ Blood RNA system - and assessed the effects of suboptimal blood volumes in collection tubes and of transportation of blood samples by standard mail. Endpoints to characterize the samples were RNA quality and yield, and the RNA transcript stability of selected genes. Findings High-quality RNA could be extracted from blood samples stabilized with both PAXgene and Tempus tubes. The RNA yields obtained from the blood samples collected in Tempus tubes were consistently higher than from PAXgene tubes. Higher RNA yields were obtained from cord blood (3 – 4 times) compared to adult blood with both types of tubes. Transportation of samples by standard mail had moderate effects on RNA quality and RNA transcript stability; the overall RNA quality of the transported samples was high. Some unexplained changes in gene expression were noted, which seemed to correlate with suboptimal blood volumes collected in the tubes. Temperature variations during transportation may also be of some importance. Conclusions Our results strongly suggest that special collection tubes are necessary for RNA stabilization and they should be used for establishing new biobanks. We also show that the 50,000 samples collected in the MoBa biobank provide RNA of high quality and in sufficient amounts to allow gene expression analyses for studying the association of disease with altered patterns of gene expression. PMID:22988904

  11. Comparison of three methods of sampling trout blood for measurements of hematocrit

    Science.gov (United States)

    Steucke, Erwin W.; Schoettger, Richard A.

    1967-01-01

    Trout blood is frequently collected for hematocrit measurements by excising the caudal fin (Snieszko, 1960), but this technique is impractical if valuable fish are to be sampled or if repeated observations are desired. Schiffman (1959) and Snieszko (1960) collected blood from the dorsal aorta and the heart, but these methods are relatively slow and require the preparation of needles and syringes. The use of pointed capillary tubes for cardiac punctures increases the speed of sampling, but body fluids may dilute the blood (Perkins, 1957; Larsen and Snieszko, 1961; and Normandau, 1962). There is need for methods of sampling which are rapid and which neither influence hematological determinations nor harm the fish.

  12. Performance of an automated solid-phase red cell adherence system compared with that of a manual gel microcolumn assay for the identification of antibodies eluted from red blood cells.

    Science.gov (United States)

    Finck, R H; Davis, R J; Teng, S; Goldfinger, D; Ziman, A F; Lu, Q; Yuan, S

    2011-01-01

    IgG antibodies coating red blood cells (RBCs) can be removed by elution procedures and their specificity determined by antibody identification studies. Although such testing is traditionally performed using the tube agglutination assay, prior studies have shown that the gel microcolumn (GMC) assay may also be used with comparable results. The purpose of this study was to compare an automated solid-phase red cell adherence (SPRCA) system with a GMC assay for the detection of antibodies eluted from RBCs. Acid eluates from 51 peripheral blood (PB) and 7 cord blood (CB) samples were evaluated by both an automated SPRCA instrument and a manual GMC assay. The concordance rate between the two systems for peripheral RBC samples was 88.2 percent (45 of 51), including cases with alloantibodies (n = 8), warm autoantibodies (n = 12), antibodies with no identifiable specificity (n = 2), and negative results (n = 23). There were six discordant cases, of which four had alloantibodies (including anti-Jka, -E, and -e) demonstrable by the SPRCA system only. In the remaining 2 cases, anti-Fya and antibodies with no identifiable specificity were demonstrable by the GMC assay only. All seven CB specimens produced concordant results, showing anti-A (n = 3), -B (n = 1), maternal anti-Jka (n = 2), or a negative result (n = 1). Automated SPRCA technology has a performance that is comparable with that of a manual GMC assay for identifying antibodies eluted from PB and CB RBCs.

  13. Evaluation of a new handheld point-of-care blood gas analyser using 100 equine blood samples.

    Science.gov (United States)

    Bardell, David; West, Eleanor; Mark Senior, J

    2017-02-22

    To determine whether the Enterprise point-of-care blood analysis system (EPOC) produces results in agreement with two other blood gas analysers in regular clinical use (i-STAT and Radiometer ABL77) and to investigate the precision of the new machine when used with equine whole blood. Prospective, randomized, non-blinded, comparative laboratory analyser study. Horses admitted to a university teaching hospital requiring arterial or venous blood gas analysis as part of their routine clinical management. One hundred equine blood samples were run immediately, consecutively and in randomized order on three blood gas analysers. Results of variables common to all three analysers were tested for agreement and compared with guidelines used in human medicine. These require 80% of results from the test analyser to fall within a defined range or percentage of results from the comparator devices to achieve acceptability. Additionally, 21 samples were run twice in quick succession on the EPOC analyser to investigate precision. Agreement targets were not met for haematocrit, haemoglobin and base excess for either i-STAT or ABL77 analysers. EPOC precision targets were not met for partial pressure of carbon dioxide, ionized calcium, haematocrit and haemoglobin. Overall comparative performance of the EPOC was good to excellent for pH, oxygen tension, potassium, bicarbonate and oxygen saturation of haemoglobin, but marginal to poor for other parameters. The EPOC may be useful in performing analysis of equine whole blood, but trend analysis of carbon dioxide tension, ionized calcium, haematocrit and haemoglobin should be interpreted with caution. The EPOC should not be used interchangeably with other blood gas analysers. Copyright © 2016 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.

  14. Pediatric blood sample collection from a pre-existing peripheral intravenous (PIV) catheter.

    Science.gov (United States)

    Braniff, Heather; DeCarlo, Ann; Haskamp, Amy Corey; Broome, Marion E

    2014-01-01

    Aiming to minimize pain in a hospitalized child, the purpose of this observational study was to describe characteristics of blood samples collected from pre-existing peripheral intravenous (PIV) catheters in pediatric patients. One hundred and fifty blood samples were reviewed for number of unusable samples requiring a specimen to be re-drawn. Success of the blood draw and prevalence of the loss of the PIV following blood collection was also measured. Findings included one clotted specimen, success rate of 91.3%, and 1.3% of PIVs becoming non-functional after collection. Obtaining blood specimens from a pre-existing PIV should be considered in a pediatric patient. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. The Effect of Pneumatic Tube Systems on the Hemolysis of Biochemistry Blood Samples.

    Science.gov (United States)

    Cakirca, Gokhan; Erdal, Huseyin

    2017-05-01

    Pneumatic tube systems (PTSs) are widely used in many hospitals because they lead to reduced turnaround times and cost efficiency. However, PTSs may affect the quality of the blood samples transported to the laboratory. The aim of this study was to investigate the effect of the PTS used in our hospital on the hemolysis of the biochemical blood samples transported to the laboratory. A total of 148 samples were manually transported to the laboratory by hospital staff, 148 samples were transported with the PTS, and 113 were transported with the PTS without use of sponge-rubber inserts (PTSws). Hemolysis rates and the levels of biochemical analytes for the different transportation methods were compared. No significant difference was found between the samples transported manually and with the PTS with regard to hemolysis rate and the levels of biochemical analytes. However, the samples transported with the PTSws showed a significant difference compared with the samples transported manually and with the PTS with regard to hemolysis rate and potassium and lactate dehydrogenase levels. The percentages of the samples that exceeded the permissible threshold for the hemolysis among the samples transported manually, with the PTS, and with the PTSws were 10%, 8%, and 47%, respectively. A PTS can be used safely for transporting biochemistry blood samples to the laboratory. However, a sponge-rubber insert that holds sample tubes must be used with the PTS to prevent the hemolysis of blood samples. Copyright © 2016 Emergency Nurses Association. Published by Elsevier Inc. All rights reserved.

  16. Chlamydia trachomatis antibody detection in home-collected blood samples for use in epidemiological studies.

    NARCIS (Netherlands)

    Hoenderboom, B M; van Ess, E F; van den Broek, I V F; van Loo, I H M; Hoebe, C J P A; Ouburg, S; Morré, S A

    Capillary blood collected in serum tubes was subjected to centrifugation delay while stored at room temperature. Chlamydia trachomatis (CT) IgG concentrations in aliquoted serum of these blood samples remained stable for seven days after collection. CT IgG concentrations can reliably be measured in

  17. Leukocyte count affects expression of reference genes in canine whole blood samples

    NARCIS (Netherlands)

    Piek, C.J.; Brinkhof, B.; Rothuizen, J.; Dekker, A.; Penning, L.C.

    2011-01-01

    Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263

  18. Evaluation of some toxic metals in blood samples of smokers in ...

    African Journals Online (AJOL)

    Purpose: To determine some toxic elements in the blood of cigarette and tobacco pipe smokers in Riyadh, Saudi Arabia. Methods: The study setting was Riyadh, the capital city of Saudi Arabia, Riyadh City. Male volunteers, aged 20 - 58 year, whose blood samples were collected, were classified into three groups of ...

  19. Quality standards in Biobanking: authentication by genetic profiling of blood spots from donor's original sample.

    Science.gov (United States)

    Cardoso, Sergio; Valverde, Laura; Odriozola, Adrian; Elcoroaristizabal, Xabier; de Pancorbo, Marian M

    2010-07-01

    The field of Biobanking requires extensive work to maintain traceability of samples. However, sometimes the necessity to authenticate a sample may arise. To address these circumstances, we herein present a method for authenticating derivatives by using a blood spot from each donor, attached to a sample authentication form, by means of genetic profiling. Blood spots are collected at the time a blood sample is donated at a health centre and before processing the blood sample at the biobank. To test the validity of our approach over time, we analyzed 26 blood spots stored at room temperature in our facilities for more than 15 years. DNA was successfully extracted from the three storage materials tested in this study and 15 STR markers plus amelogenin were subsequently analyzed. The storage of a small blood spot attached to a sample authentication form proved to be efficient for genetic profiling and, therefore, may constitute a long-lasting (at least 15 years), cost-effective and effortless approach for genetic authentication of samples in biobanks.

  20. The effects of storage temperature and duration of blood samples on DNA and RNA qualities.

    Science.gov (United States)

    Huang, Lien-Hung; Lin, Pei-Hsien; Tsai, Kuo-Wang; Wang, Liang-Jen; Huang, Ying-Hsien; Kuo, Ho-Chang; Li, Sung-Chou

    2017-01-01

    DNA and RNA samples from blood are the common examination target for non-invasive physical tests and/or biomedical studies. Since high-quality DNA and RNA samples guarantee the correctness of these tests and/or studies, we investigated the effects of storage temperature and storage duration of whole blood on DNA and RNA qualities. Subjects were enrolled to donate blood samples which were stored for different durations and at different temperatures, followed by the examinations on RNA quality, qPCR, DNA quality and DNA methylation. For RNA, we observed obvious quality decline with storage duration longer than 24 hours. Storage at low temperature does not keep RNA samples from degradation. And, storing whole blood samples in freezer dramatically damage RNA. For DNA, quality decline was not observed even with storage duration for 15 days. However, DNA methylation significantly altered with storage duration longer than three days. Storage duration within 24 hours is critical for collecting high-quality RNA samples for next-generation sequencing (NGS) assays (RIN≧8). If microarray assays are expected (RIN≧7), storage duration within 32 hours is acceptable. Although DNA is resistant within 15 days when kept in whole blood, DNA quantity dramatically decreases owing to WBC lysis. In addition, duration for more than three days significantly alter DNA methylation status, globally and locally. Our result provides a reference for dealing with blood samples.

  1. Effects of Transport and Storage Conditions on Gene Expression in Blood Samples.

    Science.gov (United States)

    Malentacchi, Francesca; Pizzamiglio, Sara; Wyrich, Ralf; Verderio, Paolo; Ciniselli, Chiara; Pazzagli, Mario; Gelmini, Stefania

    2016-04-01

    Inappropriate handling of blood samples might induce or repress gene expression and/or lead to RNA degradation affecting downstream analysis. In particular, sample transport is a critical step for biobanking or multicenter studies because of uncontrolled variables (i.e., unstable temperature). We report the results of a pilot study implemented within the EC funded SPIDIA project, aimed to investigate the role of transport and storage of blood samples containing and not containing an RNA stabilizer. Blood was collected from a single donor both in EDTA and in PAXgene Blood RNA tubes. Half of the samples were sent to a second laboratory both at room temperature and at 4°C, whereas the remaining samples were stored at room temperature and at 4°C. Gene expression of selected genes (c-FOS, IL-1β, IL-8, and GAPDH) known to be induced or repressed by ex vivo blood handling and of blood-mRNA quality biomarkers identified and validated within the SPIDIA project, which allow for monitoring changes in unstabilized blood samples after collection and during transport and storage, were analyzed by RT-qPCR. If the shipment of blood in tubes not containing RNA stabilizer is not performed under a stable condition, gene profile studies can be affected by the effects of transport. Moreover, also controlled temperature shipment (4°C) can influence the expression of specific genes if blood is collected in tubes not containing a stabilizer. The use of dedicated biomarkers or time course experiments should be performed in order to verify potential bias on gene expression analysis due to sample shipment and storage conditions. Alternatively, the use of RNA stabilizer containing tubes can represent a reliable option to avoid ex vivo RNA changes.

  2. A method for estimating radioactive cesium concentrations in cattle blood using urine samples.

    Science.gov (United States)

    Sato, Itaru; Yamagishi, Ryoma; Sasaki, Jun; Satoh, Hiroshi; Miura, Kiyoshi; Kikuchi, Kaoru; Otani, Kumiko; Okada, Keiji

    2017-12-01

    In the region contaminated by the Fukushima nuclear accident, radioactive contamination of live cattle should be checked before slaughter. In this study, we establish a precise method for estimating radioactive cesium concentrations in cattle blood using urine samples. Blood and urine samples were collected from a total of 71 cattle on two farms in the 'difficult-to-return zone'. Urine 137 Cs, specific gravity, electrical conductivity, pH, sodium, potassium, calcium, and creatinine were measured and various estimation methods for blood 137 Cs were tested. The average error rate of the estimation was 54.2% without correction. Correcting for urine creatinine, specific gravity, electrical conductivity, or potassium improved the precision of the estimation. Correcting for specific gravity using the following formula gave the most precise estimate (average error rate = 16.9%): [blood 137 Cs] = [urinary 137 Cs]/([specific gravity] - 1)/329. Urine samples are faster to measure than blood samples because urine can be obtained in larger quantities and has a higher 137 Cs concentration than blood. These advantages of urine and the estimation precision demonstrated in our study, indicate that estimation of blood 137 Cs using urine samples is a practical means of monitoring radioactive contamination in live cattle. © 2017 Japanese Society of Animal Science.

  3. Na2EDTA anticoagulant impaired blood samples from the teleost Piaractus mesopotamicus

    Directory of Open Access Journals (Sweden)

    Thaís Heloisa Vaz Farias

    2016-05-01

    Full Text Available Abstract: The present study aimed to evaluate the effects of Na heparin and Na2EDTA on blood of Piaractus mesopotamicus (360.7±42.4g, 26.4±1.0cm. Twenty fishes were sampled in two experiment trials, ten for erythrocyte fragility analysis and ten for hematologic and plasma biochemical study. The blood collected by venous-caudal puncture was fractioned and stored in anticoagulants solution: Na2EDTA 10%, Na2EDTA 3%, Na heparin 5000 IU and Na heparin 100 IU. Plasmatic levels of calcium presented in the Na2EDTA stored samples were about 80% lower than both heparin groups. Blood samples of P. mesopotamicus stored with Na2EDTA demonstrated increase in the hematocrit and MCV, and decrease in MCHC. The dose-response effect was observed in this study. The results are reinforced by the higher levels of plasmatic protein and hemolysis presented in the Na2EDTA 10% stored blood, confirming the deleterious effect of this anticoagulant treatment on the quality of blood samples. Na2EDTA is not indicated to store P. mesopotamicus blood samples, but sodium heparin at 100 IU is the most recommended anticoagulant, since this treatment presented the lower rate of alterations in the stored blood.

  4. A duplex PCR for rapid and simultaneous detection of Brucella spp. in human blood samples.

    Science.gov (United States)

    Mirnejad, Reza; Mohamadi, Mozafar; Piranfar, Vahbeh; Mortazavi, Seied Mojtaba; Kachuei, Reza

    2013-06-01

    To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus 39 (B. abortus 39) (23%), 13 for Brucella melitensis 39 (B. melitensis 39) (25%) and 0 for Brucella ovis 39 (B. ovis 39) (0%). This work demonstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples. Copyright © 2013 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

  5. Perfluoroalkyl Acid Concentrations in Blood Samples Subjected to Transportation and Processing Delay

    DEFF Research Database (Denmark)

    Bach, Cathrine Carlsen; Henriksen, Tine Brink; Bossi, Rossana

    2015-01-01

    BACKGROUND: In studies of perfluoroalkyl acids, the validity and comparability of measured concentrations may be affected by differences in the handling of biospecimens. We aimed to investigate whether measured plasma levels of perfluoroalkyl acids differed between blood samples subjected to delay...... and transportation prior to processing and samples with immediate processing and freezing. METHODS: Pregnant women recruited at Aarhus University Hospital, Denmark, (n = 88) provided paired blood samples. For each pair of samples, one was immediately processed and plasma was frozen, and the other was delayed...... and transported as whole blood before processing and freezing of plasma (similar to the Danish National Birth Cohort). We measured 12 perfluoroalkyl acids and present results for compounds with more than 50% of samples above the lower limit of quantification. RESULTS: For samples taken in the winter, relative...

  6. Evaluation of Glucose-6-Phosphate Dehydrogenase stability in stored blood samples.

    Science.gov (United States)

    Jalil, Norunaluwar; Azma, Raja Zahratul; Mohamed, Emida; Ithnin, Azlin; Alauddin, Hafiza; Baya, Siti Noor; Othman, Ainoon

    2016-01-01

    Glucose-6-Phosphate Dehydrogenase (G6PD) deficiency is the commonest cause of neonatal jaundice in Malaysia. Recently, OSMMR2000-D G6PD Assay Kit has been introduced to quantitate the level of G6PD activity in newborns delivered in Universiti Kebangsaan Malaysia Medical Centre (UKMMC). As duration of sample storage prior to analysis is one of the matters of concern, this study was conducted to identify the stability of G6PD enzyme during storage. A total of 188 cord blood samples from normal term newborns delivered at UKMMC were selected for this study. The cord bloods samples were collected in ethylene-diamine-tetra-acetic acid (EDTA) tubes and refrigerated at 2-8 °C. In addition, 32 out of 188 cord blood samples were spotted on chromatography paper, air-dried and stored at room temperature. G6PD enzyme activities were measured daily for 7 days using the OSMMR2000-D G6PD Assay Kit on both the EDTA blood and dried blood samples. The mean value for G6PD activity was compared between days of analysis using Student Paired T-Test. In this study, 172 out of 188 cord blood samples showed normal enzyme levels while 16 had levels corresponding to severe enzyme deficiency. The daily mean G6PD activity for EDTA blood samples of newborns with normal G6PD activity showed a significant drop on the fourth day of storage (p samples with severely deficient G6PD activity, significant drop was seen on third day of storage (p = 0.002). Analysis of dried cord blood showed a significant reduction in enzyme activity as early as the second day of storage (p = 0.001). It was also noted that mean G6PD activity for spotted blood samples were lower compared to those in EDTA tubes for all days (p = 0.001). Thus, EDTA blood samples stored at 2-8 °C appeared to have better stability in terms of their G6PD enzyme level as compared to dried blood samples on filter paper, giving a storage time of up to 3 days.

  7. A Novel Automated Slide-Based Technology for Visualization, Counting, and Characterization of the Formed Elements of Blood: A Proof of Concept Study.

    Science.gov (United States)

    Winkelman, James W; Tanasijevic, Milenko J; Zahniser, David J

    2017-08-01

    - A novel automated slide-based approach to the complete blood count and white blood cell differential count is introduced. - To present proof of concept for an image-based approach to complete blood count, based on a new slide preparation technique. A preliminary data comparison with the current flow-based technology is shown. - A prototype instrument uses a proprietary method and technology to deposit a precise volume of undiluted peripheral whole blood in a monolayer onto a glass microscope slide so that every cell can be distinguished, counted, and imaged. The slide is stained, and then multispectral image analysis is used to measure the complete blood count parameters. Images from a 600-cell white blood cell differential count, as well as 5000 red blood cells and a variable number of platelets, that are present in 600 high-power fields are made available for a technologist to view on a computer screen. An initial comparison of the basic complete blood count parameters was performed, comparing 1857 specimens on both the new instrument and a flow-based hematology analyzer. - Excellent correlations were obtained between the prototype instrument and a flow-based system. The primary parameters of white blood cell, red blood cell, and platelet counts resulted in correlation coefficients (r) of 0.99, 0.99, and 0.98, respectively. Other indices included hemoglobin (r = 0.99), hematocrit (r = 0.99), mean cellular volume (r = 0.90), mean corpuscular hemoglobin (r = 0.97), and mean platelet volume (r = 0.87). For the automated white blood cell differential counts, r values were calculated for neutrophils (r = 0.98), lymphocytes (r = 0.97), monocytes (r = 0.76), eosinophils (r = 0.96), and basophils (r = 0.63). - Quantitative results for components of the complete blood count and automated white blood cell differential count can be developed by image analysis of a monolayer preparation of a known volume of peripheral blood.

  8. A Centrifugal Microfluidic Platform That Separates Whole Blood Samples into Multiple Removable Fractions Due to Several Discrete but Continuous Density Gradient Sections

    Science.gov (United States)

    Moen, Scott T.; Hatcher, Christopher L.; Singh, Anup K.

    2016-01-01

    We present a miniaturized centrifugal platform that uses density centrifugation for separation and analysis of biological components in small volume samples (~5 μL). We demonstrate the ability to enrich leukocytes for on-disk visualization via microscopy, as well as recovery of viable cells from each of the gradient partitions. In addition, we simplified the traditional Modified Wright-Giemsa staining by decreasing the time, volume, and expertise involved in the procedure. From a whole blood sample, we were able to extract 95.15% of leukocytes while excluding 99.8% of red blood cells. This platform has great potential in both medical diagnostics and research applications as it offers a simpler, automated, and inexpensive method for biological sample separation, analysis, and downstream culturing. PMID:27054764

  9. Nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia

    OpenAIRE

    Lenicek Krleza, Jasna

    2014-01-01

    Introduction: Capillary sampling is increasingly used to obtain blood for laboratory tests in volumes as small as necessary and as non-invasively as possible. Whether capillary blood sampling is also frequent in Croatia, and whether it is performed according to international laboratory standards is unclear. Materials and methods: All medical laboratories that participate in the Croatian National External Quality Assessment Program (N = 204) were surveyed on-line to collect information about t...

  10. Platelet-rich fibrin prepared from stored whole-blood samples.

    Science.gov (United States)

    Isobe, Kazushige; Suzuki, Masashi; Watanabe, Taisuke; Kitamura, Yutaka; Suzuki, Taiji; Kawabata, Hideo; Nakamura, Masayuki; Okudera, Toshimitsu; Okudera, Hajime; Uematsu, Kohya; Nakata, Koh; Tanaka, Takaaki; Kawase, Tomoyuki

    2017-12-01

    In regenerative therapy, self-clotted platelet concentrates, such as platelet-rich fibrin (PRF), are generally prepared on-site and are immediately used for treatment. If blood samples or prepared clots can be preserved for several days, their clinical applicability will expand. Here, we prepared PRF from stored whole-blood samples and examined their characteristics. Blood samples were collected from non-smoking, healthy male donors (aged 27-67 years, N = 6), and PRF clots were prepared immediately or after storage for 1-2 days. Fibrin fiber was examined by scanning electron microscopy. Bioactivity was evaluated by means of a bioassay system involving human periosteal cells, whereas PDGF-BB concentrations were determined by an enzyme-linked immunosorbent assay. Addition of optimal amounts of a 10% CaCl 2 solution restored the coagulative ability of whole-blood samples that contained an anticoagulant (acid citrate dextrose) and were stored for up to 2 days at ambient temperature. In PRF clots prepared from the stored whole-blood samples, the thickness and cross-links of fibrin fibers were almost identical to those of freshly prepared PRF clots. PDGF-BB concentrations in the PRF extract were significantly lower in stored whole-blood samples than in fresh samples; however, both extracts had similar stimulatory effects on periosteal-cell proliferation. Quality of PRF clots prepared from stored whole-blood samples is not reduced significantly and can be ensured for use in regenerative therapy. Therefore, the proposed method enables a more flexible treatment schedule and choice of a more suitable platelet concentrate immediately before treatment, not after blood collection.

  11. Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples.

    Science.gov (United States)

    Uyaguari-Diaz, Miguel I; Slobodan, Jared R; Nesbitt, Matthew J; Croxen, Matthew A; Isaac-Renton, Judith; Prystajecky, Natalie A; Tang, Patrick

    2015-04-17

    Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

  12. Survivability of Existing Peripheral Intravenous Access Following Blood Sampling in a Pediatric Population.

    Science.gov (United States)

    O'Neil, Sheree W; Friesen, Mary Ann; Stanger, Debra; Trickey, Amber Williams

    2018-03-07

    Although pediatric patients report venipuncture as their most feared experience during hospitalization, blood sampling from peripheral intravenous accesses (PIVs) is not standard of care. Blood sampling from PIVs has long been considered by healthcare personnel to harm the access. In an effort to minimize painful procedures, pediatric nursing staff conducted a prospective, observational study to determine if blood sampling using existing PIVs resulted in the loss of the access. The ability to obtain the sample from the PIV was measured along with patient and PIV characteristics. Specimen collection using 100 existing PIVs was attempted on pediatric inpatients. Each PIV was observed for functionality, infiltration, occlusion, and dislodgement following collection and again in 4h. Frequencies of PIV loss and successful blood sampling were calculated. Patient age, PIV gauge, access site, and PIV age were evaluated for associations with successful sampling using chi-square tests, Fisher's exact tests, and logistic regression. PIV survivability was reported at 99%. The ability to obtain a complete specimen was reported at 76% and found to be significantly related to PIV age and site. Size of PIV and patient's age were not significantly related to successful sampling. Encouraging rates of PIV survivability and collectability suggest blood sampling from PIVs to be a valuable technique to minimize painful and distressful procedures. Nursing practice was changed in this pediatric department. Patients and families are saved the pain and distress of venipuncture. Nurses reported saving time and personal distress by avoiding the venipuncture procedure. Copyright © 2018 Elsevier Inc. All rights reserved.

  13. Automation impact study of Army training management 2: Extension of sampling and collection of installation resource data

    Energy Technology Data Exchange (ETDEWEB)

    Sanquist, T.F.; McCallum, M.C.; Hunt, P.S.; Slavich, A.L.; Underwood, J.A.; Toquam, J.L.; Seaver, D.A.

    1989-05-01

    This automation impact study of Army training management (TM) was performed for the Army Development and Employment Agency (ADEA) and the Combined Arms Training Activity (CATA) by the Battelle Human Affairs Research Centers and the Pacific Northwest Laboratory. The primary objective of the study was to provide the Army with information concerning the potential costs and savings associated with automating the TM process. This study expands the sample of units surveyed in Phase I of the automation impact effort (Sanquist et al., 1988), and presents data concerning installation resource management in relation to TM. The structured interview employed in Phase I was adapted to a self-administered survey. The data collected were compatible with that of Phase I, and both were combined for analysis. Three US sites, one reserve division, one National Guard division, and one unit in the active component outside the continental US (OCONUS) (referred to in this report as forward deployed) were surveyed. The total sample size was 459, of which 337 respondents contributed the most detailed data. 20 figs., 62 tabs.

  14. Dried blood spots on carboxymethyl cellulose sheets: Rapid sample preparation based on dissolution and precipitation

    DEFF Research Database (Denmark)

    Skoglund Ask, Kristine; Pedersen-Bjergaard, Stig; Gjelstad, Astrid

    2016-01-01

    This short communication describes the use of carboxymethyl cellulose sheets as sampling material for dried blood spots. Whole blood, spiked with quetiapine, a hydrophobic and basic small molecule drug substance, was spotted on the sheet and subsequently dried. The dried spot was then almost...... completely dissolved in acidified aqueous solution. It was shown that the dissolved polymer, together with major blood components can easily be precipitated and removed with acetonitrile. The presented sampling on a water-soluble biopolymer derivative followed by precipitation resulted in a simple protocol...

  15. Elimination of heparin interference during microarray processing of fresh and biobank-archived blood samples.

    Science.gov (United States)

    Hebels, Dennie G A J; van Herwijnen, Marcel H M; Brauers, Karen J J; de Kok, Theo M C M; Chalkiadaki, Georgia; Kyrtopoulos, Soterios A; Kleinjans, Jos C S

    2014-07-01

    In the context of environmental health research, biobank blood samples have recently been identified as suitable for high-throughput omics analyses enabling the identification of new biomarkers of exposure and disease. However, blood samples containing the anti-coagulant heparin could complicate transcriptomic analysis because heparin may inhibit RNA polymerase causing inefficient cRNA synthesis and fluorophore labelling. We investigated the inhibitory effect of heparin and the influence of storage conditions (0 or 3 hr bench times, storage at room temperature or -80°C) on fluorophore labelling in heparinized fresh human buffy coat and whole blood biobank samples during the mRNA work-up protocol for microarray analysis. Subsequently, we removed heparin by lithium chloride (LiCl) treatment and performed a quality control analysis of LiCl-treated biobank sample microarrays to prove their suitability for downstream data analysis. Both fresh and biobank samples experienced varying degrees of heparin-induced inhibition of fluorophore labelling, making most samples unusable for microarray analysis. RNA derived from EDTA and citrate blood was not inhibited. No effect of bench time was observed but room temperature storage gave slightly better results. Strong correlations were observed between original blood sample RNA yield and the amount of synthesized cRNA. LiCl treatment restored sample quality to normal standards in both fresh and biobank samples and the previously identified correlations disappeared. Microarrays hybridized with LiCl-treated biobank samples were of excellent quality with no identifiable influence of heparin. We conclude that, to obtain high quality results, in most cases heparin removal is essential in blood-derived RNA samples intended for microarray analysis. Copyright © 2014 Wiley Periodicals, Inc.

  16. Validation of the Andon KD-575 automated blood pressure monitor according to the European society of hypertension international protocol in adults.

    Science.gov (United States)

    Pan, Feng; Wan, Yi; Chen, Wan; Li, Ni; Tong, Wei; Xu, Yongyong

    2010-06-01

    Blood pressure measuring devices should be subjected to an independent evaluation of accuracy before clinical use. The results of the validation of the Andon KD-575 automated blood pressure monitor according to the European Society of Hypertension International Protocol in adults are presented in this paper. Sequential same-arm measurements of systolic and diastolic blood pressure were obtained in 33 participants using the mercury sphygmomanometer (two observers) and the test device (one supervisor). According to the validation protocol, 99 pairs of test device and reference blood pressure measurements were obtained during the two phases of the study (three pairs for each of the 33 participants). In phase 1, the Andon KD-575 device produced 26, 38 and 43 measurements within 5, 10 and 15 mmHg, respectively, for systolic blood pressure and 35, 44 and 45 mmHg, respectively, for diastolic blood pressure. However, in phase 2, the test device failed to meet the criteria for systolic blood pressure but comfortably passed all of the criteria for diastolic blood pressure with a mean (standard deviation) device-observer difference of 2.00+/-7.12 mmHg for systolic and 1.47+/-4.71 mmHg for diastolic blood pressure. The Andon KD-575 automated blood pressure monitor cannot be recommended for clinical use in an adult population because it records systolic blood pressure inaccurately.

  17. Stability of heparin blood samples during transport based on defined pre-analytical quality goals

    DEFF Research Database (Denmark)

    Jensen, Esther A; Stahl, Marta; Brandslund, Ivan

    2008-01-01

    two periods (winter and summer). Transport conditions (mail, courier pick-up, or brought to hospital by public coach), storage time, storage temperature and centrifugation requirements were different in the two counties. Results were tested for deviation from a "0-sample", the blood sample taken......, centrifuged and separated at the doctor's office within 45-60 min. This sample was considered as the best estimate of a comparison value. RESULTS: The pre-set quality goals were fulfilled for all the investigated components for samples transported to hospital by courier either as whole blood or as "on gel...... whole blood if the above mentioned conditions are met. There is no need for centrifugation in the primary sector. Neither mailing of samples with plasma "on gel" nor public transport by coach bus fulfil our analytical goals....

  18. Lab on valve-multisyringe flow injection system (LOV-MSFIA) for fully automated uranium determination in environmental samples.

    Science.gov (United States)

    Avivar, Jessica; Ferrer, Laura; Casas, Montserrat; Cerdà, Víctor

    2011-06-15

    The hyphenation of lab-on-valve (LOV) and multisyringe flow analysis (MSFIA), coupled to a long path length liquid waveguide capillary cell (LWCC), allows the spectrophotometric determination of uranium in different types of environmental sample matrices, without any manual pre-treatment, and achieving high selectivity and sensitivity levels. On-line separation and preconcentration of uranium is carried out by means of UTEVA resin. The potential of the LOV-MSFIA makes possible the fully automation of the system by the in-line regeneration of the column. After elution, uranium(VI) is spectrophotometrically detected after reaction with arsenazo-III. The determination of levels of uranium present in environmental samples is required in order to establish an environmental control. Thus, we propose a rapid, cheap and fully automated method to determine uranium(VI) in environmental samples. The limit of detection reached is 1.9 ηg of uranium and depending on the preconcentrated volume; it results in ppt levels (10.3 ηg L(-1)). Different water sample matrices (seawater, well water, freshwater, tap water and mineral water) and a phosphogypsum sample (with natural uranium content) were satisfactorily analyzed. Copyright © 2010 Elsevier B.V. All rights reserved.

  19. Simplifying sample pretreatment: application of dried blood spot (DBS) method to blood samples, including postmortem, for UHPLC-MS/MS analysis of drugs of abuse.

    Science.gov (United States)

    Odoardi, Sara; Anzillotti, Luca; Strano-Rossi, Sabina

    2014-10-01

    The complexity of biological matrices, such as blood, requires the development of suitably selective and reliable sample pretreatment procedures prior to their instrumental analysis. A method has been developed for the analysis of drugs of abuse and their metabolites from different chemical classes (opiates, methadone, fentanyl and analogues, cocaine, amphetamines and amphetamine-like substances, ketamine, LSD) in human blood using dried blood spot (DBS) and subsequent UHPLC-MS/MS analysis. DBS extraction required only 100μL of sample, added with the internal standards and then three droplets (30μL each) of this solution were spotted on the card, let dry for 1h, punched and extracted with methanol with 0.1% of formic acid. The supernatant was evaporated and the residue was then reconstituted in 100μL of water with 0.1% of formic acid and injected in the UHPLC-MS/MS system. The method was validated considering the following parameters: LOD and LOQ, linearity, precision, accuracy, matrix effect and dilution integrity. LODs were 0.05-1ng/mL and LOQs were 0.2-2ng/mL. The method showed satisfactory linearity for all substances, with determination coefficients always higher than 0.99. Intra and inter day precision, accuracy, matrix effect and dilution integrity were acceptable for all the studied substances. The addition of internal standards before DBS extraction and the deposition of a fixed volume of blood on the filter cards ensured the accurate quantification of the analytes. The validated method was then applied to authentic postmortem blood samples. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  20. Effects of anesthesia and blood sampling techniques on plasma metabolites and corticosterone in the rat.

    Science.gov (United States)

    Arnold, Myrtha; Langhans, Wolfgang

    2010-04-19

    Blood is routinely sampled from laboratory animals in biomedical research, and many of the commonly applied sampling techniques require anesthesia. Acute effects of many sampling and anesthesia procedures may confound the results, but those effects are incompletely characterized. We here compare the effects of four common anesthesia procedures (inhalation anesthesia with ether (EA) or isoflurane (IA) and intraperitoneal injection anesthesia with xylazin/ketamine (XKA) or medetomidine/midazolam/fentanyl (MMFA)) on plasma concentrations of glucose, lactate, non-esterified fatty acids (NEFAs), and corticosterone in blood obtained from a previously implanted jugular vein (JV) catheter with the effect of JV blood sampling from non-anesthetized, freely-moving rats (JV-NA). Also, we included in the comparison two other blood sampling procedures usually performed without anesthesia (NA), i.e., puncture of the saphenic vein (SV) and tail incision (TI). Whereas the control procedure (JV-NA) did not significantly affect any of the target parameters, plasma glucose increased from 14 (JV-IA) to 44 (JV-MMFA) % (all Ps=0.05 when compared with the control procedure) in all blood samples collected in anesthesia and was 12 and 14% lower (both Psprocedures except for JV-XKA and JV-MMF. Plasma NEFAs increased to 52% (Pprocedure and appeared to decrease with the JV-IA and JV-MMFA procedures (both Ps>0.05). Finally, only the JV-EA and the JV-MMFA procedures increased plasma corticosterone (+525 and +353%, respectively, both Psprocedures appeared to increase it as well, but these differences did not reach statistical significance. Thus, anesthesia and blood sampling procedures can have profound acute effects on plasma metabolite and hormone concentrations. This must be considered for the design and interpretation of blood sampling experiments in laboratory animals. (c) 2010. Published by Elsevier Inc.

  1. The prevalence of toxoplasmosis in Imam Reza Hospital blood bank samples, Tehran, Iran.

    Science.gov (United States)

    Shaddel, M; Mirzaii Dizgah, I; Sharif, F

    2014-10-01

    The prevalence of toxoplasma gondii (T.g) infection in blood donors has been poorly studied. The aim of this study was to assess the prevalence of acute and chronic toxoplasmosis in blood products. A total of 223 blood products (101 fresh frozen plasma (FFP) and 122 packed cells (PC)) in Imam Reza hospital blood bank, Tehran, Iran were tested for specific T.g antibodies (IgG and IgM) by ELISA method. Positive IgG anti-T.g samples were further tested for IgM anti-T.g. A positive IgG test with the negative and positive IgM test was interpreted as a chronic and acute toxoplasmosis respectively. Of 223 samples 38.6% and 0.45% were positive for IgG anti-T.g and IgM anti-T.g levels respectively. Therefore, one and 85 samples were involved acute and chronic toxoplasmosis respectively. Twenty-six of fresh frozen plasma samples were positive for IgG anti-T.g and one of them was positive for IgM anti-T.g. Sixty packed cell samples were positive for IgG anti-T.g. Our study showed that there were chronic and acute toxoplasmosis in blood products and the prevalence of toxoplasmosis especially chronic form was high. Therefore screening of blood for T.g antibodies may be considered. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Perfluoroalkyl Acid Concentrations in Blood Samples Subjected to Transportation and Processing Delay.

    Science.gov (United States)

    Bach, Cathrine Carlsen; Henriksen, Tine Brink; Bossi, Rossana; Bech, Bodil Hammer; Fuglsang, Jens; Olsen, Jørn; Nohr, Ellen Aagaard

    2015-01-01

    In studies of perfluoroalkyl acids, the validity and comparability of measured concentrations may be affected by differences in the handling of biospecimens. We aimed to investigate whether measured plasma levels of perfluoroalkyl acids differed between blood samples subjected to delay and transportation prior to processing and samples with immediate processing and freezing. Pregnant women recruited at Aarhus University Hospital, Denmark, (n = 88) provided paired blood samples. For each pair of samples, one was immediately processed and plasma was frozen, and the other was delayed and transported as whole blood before processing and freezing of plasma (similar to the Danish National Birth Cohort). We measured 12 perfluoroalkyl acids and present results for compounds with more than 50% of samples above the lower limit of quantification. For samples taken in the winter, relative differences between the paired samples ranged between -77 and +38% for individual perfluoroalkyl acids. In most cases concentrations were lower in the delayed and transported samples, e.g. the relative difference was -29% (95% confidence interval -30; -27) for perfluorooctane sulfonate. For perfluorooctanoate there was no difference between the two setups [corresponding estimate 1% (0, 3)]. Differences were negligible in the summer for all compounds. Transport of blood samples and processing delay, similar to conditions applied in some large, population-based studies, may affect measured perfluoroalkyl acid concentrations, mainly when outdoor temperatures are low. Attention to processing conditions is needed in studies of perfluoroalkyl acid exposure in humans.

  3. Feasibility of self-sampled dried blood spot and saliva samples sent by mail in a population-based study.

    Science.gov (United States)

    Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

    2015-04-11

    In large epidemiological studies it is often challenging to obtain biological samples. Self-sampling by study participants using dried blood spots (DBS) technique has been suggested to overcome this challenge. DBS is a type of biosampling where blood samples are obtained by a finger-prick lancet, blotted and dried on filter paper. However, the feasibility and efficacy of collecting DBS samples from study participants in large-scale epidemiological studies is not known. The aim of the present study was to test the feasibility and response rate of collecting self-sampled DBS and saliva samples in a population-based study of women above 50 years of age. We determined response proportions, number of phone calls to the study center with questions about sampling, and quality of the DBS. We recruited women through a study conducted within the Norwegian Breast Cancer Screening Program. Invitations, instructions and materials were sent to 4,597 women. The data collection took place over a 3 month period in the spring of 2009. Response proportions for the collection of DBS and saliva samples were 71.0% (3,263) and 70.9% (3,258), respectively. We received 312 phone calls (7% of the 4,597 women) with questions regarding sampling. Of the 3,263 individuals that returned DBS cards, 3,038 (93.1%) had been packaged and shipped according to instructions. A total of 3,032 DBS samples were sufficient for at least one biomarker analysis (i.e. 92.9% of DBS samples received by the laboratory). 2,418 (74.1%) of the DBS cards received by the laboratory were filled with blood according to the instructions (i.e. 10 completely filled spots with up to 7 punches per spot for up to 70 separate analyses). To assess the quality of the samples, we selected and measured two biomarkers (carotenoids and vitamin D). The biomarker levels were consistent with previous reports. Collecting self-sampled DBS and saliva samples through the postal services provides a low cost, effective and feasible

  4. Feasibility of self-sampled dried blood spot and saliva samples sent by mail in a population-based study

    International Nuclear Information System (INIS)

    Sakhi, Amrit Kaur; Bastani, Nasser Ezzatkhah; Ellingjord-Dale, Merete; Gundersen, Thomas Erik; Blomhoff, Rune; Ursin, Giske

    2015-01-01

    In large epidemiological studies it is often challenging to obtain biological samples. Self-sampling by study participants using dried blood spots (DBS) technique has been suggested to overcome this challenge. DBS is a type of biosampling where blood samples are obtained by a finger-prick lancet, blotted and dried on filter paper. However, the feasibility and efficacy of collecting DBS samples from study participants in large-scale epidemiological studies is not known. The aim of the present study was to test the feasibility and response rate of collecting self-sampled DBS and saliva samples in a population–based study of women above 50 years of age. We determined response proportions, number of phone calls to the study center with questions about sampling, and quality of the DBS. We recruited women through a study conducted within the Norwegian Breast Cancer Screening Program. Invitations, instructions and materials were sent to 4,597 women. The data collection took place over a 3 month period in the spring of 2009. Response proportions for the collection of DBS and saliva samples were 71.0% (3,263) and 70.9% (3,258), respectively. We received 312 phone calls (7% of the 4,597 women) with questions regarding sampling. Of the 3,263 individuals that returned DBS cards, 3,038 (93.1%) had been packaged and shipped according to instructions. A total of 3,032 DBS samples were sufficient for at least one biomarker analysis (i.e. 92.9% of DBS samples received by the laboratory). 2,418 (74.1%) of the DBS cards received by the laboratory were filled with blood according to the instructions (i.e. 10 completely filled spots with up to 7 punches per spot for up to 70 separate analyses). To assess the quality of the samples, we selected and measured two biomarkers (carotenoids and vitamin D). The biomarker levels were consistent with previous reports. Collecting self-sampled DBS and saliva samples through the postal services provides a low cost, effective and feasible

  5. A simple method for measurement of cerebral blood flow using 123I-IMP SPECT with calibrated standard input function by one point blood sampling. Validation of calibration by one point venous blood sampling as a substitute for arterial blood sampling

    International Nuclear Information System (INIS)

    Ito, Hiroshi; Akaizawa, Takashi; Goto, Ryoui

    1994-01-01

    In a simplified method for measurement of cerebral blood flow using one 123 I-IMP SPECT scan and one point arterial blood sampling (Autoradiography method), input function is obtained by calibrating a standard input function by one point arterial blood sampling. A purpose of this study is validation of calibration by one point venous blood sampling as a substitute for one point arterial blood sampling. After intravenous infusion of 123 I-IMP, frequent arterial and venous blood sampling were simultaneously performed on 12 patients of CNS disease without any heart and lung disease and 5 normal volunteers. The radioactivity ratio of venous whole blood which obtained from cutaneous cubital vein to arterial whole blood were 0.76±0.08, 0.80±0.05, 0.81±0.06, 0.83±0.11 at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivities were always 20% lower than those of arterial blood radioactivity during 50 min. However, the ratio which obtained from cutaneous dorsal hand vein to artery were 0.93±0.02, 0.94±0.05, 0.98±0.04, 0.98±0.03, at 10, 20, 30, 50 min after 123 I-IMP infusion, respectively. The venous blood radioactivity was consistent with artery. These indicate that arterio-venous difference of radioactivity in a peripheral cutaneous vein like a dorsal hand vein is minimal due to arteriovenous shunt in palm. Therefore, a substitution by blood sampling from cutaneous dorsal hand vein for artery will be possible. Optimized time for venous blood sampling evaluated by error analysis was 20 min after 123 I-IMP infusion, which is 10 min later than that of arterial blood sampling. (author)

  6. Automated Glycan Assembly of Complex Oligosaccharides Related to Blood Group Determinants.

    Science.gov (United States)

    Hahm, Heung Sik; Liang, Chien-Fu; Lai, Chian-Hui; Fair, Richard J; Schuhmacher, Frank; Seeberger, Peter H

    2016-07-15

    Lactotetraosyl (Lc4) and neo-lactotetraosyl (nLc4) are backbones that are common to many glycans. Using automated glycan assembly, these common core structures were constructed and elaborated to access synthetically challenging glycans of biological relevance. The incorporation of α-fucoses is demonstrated for H-type I and II; α(1,3)-galactose epitopes were prepared, and the pentasaccharide HNK-1 required incorporation of a 3-O-sulfate. In addition to preparing the target structures, essential insights were gained regarding the relationships of glycosylating agents and nucleophiles as well as the linker stability.

  7. Maternal red blood cell alloantibodies identified in blood samples obtained from Iranian pregnant women: the first population study in Iran.

    Science.gov (United States)

    Shahverdi, Ehsan; Moghaddam, Mostafa; Gorzin, Fateme

    2017-01-01

    The objective was to determine the frequency of occurrence of alloantibodies among pregnant women in Iran. This was a prospective cross-sectional study, which was carried out in the immunohematology reference laboratory of the Iranian Blood Transfusion Organization in Tehran, Iran, in 2008 to 2015. Screening and identification of red blood cell (RBC) alloantibodies was done on the sera of 7340 pregnant females using the standard tube method and gel column agglutination technique. Alloantibodies were identified in the serum of 332 of the 7340 (4.5%) pregnant women. A total of 410 antibodies were detected in 332 positive maternal serum samples with no previous history of blood transfusion. Anti-D was the most common antibody accounting for 70.5% of all the antibodies formed in D- women. The incidence of specific alloimmunization other than Rh group was 14.4%. We concluded that the alloimmunization rate was high in comparison with wide pattern in previous studies. In Iran, like other developing countries, alloimmunization screening tests are performed only to detect anti-D in pregnant D- women. This high rate of alloimmunization, quite possibly, is due to the fact that the majority of blood samples came from pregnant women known to have previous obstetric problems. However, we suggest that RBC antibody screening tests should be extended to all D+ women. © 2016 AABB.

  8. Intraosseous blood samples for point-of-care analysis: agreement between intraosseous and arterial analyses.

    Science.gov (United States)

    Jousi, Milla; Saikko, Simo; Nurmi, Jouni

    2017-09-11

    Point-of-care (POC) testing is highly useful when treating critically ill patients. In case of difficult vascular access, the intraosseous (IO) route is commonly used, and blood is aspirated to confirm the correct position of the IO-needle. Thus, IO blood samples could be easily accessed for POC analyses in emergency situations. The aim of this study was to determine whether IO values agree sufficiently with arterial values to be used for clinical decision making. Two samples of IO blood were drawn from 31 healthy volunteers and compared with arterial samples. The samples were analysed for sodium, potassium, ionized calcium, glucose, haemoglobin, haematocrit, pH, blood gases, base excess, bicarbonate, and lactate using the i-STAT® POC device. Agreement and reliability were estimated by using the Bland-Altman method and intraclass correlation coefficient calculations. Good agreement was evident between the IO and arterial samples for pH, glucose, and lactate. Potassium levels were clearly higher in the IO samples than those from arterial blood. Base excess and bicarbonate were slightly higher, and sodium and ionised calcium values were slightly lower, in the IO samples compared with the arterial values. The blood gases in the IO samples were between arterial and venous values. Haemoglobin and haematocrit showed remarkable variation in agreement. POC diagnostics of IO blood can be a useful tool to guide treatment in critical emergency care. Seeking out the reversible causes of cardiac arrest or assessing the severity of shock are examples of situations in which obtaining vascular access and blood samples can be difficult, though information about the electrolytes, acid-base balance, and lactate could guide clinical decision making. The analysis of IO samples should though be limited to situations in which no other option is available, and the results should be interpreted with caution, because there is not yet enough scientific evidence regarding the agreement of IO

  9. Attended and Unattended Automated Office Blood Pressure Measurements Have Better Agreement With Ambulatory Monitoring Than Conventional Office Readings.

    Science.gov (United States)

    Andreadis, Emmanuel A; Geladari, Charalampia V; Angelopoulos, Epameinondas T; Savva, Florentia S; Georgantoni, Anna I; Papademetriou, Vasilios

    2018-04-07

    Automated office blood pressure (AOBP) measurement is superior to conventional office blood pressure (OBP) because it eliminates the "white coat effect" and shows a strong association with ambulatory blood pressure. We conducted a cross-sectional study in 146 participants with office hypertension, and we compared AOBP readings, taken with or without the presence of study personnel, before and after the conventional office readings to determine whether their variation in blood pressure showed a difference in blood pressure values. We also compared AOBP measurements with daytime ambulatory blood pressure monitoring and conventional office readings. The mean age of the studied population was 56±12 years, and 53.4% of participants were male. Bland-Altman analysis revealed a bias (ie, mean of the differences) of 0.6±6 mm Hg systolic for attended AOBP compared with unattended and 1.4±6 and 0.1±6 mm Hg bias for attended compared with unattended systolic AOBP when measurements were performed before and after conventional readings, respectively. A small bias was observed when unattended and attended systolic AOBP measurements were compared with daytime ambulatory blood pressure monitoring (1.3±13 and 0.6±13 mm Hg, respectively). Biases were higher for conventional OBP readings compared with unattended AOBP (-5.6±15 mm Hg for unattended AOBP and oscillometric OBP measured by a physician, -6.8±14 mm Hg for unattended AOBP and oscillometric OBP measured by a nurse, and -2.1±12 mm Hg for unattended AOBP and auscultatory OBP measured by a second physician). Our findings showed that independent of the presence or absence of medical staff, AOBP readings revealed similar values that were closer to daytime ambulatory blood pressure monitoring than conventional office readings, further supporting the use of AOBP in the clinical setting. © 2018 The Authors. Published on behalf of the American Heart Association, Inc., by Wiley.

  10. Plasmodium falciparum HRP2 ELISA for analysis of dried blood spot samples in rural Zambia.

    Science.gov (United States)

    Gibson, Lauren E; Markwalter, Christine F; Kimmel, Danielle W; Mudenda, Lwiindi; Mbambara, Saidon; Thuma, Philip E; Wright, David W

    2017-08-23

    Dried blood spots are commonly used for sample collection in clinical and non-clinical settings. This method is simple, and biomolecules in the samples remain stable for months at room temperature. In the field, blood samples for the study and diagnosis of malaria are often collected on dried blood spot cards, so development of a biomarker extraction and analysis method is needed. A simple extraction procedure for the malarial biomarker Plasmodium falciparum histidine-rich protein 2 (HRP2) from dried blood spots was optimized to achieve maximum extraction efficiency. This method was used to assess the stability of HRP2 in dried blood spots. Furthermore, 328 patient samples made available from rural Zambia were analysed for HRP2 using the developed method. These samples were collected at the initial administration of artemisinin-based combination therapy and at several points following treatment. An average extraction efficiency of 70% HRP2 with a low picomolar detection limit was achieved. In specific storage conditions HRP2 was found to be stable in dried blood spots for at least 6 months. Analysis of patient samples showed the method to have a sensitivity of 94% and a specificity of 89% when compared with microscopy, and trends in HRP2 clearance after treatment were observed. The dried blood spot ELISA for HRP2 was found to be sensitive, specific and accurate. The method was effectively used to assess biomarker clearance characteristics in patient samples, which prove it to be ideal for gaining further insight into the disease and epidemiological applications.

  11. Identification of clinical biomarkers for pre-analytical quality control of blood samples.

    Science.gov (United States)

    Kang, Hyun Ju; Jeon, Soon Young; Park, Jae-Sun; Yun, Ji Young; Kil, Han Na; Hong, Won Kyung; Lee, Mee-Hee; Kim, Jun-Woo; Jeon, Jae-Pil; Han, Bok Ghee

    2013-04-01

    Pre-analytical conditions are key factors in maintaining the high quality of biospecimens. They are necessary for accurate reproducibility of experiments in the field of biomarker discovery as well as achieving optimal specificity of laboratory tests for clinical diagnosis. In research at the National Biobank of Korea, we evaluated the impact of pre-analytical conditions on the stability of biobanked blood samples by measuring biochemical analytes commonly used in clinical laboratory tests. We measured 10 routine laboratory analytes in serum and plasma samples from healthy donors (n = 50) with a chemistry autoanalyzer (Hitachi 7600-110). The analyte measurements were made at different time courses based on delay of blood fractionation, freezing delay of fractionated serum and plasma samples, and at different cycles (0, 1, 3, 6, 9) of freeze-thawing. Statistically significant changes from the reference sample mean were determined using the repeated-measures ANOVA and the significant change limit (SCL). The serum levels of GGT and LDH were changed significantly depending on both the time interval between blood collection and fractionation and the time interval between fractionation and freezing of serum and plasma samples. The glucose level was most sensitive only to the elapsed time between blood collection and centrifugation for blood fractionation. Based on these findings, a simple formula (glucose decrease by 1.387 mg/dL per hour) was derived to estimate the length of time delay after blood collection. In addition, AST, BUN, GGT, and LDH showed sensitive responses to repeated freeze-thaw cycles of serum and plasma samples. These results suggest that GGT and LDH measurements can be used as quality control markers for certain pre-analytical conditions (eg, delayed processing or repeated freeze-thawing) of blood samples which are either directly used in the laboratory tests or stored for future research in the biobank.

  12. A content validated questionnaire for assessment of self reported venous blood sampling practices

    Directory of Open Access Journals (Sweden)

    Bölenius Karin

    2012-01-01

    Full Text Available Abstract Background Venous blood sampling is a common procedure in health care. It is strictly regulated by national and international guidelines. Deviations from guidelines due to human mistakes can cause patient harm. Validated questionnaires for health care personnel can be used to assess preventable "near misses"--i.e. potential errors and nonconformities during venous blood sampling practices that could transform into adverse events. However, no validated questionnaire that assesses nonconformities in venous blood sampling has previously been presented. The aim was to test a recently developed questionnaire in self reported venous blood sampling practices for validity and reliability. Findings We developed a questionnaire to assess deviations from best practices during venous blood sampling. The questionnaire contained questions about patient identification, test request management, test tube labeling, test tube handling, information search procedures and frequencies of error reporting. For content validity, the questionnaire was confirmed by experts on questionnaires and venous blood sampling. For reliability, test-retest statistics were used on the questionnaire answered twice. The final venous blood sampling questionnaire included 19 questions out of which 9 had in total 34 underlying items. It was found to have content validity. The test-retest analysis demonstrated that the items were generally stable. In total, 82% of the items fulfilled the reliability acceptance criteria. Conclusions The questionnaire could be used for assessment of "near miss" practices that could jeopardize patient safety and gives several benefits instead of assessing rare adverse events only. The higher frequencies of "near miss" practices allows for quantitative analysis of the effect of corrective interventions and to benchmark preanalytical quality not only at the laboratory/hospital level but also at the health care unit/hospital ward.

  13. Design and building of a homemade sample changer for automation of the irradiation in neutron activation analysis technique

    International Nuclear Information System (INIS)

    Gago, Javier; Hernandez, Yuri; Baltuano, Oscar; Bedregal, Patricia; Lopez, Yon; Urquizo, Rafael

    2014-01-01

    Because the RP-10 research reactor operates during weekends, it was necessary to design and build a sample changer for irradiation as part of the automation process of neutron activation analysis technique. The device is formed by an aluminum turntable disk which can accommodate 19 polyethylene capsules, containing samples to be sent using the pneumatic transfer system from the laboratory to the irradiation position. The system is operate by a control switchboard to send and return capsules in a variable preset time and by two different ways, allowing the determination of short, medium and long lived radionuclides. Also another mechanism is designed called 'exchange valve' for changing travel paths (pipelines) allowing the irradiated samples to be stored for a longer time in the reactor hall. The system design has allowed complete automation of this technique, enabling the irradiation of samples without the presence of an analyst. The design, construction and operation of the device is described and presented in this article. (authors).

  14. HIV-1 subtype and viral tropism determination for evaluating antiretroviral therapy options: an analysis of archived Kenyan blood samples

    Directory of Open Access Journals (Sweden)

    Lagat Nancy J

    2009-12-01

    Full Text Available Abstract Background Infection with HIV-1 is characterized by genetic diversity such that specific viral subtypes are predominant in specific geographical areas. The genetic variation in HIV-1 pol and env genes is responsible for rapid development of resistance to current drugs. This variation has influenced disease progression among the infected and necessitated the search for alternative drugs with novel targets. Though successfully used in developed countries, these novel drugs are still limited in resource-poor countries. The aim of this study was to determine HIV-1 subtypes, recombination, dual infections and viral tropism of HIV-1 among Kenyan patients prior to widespread use of antiretroviral drugs. Methods Remnant blood samples from consenting sexually transmitted infection (STI patients in Nairobi were collected between February and May 2001 and stored. Polymerase chain reaction and cloning of portions of HIV-1 gag, pol and env genes was carried out followed by automated DNA sequencing. Results Twenty HIV-1 positive samples (from 11 females and 9 males were analyzed. The average age of males (32.5 years and females (26.5 years was significantly different (p value Conclusion HIV-1 subtype A accounted for majority of the infections. Though perceived to be a high risk population, the prevalence of recombination in this sample was low with no dual infections detected. Genotypic co-receptor analysis showed that most patients harbored viruses that are predicted to use CCR5.

  15. Extended automated separation techniques in destructive neutron activation analysis; application to various biological materials, including human tissues and blood

    International Nuclear Information System (INIS)

    Tjioe, P.S.; Goeij, J.J.M. de; Houtman, J.P.W.

    1976-09-01

    Neutron activation analysis may be performed as a multi-element and low-level technique for many important trace elements in biological materials, provided that post-irradiation chemical separations are applied. This paper describes a chemical separation consisting of automated procedures for destruction, distillation, and anion-chromatography. The system developed enables the determination of 14 trace elements in biological materials, viz. antimony, arsenic, bromine, cadmium, chromium, cobalt, copper, gold, iron, mercury, molybdenum, nickel, selenium, and zinc. The aspects of sample preparation, neutron irradiation, gamma-spectrum evaluation, and blank-value contribution are also discussed

  16. Sampling methods to the statistical control of the production of blood components.

    Science.gov (United States)

    Pereira, Paulo; Seghatchian, Jerard; Caldeira, Beatriz; Santos, Paula; Castro, Rosa; Fernandes, Teresa; Xavier, Sandra; de Sousa, Gracinda; de Almeida E Sousa, João Paulo

    2017-12-01

    The control of blood components specifications is a requirement generalized in Europe by the European Commission Directives and in the US by the AABB standards. The use of a statistical process control methodology is recommended in the related literature, including the EDQM guideline. The control reliability is dependent of the sampling. However, a correct sampling methodology seems not to be systematically applied. Commonly, the sampling is intended to comply uniquely with the 1% specification to the produced blood components. Nevertheless, on a purely statistical viewpoint, this model could be argued not to be related to a consistent sampling technique. This could be a severe limitation to detect abnormal patterns and to assure that the production has a non-significant probability of producing nonconforming components. This article discusses what is happening in blood establishments. Three statistical methodologies are proposed: simple random sampling, sampling based on the proportion of a finite population, and sampling based on the inspection level. The empirical results demonstrate that these models are practicable in blood establishments contributing to the robustness of sampling and related statistical process control decisions for the purpose they are suggested for. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Analysis of multiple single nucleotide polymorphisms (SNP) on DNA traces from plasma and dried blood samples

    NARCIS (Netherlands)

    Catsburg, Arnold; van der Zwet, Wil C.; Morre, Servaas A.; Ouburg, Sander; Vandenbroucke-Grauls, Christina M. J. E.; Savelkoul, Paul H. M.

    2007-01-01

    Reliable analysis of single nucleotide polymorphisms (SNPs) in DNA derived from samples containing low numbers of cells or from suboptimal sources can be difficult. A new procedure to characterize multiple SNPs in traces of DNA from plasma and old dried blood samples was developed. Six SNPs in the

  18. Measurement of cerebral blood flow the blood sampling method using 99mTc-ECD. Simultaneous scintigram scanning of arterial blood samples and the brain with a gamma camera

    International Nuclear Information System (INIS)

    Hachiya, Takenori; Inugami, Atsushi; Iida, Hidehiro; Mizuta, Yoshihiko; Kawakami, Takeshi; Inoue, Minoru

    1999-01-01

    To measure regional cerebral blood flow (rCBF) by blood sampling using 99m Tc-ECD we devised a method of measuring the radioactive concentration in arterial blood sample with a gamma camera. In this method the head and a blood sample are placed within the same visual field to record the SPECT data of both specimens simultaneously. The results of an evaluation of the counting rate performance, applying the 30 hours decaying method using 99m Tc solution showed that this method is not comparable to the well-type scintillation counter and in clinical cases the active concentration in arterial blood sample remained well within the dynamic range. In addition, examination of the influence of scattered radiation from the brain by the dilution method showed that it was negligible at a distance of more than 7.5 cm between the brain and the arterial blood sample. In the present study we placed a head-shaped phantom next to the sample. The results of the examinations suggested that this method is suitable for clinical application, and because it does not require a well-type scintillation counter, it is expected to find wide application. (author)

  19. Paper membrane-based SERS platform for the determination of glucose in blood samples.

    Science.gov (United States)

    Torul, Hilal; Çiftçi, Hakan; Çetin, Demet; Suludere, Zekiye; Boyacı, Ismail Hakkı; Tamer, Uğur

    2015-11-01

    In this report, we present a paper membrane-based surface-enhanced Raman scattering (SERS) platform for the determination of blood glucose level using a nitrocellulose membrane as substrate paper, and the microfluidic channel was simply constructed by wax-printing method. The rod-shaped gold nanorod particles were modified with 4-mercaptophenylboronic acid (4-MBA) and 1-decanethiol (1-DT) molecules and used as embedded SERS probe for paper-based microfluidics. The SERS measurement area was simply constructed by dropping gold nanoparticles on nitrocellulose membrane, and the blood sample was dropped on the membrane hydrophilic channel. While the blood cells and proteins were held on nitrocellulose membrane, glucose molecules were moved through the channel toward the SERS measurement area. Scanning electron microscopy (SEM) was used to confirm the effective separation of blood matrix, and total analysis is completed in 5 min. In SERS measurements, the intensity of the band at 1070 cm(-1) which is attributed to B-OH vibration decreased depending on the rise in glucose concentration in the blood sample. The glucose concentration was found to be 5.43 ± 0.51 mM in the reference blood sample by using a calibration equation, and the certified value for glucose was 6.17 ± 0.11 mM. The recovery of the glucose in the reference blood sample was about 88 %. According to these results, the developed paper-based microfluidic SERS platform has been found to be suitable for use for the detection of glucose in blood samples without any pretreatment procedure. We believe that paper-based microfluidic systems may provide a wide field of usage for paper-based applications.

  20. Multispectral Imaging Analysis of Circulating Tumor Cells in Negatively Enriched Peripheral Blood Samples.

    Science.gov (United States)

    Miller, Brandon; Lustberg, Maryam; Summers, Thomas A; Chalmers, Jeffrey J

    2017-01-01

    A variety of biomarkers are present on cells in peripheral blood of patients with a variety of disorders, including solid tumor malignancies. While rare, characterization of these cells for specific protein levels with the advanced technology proposed, will lead to future validation studies of blood samples as "liquid biopsies" for the evaluation of disease status and therapeutic response. While circulating tumor cells (CTCs) have been isolated in the blood samples of patients with solid tumors, the exact role of CTCs as clinically useful predictive markers is still debated. Current commercial technology has significant bias in that a positive selection technology is used that preassumes specific cell surface markers (such as EpCAM) are present on CTCs. However, CTCs with low EpCAM expression have been experimentally demonstrated to be more likely to be missed by this method. In contrast, this application uses a previously developed, technology that performs a purely negative enrichment methodology on peripheral blood, yielding highly enriched blood samples that contain CTCs as well as other, undefined cell types. The focus of this contribution is the use of multispectral imaging of epifluorescent, microscopic images of these enriched cells in order to help develop clinically relevant liquid biopsies from peripheral blood samples.

  1. The effectiveness of cooling conditions on temperature of canine EDTA whole blood samples

    Directory of Open Access Journals (Sweden)

    Karen M. Tobias

    2016-11-01

    Full Text Available Background Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Methods Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia. Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Results Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature, while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice

  2. The effectiveness of cooling conditions on temperature of canine EDTA whole blood samples.

    Science.gov (United States)

    Tobias, Karen M; Serrano, Leslie; Sun, Xiaocun; Flatland, Bente

    2016-01-01

    Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV) of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia). Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature), while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice. Canine EDTA whole blood samples cool most

  3. Noise contamination from PET blood sampling pump: Effects on structural MRI image quality in simultaneous PET/MR studies.

    Science.gov (United States)

    Bartlett, Elizabeth; DeLorenzo, Christine; Parsey, Ramin; Huang, Chuan

    2018-02-01

    To fully quantify PET imaging outcome measures, a blood sampling pump is often used during the PET acquisition. With simultaneous PET/MR studies, a structural magnetization-prepared rapid gradient-echo (MP-RAGE) may also be acquired while the pump is generating electromagnetic noise. This study investigated whether this noise contamination would be detrimental to the quantification of volume and cortical thickness measures obtained from automated segmentation of the MP-RAGE image. MP-RAGE T1w structural images were acquired for a phantom and 10 healthy volunteers (five female, 27.2 ± 5.1 y old) with the blood sampling pump and without. The white matter signal-to-noise ratio (SNR) was computed for all images. Region-wise cortical thickness and volume were extracted with Freesurfer 5.3.0. The phantom SNR and the white matter human subject SNR was degraded in the MP-RAGE images acquired with the pump (P = 0.005; white matter SNR: 43.9 and 50.8 with the pump and without). Intrasession, region-wise volume and cortical thickness estimates were significantly overestimated with the pump (percent difference: 1.14 ± 2.67% for volume (P = 0.0003) and 0.34 ± 1.59% (P = 0.02) for cortical thickness). Regions with percent differences greater than 5% between pump conditions were those close to tissue-air interfaces: entorhinal, frontal pole, parsorbitalis, temporal pole, and medial orbitofrontal. Synthetically adding Gaussian noise to the without pump MP-RAGE images yielded similar, significant detriments to cortical morphometry compared to without the pump. This study provides evidence that the use of PET blood sampling pumps may generate unstructured, Gaussian-distributed noise in MP-RAGE images that significantly alters the accuracy of Freesurfer-derived volume and cortical thickness estimates. While many cortical regions showed a percent difference of less than 1% with the pump, regions close to tissue-air interfaces, subject to larger susceptibility artifacts

  4. Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia.

    Science.gov (United States)

    Nordentoft, Merete; Larsen, Janne Tidselbak; Pedersen, Carsten Bøcker; Sørensen, Holger Jelling; Hollegaard, Mads Villiam; Hougaard, David Michael; Mortensen, Preben Bo; Petersen, Liselotte

    2015-03-01

    The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have found that early and late blood sampling, compared to sampling at day 5, was associated with increased risk of schizophrenia. As delay in sampling of blood for neonatal screening cannot in itself influence the risk of schizophrenia, it must be seen as a proxy for unknown underlying causes responsible for this association. Therefore, we investigated whether the increased risk can be explained by other risk factors for schizophrenia. A case-control design was applied. A total of 846 cases with schizophrenia were selected from the Danish Psychiatric Case Register. One control was selected for each case, matched on sex and exact date of birth. Both early and late blood sampling was associated with increased risk for schizophrenia. Compared to blood sampling at day 5, sampling at days 0 to 4 after birth was associated with an incidence rate ratio (IRR) of 1.46 (95% CI 1.15-1.87) for development of schizophrenia, and sampling at days 6 to 9 and at days 10 to 53 was associated with an IRR of 1.5 (95% CI 1.13-1.98) and 3.00 (95% CI 1.59-5.67), respectively. After adjusting the estimates for place of birth, both parents' psychiatric illness, maternal and paternal age, parents' country of origin, child admission, and parental education and income, the estimates were slightly different. Thus, blood collection at 0-4days was associated with an IRR of 1.27 (95% CI 0.94-1.71), 6-9days 1.31 (95% CI 0.94-1.84) and 10+days 3.52 (95% CI 1.50 to 8.24). After adjusting risk estimates for well-known risk factors, delay in sampling of blood for neonatal screening was associated with unexplained increased risk of schizophrenia. Thus, a key finding is that age at test is a proxy for unobserved risk factors

  5. Stability and reliability of glycated haemoglobin measurements in blood samples stored at -20°C.

    Science.gov (United States)

    Venkataraman, Vijayachandrika; Anjana, Ranjit Mohan; Pradeepa, Rajendra; Deepa, Mohan; Jayashri, Ramamoorthy; Anbalagan, Viknesh Prabu; Akila, Bridgitte; Madhu, Sri Venkata; Lakshmy, Ramakrishnan; Mohan, Viswanathan

    2016-01-01

    To validate the stability of glycated haemoglobin (HbA1c) measurements in blood samples stored at -20°C for up to one month. The study group comprised 142 type 2 diabetic subjects visiting a tertiary centre for diabetes at Chennai city in south India. The HbA1c assay was done on a fasting blood sample using the Bio-Rad Variant machine on Day 0 (day of blood sample collection). Several aliquots were stored at -20°C and the assay was repeated on the 3rd, 7th, 15th, and 30th day after the sample collection. Bland-Altman plots were constructed and variation in the HbA1c levels on the different days was compared with the day 0 level. The median differences between HbA1c levels measured on Day 0 and the 3rd, 7th, 15th, and 30th day after blood collection were 0.0%, 0.2%, 0.3% and 0.5% respectively. Bland-Altman plot analysis showed that the differences between the day '0' and the different time points tend to get larger with time, but these were not clinically significant. HbA1c levels are relatively stable up to 2weeks, if blood samples are stored at -20°C. Copyright © 2016 Elsevier Inc. All rights reserved.

  6. A simplified method for determination of radioactive iron in whole-blood samples

    DEFF Research Database (Denmark)

    Bukhave, Klaus; Sørensen, Anne Dorthe; Hansen, M.

    2001-01-01

    in humans. The overall recovery of radioiron from blood is more than 90%, and the coefficient of variation, as judged by the variation in the ratio Fe-55/Fe-59 is in the order of 4%. Combined with whole-body counting of 59Fe and direct gamma -counting of Fe-59 on blood samples, this method represents......For studies on iron absorption in man radioisotopes represent an easy and simple tool. However, measurement of the orbital electron emitting radioiron, Fe-55, in blood is difficult and insufficiently described in the literature. The present study describes a relatively simple method...... for simultaneous determination of Fe-55 and Fe-59 in blood, using a dry-ashing procedure and recrystallization of the remaining iron. The detection Limit of the method permits measurements of 0.1 Bq/ml blood thus allowing detection of Less than 1% absorption from a 40 kBq dose, which is ethically acceptable...

  7. Development and validation of an automated liquid-liquid extraction GC/MS method for the determination of THC, 11-OH-THC, and free THC-carboxylic acid (THC-COOH) from blood serum.

    Science.gov (United States)

    Purschke, Kirsten; Heinl, Sonja; Lerch, Oliver; Erdmann, Freidoon; Veit, Florian

    2016-06-01

    The analysis of Δ(9)-tetrahydrocannabinol (THC) and its metabolites 11-hydroxy-Δ(9)-tetrahydrocannabinol (11-OH-THC), and 11-nor-9-carboxy-Δ(9)-tetrahydrocannabinol (THC-COOH) from blood serum is a routine task in forensic toxicology laboratories. For examination of consumption habits, the concentration of the phase I metabolite THC-COOH is used. Recommendations for interpretation of analysis values in medical-psychological assessments (regranting of driver's licenses, Germany) include threshold values for the free, unconjugated THC-COOH. Using a fully automated two-step liquid-liquid extraction, THC, 11-OH-THC, and free, unconjugated THC-COOH were extracted from blood serum, silylated with N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), and analyzed by GC/MS. The automation was carried out by an x-y-z sample robot equipped with modules for shaking, centrifugation, and solvent evaporation. This method was based on a previously developed manual sample preparation method. Validation guidelines of the Society of Toxicological and Forensic Chemistry (GTFCh) were fulfilled for both methods, at which the focus of this article is the automated one. Limits of detection and quantification for THC were 0.3 and 0.6 μg/L, for 11-OH-THC were 0.1 and 0.8 μg/L, and for THC-COOH were 0.3 and 1.1 μg/L, when extracting only 0.5 mL of blood serum. Therefore, the required limit of quantification for THC of 1 μg/L in driving under the influence of cannabis cases in Germany (and other countries) can be reached and the method can be employed in that context. Real and external control samples were analyzed, and a round robin test was passed successfully. To date, the method is employed in the Institute of Legal Medicine in Giessen, Germany, in daily routine. Automation helps in avoiding errors during sample preparation and reduces the workload of the laboratory personnel. Due to its flexibility, the analysis system can be employed for other liquid-liquid extractions as

  8. EXPERIMENTS TOWARDS DETERMINING BEST TRAINING SAMPLE SIZE FOR AUTOMATED EVALUATION OF DESCRIPTIVE ANSWERS THROUGH SEQUENTIAL MINIMAL OPTIMIZATION

    Directory of Open Access Journals (Sweden)

    Sunil Kumar C

    2014-01-01

    Full Text Available With number of students growing each year there is a strong need to automate systems capable of evaluating descriptive answers. Unfortunately, there aren’t many systems capable of performing this task. In this paper, we use a machine learning tool called LightSIDE to accomplish auto evaluation and scoring of descriptive answers. Our experiments are designed to cater to our primary goal of identifying the optimum training sample size so as to get optimum auto scoring. Besides the technical overview and the experiments design, the paper also covers challenges, benefits of the system. We also discussed interdisciplinary areas for future research on this topic.

  9. High-frequency, long-duration water sampling in acid mine drainage studies: a short review of current methods and recent advances in automated water samplers

    Science.gov (United States)

    Chapin, Thomas

    2015-01-01

    Hand-collected grab samples are the most common water sampling method but using grab sampling to monitor temporally variable aquatic processes such as diel metal cycling or episodic events is rarely feasible or cost-effective. Currently available automated samplers are a proven, widely used technology and typically collect up to 24 samples during a deployment. However, these automated samplers are not well suited for long-term sampling in remote areas or in freezing conditions. There is a critical need for low-cost, long-duration, high-frequency water sampling technology to improve our understanding of the geochemical response to temporally variable processes. This review article will examine recent developments in automated water sampler technology and utilize selected field data from acid mine drainage studies to illustrate the utility of high-frequency, long-duration water sampling.

  10. A nonlethal sampling method to obtain, generate and assemble whole blood transcriptomes from small, wild mammals.

    Science.gov (United States)

    Huang, Zixia; Gallot, Aurore; Lao, Nga T; Puechmaille, Sébastien J; Foley, Nicole M; Jebb, David; Bekaert, Michaël; Teeling, Emma C

    2016-01-01

    The acquisition of tissue samples from wild populations is a constant challenge in conservation biology, especially for endangered species and protected species where nonlethal sampling is the only option. Whole blood has been suggested as a nonlethal sample type that contains a high percentage of bodywide and genomewide transcripts and therefore can be used to assess the transcriptional status of an individual, and to infer a high percentage of the genome. However, only limited quantities of blood can be nonlethally sampled from small species and it is not known if enough genetic material is contained in only a few drops of blood, which represents the upper limit of sample collection for some small species. In this study, we developed a nonlethal sampling method, the laboratory protocols and a bioinformatic pipeline to sequence and assemble the whole blood transcriptome, using Illumina RNA-Seq, from wild greater mouse-eared bats (Myotis myotis). For optimal results, both ribosomal and globin RNAs must be removed before library construction. Treatment of DNase is recommended but not required enabling the use of smaller amounts of starting RNA. A large proportion of protein-coding genes (61%) in the genome were expressed in the blood transcriptome, comparable to brain (65%), kidney (63%) and liver (58%) transcriptomes, and up to 99% of the mitogenome (excluding D-loop) was recovered in the RNA-Seq data. In conclusion, this nonlethal blood sampling method provides an opportunity for a genomewide transcriptomic study of small, endangered or critically protected species, without sacrificing any individuals. © 2015 John Wiley & Sons Ltd.

  11. From pioneering to implementing automated blood pressure measurement in clinical practice: Thomas Pickering's legacy

    DEFF Research Database (Denmark)

    Stolarz-Skrzypek, Katarzyna; Thijs, Lutgarde; Wizner, Barbara

    2010-01-01

    Thomas G. Pickering spent most of his scientific career in carrying out research on clinical hypertension and blood pressure (BP) measurement. In our review of Pickering's seminal work, we first focused on white-coat hypertension and masked hypertension, two terms that he had introduced. Next, we...

  12. A large-capacity sample-changer for automated gamma-ray spectroscopy

    International Nuclear Information System (INIS)

    Andeweg, A.H.

    1980-01-01

    An automatic sample-changer has been developed at the National Institute for Metallurgy for use in gamma-ray spectroscopy with a lithium-drifted germanium detector. The sample-changer features remote storage, which prevents cross-talk and reduces background. It has a capacity for 200 samples and a sample container that takes liquid or solid samples. The rotation and vibration of samples during counting ensure that powdered samples are compacted, and improve the precision and reproducibility of the counting geometry [af

  13. Sample registration software for process automation in the Neutron Activation Analysis (NAA) Facility in Malaysia nuclear agency

    Energy Technology Data Exchange (ETDEWEB)

    Rahman, Nur Aira Abd, E-mail: nur-aira@nuclearmalaysia.gov.my; Yussup, Nolida; Ibrahim, Maslina Bt. Mohd; Mokhtar, Mukhlis B.; Soh Shaari, Syirrazie Bin Che; Azman, Azraf B. [Technical Support Division, Malaysian Nuclear Agency, 43000, Kajang, Selangor (Malaysia); Salim, Nazaratul Ashifa Bt. Abdullah [Division of Waste and Environmental Technology, Malaysian Nuclear Agency, 43000, Kajang, Selangor (Malaysia); Ismail, Nadiah Binti [Fakulti Kejuruteraan Elektrik, UiTM Pulau Pinang, 13500 Permatang Pauh, Pulau Pinang (Malaysia)

    2015-04-29

    Neutron Activation Analysis (NAA) had been established in Nuclear Malaysia since 1980s. Most of the procedures established were done manually including sample registration. The samples were recorded manually in a logbook and given ID number. Then all samples, standards, SRM and blank were recorded on the irradiation vial and several forms prior to irradiation. These manual procedures carried out by the NAA laboratory personnel were time consuming and not efficient. Sample registration software is developed as part of IAEA/CRP project on ‘Development of Process Automation in the Neutron Activation Analysis (NAA) Facility in Malaysia Nuclear Agency (RC17399)’. The objective of the project is to create a pc-based data entry software during sample preparation stage. This is an effective method to replace redundant manual data entries that needs to be completed by laboratory personnel. The software developed will automatically generate sample code for each sample in one batch, create printable registration forms for administration purpose, and store selected parameters that will be passed to sample analysis program. The software is developed by using National Instruments Labview 8.6.

  14. A comparison of various methods of blood sampling in mice and rats: Effects on animal welfare

    DEFF Research Database (Denmark)

    Harikrishnan, Vs; Hansen, Axel K; Abelson, Klas Sp

    2018-01-01

    -puncture activity and anxiety levels of rats and mice were measured using an elevated plus maze test and an open field test. Stress levels 24 h post-puncture were assessed by analysing faecal corticosteroid metabolites. Sucrose intake and faecal corticosteroid levels were not affected by the blood sampling...... procedures. Rats showed reduced activity in the open field test and an increased level of anxiety in the elevated plus maze test following retrobulbar plexus puncture and isoflurane anaesthesia. In mice, nest building activity was affected in all the groups compared with the control group, except for animals......This study was conducted to investigate the effects of blood sampling on animal welfare in a total of 60 NTac:SD rats and 72 C57BL/6NTac mice of both sexes. Blood was sampled either by sublingual vein puncture, tail vein puncture or by retrobulbar plexus/sinus puncture under light isoflurane...

  15. Radioimmunoassay screening and GC/MS confirmation of whole blood samples for drugs of abuse

    Energy Technology Data Exchange (ETDEWEB)

    Spiehler, V.R.; Sedgwick, P.

    From 1981 to 1984, an average of 300 radioimmunoassay screens on whole blood were performed each week in the authors laboratory. Most samples were screened for opiates phencyclidine and its analogs, barbiturates, and cocaine or its metabolite benzoylecgonine. A commercially available radioimmunoassay was used with modifications to facilitate screening of whole blood. Increasing sample size increased the sensitivity of the assay. Changing reagent concentration (1:1 dilution), incubation time, sample matrix (water, urine, or blood), or fraction counted (precipitate or supernatant) did not affect the utility of the standard curve or the sensitivity of the assay. All positive results for phencyclidine, opiates, cocaine, and related compounds were confirmed by GC/MA. Barbiturate positives were confirmed by UV spectrophotometry.

  16. Genome-wide scans using archived neonatal dried blood spot samples

    Directory of Open Access Journals (Sweden)

    Wiuf Carsten

    2009-07-01

    Full Text Available Abstract Background Identification of disease susceptible genes requires access to DNA from numerous well-characterised subjects. Archived residual dried blood spot samples from national newborn screening programs may provide DNA from entire populations and medical registries the corresponding clinical information. The amount of DNA available in these samples is however rarely sufficient for reliable genome-wide scans, and whole-genome amplification may thus be necessary. This study assess the quality of DNA obtained from different amplification protocols by evaluating fidelity and robustness of the genotyping of 610,000 single nucleotide polymorphisms, using the Illumina Infinium HD Human610-Quad BeadChip. Whole-genome amplified DNA from 24 neonatal dried blood spot samples stored between 15 to 25 years was tested, and high-quality genomic DNA from 8 of the same individuals was used as reference. Results Using 3.2 mm disks from dried blood spot samples the optimal DNA-extraction and amplification protocol resulted in call-rates between 99.15% – 99.73% (mean 99.56%, N = 16, and conflicts with reference DNA in only three per 10,000 genotype calls. Conclusion Whole-genome amplified DNA from archived neonatal dried blood spot samples can be used for reliable genome-wide scans and is a cost-efficient alternative to collecting new samples.

  17. Identifying the potential of changes to blood sample logistics using simulation.

    Science.gov (United States)

    Jørgensen, Pelle; Jacobsen, Peter; Poulsen, Jørgen Hjelm

    2013-01-01

    Using simulation as an approach to display and improve internal logistics at hospitals has great potential. This study shows how a simulation model displaying the morning blood-taking round at a Danish public hospital can be developed and utilized with the aim of improving the logistics. The focus of the simulation was to evaluate changes made to the transportation of blood samples between wards and the laboratory. The average- (AWT) and maximum waiting time (MWT) from a blood sample was drawn at the ward until it was received at the laboratory, and the distribution of arrivals of blood samples in the laboratory were used as the evaluation criteria. Four different scenarios were tested and compared with the current approach: (1) Using AGVs (mobile robots), (2) using a pneumatic tube system, (3) using porters that are called upon, or (4) using porters that come to the wards every 45 minutes. Furthermore, each of the scenarios was tested in terms of what amount of resources would give the optimal result. The simulations showed a big improvement potential in implementing a new technology/mean for transporting the blood samples. The pneumatic tube system showed the biggest potential lowering the AWT and MWT with approx. 36% and 18%, respectively. Additionally, all of the scenarios had a more even distribution of arrivals except for porters coming to the wards every 45 min. As a consequence of the results obtained in the study, the hospital decided to implement a pneumatic tube system.

  18. Rapid microbial sample preparation from blood using a novel concentration device.

    Directory of Open Access Journals (Sweden)

    Anna K Boardman

    Full Text Available Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1-100 CFU/mL, traditionally requiring growth (blood culture or amplification (e.g., PCR for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components. Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms' viability, giving a 30-μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40-50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSAto prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample.

  19. Hybrid image and blood sampling input function for quantification of small animal dynamic PET data

    International Nuclear Information System (INIS)

    Shoghi, Kooresh I.; Welch, Michael J.

    2007-01-01

    We describe and validate a hybrid image and blood sampling (HIBS) method to derive the input function for quantification of microPET mice data. The HIBS algorithm derives the peak of the input function from the image, which is corrected for recovery, while the tail is derived from 5 to 6 optimally placed blood sampling points. A Bezier interpolation algorithm is used to link the rightmost image peak data point to the leftmost blood sampling point. To assess the performance of HIBS, 4 mice underwent 60-min microPET imaging sessions following a 0.40-0.50-mCi bolus administration of 18 FDG. In total, 21 blood samples (blood-sampled plasma time-activity curve, bsPTAC) were obtained throughout the imaging session to compare against the proposed HIBS method. MicroPET images were reconstructed using filtered back projection with a zoom of 2.75 on the heart. Volumetric regions of interest (ROIs) were composed by drawing circular ROIs 3 pixels in diameter on 3-4 transverse planes of the left ventricle. Performance was characterized by kinetic simulations in terms of bias in parameter estimates when bsPTAC and HIBS are used as input functions. The peak of the bsPTAC curve was distorted in comparison to the HIBS-derived curve due to temporal limitations and delay in blood sampling, which affected the rates of bidirectional exchange between plasma and tissue. The results highlight limitations in using bsPTAC. The HIBS method, however, yields consistent results, and thus, is a substitute for bsPTAC

  20. Sources of Pre-Analytical Variations in Yield of DNA Extracted from Blood Samples: Analysis of 50,000 DNA Samples in EPIC

    Science.gov (United States)

    Caboux, Elodie; Lallemand, Christophe; Ferro, Gilles; Hémon, Bertrand; Mendy, Maimuna; Biessy, Carine; Sims, Matt; Wareham, Nick; Britten, Abigail; Boland, Anne; Hutchinson, Amy; Siddiq, Afshan; Vineis, Paolo; Riboli, Elio; Romieu, Isabelle; Rinaldi, Sabina; Gunter, Marc J.; Peeters, Petra H. M.; van der Schouw, Yvonne T.; Travis, Ruth; Bueno-de-Mesquita, H. Bas; Canzian, Federico; Sánchez, Maria-José; Skeie, Guri; Olsen, Karina Standahl; Lund, Eiliv; Bilbao, Roberto; Sala, Núria; Barricarte, Aurelio; Palli, Domenico; Navarro, Carmen; Panico, Salvatore; Redondo, Maria Luisa; Polidoro, Silvia; Dossus, Laure; Boutron-Ruault, Marie Christine; Clavel-Chapelon, Françoise; Trichopoulou, Antonia; Trichopoulos, Dimitrios; Lagiou, Pagona; Boeing, Heiner; Fisher, Eva; Tumino, Rosario; Agnoli, Claudia; Hainaut, Pierre

    2012-01-01

    The European Prospective Investigation into Cancer and nutrition (EPIC) is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS) and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88%) performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies. PMID:22808065

  1. Sources of pre-analytical variations in yield of DNA extracted from blood samples: analysis of 50,000 DNA samples in EPIC.

    Directory of Open Access Journals (Sweden)

    Elodie Caboux

    Full Text Available The European Prospective Investigation into Cancer and nutrition (EPIC is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88% performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies.

  2. Pneumatic tube-transported blood samples in lithium heparinate gel separator tubes may be more susceptible to haemolysis than blood samples in serum tubes.

    Science.gov (United States)

    Böckel-Frohnhöfer, Nicole; Hübner, Ulrich; Hummel, Björn; Geisel, Jürgen

    2014-10-01

    Pneumatic tube systems are widely used in hospitals. Advantages are high speed and rapid availability of the samples. However, the transportation by pneumatic tube promotes haemolysis. Haemolysis interferes with many spectrophotometric assays and is a common problem in clinical laboratories. The haemolysis index (HI) as a semi-quantitative representation of the level of haemolysis was compared in unpaired tube-transported and hand-delivered routine lithium heparinate plasma samples (n = 1368 and n = 837, respectively). Additionally, the HI distribution was measured in lithium heparinate plasma samples with a HI above the threshold value of 20 and in paired serum samples after transportation by pneumatic tube system. HI values above 20 can interfere with the selected assays: Creatine kinase (CK), creatine kinase-MB (CK-MB) and alanine aminotransferase (ALT) activities. These parameters were determined to demonstrate how haemolysis affects the results. 17.5% of the tube-transported plasma samples and 2.6% of the hand-delivered plasma samples had a HI above 20. The median HI in pneumatic tube-transported lithium heparinate plasma was 85 and 33 in the paired serum samples. The median HI difference between paired plasma and serum was 46. Blood samples in lithium heparinate tubes may be substantially more susceptible to haemolysis by pneumatic tube transportation than serum tube samples. Although our results cannot be universally applied to laboratories with different pneumatic tube systems, it is recommended that each laboratory evaluate carefully the degree of haemolysis after the transportation by the own pneumatic tube system and in terms of the sample type.

  3. Large-scale subject-specific cerebral arterial tree modeling using automated parametric mesh generation for blood flow simulation.

    Science.gov (United States)

    Ghaffari, Mahsa; Tangen, Kevin; Alaraj, Ali; Du, Xinjian; Charbel, Fady T; Linninger, Andreas A

    2017-12-01

    In this paper, we present a novel technique for automatic parametric mesh generation of subject-specific cerebral arterial trees. This technique generates high-quality and anatomically accurate computational meshes for fast blood flow simulations extending the scope of 3D vascular modeling to a large portion of cerebral arterial trees. For this purpose, a parametric meshing procedure was developed to automatically decompose the vascular skeleton, extract geometric features and generate hexahedral meshes using a body-fitted coordinate system that optimally follows the vascular network topology. To validate the anatomical accuracy of the reconstructed vasculature, we performed statistical analysis to quantify the alignment between parametric meshes and raw vascular images using receiver operating characteristic curve. Geometric accuracy evaluation showed an agreement with area under the curves value of 0.87 between the constructed mesh and raw MRA data sets. Parametric meshing yielded on-average, 36.6% and 21.7% orthogonal and equiangular skew quality improvement over the unstructured tetrahedral meshes. The parametric meshing and processing pipeline constitutes an automated technique to reconstruct and simulate blood flow throughout a large portion of the cerebral arterial tree down to the level of pial vessels. This study is the first step towards fast large-scale subject-specific hemodynamic analysis for clinical applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

  4. Application of gelatin zymography for evaluating low levels of contaminating neutrophils in red blood cell samples.

    Science.gov (United States)

    Achilli, Cesare; Ciana, Annarita; Balduini, Cesare; Risso, Angela; Minetti, Giampaolo

    2011-02-15

    Supposedly "homogeneous" red blood cell (RBC) samples are commonly obtained by "washing" whole blood free of plasma, platelets, and white cells with physiological solutions, a procedure that does not result, however, in sufficient removal of polymorphonuclear neutrophils (PMNs), leading to possible artifactual results. Pure RBC samples can be obtained only by leukodepletion procedures. Proposed here is a version of gelatin zymography adapted to detect matrix metalloproteinase 9 (MMP-9), selectively expressed by PMNs, in heterogeneous mixtures of RBCs and PMNs that can reveal contamination at levels as low as 1 PMN/10⁶ RBCs. Copyright © 2010 Elsevier Inc. All rights reserved.

  5. Eosinophilia in routine blood samples as a biomarker for solid tumor development

    DEFF Research Database (Denmark)

    Andersen, Christen Bertel L; Siersma, V.D.; Hasselbalch, H.C.

    2014-01-01

    eosinophilia in routine blood samples as a potential biomarker of solid tumor development in a prospective design. MATERIAL AND METHODS: From the Copenhagen Primary Care Differential Count (CopDiff) Database, we identified 356 196 individuals with at least one differential cell count (DIFF) encompassing...... was increased with mild eosinophilia [OR 1.93 (CI 1.29-2.89), p = 0.0013]. No associations with eosinophilia were observed for the remaining solid cancers. CONCLUSION: We demonstrate that eosinophilia in routine blood samples associates with an increased risk of bladder cancer. Our data emphasize...

  6. Blood venous sample collection: Recommendations overview and a checklist to improve quality.

    Science.gov (United States)

    Giavarina, Davide; Lippi, Giuseppe

    2017-07-01

    The extra-analytical phases of the total testing process have substantial impact on managed care, as well as an inherent high risk of vulnerability to errors which is often greater than that of the analytical phase. The collection of biological samples is a crucial preanalytical activity. Problems or errors occurring shortly before, or soon after, this preanalytical step may impair sample quality and characteristics, or else modify the final results of testing. The standardization of fasting requirements, rest, patient position and psychological state of the patient are therefore crucial for mitigating the impact of preanalytical variability. Moreover, the quality of materials used for collecting specimens, along with their compatibility, can guarantee sample quality and persistence of chemical and physical characteristics of the analytes over time, so safeguarding the reliability of testing. Appropriate techniques and sampling procedures are effective to prevent problems such as hemolysis, undue clotting in the blood tube, draw of insufficient sample volume and modification of analyte concentration. An accurate identification of both patient and blood samples is a key priority as for other healthcare activities. Good laboratory practice and appropriate training of operators, by specifically targeting collection of biological samples, blood in particular, may greatly improve this issue, thus lowering the risk of errors and their adverse clinical consequences. The implementation of a simple and rapid check-list, including verification of blood collection devices, patient preparation and sampling techniques, was found to be effective for enhancing sample quality and reducing some preanalytical errors associated with these procedures. The use of this tool, along with implementation of objective and standardized systems for detecting non-conformities related to unsuitable samples, can be helpful for standardizing preanalytical activities and improving the quality of

  7. Client-server architecture applied to system automation radioactivity sampling in atmosphere

    Science.gov (United States)

    Hubbard, C. W.; McKinnon, A. D.

    1997-06-01

    Control software for an automated particulate air sampler is described. The software is divided into a number of small, cooperating server processes, each of which is responsible for the control of a particular device or subsystem. For each process, an effort was made to isolate the details of the underlying device or subsystem from the server interface. This made it possible to change the hardware without making changes to any of the server's client processes. A single supervisor process is responsible for overall system control. The design of the control algorithm was facilitated by employing a state machine model. Such a model is easy to study, easy to modify, and provides a clear understanding of the control mechanism to programmers and non-programmers alike. A state machine library was developed which greatly eased the task of implementing the design and ensured that the control algorithm detailed by the state machine model was the same algorithm that was actually employed.

  8. Preanalytical aspects and sample quality assessment in metabolomics studies of human blood.

    Science.gov (United States)

    Yin, Peiyuan; Peter, Andreas; Franken, Holger; Zhao, Xinjie; Neukamm, Sabine S; Rosenbaum, Lars; Lucio, Marianna; Zell, Andreas; Häring, Hans-Ulrich; Xu, Guowang; Lehmann, Rainer

    2013-05-01

    Metabolomics is a powerful tool that is increasingly used in clinical research. Although excellent sample quality is essential, it can easily be compromised by undetected preanalytical errors. We set out to identify critical preanalytical steps and biomarkers that reflect preanalytical inaccuracies. We systematically investigated the effects of preanalytical variables (blood collection tubes, hemolysis, temperature and time before further processing, and number of freeze-thaw cycles) on metabolomics studies of clinical blood and plasma samples using a nontargeted LC-MS approach. Serum and heparinate blood collection tubes led to chemical noise in the mass spectra. Distinct, significant changes of 64 features in the EDTA-plasma metabolome were detected when blood was exposed to room temperature for 2, 4, 8, and 24 h. The resulting pattern was characterized by increases in hypoxanthine and sphingosine 1-phosphate (800% and 380%, respectively, at 2 h). In contrast, the plasma metabolome was stable for up to 4 h when EDTA blood samples were immediately placed in iced water. Hemolysis also caused numerous changes in the metabolic profile. Unexpectedly, up to 4 freeze-thaw cycles only slightly changed the EDTA-plasma metabolome, but increased the individual variability. Nontargeted metabolomics investigations led to the following recommendations for the preanalytical phase: test the blood collection tubes, avoid hemolysis, place whole blood immediately in ice water, use EDTA plasma, and preferably use nonrefrozen biobank samples. To exclude outliers due to preanalytical errors, inspect the biomarker signal intensities reflecting systematic as well as accidental and preanalytical inaccuracies before processing the bioinformatics data. © 2013 American Association for Clinical Chemistry.

  9. Comparative evaluation of blood and serum samples in rapid immunochromatographic tests for visceral leishmaniasis.

    Science.gov (United States)

    Kumar, Dinesh; Khanal, Basudha; Tiwary, Puja; Mudavath, Shyam Lal; Tiwary, Narendra K; Singh, Rupa; Koirala, Kanika; Boelaert, Marleen; Rijal, Suman; Sundar, Shyam

    2013-12-01

    Rapid diagnostic tests (RDTs) based on the detection of specific antibodies in serum are commonly used for the diagnosis of visceral leishmaniasis (VL). Several commercial kits are available, and some of them allow the use of whole-blood samples instead of serum. An RDT is much more user-friendly for blood samples than for serum samples. In this study, we examined the sensitivities and specificities of six different commercially available immunochromatographic tests for their accuracy in detecting Leishmania infection in whole blood and serum of parasitologically confirmed VL cases. This study was performed in areas of India and Nepal where VL is endemic. A total of 177 confirmed VL cases, 208 healthy controls from areas of endemicity (EHCs), 26 malaria patients (MP), and 37 tuberculosis (TB) patients were enrolled. The reproducibilities of the blood and serum results and between-reader and between-laboratory results were tested. In India, the sensitivities of all the RDTs ranged between 94.7 and 100.0%, with no significant differences between whole blood and serum. The specificities ranged between 92.4 and 100.0%, except for the specificity of the Onsite Leishmania Ab RevB kit, which was lower (33.6 to 42.0%). No differences in specificities were observed for blood and serum. In Nepal, the sensitivities of all the test kits, for whole-blood as well as serum samples, ranged between 96.3 and 100.0%, and the specificities ranged between 90.1 and 96.1%, again with the exception of that of the Onsite Leishmania Ab RevB test, which was markedly lower (48.7 to 49.3%). The diagnostic accuracies of all the tests, except for one brand, were excellent for the whole-blood and serum samples. We conclude that whole blood is an adequate alternative for serum in RDTs for VL, with sensitivities and specificities comparable to those obtained in serum samples, provided that the test kit is of overall good quality.

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

    Science.gov (United States)

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

    2016-05-06

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

  11. Automated method for simultaneous lead and strontium isotopic analysis applied to rainwater samples and airborne particulate filters (PM10).

    Science.gov (United States)

    Beltrán, Blanca; Avivar, Jessica; Mola, Montserrat; Ferrer, Laura; Cerdà, Víctor; Leal, Luz O

    2013-09-03

    A new automated, sensitive, and fast system for the simultaneous online isolation and preconcentration of lead and strontium by sorption on a microcolumn packed with Sr-resin using an inductively coupled plasma mass spectrometry (ICP-MS) detector was developed, hyphenating lab-on-valve (LOV) and multisyringe flow injection analysis (MSFIA). Pb and Sr are directly retained on the sorbent column and eluted with a solution of 0.05 mol L(-1) ammonium oxalate. The detection limits achieved were 0.04 ng for lead and 0.03 ng for strontium. Mass calibration curves were used since the proposed system allows the use of different sample volumes for preconcentration. Mass linear working ranges were between 0.13 and 50 ng and 0.1 and 50 ng for lead and strontium, respectively. The repeatability of the method, expressed as RSD, was 2.1% and 2.7% for Pb and Sr, respectively. Environmental samples such as rainwater and airborne particulate (PM10) filters as well as a certified reference material SLRS-4 (river water) were satisfactorily analyzed obtaining recoveries between 90 and 110% for both elements. The main features of the LOV-MSFIA-ICP-MS system proposed are the capability to renew solid phase extraction at will in a fully automated way, the remarkable stability of the column which can be reused up to 160 times, and the potential to perform isotopic analysis.

  12. 1-Hydroxypyrene Levels in Blood Samples of Rats After Exposure to Generator Fumes

    Science.gov (United States)

    Ifegwu, Clinton; Igwo-Ezikpe, Miriam N.; Anyakora, Chimezie; Osuntoki, Akinniyi; Oseni, Kafayat A.; Alao, Eragbae O.

    2013-01-01

    Polynuclear Aromatic Hydrocarbons (PAHs) are a major component of fuel generator fumes. Carcinogenicity of these compounds has long been established. In this study, 37 Swiss albino rats were exposed to generator fumes at varied distances for 8 hours per day for a period of 42 days and the level of 1-hydroxypyrene in their blood was evaluated. This study also tried to correlate the level of blood 1-hyroxypyrene with the distance from the source of pollution. Plasma was collected by centrifuging the whole blood sample followed by complete hydrolysis of the conjugated 1-hydroxypyrene glucuronide to yield the analyte of interest, 1-hydroxypyrene, which was achieved using beta glucuronidase. High performance liquid chromatography (HPLC) with UV detector was used to determine the 1-hydroxypyrene concentrations in the blood samples. The mobile phase was water:methanol (12:88 v/v) isocratic run at the flow rate of 1.2 mL/min with CI8 stationary phase at 250 nm. After 42 days of exposure, blood concentration level of 1-hydroxypyrene ranged from 34 μg/mL to 26.29 μg/mL depending on the distance from source of exposure. The control group had no 1-hydroxypyrene in their blood. After the period of exposure, percentage of death correlated with the distance from the source of exposure. Percentage of death ranged from 56% to zero depending on the proximity to source of pollution. PMID:24179393

  13. Surveillance cultures of samples obtained from biopsy channels and automated endoscope reprocessors after high-level disinfection of gastrointestinal endoscopes

    Directory of Open Access Journals (Sweden)

    Chiu King-Wah

    2012-09-01

    Full Text Available Abstract Background The instrument channels of gastrointestinal (GI endoscopes may be heavily contaminated with bacteria even after high-level disinfection (HLD. The British Society of Gastroenterology guidelines emphasize the benefits of manually brushing endoscope channels and using automated endoscope reprocessors (AERs for disinfecting endoscopes. In this study, we aimed to assess the effectiveness of decontamination using reprocessors after HLD by comparing the cultured samples obtained from biopsy channels (BCs of GI endoscopes and the internal surfaces of AERs. Methods We conducted a 5-year prospective study. Every month random consecutive sampling was carried out after a complete reprocessing cycle; 420 rinse and swabs samples were collected from BCs and internal surface of AERs, respectively. Of the 420 rinse samples collected from the BC of the GI endoscopes, 300 were obtained from the BCs of gastroscopes and 120 from BCs of colonoscopes. Samples were collected by flushing the BCs with sterile distilled water, and swabbing the residual water from the AERs after reprocessing. These samples were cultured to detect the presence of aerobic and anaerobic bacteria and mycobacteria. Results The number of culture-positive samples obtained from BCs (13.6%, 57/420 was significantly higher than that obtained from AERs (1.7%, 7/420. In addition, the number of culture-positive samples obtained from the BCs of gastroscopes (10.7%, 32/300 and colonoscopes (20.8%, 25/120 were significantly higher than that obtained from AER reprocess to gastroscopes (2.0%, 6/300 and AER reprocess to colonoscopes (0.8%, 1/120. Conclusions Culturing rinse samples obtained from BCs provides a better indication of the effectiveness of the decontamination of GI endoscopes after HLD than culturing the swab samples obtained from the inner surfaces of AERs as the swab samples only indicate whether the AERs are free from microbial contamination or not.

  14. An integrative pharmacological approach to radio telemetry and blood sampling in pharmaceutical drug discovery and safety assessment

    Directory of Open Access Journals (Sweden)

    Kamendi Harriet W

    2011-01-01

    Full Text Available Abstract Background A successful integration of the automated blood sampling (ABS and telemetry (ABST system is described. The new ABST system facilitates concomitant collection of physiological variables with blood and urine samples for determination of drug concentrations and other biochemical measures in the same rat without handling artifact. Method Integration was achieved by designing a 13 inch circular receiving antenna that operates as a plug-in replacement for the existing pair of DSI's orthogonal antennas which is compatible with the rotating cage and open floor design of the BASi Culex® ABS system. The circular receiving antenna's electrical configuration consists of a pair of electrically orthogonal half-toroids that reinforce reception of a dipole transmitter operating within the coil's interior while reducing both external noise pickup and interference from other adjacent dipole transmitters. Results For validation, measured baclofen concentration (ABST vs. satellite (μM: 69.6 ± 23.8 vs. 76.6 ± 19.5, p = NS and mean arterial pressure (ABST vs. traditional DSI telemetry (mm Hg: 150 ± 5 vs.147 ± 4, p = NS variables were quantitatively and qualitatively similar between rats housed in the ABST system and traditional home cage approaches. Conclusion The ABST system offers unique advantages over traditional between-group study paradigms that include improved data quality and significantly reduced animal use. The superior within-group model facilitates assessment of multiple physiological and biochemical responses to test compounds in the same animal. The ABST also provides opportunities to evaluate temporal relations between parameters and to investigate anomalous outlier events because drug concentrations, physiological and biochemical measures for each animal are available for comparisons.

  15. Electronic remote blood issue combined with a computer-controlled, automated refrigerator for major surgery in operating theatres at a distance from the transfusion service.

    Science.gov (United States)

    Verlicchi, Franco; Pacilli, Pasqua; Bragliani, Arianna; Rapuano, Silvia; Dini, Daniele; Vincenzi, Daniele

    2018-02-01

    The difficulty of supplying red blood cells within an adequate time to patients undergoing surgery is a known problem for transfusion services, particularly if the operating theater is located at some distance from the blood bank. The consequences frequently are that more blood is ordered than required; several units are allocated and issued; and unused units must be returned to the blood bank. Some sparse reports have demonstrated that remote blood issue systems can improve the efficiency of issuing blood. This study describes a computer-controlled, self-service, remote blood-release system, combined with an automated refrigerator, installed in a hospital at which major surgery was performed, located 5 kilometers away from the transfusion service. With this system, red blood cell units were electronically allocated to patients immediately before release, when the units actually were needed. Two 2-year periods, before and after implementation of the system, were compared. After implementation of the system, the ratio of red blood cell units returned to the transfusion service was reduced from 48.9% to 1.6% of the issued units (8852 of 18,090 vs. 182 of 11,152 units; p blood cell units was observed, probably mainly due to changes in the number and complexity of surgical procedures. No transfusion errors occurred in the two periods. The current results demonstrate that the remote blood-release system is safe and useful for improving the efficiency of blood issue for patients in remote operating theatres. © 2017 AABB.

  16. Detection of Merkel Cell Polyomavirus DNA in Serum Samples of Healthy Blood Donors

    Directory of Open Access Journals (Sweden)

    Elisa Mazzoni

    2017-11-01

    Full Text Available Merkel cell polyomavirus (MCPyV has been detected in 80% of Merkel cell carcinomas (MCC. In the host, the MCPyV reservoir remains elusive. MCPyV DNA sequences were revealed in blood donor buffy coats. In this study, MCPyV DNA sequences were investigated in the sera (n = 190 of healthy blood donors. Two MCPyV DNA sequences, coding for the viral oncoprotein large T antigen (LT, were investigated using polymerase chain reaction (PCR methods and DNA sequencing. Circulating MCPyV sequences were detected in sera with a prevalence of 2.6% (5/190, at low-DNA viral load, which is in the range of 1–4 and 1–5 copies/μl by real-time PCR and droplet digital PCR, respectively. DNA sequencing carried out in the five MCPyV-positive samples indicated that the two MCPyV LT sequences which were analyzed belong to the MKL-1 strain. Circulating MCPyV LT sequences are present in blood donor sera. MCPyV-positive samples from blood donors could represent a potential vehicle for MCPyV infection in receivers, whereas an increase in viral load may occur with multiple blood transfusions. In certain patient conditions, such as immune-depression/suppression, additional disease or old age, transfusion of MCPyV-positive samples could be an additional risk factor for MCC onset.

  17. Detection of the BLV provirus from nasal secretion and saliva samples using BLV-CoCoMo-qPCR-2: Comparison with blood samples from the same cattle.

    Science.gov (United States)

    Yuan, Yuan; Kitamura-Muramatsu, Yuri; Saito, Susumu; Ishizaki, Hiroshi; Nakano, Miwa; Haga, Satoshi; Matoba, Kazuhiro; Ohno, Ayumu; Murakami, Hironobu; Takeshima, Shin-Nosuke; Aida, Yoko

    2015-12-02

    Bovine leukemia virus (BLV) induces enzootic bovine leukosis, which is the most common neoplastic disease in cattle. Sero-epidemiological studies show that BLV infection occurs worldwide. Direct contact between infected and uninfected cattle is thought to be one of the risk factors for BLV transmission. Contact transmission occurs via a mixture of natural sources, blood, and exudates. To confirm that BLV provirus is detectable in these samples, matched blood, nasal secretion, and saliva samples were collected from 50 cattle, and genomic DNA was extracted. BLV-CoCoMo-qPCR-2, an assay developed for the highly sensitive detection of BLV, was then used to measure the proviral load in blood (n=50), nasal secretions (n=48), and saliva (n=47) samples. The results showed that 35 blood samples, 14 nasal secretion samples, and 6 saliva samples were positive for the BLV provirus. Matched blood samples from cattle that were positive for the BLV provirus (either in nasal secretion or saliva samples) were also positive in their blood. The proviral load in the positive blood samples was >14,000 (copies/1×10(5) cells). Thus, even though the proviral load in the nasal secretion and saliva samples was much lower (blood, prolonged direct contact between infected and healthy cattle may be considered as a risk factor for BLV transmission. Copyright © 2015 Elsevier B.V. All rights reserved.

  18. Large-scale prospective T cell function assays in shipped, unfrozen blood samples

    DEFF Research Database (Denmark)

    Hadley, David; Cheung, Roy K; Becker, Dorothy J

    2014-01-01

    , for measuring core T cell functions. The Trial to Reduce Insulin-dependent diabetes mellitus in the Genetically at Risk (TRIGR) type 1 diabetes prevention trial used consecutive measurements of T cell proliferative responses in prospectively collected fresh heparinized blood samples shipped by courier within...... cell immunocompetence. We have found that the vast majority of the samples were viable up to 3 days from the blood draw, yet meaningful responses were found in a proportion of those with longer travel times. Furthermore, the shipping time of uncooled samples significantly decreased both the viabilities...... North America. In this article, we report on the quality control implications of this simple and pragmatic shipping practice and the interpretation of positive- and negative-control analytes in our assay. We used polyclonal and postvaccination responses in 4,919 samples to analyze the development of T...

  19. Possibilities of radioisotope renal clearances without blood sampling in patients with chronic glomerulonephritis

    International Nuclear Information System (INIS)

    Shejretova, E.; Velikov, P.

    1987-01-01

    Two clearance methods are proposed: 1) Effective renal plasma flow determination with 131 J-hippuran, and 2) glomeruli filtration determination with 169 Yb-EDTA. Thirty one adult patients with chronic glomerulonephritis and 59 without clinical and laboratory evidence of urinary tract disease comprised the study group. Statistically significant decrease in the effective renal plasma flow (p<0,001) and of glomerular filtration (p<0,01) was recorded in comparison to the control group, which is an evidence of the good diagnostic possibilities of the methods. The significant difference between the group of patients with elevated blood urea levels or pathologic Zimnincki's test and reduced glomerular filtration, compared to those with normal glomerular filtration, is an indication for the high diagnostic value of the glomerular filtration method. The clearance methods without blood sampling are particularly useful in the diagnosis and follow-up of patients with chronic glomerulonephritis, because of the burdenless technique, lack of blood sampling, and sufficient accuracy

  20. The effect of delayed transportation of blood samples on serum bilirubin values in neonates.

    Science.gov (United States)

    Saththasivam, Poovendran; Voralu, Kirtanaa; Ramli, Noraida; Mustapha, Mohd Rafi; Omar, Julia; Van Rostenberghe, Hans

    2010-07-01

    Delays in transporting blood samples may cause inaccurate results. Samples may be exposed to light or heat during delays, resulting in the degradation of analytes, for example, bilirubin. This study was done to determine the effect of delays in the transportation of blood samples on serum bilirubin test results. Samples taken from neonates admitted to a tertiary hospital with jaundice were included in the study. The samples were collected through venipuncture in 3 labelled containers. The first container was sent immediately to the laboratory, while the second and third containers were sent after being kept in the ward for 1 and 3 hours, respectively. Bilirubin values were measured colourimetrically at a wavelength of 578 nm using a Roche Hitachi 912 Chemistry Analyser upon arrival in the laboratory. A total of 36 serum samples were studied. The mean of the indirect bilirubin measurements for 0-, 1-, and 3-hour samples were 174 (SD 68.65), 186.97 (SD 60.47), and 184.56 (SD 66.93), respectively. There was a significant difference in the mean indirect bilirubin measurement of 1-hour samples (P = 0.047, 95% CI -24.66 to -1.18) and 3-hour samples (P = 0.045, 95% CI -19.77 to -0.23) compared with 0-hour samples. There were no significant differences observed in either the mean total bilirubin or the mean direct bilirubin measurements of different time intervals. This study confirms that delays in the transportation of blood samples influence the bilirubin test results.

  1. Comparison of three methods for recovery of Brucella canis DNA from canine blood samples.

    Science.gov (United States)

    Batinga, Maria Cryskely A; Dos Santos, Jaíne C; Lima, Julia T R; Bigotto, Maria Fernanda D; Muner, Kerstin; Faita, Thalita; Soares, Rodrigo M; da Silva, David A V; Oliveira, Trícia M F S; Ferreira, Helena L; Diniz, Jaqueline A; Keid, Lara B

    2017-12-01

    Brucella canis, a gram-negative, facultative intracellular and zoonotic bacterium causes canine brucellosis. Direct methods are the most appropriate for the detection of canine brucellosis and bacterial isolation from blood samples has been employed as gold-standard method. However, due to the delay in obtaining results and the biological risk of the bacterial culturing, the polymerase chain reaction (PCR) has been successfully used as an alternative method for the diagnosis of the infection. Sample preparation is a key step for successful PCR and protocols that provide high DNA yield and purity are recommended to ensure high diagnostic sensitivity. The objective of this study was to evaluate the performance of PCR for the diagnosis of B. canis infection in 36 dogs by testing DNA of whole blood obtained through different extraction and purification protocols. Methods 1 and 2 were based on a commercial kit, using protocols recommended for DNA purification of whole blood and tissue samples, respectively. Method 3 was an in-house method based on enzymatic lysis and purification using organic solvents. The results of the PCR on samples obtained through three different DNA extraction protocols were compared to the blood culture. Of the 36 dogs, 13 (36.1%) were positive by blood culturing, while nine (25.0%), 14 (38.8%), and 15 (41.6%) were positive by PCR after DNA extraction using methods 1, 2 and 3, respectively. PCR performed on DNA purified by Method 2 was as efficient as blood culturing and PCR performed on DNA purified with in-house method, but had the advantage of being less laborious and, therefore, a suitable alternative for the direct B. canis detection in dogs. Copyright © 2017. Published by Elsevier B.V.

  2. Delay in blood sampling for routine newborn screening is associated with increased risk of schizophrenia

    DEFF Research Database (Denmark)

    Nordentoft, Merete; Tidselbak Larsen, Janne; Pedersen, Carsten Bøcker

    2015-01-01

    BACKGROUND: The Danish Neonatal Screening Biobank, containing dried blood spot samples from all newborn in Denmark, is a unique source of data that can be utilized for analyses of genetic and environmental exposures related to schizophrenia and other mental disorders. In previous analyses, we have......, matched on sex and exact date of birth. RESULTS: Both early and late blood sampling was associated with increased risk for schizophrenia. Compared to blood sampling at day 5, sampling at days 0 to 4 after birth was associated with an incidence rate ratio (IRR) of 1.46 (95% CI 1.15-1.87) for development...... admission, and parental education and income, the estimates were slightly different. Thus, blood collection at 0-4days was associated with an IRR of 1.27 (95% CI 0.94-1.71), 6-9days 1.31 (95% CI 0.94-1.84) and 10+days 3.52 (95% CI 1.50 to 8.24). DISCUSSION: After adjusting risk estimates for well-known risk...

  3. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status

    Directory of Open Access Journals (Sweden)

    Bonelli Patrizia

    2016-04-01

    Full Text Available Background: The measurement of vitamin D is now commonplace for preventing osteoporosis and restoring an appropriate concentration that would be effective to counteract the occurrence of other human disorders. The aim of this study was to establish whether blood sampling seasonality may influence total vitamin D concentration in a general population of Italian unselected outpatients.

  4. Uranium concentration in blood samples of Southern Iraqi leukemia patients using CR-39 track detector

    International Nuclear Information System (INIS)

    Al-Hamzawi, A.A.; Al-Qadisiyah University, Qadisiyah; Jaafar, M.S.; Tawfiq, N.F.

    2014-01-01

    The simple and effective technique of fission track etch has been applied to determine trace concentration of uranium in human blood samples taken from two groups of male and female participants: leukemia patients and healthy subjects group. The blood samples of leukemia patients and healthy subjects were collected from three key southern governorates namely, Basrah, Muthanna and Dhi-Qar. These governorates were the centers of intensive military activities during the 1991 and 2003 Gulf wars, and the discarded weapons are still lying around in these regions. CR-39 track detector was used for registration of induced fission tracks. The results show that the highest recorded uranium concentration in the blood samples of leukemia patients was 4.71 ppb (female, 45 years old, from Basrah) and the minimum concentration was 1.91 ppb (male, 3 years old, from Muthanna). For healthy group, the maximum uranium concentration was 2.15 ppb (female, 55 years old, from Basrah) and the minimum concentration was 0.86 ppb (male, 5 years old, from Dhi-Qar). It has been found that the uranium concentrations in human blood samples of leukemia patients are higher than those of the healthy group. These uranium concentrations in the leukemia patients group were significantly different (P < 0.001) from those in the healthy group. (author)

  5. Sample to answer visualization pipeline for low-cost point-of-care blood cell counting

    CSIR Research Space (South Africa)

    Smith, S

    2015-02-01

    Full Text Available We present a visualization pipeline from sample to answer for point-of-care blood cell counting applications. Effective and low-cost point-of-care medical diagnostic tests provide developing countries and rural communities with accessible healthcare...

  6. Workup of Human Blood Samples for Deep Sequencing of HIV-1 Genomes

    NARCIS (Netherlands)

    Cornelissen, Marion; Gall, Astrid; van der Kuyl, Antoinette; Wymant, Chris; Blanquart, François; Fraser, Christophe; Berkhout, Ben

    2018-01-01

    We describe a detailed protocol for the manual workup of blood (plasma/serum) samples from individuals infected with the human immunodeficiency virus type 1 (HIV-1) for deep sequence analysis of the viral genome. The study optimizing the assay was performed in the context of the BEEHIVE (Bridging

  7. Standardised Resting Time Prior to Blood Sampling and Diurnal Variation Associated with Risk of Patient Misclassification

    DEFF Research Database (Denmark)

    Bøgh Andersen, Ida; Brasen, Claus L.; Christensen, Henry

    2015-01-01

    BACKGROUND: According to current recommendations, blood samples should be taken in the morning after 15 minutes' resting time. Some components exhibit diurnal variation and in response to pressures to expand opening hours and reduce waiting time, the aims of this study were to investigate the imp...

  8. Automated spectrometer interface for measurement of short half-life samples for neutron activation analysis

    Energy Technology Data Exchange (ETDEWEB)

    Lapolli, André L.; Secco, Marcello; Genezini, Frederico A.; Zahn, Guilherme S.; Moreira, Edson G., E-mail: alapolli@ipen.br [Instituto de Pesquisas Energéticas e Nucleares (IPEN/CNEN-SP), São Paulo, SP (Brazil)

    2017-07-01

    In this paper a source positioning system was developed, based on a HPGe detector coupled to a Canberra DAS 1000 data acquisition system and Canberra's GENIE2K software and libraries. The system is composed of a step motor coupled to an Arduino Uno microcontroller, which is programmed using C language to allow for a source-detector distance between 0.3 and 20 cm - both components are coupled to a PC computer using the USB interface. In order to allow automated data acquisition, two additional pieces of software were developed. The first one, a Human-Machine Interface (HMI) programmed in Visual Basic 6, allows the programming and monitoring of the data acquisition process, and the other, in REXX language, controls the data acquisition process in the background. The HMI is user-friendly and versatile, so that the even rather complex data acquisition processes may be easily programmed. When the experiment scheme is saved, two files are created and used by the REXX code to control the acquisition process so that the data acquisition is automatically stopped and saved after a user-defined time, then the source is repositioned and data acquisition is cleared and restarted. While in the present stage the system only offers three distinct source positions, finer source-position adjusting is under development. In its present configuration the system has been tested for stability and repeatability in all three positions, with an an excellent performance (author)

  9. Automated Sample Preparation Platform for Mass Spectrometry-Based Plasma Proteomics and Biomarker Discovery

    Directory of Open Access Journals (Sweden)

    Vilém Guryča

    2014-03-01

    Full Text Available The identification of novel biomarkers from human plasma remains a critical need in order to develop and monitor drug therapies for nearly all disease areas. The discovery of novel plasma biomarkers is, however, significantly hampered by the complexity and dynamic range of proteins within plasma, as well as the inherent variability in composition from patient to patient. In addition, it is widely accepted that most soluble plasma biomarkers for diseases such as cancer will be represented by tissue leakage products, circulating in plasma at low levels. It is therefore necessary to find approaches with the prerequisite level of sensitivity in such a complex biological matrix. Strategies for fractionating the plasma proteome have been suggested, but improvements in sensitivity are often negated by the resultant process variability. Here we describe an approach using multidimensional chromatography and on-line protein derivatization, which allows for higher sensitivity, whilst minimizing the process variability. In order to evaluate this automated process fully, we demonstrate three levels of processing and compare sensitivity, throughput and reproducibility. We demonstrate that high sensitivity analysis of the human plasma proteome is possible down to the low ng/mL or even high pg/mL level with a high degree of technical reproducibility.

  10. Automated spectrometer interface for measurement of short half-life samples for neutron activation analysis

    International Nuclear Information System (INIS)

    Lapolli, André L.; Secco, Marcello; Genezini, Frederico A.; Zahn, Guilherme S.; Moreira, Edson G.

    2017-01-01

    In this paper a source positioning system was developed, based on a HPGe detector coupled to a Canberra DAS 1000 data acquisition system and Canberra's GENIE2K software and libraries. The system is composed of a step motor coupled to an Arduino Uno microcontroller, which is programmed using C language to allow for a source-detector distance between 0.3 and 20 cm - both components are coupled to a PC computer using the USB interface. In order to allow automated data acquisition, two additional pieces of software were developed. The first one, a Human-Machine Interface (HMI) programmed in Visual Basic 6, allows the programming and monitoring of the data acquisition process, and the other, in REXX language, controls the data acquisition process in the background. The HMI is user-friendly and versatile, so that the even rather complex data acquisition processes may be easily programmed. When the experiment scheme is saved, two files are created and used by the REXX code to control the acquisition process so that the data acquisition is automatically stopped and saved after a user-defined time, then the source is repositioned and data acquisition is cleared and restarted. While in the present stage the system only offers three distinct source positions, finer source-position adjusting is under development. In its present configuration the system has been tested for stability and repeatability in all three positions, with an an excellent performance (author)

  11. Solid recovered fuels in the cement industry--semi-automated sample preparation unit as a means for facilitated practical application.

    Science.gov (United States)

    Aldrian, Alexia; Sarc, Renato; Pomberger, Roland; Lorber, Karl E; Sipple, Ernst-Michael

    2016-03-01

    One of the challenges for the cement industry is the quality assurance of alternative fuel (e.g., solid recovered fuel, SRF) in co-incineration plants--especially for inhomogeneous alternative fuels with large particle sizes (d95⩾100 mm), which will gain even more importance in the substitution of conventional fuels due to low production costs. Existing standards for sampling and sample preparation do not cover the challenges resulting from these kinds of materials. A possible approach to ensure quality monitoring is shown in the present contribution. For this, a specially manufactured, automated comminution and sample divider device was installed at a cement plant in Rohožnik. In order to prove its practical suitability with methods according to current standards, the sampling and sample preparation process were validated for alternative fuel with a grain size >30 mm (i.e., d95=approximately 100 mm), so-called 'Hotdisc SRF'. Therefore, series of samples were taken and analysed. A comparison of the analysis results with the yearly average values obtained through a reference investigation route showed good accordance. Further investigations during the validation process also showed that segregation or enrichment of material throughout the comminution plant does not occur. The results also demonstrate that compliance with legal standards regarding the minimum sample amount is not sufficient for inhomogeneous and coarse particle size alternative fuels. Instead, higher sample amounts after the first particle size reduction step are strongly recommended in order to gain a representative laboratory sample. © The Author(s) 2016.

  12. Automated blood glucose control in type 1 diabetes: A review of progress and challenges.

    Science.gov (United States)

    Bertachi, Arthur; Ramkissoon, Charrise M; Bondia, Jorge; Vehí, Josep

    2018-03-01

    Since the 2000s, research teams worldwide have been working to develop closed-loop (CL) systems able to automatically control blood glucose (BG) levels in patients with type 1 diabetes. This emerging technology is known as artificial pancreas (AP), and its first commercial version just arrived in the market. The main objective of this paper is to present an extensive review of the clinical trials conducted since 2011, which tested various implementations of the AP for different durations under varying conditions. A comprehensive table that contains key information from the selected publications is provided, and the main challenges in AP development and the mitigation strategies used are discussed. The development timelines for different AP systems are also included, highlighting the main evolutions over the clinical trials for each system. Copyright © 2017 SEEN y SED. Publicado por Elsevier España, S.L.U. All rights reserved.

  13. Automated Microfluidic Droplet-Based Sample Chopper for Detection of Small Fluorescence Differences Using Lock-In Analysis.

    Science.gov (United States)

    Negou, Jean T; Avila, L Adriana; Li, Xiangpeng; Hagos, Tesfagebriel M; Easley, Christopher J

    2017-06-06

    Fluorescence is widely used for small-volume analysis and is a primary tool for on-chip detection in microfluidic devices, yet additional expertise, more elaborate optics, and phase-locked detectors are needed for ultrasensitive measurements. Recently, we designed a microfluidic analog to an optical beam chopper (μChopper) that alternated formation of picoliter volume sample and reference droplets. Without complex optics, the device negated large signal drifts (1/f noise), allowing absorbance detection in a mere 27 μm optical path. Here, we extend the μChopper concept to fluorescence detection with standard wide-field microscope optics. Precision of droplet control in the μChopper was improved by automation with pneumatic valves, allowing fluorescence measurements to be strictly phase locked at 0.04 Hz bandwidth to droplets generated at 3.50 Hz. A detection limit of 12 pM fluorescein was achieved when sampling 20 droplets, and as few as 310 zeptomoles (3.1 × 10 -19 mol) were detectable in single droplets (8.8 nL). When applied to free fatty acid (FFA) uptake in 3T3-L1 adipocytes, this μChopper permitted single-cell FFA uptake rates to be quantified at 3.5 ± 0.2 × 10 -15 mol cell -1 for the first time. Additionally, homogeneous immunoassays in droplets exhibited insulin detection limits of 9.3 nM or 190 amol (1.9 × 10 -16 mol). The combination of this novel, automated μChopper with lock-in detection provides a high-performance platform for detecting small differences with standard fluorescence optics, particularly in situations where sample volume is limited. The technique should be simple to implement into a variety of other droplet fluidics devices.

  14. Direct RNA-based detection of CTX-M β-lactamases in human blood samples.

    Science.gov (United States)

    Stein, Claudia; Makarewicz, Oliwia; Pfeifer, Yvonne; Brandt, Christian; Pletz, Mathias W

    2015-05-01

    Bloodstream infections with ESBL-producers are associated with increased mortality, which is due to delayed appropriate treatment resulting in clinical failure. Current routine diagnostics for detection of bloodstream infections consists of blood culture followed by species identification and susceptibility testing. In attempts to improve and accelerate diagnostic procedures, PCR-based methods have been developed. These methods focus on species identification covering only a limited number of ESBL coding genes. Therefore, they fail to cover the steadily further evolving genetic diversity of clinically relevant β-lactamases. We have recently designed a fast and novel RNA targeting method to detect and specify CTX-M alleles from bacterial cultures, based on an amplification-pyrosequencing approach. We further developed this assay towards a diagnostic tool for clinical use and evaluated its sensitivity and specificity when applied directly to human blood samples. An optimized protocol for mRNA isolation allows detection of specific CTX-M groups from as little as 100 CFU/mL blood via reverse transcription, amplification, and pyrosequencing directly from human EDTA blood samples as well as from pre-incubated human blood cultures with a turnaround time for test results of <7 h. Copyright © 2015 Elsevier GmbH. All rights reserved.

  15. Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study

    Directory of Open Access Journals (Sweden)

    Williams Adam R

    2009-12-01

    Full Text Available Abstract Background Whole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a venous blood sample of at least 2.5 mL. While fingerstick blood collection has been used for many different assays, there has yet to be a kit developed to isolate high quality RNA for use in gene expression studies from such small human samples. The clinical and field testing advantages of obtaining reliable and reproducible gene expression data from a fingerstick are many; it is less invasive, time saving, more mobile, and eliminates the need of a trained phlebotomist. Furthermore, this method could also be employed in small animal studies, i.e. mice, where larger sample collections often require sacrificing the animal. In this study, we offer a rapid and simple method to extract sufficient amounts of high quality total RNA from approximately 70 μl of whole blood collected via a fingerstick using a modified protocol of the commercially available Qiagen PAXgene RNA Blood Kit. Results From two sets of fingerstick collections, about 70 uL whole blood collected via finger lancet and capillary tube, we recovered an average of 252.6 ng total RNA with an average RIN of 9.3. The post-amplification yields for 50 ng of total RNA averaged at 7.0 ug cDNA. The cDNA hybridized to Affymetrix HG-U133 Plus 2.0 GeneChips had an average % Present call of 52.5%. Both fingerstick collections were highly correlated with r2 values ranging from 0.94 to 0.97. Similarly both fingerstick collections were highly correlated to the venous collection with r2 values ranging from 0.88 to 0

  16. A simple method for regional cerebral blood flow measurement by one-point arterial blood sampling and 123I-IMP microsphere model (part 2). A study of time correction of one-point blood sample count

    International Nuclear Information System (INIS)

    Masuda, Yasuhiko; Makino, Kenichi; Gotoh, Satoshi

    1999-01-01

    In our previous paper regarding determination of the regional cerebral blood flow (rCBF) using the 123 I-IMP microsphere model, we reported that the accuracy of determination of the integrated value of the input function from one-point arterial blood sampling can be increased by performing correction using the 5 min: 29 min ratio for the whole-brain count. However, failure to carry out the arterial blood collection at exactly 5 minutes after 123 I-IMP injection causes errors with this method, and there is thus a time limitation. We have now revised out method so that the one-point arterial blood sampling can be performed at any time during the interval between 5 minutes and 20 minutes after 123 I-IMP injection, with addition of a correction step for the sampling time. This revised method permits more accurate estimation of the integral of the input functions. This method was then applied to 174 experimental subjects: one-point blood samples collected at random times between 5 and 20 minutes, and the estimated values for the continuous arterial octanol extraction count (COC) were determined. The mean error rate between the COC and the actual measured continuous arterial octanol extraction count (OC) was 3.6%, and the standard deviation was 12.7%. Accordingly, in 70% of the cases, the rCBF was able to be estimated within an error rate of 13%, while estimation was possible in 95% of the cases within an error rate of 25%. This improved method is a simple technique for determination of the rCBF by 123 I-IMP microsphere model and one-point arterial blood sampling which no longer shows a time limitation and does not require any octanol extraction step. (author)

  17. Midazolam sedates Passeriformes for field sampling but affects multiple venous blood analytes

    Directory of Open Access Journals (Sweden)

    Heatley JJ

    2015-01-01

    Full Text Available J Jill Heatley,1 Jennifer Cary,2,3 Lyndsey Kingsley,1 Hughes Beaufrere,4 Karen E Russell,5 Gary Voelker2,3 1Department of Small Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, 2Department of Wildlife and Fisheries Sciences, 3Texas A&M Biodiversity Research and Teaching Collections, Texas A&M University, College Station, TX, USA; 4Health Sciences Centre, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada; 5Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, College Station, TX, USA Abstract: Feasibility and effect of midazolam administration on blood analytes and for sedation of Passeriformes being collected in a larger study of genetic biodiversity was assessed. Midazolam (5.6±2.7 mg/kg was administered intranasally prior to sampling, euthanasia, and specimen preparation of 104 passerine birds. Each bird was assessed for sedation score and then multiple analytes were determined from jugular blood samples using the i-STAT® point of care analyzer at “bird side”. Most birds were acceptably sedated, sedation became more pronounced as midazolam dose increased, and only a single bird died. Electrolyte concentrations and venous blood gas analytes were affected by midazolam administration while blood pH, packed cell volume, hemoglobin, and calculated hematocrit were not. Intranasal midazolam gives adequate sedation and is safe for short-term use in free-living Passeriformes. Based on venous blood analyte data, sedation of Passeriformes prior to handling appears to reduce stress but also produces venous blood gas differences consistent with hypoventilation relative to birds which were not given midazolam. Further study is recommended to investigate midazolam's continued use in free-living avian species. Studies should include safety, reversal and recovery, effect upon additional endogenous analytes, and compatibility with studies of ecology and toxicology

  18. Fabrication and Characterization of a Microfluidic Device to Ultrapurify Blood Samples

    KAUST Repository

    Tallerico, Marco

    2015-05-04

    The improvement of blood cell sorting techniques in recent years have attracted the attention of many researchers due to the possible benefits that these methods can lead in biology, regenerative medicine, materials science and therapeutic area. In this work a cell sorting technique based on filtration is described. The separation occurs by means of a microfluidic device, suitably designed, manufactured and tested, that is connected to an external experimental set-up. The fabrication process can be divided in two parts: at first it is described the manufacturing process of a filtering membrane, with holes of specific size that allow the passage of only certain cell types. Following the microfluidic device is fabricated through the mechanical micromilling. The membrane and the microdevice are suitably bonded and tested by means of an external connection with syringe pumps that inject blood samples at specific flow rates. The device is designed to separate blood cells and tumor cells only by using differences in size and shape. In particular during the first experiments red blood cells and platelets are sorted from white blood cells; in the other experiments red blood cells and platelets are separated from white blood cells and tumor cells. The microdevice has proven to be very efficient, in fact a capture efficiency of 99% is achieved. For this reason it could be used in identification and isolation of circulating tumor cells, a very rare cancer cell type whose presence in the bloodstream could be symptom of future solid tumor formation. The various experiments have also demonstrated that tumor cells survive even after the separation treatment, and then the suffered stress during the sorting process does not harm the biological sample.

  19. Automated radioanalytical system incorporating microwave-assisted sample preparation, chemical separation, and online radiometric detection for the monitoring of total 99Tc in nuclear waste processing streams.

    Science.gov (United States)

    Egorov, Oleg B; O'Hara, Matthew J; Grate, Jay W

    2012-04-03

    An automated fluidic instrument is described that rapidly determines the total (99)Tc content of aged nuclear waste samples, where the matrix is chemically and radiologically complex and the existing speciation of the (99)Tc is variable. The monitor links microwave-assisted sample preparation with an automated anion exchange column separation and detection using a flow-through solid scintillator detector. The sample preparation steps acidify the sample, decompose organics, and convert all Tc species to the pertechnetate anion. The column-based anion exchange procedure separates the pertechnetate from the complex sample matrix, so that radiometric detection can provide accurate measurement of (99)Tc. We developed a preprogrammed spike addition procedure to automatically determine matrix-matched calibration. The overall measurement efficiency that is determined simultaneously provides a self-diagnostic parameter for the radiochemical separation and overall instrument function. Continuous, automated operation was demonstrated over the course of 54 h, which resulted in the analysis of 215 samples plus 54 hly spike-addition samples, with consistent overall measurement efficiency for the operation of the monitor. A sample can be processed and measured automatically in just 12.5 min with a detection limit of 23.5 Bq/mL of (99)Tc in low activity waste (0.495 mL sample volume), with better than 10% RSD precision at concentrations above the quantification limit. This rapid automated analysis method was developed to support nuclear waste processing operations planned for the Hanford nuclear site.

  20. Accelerated Evaluation of Automated Vehicles Safety in Lane-Change Scenarios Based on Importance Sampling Techniques.

    Science.gov (United States)

    Zhao, Ding; Lam, Henry; Peng, Huei; Bao, Shan; LeBlanc, David J; Nobukawa, Kazutoshi; Pan, Christopher S

    2017-03-01

    Automated vehicles (AVs) must be thoroughly evaluated before their release and deployment. A widely used evaluation approach is the Naturalistic-Field Operational Test (N-FOT), which tests prototype vehicles directly on the public roads. Due to the low exposure to safety-critical scenarios, N-FOTs are time consuming and expensive to conduct. In this paper, we propose an accelerated evaluation approach for AVs. The results can be used to generate motions of the other primary vehicles to accelerate the verification of AVs in simulations and controlled experiments. Frontal collision due to unsafe cut-ins is the target crash type of this paper. Human-controlled vehicles making unsafe lane changes are modeled as the primary disturbance to AVs based on data collected by the University of Michigan Safety Pilot Model Deployment Program. The cut-in scenarios are generated based on skewed statistics of collected human driver behaviors, which generate risky testing scenarios while preserving the statistical information so that the safety benefits of AVs in nonaccelerated cases can be accurately estimated. The cross-entropy method is used to recursively search for the optimal skewing parameters. The frequencies of the occurrences of conflicts, crashes, and injuries are estimated for a modeled AV, and the achieved accelerated rate is around 2000 to 20 000. In other words, in the accelerated simulations, driving for 1000 miles will expose the AV with challenging scenarios that will take about 2 to 20 million miles of real-world driving to encounter. This technique thus has the potential to greatly reduce the development and validation time for AVs.

  1. Using improved serial blood sampling method of mice to study pharmacokinetics and drug-drug interaction.

    Science.gov (United States)

    Watanabe, Ayahisa; Watari, Ryosuke; Ogawa, Keiko; Shimizu, Ryosuke; Tanaka, Yukari; Takai, Nozomi; Nezasa, Ken-ichi; Yamaguchi, Yoshitaka

    2015-03-01

    In pharmacokinetic evaluation of mice, using serial sampling methods rather than a terminal blood sampling method could reduce the number of animals needed and lead to more reliable data by excluding individual differences. In addition, using serial sampling methods can be valuable for evaluation of the drug-drug interaction (DDI) potential of drug candidates. In this study, we established an improved method for serially sampling the blood from one mouse by only one incision of the lateral tail vein, and investigated whether our method could be adapted to pharmacokinetic and DDI studies. After intravenous and oral administration of ibuprofen and fexofenadine (BCS class II and III), the plasma concentration and pharmacokinetic parameters were evaluated by our method and a terminal blood sampling method, with the result that both methods gave comparable results (ibuprofen: 63.8 ± 4.0% and 64.4%, fexofenadine: 6.5 ± 0.7% and 7.9%, respectively, in bioavailability). In addition, our method could be adapted to DDI study for cytochrome P450 and organic anion transporting polypeptide inhibition. These results demonstrate that our method can be useful for pharmacokinetic evaluation from the perspective of reliable data acquisition as well as easy handling and low stress to mice and improve the quality of pharmacokinetic and DDI studies. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

  2. Effects of sampling rate on automated fatigue recognition in surface EMG signals

    Directory of Open Access Journals (Sweden)

    Kahl Lorenz

    2015-09-01

    Full Text Available This study investigated the effects different sampling rates may produce on the quality of muscle fatigue detection algorithms. sEMG signals were obtained from isometric contractions of the arm. Subsampled signals resulting in technically relevant sampling rates were computationally deduced from the original recordings. The spectral based fatigue recognition methods mean and median frequency as well as spectral moment ratio were included in this investigation, as well as the sample and the fuzzy approximate entropy. The resulting fatigue indices were evaluated with respect to noise and separability of different load levels. We concluded that the spectral moment ratio provides the best results in fatigue detection over a wide range of sampling rates.

  3. Alternative sampling site for blood glucose testing in cats: giving the ears a rest.

    Science.gov (United States)

    Zeugswetter, Florian K; Rebuzzi, Laura; Karlovits, Sonja

    2010-09-01

    BACKGROUND AND STUDY RATIONALE: Home monitoring is an important part of the long-term management of diabetic cats. Despite the extensive use of glucometers in this species, up until now only the pinna of the ear has been validated as a testing site. This cross-sectional study investigated the feasibility and validity of sampling from the metacarpal/metatarsal pads in hospitalised cats with various diseases. The large pads were compared with the ear as a sampling site in 75 cats. Lancing the pads was tolerated very well. If the initial drop of blood was too small, an adequate volume of blood was almost always achieved by squeezing the pads. No significant differences were observed in first-attempt success rate or glucose values between the two sites. Due to the inability to obtain an adequate volume of blood or struggling, no measurement was possible in four cats. While further work is necessary to assess the utility of this technique, especially in the home environment, the results indicate that the metacarpal pads, in particular, may offer a viable alternative testing site for the measurement of blood glucose concentrations, especially if ear sampling fails. Copyright 2010 ISFM and AAFP. Published by Elsevier Ltd. All rights reserved.

  4. Determination of Se and Zn elements in blood serum samples by neutron activation analysis

    International Nuclear Information System (INIS)

    Indah Kusmartini; Rukruk Rukayah; Woro Yatu NS; Muhayatun; Rochestry Sofyan

    2010-01-01

    Se and Zn are essential elements being required for activity several enzyme systems in human metabolism. The elements in blood serum as well as to obtain information regarding the health status of individuals, so that important to investigated. Commonly Se and Zn elements in blood serum are low in concentration with limited samples weight, therefore high sensitive and accurate analysis method like NAA are needed. This study aims to determine the content of the elements Se and Zn in blood serum of employee using NAA technique. The samples were freeze dried then irradiated at rabbit system facility of G.A. Siwabessy Serpong reactor with neutron flux ~10 13 n.cm -2 .s -1 for 2 hours. Then samples were counted for 50000 s by HPGe spectrometer gamma and analyzed by software GENIE 2000. Method validation was also applied by analyzing the Reference Material IAEA Animal Blood RM A-13. The range concentration of elements Se and Zn were 0.06-0.17 μg/mL and 0.56-1.37 μg/mL with overall mean 0.10 ± 0.01 μg/mL and 0.97 ± 0.07 μg/mL. The value showed that appropriate with last researcher and another country. (author)

  5. High-throughput automated microfluidic sample preparation for accurate microbial genomics.

    Science.gov (United States)

    Kim, Soohong; De Jonghe, Joachim; Kulesa, Anthony B; Feldman, David; Vatanen, Tommi; Bhattacharyya, Roby P; Berdy, Brittany; Gomez, James; Nolan, Jill; Epstein, Slava; Blainey, Paul C

    2017-01-27

    Low-cost shotgun DNA sequencing is transforming the microbial sciences. Sequencing instruments are so effective that sample preparation is now the key limiting factor. Here, we introduce a microfluidic sample preparation platform that integrates the key steps in cells to sequence library sample preparation for up to 96 samples and reduces DNA input requirements 100-fold while maintaining or improving data quality. The general-purpose microarchitecture we demonstrate supports workflows with arbitrary numbers of reaction and clean-up or capture steps. By reducing the sample quantity requirements, we enabled low-input (∼10,000 cells) whole-genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil micro-colonies with superior results. We also leveraged the enhanced throughput to sequence ∼400 clinical Pseudomonas aeruginosa libraries and demonstrate excellent single-nucleotide polymorphism detection performance that explained phenotypically observed antibiotic resistance. Fully-integrated lab-on-chip sample preparation overcomes technical barriers to enable broader deployment of genomics across many basic research and translational applications.

  6. Development of an automated method for determination of thorium in soil samples and aerosols

    International Nuclear Information System (INIS)

    Stuart, J.E.; Robertson, R.

    1986-09-01

    Methodology for determining trace thorium levels in a variety of sample types was further developed. Thorium in filtered water samples is concentrated by ferric hydroxide precipitation followed by dissolution and co-precipitation with lanthanum fluoride. Aerosols on glass fibre, cellulose ester, or teflon filters and solid soil and sediment samples are acid digested. Subsequently thorium is concentrated by lanthanum fluoride co-precipitation. Chemical separation and measurement is then done on a Technicon AA11-C autoanalyzer, using solvent extraction into thenoyltrifuoroacetone in kerosene followed by back extraction into 2 N H NO 3 , and colourometric measurement of the thorium arsenazo III complex. Chemical yields are determined by the addition of thorium-234 tracer using gamma-ray spectrometry. The sensitivities of the methods for water, aerosol and solid samples are approximately 1.0 μg/L, 0.5 μg/g and 1.0 μg/g respectively. At thorium levels about ten times the detection limit, accuracy is estimated to be ± 10% for liquids and aerosols and ± 15% for solid samples, and precision ± 5% for all samples

  7. Interlaboratory comparison of PCR-based identification of Candida and Aspergillus DNA in spiked blood samples.

    Science.gov (United States)

    Reichard, Utz; Buchheidt, Dieter; Lass-Flörl, Cornelia; Loeffler, Juergen; Lugert, Raimond; Ruhnke, Markus; Tintelnot, Kathrin; Weig, Michael; Groß, Uwe

    2012-09-01

    Despite PCR per se being a powerful and sensitive technique, regarding the detection of fungi in patients' blood, no consensus for a standardised PCR protocol yet exists. To complement other ongoing or accomplished studies which tackle this problem, the German Reference Center for Systemic Mycoses conducted an interlaboratory comparison starting with blood samples spiked with fungal cell elements. Altogether, six laboratories using in-house PCR-protocols from Germany and Austria participated in the trial. Blood samples were spiked with vital cells of Candida albicans or Aspergillus fumigatus. Candida was used in the yeast form, whereas Aspergillus cells were either spiked as conidia or as very young germlings, also known as smoo cells. Spiked blood samples contained between 10 and 10 000 cells ml(-1). Depending on the techniques used for fungal cell disruption and DNA-amplification, detection quality was variable between laboratories, but also differed within single laboratories in different trials particularly for samples spiked with less than 100 cells ml(-1). Altogether, at least regarding the detection of A. fumigatus, two of six laboratories showed constant reliable test results also with low fungal cell number spiked samples. Protocols used by these labs do not differ substantially from others. However, as particularities, one protocol included a conventional phenol chloroform extraction during the DNA preparation process and the other included a real time PCR-protocol based on FRET probes. Other laboratory comparisons on the basis of clinical samples should follow to further evaluate the procedures. The difficulties and problems of such trials in general are discussed. © 2012 Blackwell Verlag GmbH.

  8. Suitability of small diagnostic peripheral-blood samples for cell-therapy studies.

    Science.gov (United States)

    Stephanou, Coralea; Papasavva, Panayiota; Zachariou, Myria; Patsali, Petros; Epitropou, Marilena; Ladas, Petros; Al-Abdulla, Ruba; Christou, Soteroulla; Antoniou, Michael N; Lederer, Carsten W; Kleanthous, Marina

    2017-02-01

    Primary hematopoietic stem and progenitor cells (HSPCs) are key components of cell-based therapies for blood disorders and are thus the authentic substrate for related research. We propose that ubiquitous small-volume diagnostic samples represent a readily available and as yet untapped resource of primary patient-derived cells for cell- and gene-therapy studies. In the present study we compare isolation and storage methods for HSPCs from normal and thalassemic small-volume blood samples, considering genotype, density-gradient versus lysis-based cell isolation and cryostorage media with different serum contents. Downstream analyses include viability, recovery, differentiation in semi-solid media and performance in liquid cultures and viral transductions. We demonstrate that HSPCs isolated either by ammonium-chloride potassium (ACK)-based lysis or by gradient isolation are suitable for functional analyses in clonogenic assays, high-level HSPC expansion and efficient lentiviral transduction. For cryostorage of cells, gradient isolation is superior to ACK lysis, and cryostorage in freezing media containing 50% fetal bovine serum demonstrated good results across all tested criteria. For assays on freshly isolated cells, ACK lysis performed similar to, and for thalassemic samples better than, gradient isolation, at a fraction of the cost and hands-on time. All isolation and storage methods show considerable variation within sample groups, but this is particularly acute for density gradient isolation of thalassemic samples. This study demonstrates the suitability of small-volume blood samples for storage and preclinical studies, opening up the research field of HSPC and gene therapy to any blood diagnostic laboratory with corresponding bioethics approval for experimental use of surplus material. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

  9. Confocal backscattering-based detection of leukemic cells in flowing blood samples.

    Science.gov (United States)

    Greiner, Cherry; Hunter, Martin; Rius, Francisca; Huang, Peter; Georgakoudi, Irene

    2011-10-01

    The prognostic value of assessing minimal residual disease (MRD) in leukemia has been established with advancements in flow cytometry and PCR. Nonetheless, these techniques are limited by high equipment costs, complex, and costly cell processing and the need for highly trained personnel. Here, we demonstrate the potential of exploiting differences in the relative intensities of backscattered light at three wavelengths to detect the presence of leukemic cells in samples containing varying mixtures of white blood cells (WBCs) and leukemic cells flowing through microfluidic channels. Using 405, 488, and 633 nm illumination, we identify distinct light scattering intensity distributions for Nalm-6 leukemic cells, normal mononuclear (PBMC) and polymorphonuclear (PMN) white blood cells and red blood cells. We exploit these differences to develop cell classification algorithms, whose performance is evaluated based on simultaneous acquisition of light scattering and fluorescence flow cytometry data. When this algorithm is used prospectively for the analysis of samples consisting of mixtures of PBMCs and leukemic cells, we achieve an average specificity and sensitivity of leukemic cell detection of 99.6 and 45.2%, respectively. When we consider samples that include leukemic cells along with PMNs and PBMCs, which can be acquired using a simple red blood cell lysis step following venipuncture, the specificity and sensitivity of the approach decreases to 91.6 and 39.5%, respectively. On the basis of the performance of these algorithms, we estimate that 42 or 71 μL of blood would be adequate to confirm the presence of leukemia at an 80% power level in samples containing 0.01% leukemia to either PBMCs or PBMCs and PMNs, respectively. Therefore, light scattering-based flow cytometry in a microfluidic platform could provide a low cost, highly portable, minimally invasive approach for detection and monitoring of leukemic patients. This could offer significant improvements

  10. The T-lock: automated compensation of radio-frequency induced sample heating

    International Nuclear Information System (INIS)

    Hiller, Sebastian; Arthanari, Haribabu; Wagner, Gerhard

    2009-01-01

    Modern high-field NMR spectrometers can stabilize the nominal sample temperature at a precision of less than 0.1 K. However, the actual sample temperature may differ from the nominal value by several degrees because the sample heating caused by high-power radio frequency pulses is not readily detected by the temperature sensors. Without correction, transfer of chemical shifts between different experiments causes problems in the data analysis. In principle, the temperature differences can be corrected by manual procedures but this is cumbersome and not fully reliable. Here, we introduce the concept of a 'T-lock', which automatically maintains the sample at the same reference temperature over the course of different NMR experiments. The T-lock works by continuously measuring the resonance frequency of a suitable spin and simultaneously adjusting the temperature control, thus locking the sample temperature at the reference value. For three different nuclei, 13 C, 17 O and 31 P in the compounds alanine, water, and phosphate, respectively, the T-lock accuracy was found to be <0.1 K. The use of dummy scan periods with variable lengths allows a reliable establishment of the thermal equilibrium before the acquisition of an experiment starts

  11. Evaluation of the effects of insufficient blood volume samples on the performance of blood glucose self-test meters.

    Science.gov (United States)

    Pfützner, Andreas; Schipper, Christina; Ramljak, Sanja; Flacke, Frank; Sieber, Jochen; Forst, Thomas; Musholt, Petra B

    2013-11-01

    Accuracy of blood glucose readings is (among other things) dependent on the test strip being completely filled with sufficient sample volume. The devices are supposed to display an error message in case of incomplete filling. This laboratory study was performed to test the performance of 31 commercially available devices in case of incomplete strip filling. Samples with two different glucose levels (60-90 and 300-350 mg/dl) were used to generate three different sample volumes: 0.20 µl (too low volume for any device), 0.32 µl (borderline volume), and 1.20 µl (low but supposedly sufficient volume for all devices). After a point-of-care capillary reference measurement (StatStrip, NovaBiomedical), the meter strip was filled (6x) with the respective volume, and the response of the meters (two devices) was documented (72 determinations/meter type). Correct response was defined as either an error message indicating incomplete filling or a correct reading (±20% compared with reference reading). Only five meters showed 100% correct responses [BGStar and iBGStar (both Sanofi), ACCU-CHEK Compact+ and ACCU-CHEK Mobile (both Roche Diagnostics), OneTouch Verio (LifeScan)]. The majority of the meters (17) had up to 10% incorrect reactions [predominantly incorrect readings with sufficient volume; Precision Xceed and Xtra, FreeStyle Lite, and Freedom Lite (all Abbott); GlucoCard+ and GlucoMen GM (both Menarini); Contour, Contour USB, and Breeze2 (all Bayer); OneTouch Ultra Easy, Ultra 2, and Ultra Smart (all LifeScan); Wellion Dialog and Premium (both MedTrust); FineTouch (Terumo); ACCU-CHEK Aviva (Roche); and GlucoTalk (Axis-Shield)]. Ten percent to 20% incorrect reactions were seen with OneTouch Vita (LifeScan), ACCU-CHEK Aviva Nano (Roche), OmniTest+ (BBraun), and AlphaChek+ (Berger Med). More than 20% incorrect reactions were obtained with Pura (Ypsomed), GlucoCard Meter and GlucoMen LX (both Menarini), Elite (Bayer), and MediTouch (Medisana). In summary, partial and

  12. Low-Cost 3D Printers Enable High-Quality and Automated Sample Preparation and Molecular Detection.

    Directory of Open Access Journals (Sweden)

    Kamfai Chan

    Full Text Available Most molecular diagnostic assays require upfront sample preparation steps to isolate the target's nucleic acids, followed by its amplification and detection using various nucleic acid amplification techniques. Because molecular diagnostic methods are generally rather difficult to perform manually without highly trained users, automated and integrated systems are highly desirable but too costly for use at point-of-care or low-resource settings. Here, we showcase the development of a low-cost and rapid nucleic acid isolation and amplification platform by modifying entry-level 3D printers that cost between $400 and $750. Our modifications consisted of replacing the extruder with a tip-comb attachment that houses magnets to conduct magnetic particle-based nucleic acid extraction. We then programmed the 3D printer to conduct motions that can perform high-quality extraction protocols. Up to 12 samples can be processed simultaneously in under 13 minutes and the efficiency of nucleic acid isolation matches well against gold-standard spin-column-based extraction technology. Additionally, we used the 3D printer's heated bed to supply heat to perform water bath-based polymerase chain reactions (PCRs. Using another attachment to hold PCR tubes, the 3D printer was programmed to automate the process of shuttling PCR tubes between water baths. By eliminating the temperature ramping needed in most commercial thermal cyclers, the run time of a 35-cycle PCR protocol was shortened by 33%. This article demonstrates that for applications in resource-limited settings, expensive nucleic acid extraction devices and thermal cyclers that are used in many central laboratories can be potentially replaced by a device modified from inexpensive entry-level 3D printers.

  13. Low-Cost 3D Printers Enable High-Quality and Automated Sample Preparation and Molecular Detection.

    Science.gov (United States)

    Chan, Kamfai; Coen, Mauricio; Hardick, Justin; Gaydos, Charlotte A; Wong, Kah-Yat; Smith, Clayton; Wilson, Scott A; Vayugundla, Siva Praneeth; Wong, Season

    2016-01-01

    Most molecular diagnostic assays require upfront sample preparation steps to isolate the target's nucleic acids, followed by its amplification and detection using various nucleic acid amplification techniques. Because molecular diagnostic methods are generally rather difficult to perform manually without highly trained users, automated and integrated systems are highly desirable but too costly for use at point-of-care or low-resource settings. Here, we showcase the development of a low-cost and rapid nucleic acid isolation and amplification platform by modifying entry-level 3D printers that cost between $400 and $750. Our modifications consisted of replacing the extruder with a tip-comb attachment that houses magnets to conduct magnetic particle-based nucleic acid extraction. We then programmed the 3D printer to conduct motions that can perform high-quality extraction protocols. Up to 12 samples can be processed simultaneously in under 13 minutes and the efficiency of nucleic acid isolation matches well against gold-standard spin-column-based extraction technology. Additionally, we used the 3D printer's heated bed to supply heat to perform water bath-based polymerase chain reactions (PCRs). Using another attachment to hold PCR tubes, the 3D printer was programmed to automate the process of shuttling PCR tubes between water baths. By eliminating the temperature ramping needed in most commercial thermal cyclers, the run time of a 35-cycle PCR protocol was shortened by 33%. This article demonstrates that for applications in resource-limited settings, expensive nucleic acid extraction devices and thermal cyclers that are used in many central laboratories can be potentially replaced by a device modified from inexpensive entry-level 3D printers.

  14. Automation of the radiation measuring facilities for samples in health physics - MA 9

    International Nuclear Information System (INIS)

    Martini, M.

    1980-12-01

    Routine radation measurements of samples are performed by the HMI health physics department by means of test stations for individual samples and multiple samples (using a changing equipment). The basic device of these test stations is a SCALER/TIMER system (BF 22/25, BERTHOLD Corp.). This measuring facility has been extended by a CAMAC intrumentation which incorporates an autonomous CAMAC processor (CAPRO-1, INCAA B.V.) for monitoring an automatic control of the system. The programming language is BASIC. A DECwriter (LA 34) is used for user interaction and for printing the measurement results. This report describes the features of this system and present some examples of, the dialogue with the system and the printout of data. (orig.) [de

  15. Gene methylation parallelisms between peripheral blood cells and oral mucosa samples in relation to overweight.

    Science.gov (United States)

    San-Cristobal, Rodrigo; Navas-Carretero, Santiago; Milagro, Fermín I; Riezu-Boj, J Ignacio; Guruceaga, Elizabeth; Celis-Morales, Carlos; Livingstone, Katherine M; Brennan, Lorraine; Lovegrove, Julie A; Daniel, Hannelore; Saris, Wim H; Traczyk, Iwonna; Manios, Yannis; Gibney, Eileen R; Gibney, Michael J; Mathers, John C; Martinez, J Alfredo

    2016-08-01

    Epigenetics has an important role in the regulation of metabolic adaptation to environmental modifications. In this sense, the determination of epigenetic changes in non-invasive samples during the development of metabolic diseases could play an important role in the procedures in primary healthcare practice. To help translate the knowledge of epigenetics to public health practice, the present study aims to explore the parallelism of methylation levels between white blood cells and buccal samples in relation to obesity and associated disorders. Blood and buccal swap samples were collected from a subsample of the Spanish cohort of the Food4Me study. Infinium HumanMethylation450 DNA Analysis was carried out for the determination of methylation levels. Standard deviation for β values method and concordance correlation analysis were used to select those CpG which showed best parallelism between samples. A total of 277 CpGs met the criteria and were selected for an enrichment analysis and a correlation analysis with anthropometrical and clinical parameters. From those selected CpGs, four presented high associations with BMI (cg01055691 in GAP43; r = -0.92 and rho = -0.84 for blood; r = -0.89 and rho = -0.83 for buccal sample), HOMA-IR (cg00095677 in ATP2A3; r = 0.82 and rho = -0.84 for blood; r = -0.8 and rho = -0.83 for buccal sample) and leptin (cg14464133 in ADARB2; r = -0.9182 and rho = -0.94 for blood; r = -0.893 and rho = -0.79 for buccal sample). These findings demonstrate the potential application of non-invasive buccal samples in the identification of surrogate epigenetic biomarkers and identify methylation sites in GAP43, ATP2A3 and ADARB2 genes as potential targets in relation to overweight management and insulin sensibility.

  16. Highly Effective DNA Extraction Method from Fresh, Frozen, Dried and Clotted Blood Samples

    Directory of Open Access Journals (Sweden)

    Jaleh Barar

    2011-09-01

    Full Text Available Introduction: Today, with the tremendous potential of genomics and other recent advances in science, the role of science to improve reliable DNA extraction methods is more relevant than ever before. The ideal process for genomic DNA extraction demands high quantities of pure, integral and intact genomic DNA (gDNA from the sample with minimal co-extraction of inhibitors of downstream processes. Here, we report the development of a very rapid, less-hazardous, and high throughput protocol for extracting of high quality DNA from blood samples. Methods: Dried, clotted and ethylene diamine tetra-acetic acid (EDTA treated fresh and frozen blood samples were extracted using this method in which the quality and integrity of the extracted DNA were corroborated by agarose gel electrophoresis, PCR reaction and DNA digestion using restricted enzyme. The UV spectrophotometric and gel electrophoresis analysis resulted in high A260/A280 ratio (>1.8 with high intactness of DNA. Results: PCR and DNA digestion experiments indicated that the final solutions of extracted DNA contained no inhibitory substances, which confirms that the isolated DNA is of good quality. Conclusion: The high quality and quantity of current method, no enzymatic processing and accordingly its low cost, make it appropriate for DNA extraction not only from human but also from animal blood samples in any molecular biology labs.

  17. Detection of Candida albicans DNA from blood samples using a novel electrochemical assay.

    Science.gov (United States)

    Muir, Alastair; Forrest, Gordon; Clarkson, John; Wheals, Alan

    2011-04-01

    The genus Candida contains a number of yeast species which are opportunistic pathogens and are associated with life-threatening infections in immunocompromised individuals. Provision of appropriate therapy relies on the rapid identification of the infecting species, and existing methods of identifying Candida species in clinical samples are time and resource intensive and are not always specific enough to differentiate between drug-susceptible and drug-resistant species. We have previously developed a system for the rapid detection of yeast pathogens in clinical samples using PCR followed by hybridization with a suite of five species-specific, electrochemically labelled DNA probes. The limit of detection of the assay was shown to be 37 fg (∼1 genome) per reaction using extracted genomic DNA. We carried out a study to test the limit of detection of one of the probes, CA PR3, using blood samples from a healthy donor that were spiked with genomic DNA or with C. albicans cells. Our results demonstrated a limit of detection of 37 fg (ml blood)(-1) (∼1 genome ml(-1)) using extracted DNA or 10 c.f.u. (ml blood)(-1) using C. albicans cells, indicating that the assay is capable of detecting C. albicans nucleic acid at levels that are encountered in clinical samples.

  18. Rapid diagnosis of candidaemia by real-time PCR detection of Candida DNA in blood samples.

    Science.gov (United States)

    Wellinghausen, Nele; Siegel, Dunja; Winter, Juliane; Gebert, Susanne

    2009-08-01

    This study prospectively evaluated an 18S rRNA gene-targeted real-time PCR approach in comparison with standard blood culture (BC) diagnostics for rapid diagnosis of candidaemia in a large study population of 384 patients, including 902 whole blood samples from 468 infectious episodes (IEs) of 329 adults and 55 children with haematological malignancies and various forms of immunodeficiency, and intensive care unit patients. Seven out of eight BC-proven cases (87.5 %) of candidaemia and seven out of twelve BC-positive samples (58.3 %) were positive by the Candida-specific PCR. A positive PCR result was also obtained for 28/460 BC-negative samples from IEs, including 8 patients with culture-confirmed Candida infection at primary sterile body sites. Of the PCR-positive, culture-negative patients, more than 50 % received systemic antifungal therapy. In 432/460 BC-negative IEs, the Candida specific-PCR was negative, resulting in a negative predictive value of 99.8 %. In conclusion, the Candida specific-PCR approach facilitates rapid detection of Candida DNA in blood samples of patients at risk of candidaemia within a few hours. Although standard BC diagnostics appear to remain indispensable for the detection of all cases of candidaemia, this PCR assay allowed the detection of candidaemia at a mean of 3 days earlier than BC diagnostics. Thus, it enables earlier antifungal therapy for patients with suspected candidaemia and may prevent further complications.

  19. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status.

    Science.gov (United States)

    Bonelli, Patrizia; Buonocore, Ruggero; Aloe, Rosalia; Lippi, Giuseppe

    2016-04-01

    The measurement of vitamin D is now commonplace for preventing osteoporosis and restoring an appropriate concentration that would be effective to counteract the occurrence of other human disorders. The aim of this study was to establish whether blood sampling seasonality may influence total vitamin D concentration in a general population of Italian unselected outpatients. We performed a retrospective search in the laboratory information system of the University Hospital of Parma (Italy, temperate climate), to identify the values of total serum vitamin D (25-hydroxyvitamin D) measured in outpatients aged 18 years and older, who were referred for routine health check-up during the entire year 2014. The study population consisted in 11,150 outpatients (median age 62 years; 8592 women and 2558 men). The concentration of vitamin D was consistently lower in samples collected in Winter than in the other three seasons. The frequency of subjects with vitamin D deficiency was approximately double in samples drawn in Winter and Spring than in Summer and Autumn. In the multivariate analysis, the concentration of total vitamin D was found to be independently associated with sex and season of blood testing, but not with the age of the patients. According to these findings, blood sampling seasonality should be regarded as an important preanalytical factor in vitamin D assessment. It is also reasonable to suggest that the amount of total vitamin D synthesized during the summer should be high enough to maintain the levels > 50 nmol/L throughout the remaining part of the year.

  20. Automated on-line liquid–liquid extraction system for temporal mass spectrometric analysis of dynamic samples

    Energy Technology Data Exchange (ETDEWEB)

    Hsieh, Kai-Ta; Liu, Pei-Han [Department of Applied Chemistry, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan (China); Urban, Pawel L. [Department of Applied Chemistry, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan (China); Institute of Molecular Science, National Chiao Tung University, 1001 University Rd, Hsinchu, 300, Taiwan (China)

    2015-09-24

    Most real samples cannot directly be infused to mass spectrometers because they could contaminate delicate parts of ion source and guides, or cause ion suppression. Conventional sample preparation procedures limit temporal resolution of analysis. We have developed an automated liquid–liquid extraction system that enables unsupervised repetitive treatment of dynamic samples and instantaneous analysis by mass spectrometry (MS). It incorporates inexpensive open-source microcontroller boards (Arduino and Netduino) to guide the extraction and analysis process. Duration of every extraction cycle is 17 min. The system enables monitoring of dynamic processes over many hours. The extracts are automatically transferred to the ion source incorporating a Venturi pump. Operation of the device has been characterized (repeatability, RSD = 15%, n = 20; concentration range for ibuprofen, 0.053–2.000 mM; LOD for ibuprofen, ∼0.005 mM; including extraction and detection). To exemplify its usefulness in real-world applications, we implemented this device in chemical profiling of pharmaceutical formulation dissolution process. Temporal dissolution profiles of commercial ibuprofen and acetaminophen tablets were recorded during 10 h. The extraction-MS datasets were fitted with exponential functions to characterize the rates of release of the main and auxiliary ingredients (e.g. ibuprofen, k = 0.43 ± 0.01 h{sup −1}). The electronic control unit of this system interacts with the operator via touch screen, internet, voice, and short text messages sent to the mobile phone, which is helpful when launching long-term (e.g. overnight) measurements. Due to these interactive features, the platform brings the concept of the Internet-of-Things (IoT) to the chemistry laboratory environment. - Highlights: • Mass spectrometric analysis normally requires sample preparation. • Liquid–liquid extraction can isolate analytes from complex matrices. • The proposed system automates

  1. Automated on-line liquid–liquid extraction system for temporal mass spectrometric analysis of dynamic samples

    International Nuclear Information System (INIS)

    Hsieh, Kai-Ta; Liu, Pei-Han; Urban, Pawel L.

    2015-01-01

    Most real samples cannot directly be infused to mass spectrometers because they could contaminate delicate parts of ion source and guides, or cause ion suppression. Conventional sample preparation procedures limit temporal resolution of analysis. We have developed an automated liquid–liquid extraction system that enables unsupervised repetitive treatment of dynamic samples and instantaneous analysis by mass spectrometry (MS). It incorporates inexpensive open-source microcontroller boards (Arduino and Netduino) to guide the extraction and analysis process. Duration of every extraction cycle is 17 min. The system enables monitoring of dynamic processes over many hours. The extracts are automatically transferred to the ion source incorporating a Venturi pump. Operation of the device has been characterized (repeatability, RSD = 15%, n = 20; concentration range for ibuprofen, 0.053–2.000 mM; LOD for ibuprofen, ∼0.005 mM; including extraction and detection). To exemplify its usefulness in real-world applications, we implemented this device in chemical profiling of pharmaceutical formulation dissolution process. Temporal dissolution profiles of commercial ibuprofen and acetaminophen tablets were recorded during 10 h. The extraction-MS datasets were fitted with exponential functions to characterize the rates of release of the main and auxiliary ingredients (e.g. ibuprofen, k = 0.43 ± 0.01 h −1 ). The electronic control unit of this system interacts with the operator via touch screen, internet, voice, and short text messages sent to the mobile phone, which is helpful when launching long-term (e.g. overnight) measurements. Due to these interactive features, the platform brings the concept of the Internet-of-Things (IoT) to the chemistry laboratory environment. - Highlights: • Mass spectrometric analysis normally requires sample preparation. • Liquid–liquid extraction can isolate analytes from complex matrices. • The proposed system automates the

  2. Distribution of blood types in a sample of 245 New Zealand non-purebred cats.

    Science.gov (United States)

    Cattin, R P

    2016-05-01

    To determine the distribution of feline blood types in a sample of non-pedigree, domestic cats in New Zealand, whether a difference exists in this distribution between domestic short haired and domestic long haired cats, and between the North and South Islands of New Zealand; and to calculate the risk of a random blood transfusion causing a severe transfusion reaction, and the risk of a random mating producing kittens susceptible to neonatal isoerythrolysis. The results of 245 blood typing tests in non-pedigree cats performed at the New Zealand Veterinary Pathology (NZVP) and Gribbles Veterinary Pathology laboratories between the beginning of 2009 and the end of 2014 were retrospectively collated and analysed. Cats that were identified as domestic short or long haired were included. For the cats tested at Gribbles Veterinary Pathology 62 were from the North Island, and 27 from the South Island. The blood type distribution differed between samples from the two laboratories (p=0.029), but not between domestic short and long haired cats (p=0.50), or between the North and South Islands (p=0.76). Of the 89 cats tested at Gribbles Veterinary Pathology, 70 (79%) were type A, 18 (20%) type B, and 1 (1%) type AB; for NZVP 139/156 (89.1%) cats were type A, 16 (10.3%) type B, and 1 (0.6%) type AB. It was estimated that 18.3-31.9% of random blood transfusions would be at risk of a transfusion reaction, and neonatal isoerythrolysis would be a risk in 9.2-16.1% of random matings between non-pedigree cats. The results from this study suggest that there is a high risk of complications for a random blood transfusion between non-purebred cats in New Zealand. Neonatal isoerythrolysis should be considered an important differential diagnosis in illness or mortality in kittens during the first days of life.

  3. High positive predictive value of Gram stain on catheter-drawn blood samples for the diagnosis of catheter-related bloodstream infection in intensive care neonates.

    Science.gov (United States)

    Deleers, M; Dodémont, M; Van Overmeire, B; Hennequin, Y; Vermeylen, D; Roisin, S; Denis, O

    2016-04-01

    Catheter-related bloodstream infections (CRBSIs) remain a leading cause of healthcare-associated infections in preterm infants. Rapid and accurate methods for the diagnosis of CRBSIs are needed in order to implement timely and appropriate treatment. A retrospective study was conducted during a 7-year period (2005-2012) in the neonatal intensive care unit of the University Hospital Erasme to assess the value of Gram stain on catheter-drawn blood samples (CDBS) to predict CRBSIs. Both peripheral samples and CDBS were obtained from neonates with clinically suspected CRBSI. Gram stain, automated culture and quantitative cultures on blood agar plates were performed for each sample. The paired quantitative blood culture was used as the standard to define CRBSI. Out of 397 episodes of suspected CRBSIs, 35 were confirmed by a positive ratio of quantitative culture (>5) or a colony count of CDBS culture >100 colony-forming units (CFU)/mL. All but two of the 30 patients who had a CDBS with a positive Gram stain were confirmed as having a CRBSI. Seven patients who had a CDBS with a negative Gram stain were diagnosed as CRBSI. The sensitivity, specificity, positive predictive value and negative predictive value of Gram stain on CDBS were 80, 99.4, 93.3 and 98.1 %, respectively. Gram staining on CDBS is a viable method for rapidly (<1 h) detecting CRBSI without catheter withdrawal.

  4. Portable automation of static chamber sample collection for quantifying soil gas flux

    Science.gov (United States)

    The collection of soil gas flux using the static chamber method is labor intensive. The number of chambers that can be sampled in a given time period is limited by the spacing between chambers and the availability of trained research technicians. However, the static chamber method can limit spatial ...

  5. Automated sample-processing and titration system for determining uranium in nuclear materials

    International Nuclear Information System (INIS)

    Harrar, J.E.; Boyle, W.G.; Breshears, J.D.; Pomernacki, C.L.; Brand, H.R.; Kray, A.M.; Sherry, R.J.; Pastrone, J.A.

    1977-01-01

    The system is designed for accurate, precise, and selective determination of from 10 to 180 mg of uranium in 2 to 12 cm 3 of solution. Samples, standards, and their solutions are handled on a weight basis. These weights, together with their appropriate identification numbers, are stored in computer memory and are used automatically in the assay calculations after each titration. The measurement technique (controlled-current coulometry) is based on the Davies-Gray and New Brunswick Laboratory method, in which U(VI) is reduced to U(IV) in strong H 3 PO 4 , followed by titration of the U(IV) with electrogenerated V(V). Solution pretreatment and titration are automatic. The analyzer is able to process 44 samples per loading of the sample changer, at a rate of 4 to 9 samples per hour. The system includes a comprehensive fault-monitoring system that detects analytical errors, guards against abnormal conditions which might cause errors, and prevents unsafe operation. A detailed description of the system, information on the reliability of the component subsystems, and a summary of its evaluation by the New Brunswick Laboratory are presented

  6. [Guidelines for electronic-data-processing-controlled serial diagnosis of donor blood samples].

    Science.gov (United States)

    Fischer-Fröhlich, C L; Stoll, T; Hanfland, P

    1990-01-01

    Increasing performance figures and the necessity to save expenses oblige transfusion services to automatize their donors' laboratory examination. Sufficient hard- and software for sample distribution and processing is now available. Following aspects should be regarded when switching to automatic serial screening: The identity of blood-donor, donation and laboratory result will be achieved by machine readable labeling and on-line communication between working-stations and central administration. Flexibility: Easy automatic selective laboratory screening will be possible using special barcodes including sample identification and working orders. A modular hardware concept with easily accessible programming control allows it to implement new devices or methods. Ergonomy: Automatic sample processing including selective screening and simultaneous operating robotic sample processors increase working quality, sample output and time benefits. Economy: Improved working conditions will result in saving reagents and compensating staff limitations.

  7. Detection of tumor-associated cells in cryopreserved peripheral blood mononuclear cell samples for retrospective analysis.

    Science.gov (United States)

    Zhu, Peixuan; Stanton, Melissa L; Castle, Erik P; Joseph, Richard W; Adams, Daniel L; Li, Shuhong; Amstutz, Platte; Tang, Cha-Mei; Ho, Thai H

    2016-07-02

    Cryopreserved peripheral blood mononuclear cells (PBMCs) are commonly collected in biobanks. However, little data exist regarding the preservation of tumor-associated cells in cryopreserved collections. The objective of this study was to determine the feasibility of using the CellSieve™ microfiltration assay for the isolation of circulating tumor cells (CTCs) and circulating cancer-associated macrophage-like cells (CAMLs) from cryopreserved PBMC samples. Blood samples spiked with breast (MCF-7), prostate (PC-3), and renal (786-O) cancer cell lines were used to establish analytical accuracy, efficiency, and reproducibility after cryopreservation. The spiked samples were processed through Ficoll separation, and cryopreservation was followed by thawing and microfiltration. MCF-7 cells were successfully retrieved with recovery efficiencies of 90.5 % without cryopreservation and 87.8 and 89.0 %, respectively, on day 7 and day 66 following cryopreservation. The corresponding recovery efficiencies of PC-3 cells were 83.3 % without cryopreservation and 85.3 and 84.7 %, respectively, after cryopreservation. Recovery efficiencies of 786-O cells were 92.7 % without cryopreservation, and 82.7 and 81.3 %, respectively, after cryopreservation. The recovered cells retained the morphologic characteristics and immunohistochemical markers that had been observed before freezing. The protocols were further validated by quantitation of CAMLs in blood samples from two patients with renal cell carcinoma (RCC). The recovery rates of CTCs and CAMLs from cryopreserved samples were not statistically significant different (P > 0.05) from matched fresh samples. To our knowledge, this is the first report that CAMLs could be cryopreserved and analyzed after thawing with microfiltration technology. The application of microfiltration technology to cryopreserved samples will enable much greater retrospective study of cancer patients in relation to long-term outcomes.

  8. Potentiating day-old blood samples for detection of interferon-gamma responses following infection with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Nielsen, Søren Saxmose; Jungersen, Gregers

    time interval from blood sampling to culture. The objective of the study was to assess options for use of day-old blood samples for early-stage diagnosis of MAP infections. Bovine interleukin 12 (IL-12) can induce, and IL-10 reduce, IFN-γ production. Therefore, addition of IL-12 and anti-IL-10 could...... result in production of IFN-γ in samples previously exposed to MAP antigens. Whole blood samples were collected from heifers in a Danish dairy herd known to be infected with MAP. The samples were collected on three sample dates, and on each date the blood samples were stimulated with PPDj and recombinant......The interferon gamma (IFN-γ) test measuring specific cell-mediated immune responses in whole blood can be used for diagnosis at an early stage of Mycobacterium avium subsp. paratuberculosis (MAP) infection. A major obstacle for the practical use of IFN-γ testing is the recommended maximum 8 hour...

  9. Performance evaluation of continuous blood sampling system for PET study. Comparison of three detector-systems

    CERN Document Server

    Matsumoto, K; Sakamoto, S; Senda, M; Yamamoto, S; Tarutani, K; Minato, K

    2002-01-01

    To measure cerebral blood flow with sup 1 sup 5 O PET, it is necessary to measure the time course of arterial blood radioactivity. We examined the performance of three different types of continuous blood sampling system. Three kinds of continuous blood sampling system were used: a plastic scintillator-based beta detector (conventional beta detector (BETA)), a bismuth germinate (BGO)-based coincidence gamma detector (Pico-count flow-through detector (COINC)) and a Phoswich detector (PD) composed by a combination of plastic scintillator and BGO scintillator. Performance of these systems was evaluated for absolute sensitivity, count rate characteristic, sensitivity to background gamnra photons, and reproducibility for nylon tube geometry. The absolute sensitivity of the PD was 0.21 cps/Bq for sup 6 sup 8 Ga positrons at the center of the detector. This was approximately three times higher than BETA, two times higher than COINC. The value measured with BETA was stable, even when background radioactivity was incre...

  10. Aflatoxin B1-lysine adduct in dried blood spot samples of animals and humans.

    Science.gov (United States)

    Xue, Kathy S; Cai, Wenjie; Tang, Lili; Wang, Jia-Sheng

    2016-12-01

    Dried blood spots (DBS) were proposed as potentially viable method for exposure assessment of environmental toxicants in infant and young children. For this study, we validated an experimental protocol to quantify AFB 1 -lysine adduct in DBS samples of AFB 1 -treated F344 rats, as well as samples from human field study. Significant dose-response relationships in AFB 1 -lysine adduct formation were found in DBS samples of rats treated with single- and repeated-dose AFB 1 . AFB 1 -lysine levels in DBS samples were highly correlated with corresponding serum sample levels. The Person coefficients were 0.997 for the single-dose exposure, and 0.996 for the repeated-dose exposure. Levels of AFB 1 -lysine adduct had also good agreement between DBS and serum samples as shown by Bland-Altman plot analysis. For human field study samples (n = 36), a Pearson correlation coefficient of 0.784 was found between AFB 1 -lysine adduct levels of DBS and corresponding serum samples. Bland-Altman plots showed the distribution of the log differences between DBS and serum AFB 1 -lysine levels are within 95% confidence intervals. These results showed AFB 1 -lysine adduct levels in DBS cards and serum samples from animals and human samples are comparable, and the DBS technique and analytical protocol is a good means to assess AFB 1 exposure in infant and children populations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

    DEFF Research Database (Denmark)

    Teilmann, Anne Charlotte; Nygaard Madsen, Andreas; Holst, Birgitte

    2014-01-01

    Retro-bulbar sinus puncture and facial vein phlebotomy are two widely used methods for blood sampling in laboratory mice. However, the animal welfare implications associated with these techniques are currently debated, and the possible physiological and pathological implications of blood sampling...... using these methods have been sparsely investigated. Therefore, this study was conducted to assess and compare the impacts of blood sampling by retro-bulbar sinus puncture and facial vein phlebotomy. Blood was obtained from either the retro-bulbar sinus or the facial vein from male C57BL/6J mice at two...... extensive tissue trauma after both facial vein phlebotomy and retro-bulbar sinus puncture. This study demonstrates that both blood sampling methods have a considerable impact on the animals' physiological condition, which should be considered whenever blood samples are obtained....

  12. Sorbent Tube Sampling and an Automated Thermal Desorption System for Halocarbon Analysis

    Directory of Open Access Journals (Sweden)

    Md. Anwar Hossain Khan

    2009-01-01

    Full Text Available Development and deployment of the analytical sys tem, ATD-GC-ECD has been established to monitor a suite of halogenated com pounds found in the atmosphere at trace concentrations. The instrument has been used to monitor urban back ground emission flux levels in Bristol, UK as well as Yellow stone National Park, USA and an in door rain forest (Wild Walk@Bristol, UK. The newly established sorbent tube sampling system is small and easily portable and has been used for large volume sample collection from remote areas. Auto mated Thermal Desorption (ATD provides routine atmospheric measurements with out cryogenic pre-concentration. The instrument provides good precision where the detection limit was _T_n3 pptv for the species of interest and the reproducibility was within 4% for all of the selected halocarbons. The results from two field experiments have also pro vided insight about natural missing sources of some ozone depleting halocarbons.

  13. Analysis of polycyclic aromatic hydrocarbons in soil: minimizing sample pretreatment using automated Soxhlet with ethyl acetate as extraction solvent.

    Science.gov (United States)

    Szolar, Oliver H J; Rost, Helmut; Braun, Rudolf; Loibner, Andreas P

    2002-05-15

    A simplified sample pretreatment method for industrially PAH-contaminated soils applying automated Soxhlet (Soxtherm) with ethyl acetate as extraction solvent is presented. Laborious pretreatment steps such as drying of samples, cleanup of crude extracts, and solvent exchange were allowed to be bypassed without notable performance impact. Moisture of the soil samples did not significantly influence recoveries of PAHs at a wide range of water content for the newly developed method. However, the opposite was true for the standard procedure using the more apolar 1:1 (v/v) n-hexane/acetone solvent mixture including postextraction treatments recommended by the U.S. EPA. Moreover, ethyl acetate crude extracts did not appreciably effect the chromatographic performance (HPLC-(3D)FLD), which was confirmed by a comparison of the purity of PAH spectra from both pretreatment methods. Up to 20% (v/v) in acetonitrile, ethyl acetate proved to be fully compatible with the mobile phase of the HPLC whereas the same concentration of n-hexane/acetone in acetonitrile resulted in significant retention time shifts. The newly developed pretreatment method was applied to three historically contaminated soils from different sources with extraction efficiencies not being significantly different compared to the standard procedure. Finally, the certified reference soil CRM 524 was subjected to the simplified procedure resulting in quantitative recoveries (>92%) for all PAHs analyzed.

  14. A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants.

    Science.gov (United States)

    Heub, Sarah; Tscharner, Noe; Kehl, Florian; Dittrich, Petra S; Follonier, Stéphane; Barbe, Laurent

    2016-01-01

    A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and enables to perform rapid, automated and simple SPE. The pre-concentrated solution is compatible with analysis by immunoassay, with a low organic solvent content. A method is described for the extraction and pre-concentration of natural hormone 17β-estradiol in 100 ml water samples. Reverse phase SPE is performed with octadecyl-silica sorbent and elution is done with 200 µl of methanol 50% v/v. Eluent is diluted by adding di-water to lower the amount of methanol. After preparing manually the SPE column, the overall procedure is performed automatically within 1 hr. At the end of the process, estradiol concentration is measured by using a commercial enzyme-linked immune-sorbent assay (ELISA). 100-fold pre-concentration is achieved and the methanol content in only 10% v/v. Full recoveries of the molecule are achieved with 1 ng/L spiked de-ionized and synthetic sea water samples.

  15. Wuchereria bancrofti in Tanzania: microfilarial periodicity and effect of blood sampling time on microfilarial intensities

    DEFF Research Database (Denmark)

    Simonsen, Poul Erik; Niemann, L.; Meyrowitsch, Dan Wolf

    1997-01-01

    The circadian periodicity of Wuchereria bancrofti microfilarial (mf) intensities in peripheral blood was analysed in a group of infected individuals from an endemic community in north-eastern Tanzania. The mf density was quantified at two-hourly intervals for 24 hours. A clear nocturnal periodic...... pattern was observed. Mathematical analysis of the data indicated a peak at 0152 h and a periodicity index of 117.5. A periodicity equation was developed describing the average relation between mf intensity and hour of the day for the study area. Based on the observed periodicity pattern, the effect...... of blood sampling before peak time is discussed, and the importance of taking sampling time into consideration when analysing data from epidemiological studies is emphasized. A simple method is devised which can be used to adjust for the influence of time on mf intensities, in studies where accurate...

  16. Optical detection of Trypanosoma cruzi in blood samples for diagnosis purpose

    Science.gov (United States)

    Alanis, Elvio; Romero, Graciela; Alvarez, Liliana; Martinez, Carlos C.; Basombrio, Miguel A.

    2004-10-01

    An optical method for detection of Trypanosoma Cruzi (T. cruzi) parasites in blood samples of mice infected with Chagas disease is presented. The method is intended for use in human blood, for diagnosis purposes. A thin layer of blood infected by T. cruzi parasites, in small concentrations, is examined in an interferometric microscope in which the images of the vision field are taken by a CCD camera and temporarily stored in the memory of a host computer. The whole sample is scanned displacing the microscope plate by means of step motors driven by the computer. Several consecutive images of the same field are taken and digitally processed by means of image temporal differentiation in order to detect if a parasite is eventually present in the field. Each field of view is processed in the same fashion, until the full area of the sample is covered or until a parasite is detected, in which case an acoustical warning is activated and the corresponding image is displayed permitting the technician to corroborate the result visually. A discussion of the reliability of the method as well as a comparison with other well established techniques are presented.

  17. Measuring Blood Glucose Concentrations in Photometric Glucometers Requiring Very Small Sample Volumes.

    Science.gov (United States)

    Demitri, Nevine; Zoubir, Abdelhak M

    2017-01-01

    Glucometers present an important self-monitoring tool for diabetes patients and, therefore, must exhibit high accuracy as well as good usability features. Based on an invasive photometric measurement principle that drastically reduces the volume of the blood sample needed from the patient, we present a framework that is capable of dealing with small blood samples, while maintaining the required accuracy. The framework consists of two major parts: 1) image segmentation; and 2) convergence detection. Step 1 is based on iterative mode-seeking methods to estimate the intensity value of the region of interest. We present several variations of these methods and give theoretical proofs of their convergence. Our approach is able to deal with changes in the number and position of clusters without any prior knowledge. Furthermore, we propose a method based on sparse approximation to decrease the computational load, while maintaining accuracy. Step 2 is achieved by employing temporal tracking and prediction, herewith decreasing the measurement time, and, thus, improving usability. Our framework is tested on several real datasets with different characteristics. We show that we are able to estimate the underlying glucose concentration from much smaller blood samples than is currently state of the art with sufficient accuracy according to the most recent ISO standards and reduce measurement time significantly compared to state-of-the-art methods.

  18. Comparison of Performance Characteristics of Aspergillus PCR in Testing a Range of Blood-Based Samples in Accordance with International Methodological Recommendations

    Science.gov (United States)

    Springer, Jan; Hamilton, Shanna; Michel, Denise; Barnes, Rosemary A.; Einsele, Hermann; Löffler, Juergen

    2016-01-01

    Standardized methodologies for the molecular detection of invasive aspergillosis (IA) have been established by the European Aspergillus PCR Initiative for the testing of whole blood, serum, and plasma. While some comparison of the performance of Aspergillus PCR when testing these different sample types has been performed, no single study has evaluated all three using the recommended protocols. Standardized Aspergillus PCR was performed on 423 whole-blood pellets (WBP), 583 plasma samples, and 419 serum samples obtained from hematology patients according to the recommendations. This analysis formed a bicenter retrospective anonymous case-control study, with diagnosis according to the revised European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) consensus definitions (11 probable cases and 36 controls). Values for clinical performance using individual and combined samples were calculated. For all samples, PCR positivity was significantly associated with cases of IA (for plasma, P = 0.0019; for serum, P = 0.0049; and for WBP, P = 0.0089). Plasma PCR generated the highest sensitivity (91%); the sensitivities for serum and WBP PCR were 80% and 55%, respectively. The highest specificity was achieved when testing WBP (96%), which was significantly superior to the specificities achieved when testing serum (69%, P = 0.0238) and plasma (53%, P = 0.0002). No cases were PCR negative in all specimen types, and no controls were PCR positive in all specimens. This study confirms that Aspergillus PCR testing of plasma provides robust performance while utilizing commercial automated DNA extraction processes. Combining PCR testing of different blood fractions allows IA to be both confidently diagnosed and excluded. A requirement for multiple PCR-positive plasma samples provides similar diagnostic utility and is technically less demanding. Time

  19. Stability of HE4 and CA125 in blood samples from patients diagnosed with ovarian cancer

    DEFF Research Database (Denmark)

    Sandhu, Noreen; Karlsen, Mona A; Høgdall, Claus

    2014-01-01

    OBJECTIVE: To investigate the influence of handling and storage on HE4 and CA125 serum and EDTA plasma levels to clarify any important consequences for a clinical setting. METHODS: Blood samples from 13 ovarian cancer (OC) patients were collected and allowed to clot or sediment for up to 72 hours.......024). No significant difference between CA125 serum and plasma levels were found (p = 0.46). Serum and EDTA plasma samples were stable during the eight cycles of freezing and thawing (CA125: all p > 0.2; HE4: all p > 0.5). CONCLUSION: No systematic difference could be demonstrated for HE4. CA125 is not dependent...

  20. Heel blood sampling in European neonatal intensive care units: compliance with pain management guidelines

    DEFF Research Database (Denmark)

    Losacco, Valentina; Cuttini, Marina; Greisen, Gorm

    2011-01-01

    Objective To describe the use of heel blood sampling and non-pharmacological analgesia in a large representative sample of neonatal intensive care units (NICUs) in eight European countries, and compare their self-reported practices with evidence-based recommendations. Methods Information on use...... and France were the most likely, and Belgium and Spain the least likely to employ recommended combinations of evidence-based pain management measures. Conclusions Heel puncture is a common procedure in preterm neonates, but pain appears inadequately treated in many units and countries. Better compliance...

  1. Reducing the cost of semi-automated in-gel tryptic digestion and GeLC sample preparation for high-throughput proteomics.

    Science.gov (United States)

    Ruelcke, Jayde E; Loo, Dorothy; Hill, Michelle M

    2016-10-21

    Peptide generation by trypsin digestion is typically the first step in mass spectrometry-based proteomics experiments, including 'bottom-up' discovery and targeted proteomics using multiple reaction monitoring. Manual tryptic digest and the subsequent clean-up steps can add variability even before the sample reaches the analytical platform. While specialized filter plates and tips have been designed for automated sample processing, the specialty reagents required may not be accessible or feasible due to their high cost. Here, we report a lower-cost semi-automated protocol for in-gel digestion and GeLC using standard 96-well microplates. Further cost savings were realized by re-using reagent tips with optimized sample ordering. To evaluate the methodology, we compared a simple mixture of 7 proteins and a complex cell-lysate sample. The results across three replicates showed that our semi-automated protocol had performance equal to or better than a manual in-gel digestion with respect to replicate variability and level of contamination. In this paper, we also provide the Agilent Bravo method file, which can be adapted to other liquid handlers. The simplicity, reproducibility, and cost-effectiveness of our semi-automated protocol make it ideal for routine in-gel and GeLC sample preparations, as well as high throughput processing of large clinical sample cohorts. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Blood gas sample spiking with total parenteral nutrition, lipid emulsion, and concentrated dextrose solutions as a model for predicting sample contamination based on glucose result.

    Science.gov (United States)

    Jara-Aguirre, Jose C; Smeets, Steven W; Wockenfus, Amy M; Karon, Brad S

    2018-03-16

    Evaluate the effects of blood gas sample contamination with total parenteral nutrition (TPN)/lipid emulsion and dextrose 50% (D50) solutions on blood gas and electrolyte measurement; and determine whether glucose concentration can predict blood gas sample contamination with TPN/lipid emulsion or D50. Residual lithium heparin arterial blood gas samples were spiked with TPN/lipid emulsion (0 to 15%) and D50 solutions (0 to 2.5%). Blood gas (pH, pCO2, pO2), electrolytes (Na+, K+ ionized calcium) and hemoglobin were measured with a Radiometer ABL90. Glucose concentration was measured in separated plasma by Roche Cobas c501. Chart review of neonatal blood gas results with glucose >300 mg/dL (>16.65 mmol/L) over a seven month period was performed to determine whether repeat (within 4 h) blood gas results suggested pre-analytical errors in blood gas results. Results were used to determine whether a glucose threshold could predict contamination resulting in blood gas and electrolyte results with greater than laboratory-defined allowable error. Samples spiked with 5% or more TPN/lipid emulsion solution or 1% D50 showed glucose concentration >500 mg/dL (>27.75 mmol/L) and produced blood gas (pH, pO 2 , pCO 2 ) results with greater than laboratory-defined allowable error. TPN/lipid emulsion, but not D50, produced greater than allowable error in electrolyte (Na + ,K + ,Ca ++ ,Hb) results at these concentrations. Based on chart review of 144 neonatal blood gas results with glucose >250 mg/dL received over seven months, four of ten neonatal intensive care unit (NICU) patients with glucose results >500 mg/dL and repeat blood gas results within 4 h had results highly suggestive of pre-analytical error. Only 3 of 36 NICU patients with glucose results 300-500 mg/dL and repeat blood gas results within 4 h had clear pre-analytical errors in blood gas results. Glucose concentration can be used as an indicator of significant blood sample contamination with either TPN

  3. TCRgamma gene rearrangement analysis in skin samples and peripheral blood of mycosis fungoides patients.

    Science.gov (United States)

    Kandolf Sekulović, L; Cikota, B; Stojadinović, O; Basanović, J; Skiljević, D; Medenica, Lj; Pavlović, M; Magić, Z

    2007-12-01

    Diagnosing mycosis fungoides (MF) can be challenging in the early stage of the disease because histopathological features may simulate a variety of benign inflammatory skin diseases. Assessment of T-cell clonality was found to be useful in diagnosis and follow-up of patients. In this study, PCR-based TCRgamma gene rearrangement analysis was performed in skin and peripheral blood samples of patients with MF treated at the two largest referral centers in Serbia, and the results obtained were correlated with clinical and follow-up data. Skin and peripheral blood samples were obtained with informed consent from 37 patients treated at the Department of Dermatology of the Military Medical Academy and the Medical Center of Serbia from 2001 to 2006. The median time of follow-up was 4 years. Multiplex PCR was used for TCRgamma gene rearrangement analysis in skin and peripheral blood samples. Clonality results were correlated with the clinical data and disease course data. Monoclonality was detected in skin samples of 30/37 patients (81%), in 2/5 patients with large-plaque parapsoriasis (LPP), in 28/32 (88%) patients with histologically proven MF, and in 1/16 (6%) patients with benign inflammatory dermatoses. A monoclonal pattern in both skin and peripheral blood was detected in 7/16 (44%) patients in the late stage of the disease, and in 1/7 (14%) patients in the early stage of the disease. A dominant clone was found in both skin and peripheral blood in 1/4 patients in remission, 2/5 with a stable disease, and 4/9 (44%) with disease progression. TCR-gamma gene rearrangement analysis can be regarded as a useful adjunct to diagnosis of epidermotropic lymphoproliferative disorders. The presence of a dominant clone in both the skin and peripheral blood was more frequently detected in late stages and in patients with disease progression, confirming the usefulness of clonality detection by TCR-gamma gene rearrangement analysis in follow-up of patients with primary cutaneous T

  4. A novel, fully-automated, chemiluminescent assay for the detection of 1,25-dihydroxyvitamin D in biological samples.

    Science.gov (United States)

    Valcour, Andre; Zierold, Claudia; Podgorski, Angela L; Olson, Gregory T; Wall, John V; DeLuca, Hector F; Bonelli, Fabrizio

    2016-11-01

    1,25-Dihydroxyvitamin D (1,25-(OH) 2 D), the hormonal form of vitamin D, is difficult to measure because of its low circulating levels (pg/mL), and similarity to more abundant metabolites. Here a fully-automated chemiluminescent assay that accurately and precisely measures 1,25-(OH) 2 D is described. The novel 1,25-(OH) 2 D assay was conceived based on four pillars: (1) the VDR's ligand binding domain (LBD) as a capture molecule; (2) reaction conditions wherein 1,25-(OH) 2 D favors binding to LBD vs. the vitamin D binding protein; (3) exploitation of liganded-LBD's conformational change; (4) a monoclonal antibody specific to liganded-LBD. This specific, conformational, sandwich approach, unique for automated measurement of haptens, is superior to more cumbersome, conventional competitive formats. Accuracy of the 1,25-(OH) 2 D assay was corroborated by its alignment against LC-MS/MS with fit Deming regression equations of y=0.98x + 1.93 (r=0.92), and y=1.07x+3.77 (r=0.94) for different methods from Endocrine Sciences, Laboratory Corporation of America ® and the University of Washington, respectively. Good analytical precision was manifested by its low estimated limit of quantitation (1.57pg/mL), average intra-assay imprecision (3.5%CV; range 1.1-4.7%), and average inter-assay imprecision (4.5%CV; range 3.4-7.2%). Expected and measured recovery values were congruent (93.4% mean). The novel 1,25-(OH) 2 D method exhibited excellent correlation with well validated LC-MS/MS assays from two laboratories. Significantly, its 65min turn-around time is quicker, and sample volume smaller (75μl) than current methods. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Automated Image Sampling and Classification Can Be Used to Explore Perceived Naturalness of Urban Spaces.

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    Roger Hyam

    Full Text Available The psychological restorative effects of exposure to nature are well established and extend to just viewing of images of nature. A previous study has shown that Perceived Naturalness (PN of images correlates with their restorative value. This study tests whether it is possible to detect degree of PN of images using an image classifier. It takes images that have been scored by humans for PN (including a subset that have been assessed for restorative value and passes them through the Google Vision API image classification service. The resulting labels are assigned to broad semantic classes to create a Calculated Semantic Naturalness (CSN metric for each image. It was found that CSN correlates with PN. CSN was then calculated for a geospatial sampling of Google Street View images across the city of Edinburgh. CSN was found to correlate with PN in this sample also indicating the technique may be useful in large scale studies. Because CSN correlates with PN which correlates with restorativeness it is suggested that CSN or a similar measure may be useful in automatically detecting restorative images and locations. In an exploratory aside CSN was not found to correlate with an indicator of socioeconomic deprivation.

  6. Sampling theory and automated simulations for vertical sections, applied to human brain.

    Science.gov (United States)

    Cruz-Orive, L M; Gelšvartas, J; Roberts, N

    2014-02-01

    In recent years, there have been substantial developments in both magnetic resonance imaging techniques and automatic image analysis software. The purpose of this paper is to develop stereological image sampling theory (i.e. unbiased sampling rules) that can be used by image analysts for estimating geometric quantities such as surface area and volume, and to illustrate its implementation. The methods will ideally be applied automatically on segmented, properly sampled 2D images - although convenient manual application is always an option - and they are of wide applicability in many disciplines. In particular, the vertical sections design to estimate surface area is described in detail and applied to estimate the area of the pial surface and of the boundary between cortex and underlying white matter (i.e. subcortical surface area). For completeness, cortical volume and mean cortical thickness are also estimated. The aforementioned surfaces were triangulated in 3D with the aid of FreeSurfer software, which provided accurate surface area measures that served as gold standards. Furthermore, a software was developed to produce digitized trace curves of the triangulated target surfaces automatically from virtual sections. From such traces, a new method (called the 'lambda method') is presented to estimate surface area automatically. In addition, with the new software, intersections could be counted automatically between the relevant surface traces and a cycloid test grid for the classical design. This capability, together with the aforementioned gold standard, enabled us to thoroughly check the performance and the variability of the different estimators by Monte Carlo simulations for studying the human brain. In particular, new methods are offered to split the total error variance into the orientations, sectioning and cycloid components. The latter prediction was hitherto unavailable--one is proposed here and checked by way of simulations on a given set of digitized

  7. Dendritic cells in blood and urine samples from bladder cancer patients undergoing BCG immunotherapy

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    Raffaella Rossi

    2013-12-01

    Full Text Available Objectives: Immunotherapy with BCG (Bacille Calmette-Guérin after transurethral resection of the bladder tumor represents a highly effective primary treatment for intermediate and high-risk superficial bladder cancer. The effectiveness of this therapy has been documented, but its mechanism of action is not clear yet. In the present study, we investigated the changes of dendritic cells (DC numbers in peripheral blood and urine of patients with superficial bladder cancer undergoing BCG intravescical therapy Material and method: We have enumerated plasmacytoid and myeloid DCs in the peripheral blood and in the urine of patients with bladder cancer in order to clarify the role of these cells in the evolution of the disease and the effect of therapy. DCs in blood and urine samples were assessed using the single-platform TruCOUNT assay with monoclonal antibodies. The study population included 37 healthy donors and 13 patients with diagnosis of primitive superficial bladder cancer. Results: At the time of diagnosis a reduction of blood DCs was found in patients as opposed to healthy donors, while DCs were not found in the urine in the same way as in healthy subjects. Six of these patients were followed before and after weekly and monthly instillations of BCG. In the peripheral blood, we observed an immunological recovery of DCs from the third weekly instillation up to the sixth. In the urine of patients, we didn’t find mDCs or pDCs at T0, but we found a statistically significant change from the third instillation up to the sixth. On the contrary, we didn’t find mDCs in urine during monthly instillation. Conclusions: DC Count could be used in the monitoring of patients undergoing BCG therapy. Immunological restoration of mDC numbers in peripheral blood and the efflux in urine could be important for confirming the effectiveness of BCG instillation.

  8. Protein expression profiling by antibody array analysis with use of dried blood spot samples on filter paper.

    Science.gov (United States)

    Jiang, Weidong; Mao, Ying Qing; Huang, Ruochun; Duan, Chaohui; Xi, Yun; Yang, Kai; Huang, Ruo-Pan

    2014-01-31

    Dried blood spot samples (DBSS) on filter paper offer several advantages compared to conventional serum/plasma samples: they do not require any phlebotomy or separation of blood by centrifugation; they are less invasive; they allow sample stability and shipment at room temperature; and they pose a negligible risk of infection with blood-borne viruses, such as HIV, HBV and HCV, to those who handle them. Therefore dried blood spot samples (DBSS) on filter paper can be a quick, convenient and inexpensive means of obtaining blood samples for biomarker discovery, disease screening, diagnosis and treatment monitoring in non-hospitalized, public health settings. In this study, we investigated for the first time the potential application of dried blood spot samples (DBSS) in protein expression profiling using antibody array technology. First, optimal conditions for array assay performance using dried blood spot samples (DBSS) was established, including sample elution buffer, elution time, elution temperature and assay blocking buffer. Second, we analyzed dried blood spot samples (DBSS) using three distinct antibody array platforms, including sandwich-based antibody arrays, quantitative antibody arrays and biotin-label-based antibody arrays. In comparison with paired serum samples, detection of circulating proteins in dried blood spot samples (DBSS) correlated well for both low- and high-abundance proteins on all three antibody array platforms. In conclusion, our study strongly indicates the novel application of multiplex antibody array platforms to analyze dried blood spot samples (DBSS) on filter paper represents a viable, cost-effective method for protein profiling, biomarker discovery and disease screening in a large, population-based survey. Copyright © 2013 Elsevier B.V. All rights reserved.

  9. Classical and additional antiphospholipid antibodies in blood samples of ischemic stroke patients and healthy controls.

    Science.gov (United States)

    Carmel-Neiderman, Narin-Nard; Tanne, David; Goren, Idan; Rotman-Pikielny, Pnina; Levy, Yair

    2017-04-01

    Classical antiphospholipid antibodies (aPLa) are found in 6-25% of blood samples from stroke patients. The frequency of novel aPLa antibodies in blood samples of CVA patients is not known. Enzyme-linked immunosorbent assays (ELISA) were performed on blood samples from 209 CVA patients (170 samples were obtained during the acute phase and 39 samples were from patients with complete carotid stenosis) and compared to 54 healthy controls. Subjects were tested for the presence of the classical aPL antibodies anticardiolipin (aCL) and anti-beta2-glycoprotein (aβ2gI), in addition to antiphosphatidylethanolamine (aPE), anti-phosphatidylserine (aPS), and Annexin V. All antibodies were tested for both IgM and IgG subclasses. Numeric analysis of the antibody titer levels (μ/ml) revealed a significantly higher subclinical titer by two standard deviations of many aPL autoantibodies among CVA patients (Pv < 0.05). However, according to the kit manufacturer's cutoff value, no positive antibodies were found except a trend toward higher percentage of positive aPS IgG titer in the CVA group compared to controls (6.2 vs. %0; P = 0.077). According to the manufacturer's cutoff, significantly higher levels of positive antibodies were not found among stroke patients. However, the absolute ELISA values of stroke patients were significantly higher. These results suggest that lower cutoff values than those used for APS diagnosis should be used for risk stratification of CVA among healthy individuals.

  10. Robustness of genome-wide scanning using archived dried blood spot samples as a DNA source

    Directory of Open Access Journals (Sweden)

    Børglum Anders D

    2011-07-01

    Full Text Available Abstract Background The search to identify disease-susceptible genes requires access to biological material from numerous well-characterized subjects. Archived residual dried blood spot (DBS samples, also known as Guthrie cards, from national newborn screening programs may provide a DNA source for entire populations. Combined with clinical information from medical registries, DBS samples could provide a rich source for productive research. However, the amounts of DNA which can be extracted from these precious samples are minute and may be prohibitive for numerous genotypings. Previously, we demonstrated that DBS DNA can be whole-genome amplified and used for reliable genetic analysis on different platforms, including genome-wide scanning arrays. However, it remains unclear whether this approach is workable on a large sample scale. We examined the robustness of using DBS samples for whole-genome amplification following genome-wide scanning, using arrays from Illumina and Affymetrix. Results This study is based on 4,641 DBS samples from the Danish Newborn Screening Biobank, extracted for three separate genome-wide association studies. The amount of amplified DNA was significantly (P Conclusion Our study indicates that archived DBS samples from the Danish Newborn Screening Biobank represent a reliable resource of DNA for whole-genome amplification and subsequent genome-wide association studies. With call-rates equivalent to high quality DNA samples, our results point to new opportunities for using the neonatal biobanks available worldwide in the hunt for genetic components of disease.

  11. Development of CT-guided biopsy sampling for time-dependent postmortem redistribution investigations in blood and alternative matrices--proof of concept and application on two cases.

    Science.gov (United States)

    Staeheli, Sandra N; Gascho, Dominic; Fornaro, Juergen; Laberke, Patrick; Ebert, Lars C; Martinez, Rosa Maria; Thali, Michael J; Kraemer, Thomas; Steuer, Andrea E

    2016-02-01

    The postmortem redistribution (PMR) phenomenon complicates interpretation in forensic toxicology. Human data on time-dependent PMR are rare and only exist for blood so far. A new method for investigation of time-dependent PMR in blood as well as in alternative body fluids and tissues was developed and evaluated using automated biopsy sampling. At admission of the bodies, introducer needles were placed in liver, lung, kidney, muscle, spleen, adipose tissue, heart, femoral vein, and lumbar spine using a robotic arm guided by a computed tomography scanner (CT). Needle placement accuracy was analyzed and found to be acceptable for the study purpose. Tissue biopsies and small volume body fluid samples were collected in triplicate through the introducer needles. At autopsy (around 24 h after admission), samples from the same body regions were collected. After mastering of the technical challenges, two authentic cases were analyzed as a proof of concept. Drug concentrations of venlafaxine, O-desmethylvenlafaxine, bromazepam, flupentixol, paroxetine, and lorazepam were determined by LC-MS/MS, and the percentage concentration changes between the two time points were calculated. Concentration changes were observed with both increases and decreases depending on analyte and matrix. While venlafaxine, flupentixol, paroxetine, and lorazepam generally showed changes above 30% and more, O-desmethylvenlafaxine and bromazepam did not undergo extensive PMR. The presented study shows that CT-controlled biopsy collection provides a valuable tool for systematic time-dependent PMR investigation, demanding only minimal sample amount and causing minimal damage to the body.

  12. Preliminary evaluation of a highly automated instrument for the selection of CD34+ cells from mobilized peripheral blood stem cell concentrates.

    Science.gov (United States)

    Stroncek, David F; Tran, Minh; Frodigh, Sue Ellen; David-Ocampo, Virginia; Ren, Jiaqiang; Larochelle, Andre; Sheikh, Virginia; Sereti, Irini; Miller, Jeffery L; Longin, Kevin; Sabatino, Marianna

    2016-02-01

    Cell selection is an important part of manufacturing cellular therapies. A new highly automated instrument, the CliniMACS Prodigy (Miltenyi Biotec), was evaluated for the selection of CD34+ cells from mobilized peripheral blood stem cell (PBSC) concentrates using monoclonal antibodies conjugated to paramagnetic particles. PBSCs were collected by apheresis from 36 healthy subjects given granulocyte-colony-stimulating factor (G-CSF) or G-CSF plus plerixafor. CD34+ cells from 11 PBSC concentrates were isolated with the automated CliniMACS Prodigy and 25 with the semiautomated CliniMACS Plus Instrument. The proportion of CD34+ cells in the selected products obtained with the two instruments was similar: 93.6 ± 2.6% for the automated and 95.7 ± 3.3% for the semiautomated instrument (p > 0.05). The recovery of CD34+ cells from PBSC concentrates was less for the automated than the semiautomated instrument (51.4 ± 8.2% vs. 65.1 ± 15.7%; p = 0.019). The selected products from both instruments contained few and similar quantities of platelets (PLTs) and red blood cells. The depletion of CD3+ cells was less with the automated instrument (4.34 ± 0.2 log depletion vs. 5.20 ± 0.35 log depletion; p Prodigy and found that the nonselected cells for the Prodigy contained more PLTs than those for the CliniMACS Plus. CD34+ cells can be effectively selected from mobilized PBSC concentrates with the CliniMAC Prodigy, but the recovery of CD34+ cells and depletion of CD3+ cells was lower than with the semiautomated CliniMACS Plus Instrument. © 2015 AABB.

  13. Campylobacter in Broiler Chicken and Broiler Meat in Sri Lanka: Influence of Semi-Automated vs. Wet Market Processing on Campylobacter Contamination of Broiler Neck Skin Samples

    Directory of Open Access Journals (Sweden)

    Kottawattage S. A. Kottawatta

    2017-11-01

    Full Text Available Broiler meat can become contaminated with Campylobacter of intestinal origin during processing. The present study aimed to identify the prevalence of Campylobacter in broiler flocks and meat contamination at retail shops, and determine the influence of semi-automated and wet market processing on Campylobacter contamination of neck skin samples. Samples were collected from semi-automated plants (n = 102 and wet markets (n = 25. From each batch of broilers, pooled caecal samples and neck skin samples were tested for Campylobacter. Broiler meat purchased from retail outlets (n = 37 was also tested. The prevalence of Campylobacter colonized broiler flocks was 67%. The contamination of meat at retail was 59%. Both semi-automated and wet market processing resulted to contaminate the broiler neck skins to the levels of 27.4% and 48%, respectively. When Campylobacter-free broiler flocks were processed in semi-automated facilities 15% (5/33 of neck skin samples became contaminated by the end of processing whereas 25% (2/8 became contaminated after wet market processing. Characterization of isolates revealed a higher proportion of C. coli compared to C. jejuni. Higher proportions of isolates were resistant to important antimicrobials. This study shows the importance of Campylobacter in poultry industry in Sri Lanka and the need for controlling antimicrobial resistance.

  14. Campylobacter in Broiler Chicken and Broiler Meat in Sri Lanka: Influence of Semi-Automated vs. Wet Market Processing on Campylobacter Contamination of Broiler Neck Skin Samples.

    Science.gov (United States)

    Kottawatta, Kottawattage S A; Van Bergen, Marcel A P; Abeynayake, Preeni; Wagenaar, Jaap A; Veldman, Kees T; Kalupahana, Ruwani S

    2017-11-29

    Broiler meat can become contaminated with Campylobacter of intestinal origin during processing. The present study aimed to identify the prevalence of Campylobacter in broiler flocks and meat contamination at retail shops, and determine the influence of semi-automated and wet market processing on Campylobacter contamination of neck skin samples. Samples were collected from semi-automated plants ( n = 102) and wet markets ( n = 25). From each batch of broilers, pooled caecal samples and neck skin samples were tested for Campylobacter . Broiler meat purchased from retail outlets ( n = 37) was also tested. The prevalence of Campylobacter colonized broiler flocks was 67%. The contamination of meat at retail was 59%. Both semi-automated and wet market processing resulted to contaminate the broiler neck skins to the levels of 27.4% and 48%, respectively. When Campylobacter -free broiler flocks were processed in semi-automated facilities 15% (5/33) of neck skin samples became contaminated by the end of processing whereas 25% (2/8) became contaminated after wet market processing. Characterization of isolates revealed a higher proportion of C. coli compared to C. jejuni . Higher proportions of isolates were resistant to important antimicrobials. This study shows the importance of Campylobacter in poultry industry in Sri Lanka and the need for controlling antimicrobial resistance.

  15. Campylobacter in Broiler Chicken and Broiler Meat in Sri Lanka: Influence of Semi-Automated vs. Wet Market Processing on Campylobacter Contamination of Broiler Neck Skin Samples

    Science.gov (United States)

    Kottawatta, Kottawattage S. A.; Van Bergen, Marcel A. P.; Abeynayake, Preeni; Wagenaar, Jaap A.; Veldman, Kees T.; Kalupahana, Ruwani S.

    2017-01-01

    Broiler meat can become contaminated with Campylobacter of intestinal origin during processing. The present study aimed to identify the prevalence of Campylobacter in broiler flocks and meat contamination at retail shops, and determine the influence of semi-automated and wet market processing on Campylobacter contamination of neck skin samples. Samples were collected from semi-automated plants (n = 102) and wet markets (n = 25). From each batch of broilers, pooled caecal samples and neck skin samples were tested for Campylobacter. Broiler meat purchased from retail outlets (n = 37) was also tested. The prevalence of Campylobacter colonized broiler flocks was 67%. The contamination of meat at retail was 59%. Both semi-automated and wet market processing resulted to contaminate the broiler neck skins to the levels of 27.4% and 48%, respectively. When Campylobacter-free broiler flocks were processed in semi-automated facilities 15% (5/33) of neck skin samples became contaminated by the end of processing whereas 25% (2/8) became contaminated after wet market processing. Characterization of isolates revealed a higher proportion of C. coli compared to C. jejuni. Higher proportions of isolates were resistant to important antimicrobials. This study shows the importance of Campylobacter in poultry industry in Sri Lanka and the need for controlling antimicrobial resistance. PMID:29186018

  16. The Upgrade Programme for the Structural Biology beamlines at the European Synchrotron Radiation Facility – High throughput sample evaluation and automation

    International Nuclear Information System (INIS)

    Theveneau, P; Baker, R; Barrett, R; Beteva, A; Bowler, M W; Carpentier, P; Caserotto, H; Sanctis, D de; Dobias, F; Flot, D; Guijarro, M; Giraud, T; Lentini, M; Leonard, G A; Mattenet, M; McSweeney, S M; Morawe, C; Nurizzo, D; McCarthy, A A; Nanao, M

    2013-01-01

    Automation and advances in technology are the key elements in addressing the steadily increasing complexity of Macromolecular Crystallography (MX) experiments. Much of this complexity is due to the inter-and intra-crystal heterogeneity in diffraction quality often observed for crystals of multi-component macromolecular assemblies or membrane proteins. Such heterogeneity makes high-throughput sample evaluation an important and necessary tool for increasing the chances of a successful structure determination. The introduction at the ESRF of automatic sample changers in 2005 dramatically increased the number of samples that were tested for diffraction quality. This 'first generation' of automation, coupled with advances in software aimed at optimising data collection strategies in MX, resulted in a three-fold increase in the number of crystal structures elucidated per year using data collected at the ESRF. In addition, sample evaluation can be further complemented using small angle scattering experiments on the newly constructed bioSAXS facility on BM29 and the micro-spectroscopy facility (ID29S). The construction of a second generation of automated facilities on the MASSIF (Massively Automated Sample Screening Integrated Facility) beam lines will build on these advances and should provide a paradigm shift in how MX experiments are carried out which will benefit the entire Structural Biology community.

  17. Sensitivity of PCR assays for murine gammaretroviruses and mouse contamination in human blood samples.

    Directory of Open Access Journals (Sweden)

    Li Ling Lee

    Full Text Available Gammaretroviruses related to murine leukemia virus (MLV have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated from whole blood or from peripheral blood mononuclear cells (PBMCs or following PBMC culture. We have also passaged the prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids for viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous and exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse DNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA preparations, we are unable to conclude that these sequences originated in the blood samples.

  18. Leukocyte count affects expression of reference genes in canine whole blood samples

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    Dekker Aldo

    2011-02-01

    Full Text Available Abstract Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 individual dogs, representing 73 different breeds and a group of 40 mixed breed dogs, categorized into healthy dogs and dogs with internal and hematological diseases, and dogs that underwent a surgical procedure. GeNorm analysis revealed that a combination of 5 to 6 of the most stably expressed genes constituted a stable normalizing factor. Evaluation of the expression revealed different ranking of reference genes in Normfinder and GeNorm. The disease category and the white blood cell count significantly affected reference gene expression. Conclusions The discrepancy between the ranking of reference genes in this study by Normfinder and Genorm can be explained by differences between the experimental groups such as "disease category" and "WBC count". This stresses the importance of assessing the expression stability of potential reference genes for gene experiments in canine whole blood anew for each specific experimental condition.

  19. Evaluation of a novel dried blood spot collection device (HemaSpot™) to test blood samples collected from dogs for antibodies to Leishmania infantum.

    Science.gov (United States)

    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

  20. Frequency and antimicrobial susceptibility of acinetobacter species isolated from blood samples of paediatric patients

    International Nuclear Information System (INIS)

    Javed, A.; Zafar, A.; Ejaz, H.; Zubair, M.

    2012-01-01

    Objective: Acinetobacter species is a major nosocomial pathogen causing serious infections in immuno-compromised and hospitalized patients. The aim of this study was to determine the frequency and antimicrobial susceptibility pattern of Acinetobacter species in blood samples of paediatric patients. Methodology: This cross sectional observational study was conducted during January to October, 2011 at The Children's Hospital and Institute of Child Health, Lahore. A total number of 12,032 blood samples were analysed during the study period. Acinetobacter species were Bauer disc diffusion method. Results: The blood cultures showed growth in 1,141 cultures out of which 46 (4.0%) were Acinetobacter species. The gender distribution of Acinetobacter species was 29 (63.0%) in males and 17 (37.0%) in females. A good antimicrobial susceptibility pattern of Acinetobacter species was seen with sulbactam-cefoperazone (93.0%), imepenem and meropenem (82.6% (30.4%) was poor. Conclusion: The results of the present study shows high rate of resistance of Acinetobacter species with cephalosporins in nosocomial infections. The sulbactam-cefoperazone, carbapenems and piperacillin-tazobactam showed effective antimicrobial susceptibility against Acinetobacter species. (author)

  1. Creatinine measurement on dry blood spot sample for chronic kidney disease screening.

    Science.gov (United States)

    Silva, Alan Castro Azevedo E; Gómez, Juan Fidel Bencomo; Lugon, Jocemir Ronaldo; Graciano, Miguel Luis

    2016-03-01

    Chronic kidney disease (CKD) screening is advisable due to its high morbidity and mortality and is usually performed by sampling blood and urine. Here we present an innovative and simpler method, by measuring creatinine on a dry blood spot on filter paper. One-hundred and six individuals at high risk for CKD were enrolled. The creatinine values obtained using both tests and the demographic data of each participant allowed us to determinate the eGFR. The adopted cutoff for CKD was an eGFR value 96%, predictive negative value 55% and accuracy 92%. By the CKD-EPI equation the sensitivity was 94%, specificity 55%, predictive positive value 94%, predictive negative value 55% and accuracy 90%. A Bland and Altman analysis showed a relatively narrow range of creatinine values differences (+ 0.68mg/dl to -0.55mg/dl) inside the ± 1.96 SD, without systematic differences. Measurement of creatinine on dry blood sample is an easily feasible non-invasive diagnostic test with good accuracy that may be useful to screen chronic kidney disease.

  2. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    Directory of Open Access Journals (Sweden)

    Ironside James W

    2007-08-01

    Full Text Available Abstract Background Sub-clinical variant Creutzfeldt-Jakob disease (vCJD infection and reports of vCJD transmission through blood transfusion emphasise the need for blood screening assays to ensure the safety of blood and transplanted tissues. Most assays aim to detect abnormal prion protein (PrPSc, although achieving required sensitivity is a challenge. Methods We have used innovative Atomic Dielectric Resonance Spectroscopy (ADRS, which determines dielectric properties of materials which are established by reflectivity and penetration of radio/micro waves, to analyse blood samples from patients and controls to identify characteristic ADR signatures unique to blood from vCJD and to sCJD patients. Initial sets of blood samples from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors were screened as training samples to determine group-specific ADR characteristics, and provided a basis for classification of blinded sets of samples. Results Blood sample groups from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors screened by ADRS were classified with 100% specificity and sensitivity, discriminating these by a co-variance expert analysis system. Conclusion ADRS appears capable of recognising and discriminating serum samples from vCJD, sCJD, non-CJD neurological diseases, and normal healthy adults, and might be developed to provide a system for primary screening or confirmatory assay complementary to other screening systems.

  3. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    Science.gov (United States)

    Fagge, Timothy J; Barclay, G Robin; Stove, G Colin; Stove, Gordon; Robinson, Michael J; Head, Mark W; Ironside, James W; Turner, Marc L

    2007-01-01

    Background Sub-clinical variant Creutzfeldt-Jakob disease (vCJD) infection and reports of vCJD transmission through blood transfusion emphasise the need for blood screening assays to ensure the safety of blood and transplanted tissues. Most assays aim to detect abnormal prion protein (PrPSc), although achieving required sensitivity is a challenge. Methods We have used innovative Atomic Dielectric Resonance Spectroscopy (ADRS), which determines dielectric properties of materials which are established by reflectivity and penetration of radio/micro waves, to analyse blood samples from patients and controls to identify characteristic ADR signatures unique to blood from vCJD and to sCJD patients. Initial sets of blood samples from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) were screened as training samples to determine group-specific ADR characteristics, and provided a basis for classification of blinded sets of samples. Results Blood sample groups from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors) screened by ADRS were classified with 100% specificity and sensitivity, discriminating these by a co-variance expert analysis system. Conclusion ADRS appears capable of recognising and discriminating serum samples from vCJD, sCJD, non-CJD neurological diseases, and normal healthy adults, and might be developed to provide a system for primary screening or confirmatory assay complementary to other screening systems. PMID:17760958

  4. Automated microfluidic sample-preparation platform for high-throughput structural investigation of proteins by small-angle X-ray scattering

    DEFF Research Database (Denmark)

    Lafleur, Josiane P.; Snakenborg, Detlef; Nielsen, Søren Skou

    2011-01-01

    A new microfluidic sample-preparation system is presented for the structural investigation of proteins using small-angle X-ray scattering (SAXS) at synchrotrons. The system includes hardware and software features for precise fluidic control, sample mixing by diffusion, automated X-ray exposure...... control, UV absorbance measurements and automated data analysis. As little as 15 l of sample is required to perform a complete analysis cycle, including sample mixing, SAXS measurement, continuous UV absorbance measurements, and cleaning of the channels and X-ray cell with buffer. The complete analysis...... cycle can be performed in less than 3 min. Bovine serum albumin was used as a model protein to characterize the mixing efficiency and sample consumption of the system. The N2 fragment of an adaptor protein (p120-RasGAP) was used to demonstrate how the device can be used to survey the structural space...

  5. Detection of HTLV-I and -II in Scottish blood donor samples and archive donations.

    Science.gov (United States)

    Davidson, F; Lycett, C; Jarvis, L M; Kerr, D; Lumley, S; Petrik, J; Dow, B C

    2006-10-01

    Positive samples identified during routine serological screening for HCV (hepatitis C virus), HBV (hepatitis B virus) and HIV (human immunodeficiency virus) are confirmed by nucleic acid testing in the SNBTS (Scottish National Blood Transfusion Service) PCR Reference laboratory. Serological screening for HTLV-I (human T-cell lymphotropic virus type I) and -II was implemented in Scotland in November 2002, at which time a PCR assay was not available for confirmation. Our aim was to develop a real-time PCR assay that could be used for the confirmation of samples showing HTLV-I serological positive or indeterminate reactivity and to investigate whether a serologically silent carrier status exists ('Tax' only) in the Scottish donor population. A real-time HTLV PCR was devised using a lymphoblastoid cell line which has HTLV-I sequence integrated in the genome (C8166 cells). These were spiked into peripheral blood mononuclear cells. The assay was evaluated on archived serologically confirmed HTLV-positive samples and new positives identified since implementation of screening. HTLV-I and -II were detected in cells and plasma from stored donations and a serological positive donation identified in routine screening. HTLV DNA can also be amplified from the plasma obtained from plasma preparation tubes. There was no evidence of a carrier status ('Tax' only) in 100 serologically negative blood donors tested. The PCR assay developed is reliable and sensitive, capable of identifying one copy of HTLV-I. The HTLV PCR is a useful addition for HTLV confirmation, especially in serologically indeterminate samples and for look-back studies. HTLV PCR confirmation will provide additional useful information for donor medical staff for counselling donors.

  6. Robustness of genome-wide scanning using archived dried blood spot samples as a DNA source.

    Science.gov (United States)

    Hollegaard, Mads V; Grove, Jakob; Grauholm, Jonas; Kreiner-Møller, Eskil; Bønnelykke, Klaus; Nørgaard, Mette; Benfield, Thomas L; Nørgaard-Pedersen, Bent; Mortensen, Preben B; Mors, Ole; Sørensen, Henrik T; Harboe, Zitta B; Børglum, Anders D; Demontis, Ditte; Ørntoft, Torben F; Bisgaard, Hans; Hougaard, David M

    2011-07-04

    The search to identify disease-susceptible genes requires access to biological material from numerous well-characterized subjects. Archived residual dried blood spot (DBS) samples, also known as Guthrie cards, from national newborn screening programs may provide a DNA source for entire populations. Combined with clinical information from medical registries, DBS samples could provide a rich source for productive research. However, the amounts of DNA which can be extracted from these precious samples are minute and may be prohibitive for numerous genotypings. Previously, we demonstrated that DBS DNA can be whole-genome amplified and used for reliable genetic analysis on different platforms, including genome-wide scanning arrays. However, it remains unclear whether this approach is workable on a large sample scale. We examined the robustness of using DBS samples for whole-genome amplification following genome-wide scanning, using arrays from Illumina and Affymetrix. This study is based on 4,641 DBS samples from the Danish Newborn Screening Biobank, extracted for three separate genome-wide association studies. The amount of amplified DNA was significantly (P Biobank represent a reliable resource of DNA for whole-genome amplification and subsequent genome-wide association studies. With call-rates equivalent to high quality DNA samples, our results point to new opportunities for using the neonatal biobanks available worldwide in the hunt for genetic components of disease.

  7. Leakage of Oxygen from Blood and Water Samples Stored in Plastic and Glass Syringes

    Science.gov (United States)

    Scott, Peter V.; Horton, J. N.; Mapleson, W. W.

    1971-01-01

    Theory and experiment showed that samples of blood and water stored in 2-ml and 5-ml syringes made of polypropylene, polystyrene, or S.A.N. co-polymer exchanged oxygen with their surroundings. In the first hour the exchange was due mainly to equilibration with the plastic of the syringe and only in small degree to permeation through the plastic. With high initial tension or with blood of low haemoglobin concentration the exchange can result in errors in Po2 of up to 6% in two minutes and 16% in 30 to 60 minutes. With all-glass syringes the exchange was much slower but, even so, after 24 hours was important in all but a few of 18 interchangeable glass syringes. Therefore unless analysis can be started immediately all-glass syringes are to be preferred, and for prolonged storage even these should be selected. PMID:5565518

  8. Role of plasmin on the double antibody radioimmunoassay of carcinoembryonic antigen in human blood samples

    International Nuclear Information System (INIS)

    Das, S.; Das, B.R.

    1976-01-01

    Double antibody radioimmunoassay (RIA) of carcinoembryonic antigen (CEA) on a series of freshly drawn out concurrent plasma and serum samples of normal human blood donors showed that the serum CEA values were invariably higher than the corresponding plasma CEA values. Extraneous addition of fibrinogen brought down the serum--CEA level to a value comparable to or less than the corresponding plasma value. The effect of certain factors associated with blood clotting, particularly Ca ++ , fibrinogen, and the fibrinolytic enzyme plasmin, was investigated. Ca ++ was shown to play no role whereas the effect of fibrinogen was shown to be indirect in that it served as a specific substrate for plasmin, thereby preventing the plasmin degradation of the primary antibody used in the RIA. The finding stresses the role of enzymes like plasmin in double antibody RIA in general and may explain some of the anomalous results obtained when testing biologic material containing plasmin-like substances

  9. Development testing of the chemical analysis automation polychlorinated biphenyl standard analysis method during surface soils sampling at the David Witherspoon 1630 site

    International Nuclear Information System (INIS)

    Hunt, M.A.; Klatt, L.N.; Thompson, D.H.

    1998-02-01

    The Chemical Analysis Automation (CAA) project is developing standardized, software-driven, site-deployable robotic laboratory systems with the objective of lowering the per-sample analysis cost, decreasing sample turnaround time, and minimizing human exposure to hazardous and radioactive materials associated with DOE remediation projects. The first integrated system developed by the CAA project is designed to determine polychlorinated biphenyls (PCB) content in soil matrices. A demonstration and development testing of this system was conducted in conjuction with surface soil characterization activities at the David Witherspoon 1630 Site in Knoxville, Tennessee. The PCB system consists of five hardware standard laboratory modules (SLMs), one software SLM, the task sequence controller (TSC), and the human-computer interface (HCI). Four of the hardware SLMs included a four-channel Soxhlet extractor, a high-volume concentrator, a column cleanup, and a gas chromatograph. These SLMs performed the sample preparation and measurement steps within the total analysis protocol. The fifth hardware module was a robot that transports samples between the SLMs and the required consumable supplies to the SLMs. The software SLM is an automated data interpretation module that receives raw data from the gas chromatograph SLM and analyzes the data to yield the analyte information. The TSC is a software system that provides the scheduling, management of system resources, and the coordination of all SLM activities. The HCI is a graphical user interface that presents the automated laboratory to the analyst in terms of the analytical procedures and methods. Human control of the automated laboratory is accomplished via the HCI. Sample information required for processing by the automated laboratory is entered through the HCI. Information related to the sample and the system status is presented to the analyst via graphical icons

  10. Validation of curve-fitting method for blood retention of {sup 99m}Tc-GSA. Comparison with blood sampling method

    Energy Technology Data Exchange (ETDEWEB)

    Ha-Kawa, Sang Kil; Suga, Yutaka; Kouda, Katsuyasu; Ikeda, Koshi; Tanaka, Yoshimasa [Kansai Medical Univ., Moriguchi, Osaka (Japan)

    1997-02-01

    We investigated a curve-fitting method for the rate of blood retention of {sup 99m}Tc-galactosyl serum albumin (GSA) as a substitute for the blood sampling method. Seven healthy volunteers and 27 patients with liver disease underwent {sup 99m}Tc-GSA scanning. After normalization of the y-intercept as 100 percent, a biexponential regression curve for the precordial time-activity curve provided the percent injected dose (%ID) of {sup 99m}Tc-GSA in the blood without blood sampling. The discrepancy between %ID obtained by the curve-fitting method and that by the multiple blood samples was minimal in normal volunteers 3.1{+-}2.1% (mean{+-}standard deviation, n=77 sampling). Slightly greater discrepancy was observed in patients with liver disease (7.5{+-}6.1%, n=135 sampling). The %ID at 15 min after injection obtained from the fitted curve was significantly greater in patients with liver cirrhosis than in the controls (53.2{+-}11.6%, n=13; vs. 31.9{+-}2.8%, n=7, p<0.0001). There was a highly linear correlation between the %IDs of {sup 99m}Tc-GSA and the plasma retention rate for indocyanine green (r=-0.869, p<0.0001, n=27). These results indicate that the curve-fitting method provides an accurate %ID of {sup 99m}Tc-GSA and could be a substitute for the blood sampling method. (author)

  11. Less invasive blood sampling in the animal laboratory: clinical chemistry and haematology of blood obtained by the Triatominae bug Dipetalogaster maximus.

    Science.gov (United States)

    Markvardsen, S N; Kjelgaard-Hansen, M; Ritz, C; Sørensen, D B

    2012-04-01

    Dipetalogaster maximus (Dipmax), a blood-sucking bug belonging to the family Reduviidae, has been used to obtain blood samples, for example for clinical chemistry and haematology, in a variety of zoo animals and wildlife. Using this bug allows stress-free blood sampling as the bug is able to draw blood without the mammal noticing the bug. In laboratory animal science, the need for blood samples from unstressed animals may arise, especially in animal behaviour research. The use of Dipmax bugs may prove a valuable tool for this purpose. To validate the method, we compared an array of standard blood parameters sampled from New Zealand White rabbits, sampled either by the use of bugs or by the conventional method; puncture of vena auricularis caudalis. The overall hypothesis was that there was no significant difference in clinical chemistry and haematological parameters between the bug method and the conventional method. A total of 17 clinical parameters as well as 12 haematological parameters were measured and compared in New Zealand White rabbits. The results showed that for 13 of these 29 analysed parameters, the bug method and the conventional method did not give significantly different results, and the obtained results were thus directly comparable. For the remaining parameters the obtained results were significantly different. However, all parameters were measurable in the bug samples. The influences of the bug metabolism on these parameters are discussed.

  12. Quantification and Optimization of Candida albicans DNA in Blood Samples Using Real- Time PCR

    Directory of Open Access Journals (Sweden)

    Mojtaba Nabili

    2013-10-01

    Full Text Available Background: Candida albicans (C. albicans is a major cause of candidaemia in people with impaired immunity. Blood culture is a “gold standard” for candidaemia detection but is time-consuming and relatively insensitive. We established a real-time PCR assay for C. albicans detection in blood by LightCycler PCR and melting curve analysis. Methods: Five milliliter blood samples from healthy volunteers were spiked with 100-106 C. albicans cells to determine the detection limit of our method. DNA was extracted from whole blood using glass beads and the QIAamp DNA Blood Mini Kit (Qiagen, Hilden Germany. DNA from C. albicans isolates were amplified with primers and inserted into Escherichia coli (E. coli DH5α.1 cells with the TA cloning vector (Invitrogen. The plasmid was used for standardization and optimization. A quantitative PCR assay with the LightCycler amplification and detection system based on fluorescence resonance energy transfer (FRET with two different specific probes was established. To assess the precision and reproducibility of real-time PCR the intra-assay precision was determined in six consecutive assays. Results: No cross-reactivity of the hybridization probes with the DNA of non-C. albicans species or human genomic DNA was observed, which confirmed its 100% specificity. The minimum limit detected was one C. albicans cell or 100 CFU/ml (10 fg per PCR reaction. The real-time PCR efficiency rate for Candida was high (E = 1.95. Melting curve analysis of C. albicans showed a specific melting peak temperature of 65.76 °C. Conclusion: The real-time PCR assay we developed is highly specific and sufficiently sensitive to detect the fungal load for early diagnosis of invasive candidiasis.

  13. Quantification and optimization of Candida albicans DNA in blood samples using Real- Time PCR.

    Science.gov (United States)

    Nabili, Mojtaba; Ashrafi, Mohsen; Janbabaie, Ghasem; Hedayati, Mohamad Taghi; Ali-Moghaddam, Kamran; Shokohi, Tahereh

    2013-10-01

    Candida albicans (C. albicans) is a major cause of candidaemia in people with impaired immunity. Blood culture is a "gold standard" for candidaemia detection but is time-consuming and relatively insensitive. We established a real-time PCR assay for C. albicans detection in blood by LightCycler PCR and melting curve analysis. Five milliliter blood samples from healthy volunteers were spiked with 10(0)-10(6) C. albicans cells to determine the detection limit of our method. DNA was extracted from whole blood using glass beads and the QIAamp DNA Blood Mini Kit (Qiagen, Hilden Germany). DNA from C. albicans isolates were amplified with primers and inserted into Escherichia coli (E. coli) DH5α.1 cells with the TA cloning vector (Invitrogen). The plasmid was used for standardization and optimization. A quantitative PCR assay with the LightCycler amplification and detection system based on fluorescence resonance energy transfer (FRET) with two different specific probes was established. To assess the precision and reproducibility of real-time PCR the intra-assay precision was determined in six consecutive assays. No cross-reactivity of the hybridization probes with the DNA of non-C. albicans species or human genomic DNA was observed, which confirmed its 100% specificity. The minimum limit detected was one C. albicans cell or 10(0) CFU/ml (10 fg) per PCR reaction. The real-time PCR efficiency rate for Candida was high (E = 1.95). Melting curve analysis of C. albicans showed a specific melting peak temperature of 65.76 °C. The real-time PCR assay we developed is highly specific and sufficiently sensitive to detect the fungal load for early diagnosis of invasive candidiasis.

  14. Automated column liquid chromatographic determination of amoxicillin and cefadroxil in bovine serum and muscle tissue using on-line dialysis for sample preparation

    NARCIS (Netherlands)

    Snippe, N; van de Merbel, N C; Ruiter, F P; Steijger, O M; Lingeman, H; Brinkman, U A

    1994-01-01

    A fully automated method is described for the determination of amoxicillin and cefadroxil in bovine serum and muscle tissue. The method is based on the on-line combination of dialysis and solid-phase extraction for sample preparation, and column liquid chromatography with ultraviolet detection. In

  15. Automated on-line liquid-liquid extraction system for temporal mass spectrometric analysis of dynamic samples.

    Science.gov (United States)

    Hsieh, Kai-Ta; Liu, Pei-Han; Urban, Pawel L

    2015-09-24

    Most real samples cannot directly be infused to mass spectrometers because they could contaminate delicate parts of ion source and guides, or cause ion suppression. Conventional sample preparation procedures limit temporal resolution of analysis. We have developed an automated liquid-liquid extraction system that enables unsupervised repetitive treatment of dynamic samples and instantaneous analysis by mass spectrometry (MS). It incorporates inexpensive open-source microcontroller boards (Arduino and Netduino) to guide the extraction and analysis process. Duration of every extraction cycle is 17 min. The system enables monitoring of dynamic processes over many hours. The extracts are automatically transferred to the ion source incorporating a Venturi pump. Operation of the device has been characterized (repeatability, RSD = 15%, n = 20; concentration range for ibuprofen, 0.053-2.000 mM; LOD for ibuprofen, ∼0.005 mM; including extraction and detection). To exemplify its usefulness in real-world applications, we implemented this device in chemical profiling of pharmaceutical formulation dissolution process. Temporal dissolution profiles of commercial ibuprofen and acetaminophen tablets were recorded during 10 h. The extraction-MS datasets were fitted with exponential functions to characterize the rates of release of the main and auxiliary ingredients (e.g. ibuprofen, k = 0.43 ± 0.01 h(-1)). The electronic control unit of this system interacts with the operator via touch screen, internet, voice, and short text messages sent to the mobile phone, which is helpful when launching long-term (e.g. overnight) measurements. Due to these interactive features, the platform brings the concept of the Internet-of-Things (IoT) to the chemistry laboratory environment. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Identification of a suitable internal control for fluorescence analysis on canine peripheral blood samples.

    Science.gov (United States)

    Riondato, F; Martini, V; Poggi, A; Rota, A; Comazzi, S; Sulce, M; Bruno, B; Borrelli, A; Miniscalco, B

    2016-04-01

    Reliable detection of fluorescence intensity (FI) by flow cytometry (FC) is fundamental. FI depends on instrument settings and sample processing procedures: thus, measurements should be done using internal controls with known FI. Commercially available beads-based standards are expensive, thus reducing their usability in the veterinary practice. Cell subsets with stable mean FI (MFI) within the population have been proposed as acceptable surrogates in human medicine. In veterinary medicine, no data exist about stability of antigen expression among different subjects or upon sample storage. The aim of the present study was to evaluate MFI variability of main lymphocytes antigens among the lymphoid cells within each subject, among different subjects, and upon 24-h storage, in order to identify the antigen most suitable as stable internal control in MFI analyses. Peripheral blood samples from 18 healthy dogs were analysed by FC within 3h from sampling to assess the expression of CD3, CD5, CD4, CD8, CD21 and cyCD79b using conjugated monoclonal antibodies. Analyses were restricted to the lymphoid population. Fluorescent microbeads were added to each tube, and antigen MFI was calculated as Relative Fluorescence Intensity RFI (CD/beads). Fluorescence histogram CV (fhCV) for each CD was regarded as an index of the variability of expression among lymphocytes within each subject (cell-to-cell variability); whereas the CV of RFI was regarded as an index of inter-subjects variability (dog-to-dog variability). In 11 cases, FC analyses were repeated after 24h storage at 4°C and RFI and CVs of fresh and stored samples were compared to assess variability linked to storage. CD4 was identified as the best antigen to be used as an internal control for MFI analyses in canine peripheral blood samples because of low cell-to-cell and dog-to-dog variability, and optimal stability upon 24-h storage. Blood samples from a second group of 21 healthy dogs were labelled only with CD4, in order

  17. Detection of Babesia DNA in blood and spleen samples from Eurasian badgers (Meles meles) in Scotland.

    Science.gov (United States)

    Bartley, Paul M; Wilson, Cari; Innes, Elisabeth A; Katzer, Frank

    2017-08-01

    Babesia are intraerythrocytic parasites of importance worldwide within the fields of human and veterinary medicine, as some Babesia sp., including Babesia microti are potentially zoonotic and can cause fatal disease in both humans and animals. The aims of this study were to use a nested PCR (amplifying the 18S rRNA gene) to determine the presence and species of Babesia parasite DNA found in blood (n = 47) and spleen (n = 47) samples collected from Eurasian badgers (Meles meles) in Scotland. The results showed 28/47 (59·6%) blood and 14/47 (29·8%) spleen samples tested positive for the presence of Babesia DNA. Initial sequence analysis of the Babesia DNA identified three distinct sequence types (submitted to GenBank KX528553, KX528554 and KX528555), which demonstrated ⩾99% identity to Babesia sp. parasites previously identified in badgers in Spain (KT223484 and KT223485). Phylogenetic analysis showed that the three isolates are closely related to Babesia annae, B. microti and other Piroplasmida species found in wildlife. Further sequence analysis of the samples demonstrated that the badgers were routinely infected with more than one parasite isolate and there was also evidence of genetic recombination between the Babesia parasite isolates (submitted to GenBank KY250472 - KY250477).

  18. Trace elements in blood samples of workers in Atbara railways foundry

    International Nuclear Information System (INIS)

    Mustafa, W. M.

    2013-09-01

    This study was conducted to determine trace elements and toxic substances in biological samples (blood samples) of humans. The aim of the current study was to determine the concentration of iron (Fe), copper (Cu), (Pb), lead, and zinc (Zn) in biological samples of workers employed in the industrial workshops in the River Nile state to assess the potential impact of exposure to the work environmental factors. For the purpose of comparison biological samples were collected from the same group of workers exposed to the elements of the work environment and workers not exposed to the elements of the work environment. The analysis of all elements in biological samples was done by x-ray fluorescence technique (X RF). There were no statistically significant differences between the analytical results for the exposed group and non-exposed group, using the same technique. The results showed that the concentrations of the four elements copper, lead, iron, and zinc in all biological samples from workers exposed were not much higher than those not exposed, it could be argued that there was a possible link between these elements with different causes of physiological disorder. The results also showed that need for an attention for improvements in hygiene practice in the workplace and industrial ventilation.(Author)

  19. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    Science.gov (United States)

    Krleza, Jasna Lenicek; Dorotic, Adrijana; Grzunov, Ana; Maradin, Miljenka

    2015-01-01

    Capillary blood sampling is a medical procedure aimed at assisting in patient diagnosis, management and treatment, and is increasingly used worldwide, in part because of the increasing availability of point-of-care testing. It is also frequently used to obtain small blood volumes for laboratory testing because it minimizes pain. The capillary blood sampling procedure can influence the quality of the sample as well as the accuracy of test results, highlighting the need for immediate, widespread standardization. A recent nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia has shown that capillary sampling procedures are not standardized and that only a small proportion of Croatian laboratories comply with guidelines from the Clinical Laboratory Standards Institute (CLSI) or the World Health Organization (WHO). The aim of this document is to provide recommendations for capillary blood sampling. This document has been produced by the Working Group for Capillary Blood Sampling within the Croatian Society of Medical Biochemistry and Laboratory Medicine. Our recommendations are based on existing available standards and recommendations (WHO Best Practices in Phlebotomy, CLSI GP42-A6 and CLSI C46-A2), which have been modified based on local logistical, cultural, legal and regulatory requirements. We hope that these recommendations will be a useful contribution to the standardization of capillary blood sampling in Croatia.

  20. Rapid mapping of compound eye visual sampling parameters with FACETS, a highly automated wide-field goniometer.

    Science.gov (United States)

    Douglass, John K; Wehling, Martin F

    2016-12-01

    A highly automated goniometer instrument (called FACETS) has been developed to facilitate rapid mapping of compound eye parameters for investigating regional visual field specializations. The instrument demonstrates the feasibility of analyzing the complete field of view of an insect eye in a fraction of the time required if using non-motorized, non-computerized methods. Faster eye mapping makes it practical for the first time to employ sample sizes appropriate for testing hypotheses about the visual significance of interspecific differences in regional specializations. Example maps of facet sizes are presented from four dipteran insects representing the Asilidae, Calliphoridae, and Stratiomyidae. These maps provide the first quantitative documentation of the frontal enlarged-facet zones (EFZs) that typify asilid eyes, which, together with the EFZs in male Calliphoridae, are likely to be correlated with high-spatial-resolution acute zones. The presence of EFZs contrasts sharply with the almost homogeneous distribution of facet sizes in the stratiomyid. Moreover, the shapes of EFZs differ among species, suggesting functional specializations that may reflect differences in visual ecology. Surveys of this nature can help identify species that should be targeted for additional studies, which will elucidate fundamental principles and constraints that govern visual field specializations and their evolution.

  1. Does volumetric absorptive microsampling eliminate the hematocrit bias for caffeine and paraxanthine in dried blood samples? A comparative study.

    Science.gov (United States)

    De Kesel, Pieter M M; Lambert, Willy E; Stove, Christophe P

    2015-06-30

    Volumetric absorptive microsampling (VAMS) is a novel sampling technique that allows the straightforward collection of an accurate volume of blood (approximately 10μL) from a drop or pool of blood by dipping an absorbent polymeric tip into it. The resulting blood microsample is dried and analyzed as a whole. The aim of this study was to evaluate the potential of VAMS to overcome the hematocrit bias, an important issue in the analysis of dried blood microsamples. An LC-MS/MS method for analysis of the model compounds caffeine and paraxanthine in VAMS samples was fully validated and fulfilled all pre-established criteria. In conjunction with previously validated procedures for dried blood spots (DBS) and blood, this allowed us to set up a meticulous comparative study in which both compounds were determined in over 80 corresponding VAMS, DBS and liquid whole blood samples. These originated from authentic human patient samples, covering a wide hematocrit range (0.21-0.50). By calculating the differences with reference whole blood concentrations, we found that analyte concentrations in VAMS samples were not affected by a bias that changed over the evaluated hematocrit range, in contrast to DBS results. However, VAMS concentrations tend to overestimate whole blood concentrations, as a consistent positive bias was observed. A different behavior of VAMS samples prepared from incurred and spiked blood, combined with a somewhat reduced recovery of caffeine and paraxanthine from VAMS tips at high hematocrit values, an effect that was not observed for DBS using a very similar extraction procedure, was found to be at the basis of the observed VAMS-whole blood deviations. Based on this study, being the first in which the validity and robustness of VAMS is evaluated by analyzing incurred human samples, it can be concluded that VAMS effectively assists in eliminating the effect of hematocrit. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  2. Weekday variation in triglyceride concentrations in 1.8 million blood samples

    DEFF Research Database (Denmark)

    Jaskolowski, Jörn; Ritz, Christian; Sjödin, Anders Mikael

    2017-01-01

    BACKGROUND: Triglyceride (TG) concentration is used as a marker of cardio-metabolic risk. However, diurnal and possibly weekday variation exists in TG concentrations. OBJECTIVE: To investigate weekday variation in TG concentrations among 1.8 million blood samples drawn between 2008 and 2015 from...... variations in TG concentrations were recorded for out-patients between the age of 9 to 26 years, with up to 20% higher values on Mondays compared to Fridays (all PTriglyceride concentrations were highest after the weekend and gradually declined during the week. We suggest that unhealthy...

  3. Concentrations of persistent organic pollutants (POPs) in human blood samples from Mexico City, Mexico.

    Science.gov (United States)

    Orta-García, Sandra; Pérez-Vázquez, Francisco; González-Vega, Carolina; Varela-Silva, José Antonio; Hernández-González, Lidia; Pérez-Maldonado, Iván

    2014-02-15

    Studies in Mexico have demonstrated exposure to persistent organic pollutants (POPs) in people living in different sites through the country. However, studies evaluating exposure to POPs in people living in Mexico City (one of most contaminated places in the world) are scarce. Therefore, the aim of this study was to assess the levels of polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite dichlorodiphenyldichloroethylene (DDE) in the blood as exposure biomarkers in people living in Mexico City. A total of 123 participants (blood donors aged 20-60 years) were recruited during 2010 in Mexico City. Quantitative analyses of blood samples were performed using gas chromatography coupled with mass spectrometry. Levels of the assessed compounds ranged from non-detectable (

  4. Thyroxine and thyrotropin radioimmunoassays using dried blood samples on filter paper for screening of neonatal hypothyroidism

    International Nuclear Information System (INIS)

    Beckers, C.; Cornette, C.; Francois, B.; Bouckaert, A.; Lechat, M.

    1977-01-01

    A routine and automatized methodology for thyroxine (T4) and thyrotropin (TSH) radioimmunoassay (RIA) using dried blood samples on filter paper is described. Five mm diameter dots were prepared. One eluted dot, corresponding to 4 μl of plasma, was used for T4-RIA while two were necessary for TSH-RIA. Reference filter papers were introduced in each assay for quality control. In a preliminary study on 1903 newborns, samples were obtained, generally between the 5th-7th day. Mean dot T4 was 7.38 +- 2.5 μg/dl. Mean dot TSH was 11.83 +- 9.1 μU/ml, the equation of the regression line between dot TSH (y) and serum TSH (x) being Y = 10.29 + 0.623x. (orig.) [de

  5. Slurry sampling in serum blood for mercury determination by CV-AFS

    International Nuclear Information System (INIS)

    Aranda, Pedro R.; Gil, Raul A.; Moyano, Susana; De Vito, Irma; Martinez, Luis D.

    2009-01-01

    The heavy metal mercury (Hg) is a neurotoxin known to have a serious health impact even at relatively low concentrations. A slurry method was developed for the sensitive and precise determination of mercury in human serum blood samples by cold vapor generation coupled to atomic fluorescence spectrometry (CV-AFS). All variables related to the slurry formation were studied. The optimal hydrochloric concentration and tin(II) chloride concentration for CV generation were evaluated. Calibration within the range 0.1-10 μg L -1 Hg was performed with the standard addition method, and compared with an external calibration. Additionally, the reliability of the results obtained was evaluated by analyzing mercury in the same samples, but submitted to microwave-assisted digestion method. The limit of detection was calculated as 25 ng L -1 and the relative standard deviation was 3.9% at levels around of 0.4 μg L -1 Hg

  6. Simple Sample Preparation Method for Direct Microbial Identification and Susceptibility Testing From Positive Blood Cultures

    Directory of Open Access Journals (Sweden)

    Hong-wei Pan

    2018-03-01

    Full Text Available Rapid identification and determination of the antibiotic susceptibility profiles of the infectious agents in patients with bloodstream infections are critical steps in choosing an effective targeted antibiotic for treatment. However, there has been minimal effort focused on developing combined methods for the simultaneous direct identification and antibiotic susceptibility determination of bacteria in positive blood cultures. In this study, we constructed a lysis-centrifugation-wash procedure to prepare a bacterial pellet from positive blood cultures, which can be used directly for identification by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS and antibiotic susceptibility testing by the Vitek 2 system. The method was evaluated using a total of 129 clinical bacteria-positive blood cultures. The whole sample preparation process could be completed in <15 min. The correct rate of direct MALDI-TOF MS identification was 96.49% for gram-negative bacteria and 97.22% for gram-positive bacteria. Vitek 2 antimicrobial susceptibility testing of gram-negative bacteria showed an agreement rate of antimicrobial categories of 96.89% with a minor error, major error, and very major error rate of 2.63, 0.24, and 0.24%, respectively. Category agreement of antimicrobials against gram-positive bacteria was 92.81%, with a minor error, major error, and very major error rate of 4.51, 1.22, and 1.46%, respectively. These results indicated that our direct antibiotic susceptibility analysis method worked well compared to the conventional culture-dependent laboratory method. Overall, this fast, easy, and accurate method can facilitate the direct identification and antibiotic susceptibility testing of bacteria in positive blood cultures.

  7. Aspects of fetal physiology from 18 to 37 weeks' gestation as assessed by blood sampling.

    Science.gov (United States)

    Nava, S; Bocconi, L; Zuliani, G; Kustermann, A; Nicolini, U

    1996-06-01

    To construct reference ranges for fetal pH, oxygen pressure (PO2), and hematologic and biochemical blood constituents, which can be used to analyze changes with gestation and differences with maternal values, thus elucidating some aspects of fetal biology and the effects of the maternal and placental environments. We assayed venous pH, PO2, hematocrit, glucose, uric acid, urea, creatinine, total protein, total and direct bilirubin, aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, alkaline phosphatase, lactic dehydrogenase, amylase, pseudocholinesterase, creatine kinase, triglycerides, and cholesterol concentrations in 157 fetuses and 134 mothers who underwent fetal blood sampling from 18 to 37 weeks' gestation. None of the fetuses was infected or had chromosomal, hematologic, or hormonal abnormalities. All the variables analyzed were similar in fetuses sampled at the placental cord insertion (n = 125) or at the intrahepatic vein (n = 32). Maternal and fetal concentrations of glucose (r = 0.79, P PO2 decreased with gestational age, whereas hematocrit increased, similar to what has been described previously. All of the other variables, with the exception of amylase and cholesterol, changed significantly during the investigated period of pregnancy. Gestational age explained at least 40% of the variance in values of fetal total protein, pseudocholinesterase, alanine aminotransferase, creatine kinase, and triglycerides, but only 3-25% of the variation in the remainder. Most enzymes were higher in the fetus than in the maternal circulation, and all except alkaline phosphatase increased with gestational age. The maternal-fetal glucose difference correlated significantly with hematocrit, pH, and PO2, independent of gestational age and independent of each other. With the exception of aspartate aminotransferase, all of the analyzed fetal variables were different from the maternal values, and most changed with gestational age. The mechanisms

  8. Environmental contaminants in Texas, USA, wetland reptiles: Evaluation using blood samples

    Science.gov (United States)

    Clark, D.R.; Bickham, J.W.; Baker, D.L.; Cowman, D.F.

    2000-01-01

    Four species of reptiles (diamondback water snake [Nerodia rhombifer], blotched water snake [N. erythrogaster], cottonmouth [Agkistrodon piscivorus], and red-eared slider [Trachemys scripta]) were collected at two contaminated and three reference sites in Texas, USA. Old River Slough has received intensive applications of agricultural chemicals since the 1950s. Municipal Lake received industrial arsenic wastes continuously from 1940 to 1993. Blood samples were analyzed for organochlorines, potentially toxic elements, genetic damage, and plasma cholinesterase (ChE). Dichlorodiphenyldichloroethylene (DDE) concentrations reached as high as 3.0 ppm (wet weight) in whole blood of a diamondback water snake at Old River Slough, a level probably roughly equivalent to the maximum concentration found in plasma of peregrine falcons (Falco peregrinus) in 1978 to 1979 when DDE peaked in this sensitive species. Possible impacts on diamondback water snakes are unknown, but at least one diamondback water snake was gravid when captured, indicating active reproduction. Arsenic was not found in red-eared sliders (only species sampled) from Municipal Lake. Red-eared sliders of both sexes at Old River Slough showed declining levels of ChE with increasing mass, suggesting a life-long decrease of ChE levels. Possible negative population consequences are unknown, but no evidence was found in body condition (mass relative to carapace length) that red-eared sliders at either contaminated site were harmed.

  9. Ionic Liquid-Based Liquid-Liquid Microextraction for Benzodiazepine Analysis in Postmortem Blood Samples.

    Science.gov (United States)

    De Boeck, Marieke; Dehaen, Wim; Tytgat, Jan; Cuypers, Eva

    2018-03-24

    Sample preparation is rapidly improving to fulfill the need for faster and more environmentally friendly alternatives. In this respect, ionic liquid-based dispersive liquid-liquid microextraction (IL-DLLME) is an interesting technique. However, it has not yet been evaluated for the analysis of postmortem samples, which are frequently analyzed in forensic toxicology. This study investigates the applicability of IL-DLLME coupled to liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the analysis of benzodiazepines in postmortem blood of 11 forensic cases. The method was compared with a validated solid-phase extraction (SPE) method. Bland-Altman analysis was performed on 24 benzodiazepine measurements. Both methods gave comparable results, except for flurazepam and temazepam (>55% difference). A feasible explanation is high postmortem matrix variability that was not considered during IL-DLLME validation experiments. Another issue could be the use of a single nondeuterated SPE internal standard. Overall, IL-DLLME has proven its usability for the analysis of postmortem blood. © 2018 American Academy of Forensic Sciences.

  10. Variation of Peripheral Blood Mononuclear Cell RNA Quality in Archived Samples.

    Science.gov (United States)

    Kozlakidis, Zisis; Mant, Christine; Abdinur, Fartun; Cope, Andrew; Steiner, Szabi; Peakman, Mark; Hayday, Adrian; Cason, John

    2011-09-01

    The Infectious Diseases BioBank (IDB) has consistently archived peripheral blood mononuclear cell (PBMNC) RNA for transcriptome analyses. RNA is particularly labile, and hence, these samples provide a sensitive indicator for assessing the IDB's quality-assurance measures. Independent analyses of 104 PBMNC RNA specimens from 26 volunteers revealed that the mean RNA integrity number (RIN) was high (9.02), although RIN ranged between scores of 7 and 10. This variation of RIN values was not associated with ischemic time, PBMNC quality, number of samples processed per day, self-medication after immunization, freezer location, donor characteristics, differential white blood cell counts, or daily variation in RNA extractions (all P>0.05). RIN values were related to the date of collection, with those processed during mid-summer having highest RIN scores (P=0.0001). Amongst specimens with the lowest RIN scores, no common feature could be identified. Thus, no technical explanation for the variation in RNA quality could be ascertained and these may represent normal physiological variations. These data provide strong evidence that current IDB protocols for the isolation and preservation PBMNC RNA are robust.

  11. Antibiogram pattern of Salmonella in blood samples of enteric fever patients at Lalitpur, Nepal

    Directory of Open Access Journals (Sweden)

    Biraj Gurung

    2017-01-01

    Full Text Available Objective: To determine the status of isolation blood stream serotypes of enteric fever pathogens and their antibiotic susceptibility patterns and to guide clinicians for appropriate therapy. Methods: Samples were examined by microbiological techniques to identify the causative agent and determine their antimicrobial susceptibility patterns by Kirby-Bauer disk diffusion methods and interpreted as per Clinical and Laboratory Standards Institute guidelines. Results: Among 403 blood samples, 76 (18.85% showed growth for Salmonella isolates. Distribution of Salmonella typhi and Salmonella paratyphi A isolates were found to be 54% and 46% respectively. Among 76 Salmonella isolates, 28 (36.84% were from male and remaining 48 (63.15% were from female belonging to all age-groups. Multidrug-resistance was found to be 17% among the Salmonella isolates. Nalidixic acid resistance was 73.68% in Salmonella with higher proportion in Salmonella paratyphi A (85.7% in comparison to Salmonella typhi (63.42%. Salmonella isolates demonstrated 100% susceptibility to azithromycin, ceftriazone, ciprofloxacin, ofloxacin and imipenem. Conclusions: The need of continual surveillance of resistance levels to guide clinicians for appropriate therapy based on the antibiotic susceptibility pattern for Salmonella isolates is sustained with discouragement in misuse of antibiotics prior to prescription as multidrugresistance-nalidixic acid resistant strains.

  12. Analytical and between-subject variation of thrombin generation measured by calibrated automated thrombography on plasma samples

    DEFF Research Database (Denmark)

    Kristensen, Anne F; Kristensen, Søren R; Falkmer, Ursula

    2018-01-01

    BACKGROUND: The Calibrated Automated Thrombography (CAT) is an in vitro thrombin generation (TG) assay that holds promise as a valuable tool within clinical diagnostics. However, the technique has a considerable analytical variation, and we therefore, investigated the analytical and between...

  13. Analysis of intraosseous blood samples using an EPOC point of care analyzer during resuscitation.

    Science.gov (United States)

    Tallman, Crystal Ives; Darracq, Michael; Young, Megann

    2017-03-01

    In the early phases of resuscitation in a critically ill patient, especially those in cardiac arrest, intravenous (IV) access can be difficult to obtain. Intraosseous (IO) access is often used in these critical situations to allow medication administration. When no IV access is available, it is difficult to obtain blood for point of care analysis, yet this information can be crucial in directing the resuscitation. We hypothesized that IO samples may be used with a point of care device to obtain useful information when seconds really do matter. Patients presenting to the emergency department requiring resuscitation and IO placement were prospectively enrolled in a convenience sample. 17 patients were enrolled. IO and IV samples obtained within five minutes of one another were analyzed using separate EPOC® point of care analyzers. Analytes were compared using Bland Altman Plots and intraclass correlation coefficients. In this analysis of convenience sampled critically ill patients, the EPOC® point of care analyzer provided results from IO samples. IO and IV samples were most comparable for pH, bicarbonate, sodium and base excess, and potentially for lactic acid; single outliers for bicarbonate, sodium and base excess were observed. Intraclass correlation coefficients were excellent for sodium and reasonable for pH, pO2, bicarbonate, and glucose. Correlations for other variables measured by the EPOC® analyzer were not as robust. IO samples can be used with a bedside point of care analyzer to rapidly obtain certain laboratory information during resuscitations when IV access is difficult. Copyright © 2016 Elsevier Inc. All rights reserved.

  14. Comparison of parasite loads in serum and blood samples from patients in acute and chronic phases of Chagas disease.

    Science.gov (United States)

    Hernández, Carolina; Teherán, Aníbal; Flórez, Carolina; Ramírez, Juan David

    2018-04-17

    Molecular methods have been developed for the detection and quantification of Trypanosoma cruzi DNA in blood samples from patients with Chagas disease. However, aspects of sample processing necessary for quantitative real-time PCR (qPCR), such as the addition of guanidine hydrochloride to whole blood samples, may limit timely access to molecular diagnosis. We analysed 169 samples from serum and guanidine-EDTA blood (GEB) obtained from patients in acute and chronic phases of Chagas disease. We applied qPCR targeted to the satellite DNA region. Finally, we compared the parasite loads and cycle of threshold values of the qPCR. The results confirmed the usefulness of serum samples for the detection and quantification of parasite DNA in patients with Chagas disease, especially in the acute phase. However, the parasite loads detected in serum samples from patients in the chronic phase were lower than those detected in GEB samples. The epidemiological implications of the findings are herein discussed.

  15. Direct Trace Element Analysis of Liquid Blood Samples by In-Air Ion Beam Analytical Techniques (PIXE-PIGE).

    Science.gov (United States)

    Huszank, Robert; Csedreki, László; Török, Zsófia

    2017-02-07

    There are various liquid materials whose elemental composition is of interest in various fields of science and technology. In many cases, sample preparation or the extraction can be complicated, or it would destroy the original environment before the analysis (for example, in the case of biological samples). However, multielement direct analysis of liquid samples can be realized by an external PIXE-PIGE measurement system. Particle-induced X-ray and gamma-ray emission spectroscopy (PIXE, PIGE) techniques were applied in external (in-air) microbeam configuration for the trace and main element determination of liquid samples. The direct analysis of standard solutions of several metal salts and human blood samples (whole blood, blood serum, blood plasma, and formed elements) was realized. From the blood samples, Na, P, S, Cl, K, Ca, Fe, Cu, Zn, and Br elemental concentrations were determined. The focused and scanned ion beam creates an opportunity to analyze very small volume samples (∼10 μL). As the sample matrix consists of light elements, the analysis is possible at ppm level. Using this external beam setup, it was found that it is possible to determine elemental composition of small-volume liquid samples routinely, while the liquid samples do not require any preparation processes, and thus, they can be analyzed directly. In the case of lower concentrations, the method is also suitable for the analysis (down to even ∼1 ppm level) but with less accuracy and longer measurement times.

  16. Quantification of rifapentine, a potent antituberculosis drug, from dried blood spot samples using liquid chromatographic-tandem mass spectrometric analysis.

    Science.gov (United States)

    Parsons, Teresa L; Marzinke, Mark A; Hoang, Thuy; Bliven-Sizemore, Erin; Weiner, Marc; Mac Kenzie, William R; Dorman, Susan E; Dooley, Kelly E

    2014-11-01

    The quantification of antituberculosis drug concentrations in multinational trials currently requires the collection of modest blood volumes, centrifugation, aliquoting of plasma, freezing, and keeping samples frozen during shipping. We prospectively enrolled healthy individuals into the Tuberculosis Trials Consortium Study 29B, a phase I dose escalation study of rifapentine, a rifamycin under evaluation in tuberculosis treatment trials. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying rifapentine in whole blood on dried blood spots (DBS) to facilitate pharmacokinetic/pharmacodynamic analyses in clinical trials. Paired plasma and whole-blood samples were collected by venipuncture, and whole blood was spotted on Whatman protein saver 903 cards. The methods were optimized for plasma and then validated for DBS. The analytical measuring range for quantification of rifapentine and its metabolite was 50 to 80,000 ng/ml in whole-blood DBS. The analyte was stable on the cards for 11 weeks with a desiccant at room temperature and protected from light. The method concordance for paired plasma and whole-blood DBS samples was determined after correcting for participant hematocrit or population-based estimates of bias from Bland-Altman plots. The application of either correction factor resulted in acceptable correlation between plasma and whole-blood DBS (Passing-Bablok regression corrected for hematocrit; y = 0.98x + 356). Concentrations of rifapentine may be determined from whole-blood DBS collected via venipuncture after normalization in order to account for the dilutional effects of red blood cells. Additional studies are focused on the application of this methodology to capillary blood collected by finger stick. The simplicity of processing, storage, shipping, and low blood volume makes whole-blood DBS attractive for rifapentine pharmacokinetic evaluations, especially in international and pediatric trials. Copyright © 2014

  17. Improvement of precision for pipetting blood serum samples into a graphite furnace

    Science.gov (United States)

    Bohrer, D.; do Nascimento, P. C.; Binotto, R.; Borges da Costa, J. A. T.; Szlachta, T.

    2002-12-01

    Graphite furnace atomic absorption spectrometry is a well-established technique for trace metal determination in blood and serum samples. For this kind of samples, a R.S.D. of up to 10% is considered acceptable, especially for those elements, the concentrations of which do not allow high dilution. Among the reasons that contribute to an increase of the S.D. is the retention of proteins (and analyte) in the sample dispenser capillary, because proteins might be adsorbed on polymeric surfaces. In the present work, the interaction protein/polymer was studied, considering the amount of protein that could be retained by the capillary, the influence of sample dilution, time of contact and the possible co-adsorption of metals. Serum proteins were retained in the capillary, depending on the material (Tygon>silicone rubber>poly(tetrafluorethylene)); dilution did not prevent the adsorption, and only 30 s of contact were enough for the adsorption to occur. Using a column filled with PTFE powder (60 mesh), it was possible to observe that metals were co-adsorbed to a large extent. Water, diluted nitric acid or aqueous solutions of Triton X-100 were not able to promote the complete desorption of the proteins retained by the polymeric materials. Total elution was achieved with methanol, and its use as rinse solution decreased the R.S.D. ( n=10) for aluminium, manganese and chromium determination in serum to <10%.

  18. T