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Sample records for blood sampling methods

  1. Quality of plasma sampled by different methods for multiple blood sampling in mice.

    PubMed

    Christensen, S D; Mikkelsen, L F; Fels, J J; Bodvarsdóttir, T B; Hansen, A K

    2009-01-01

    For oral glucose tolerance test (OGTT) in mice, multiple blood samples need to be taken within a few hours from conscious mice. Today, a number of essential parameters may be analysed on very small amounts of plasma, thus reducing the number of animals to be used. It is, however, crucial to obtain high-quality plasma or serum in order to avoid increased data variation and thereby increased group sizes. The aim of this study was to find the most valid and reproducible method for withdrawal of blood samples when performing OGTT. Four methods, i.e. amputation of the tail tip, lateral tail incision, puncture of the tail tip and periorbital puncture, were selected for testing at 21 degrees C and 30 degrees C after a pilot study. For each method, four blood samples were drawn from C57BL/6 mice at 30 min intervals. The presence of clots was registered, haemolysis was monitored spectrophotometrically at 430 nm, and it was noted whether it was possible to achieve 30-50 microL blood. Furthermore, a small amount of extra blood was sampled before and after the four samplings for testing of whether the sampling induced a blood glucose change over the 90 min test period. All methods resulted in acceptable amounts of plasma. Clots were observed in a sparse number of samples with no significant differences between the methods. Periorbital puncture did not lead to any haemolysed samples at all, and lateral tail incision resulted in only a few haemolysed samples, while puncture or amputation of the tail tip induced haemolysis in a significant number of samples. All methods, except for puncture of the tail tip, influenced blood glucose. Periorbital puncture resulted in a dramatic increase in blood glucose of up to 3.5 mmol/L indicating that it is stressful. Although lateral tail incision also had some impact on blood glucose, it seems to be the method of choice for OGTT, as it is likely to produce a clot-free non-haemolysed sample, while periorbital sampling, although producing a

  2. Comparison of blood chemistry values for samples collected from juvenile chinook salmon by three methods

    USGS Publications Warehouse

    Congleton, J.L.; LaVoie, W.J.

    2001-01-01

    Thirteen blood chemistry indices were compared for samples collected by three commonly used methods: caudal transection, heart puncture, and caudal vessel puncture. Apparent biases in blood chemistry values for samples obtained by caudal transection were consistent with dilution with tissue fluids: alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), creatine kinase (CK), triglyceride, and K+ were increased and Na+ and Cl- were decreased relative to values for samples obtained by caudal vessel puncture. Some enzyme activities (ALT, AST, LDH) and K+ concentrations were also greater in samples taken by heart puncture than in samples taken by caudal vessel puncture. Of the methods tested, caudal vessel puncture had the least effect on blood chemistry values and should be preferred for blood chemistry studies on juvenile salmonids.

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

    PubMed

    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.

  4. Device and method for automated separation of a sample of whole blood into aliquots

    DOEpatents

    Burtis, Carl A.; Johnson, Wayne F.

    1989-01-01

    A device and a method for automated processing and separation of an unmeasured sample of whole blood into multiple aliquots of plasma. Capillaries are radially oriented on a rotor, with the rotor defining a sample chamber, transfer channels, overflow chamber, overflow channel, vent channel, cell chambers, and processing chambers. A sample of whole blood is placed in the sample chamber, and when the rotor is rotated, the blood moves outward through the transfer channels to the processing chambers where the blood is centrifugally separated into a solid cellular component and a liquid plasma component. When the rotor speed is decreased, the plasma component backfills the capillaries resulting in uniform aliquots of plasma which may be used for subsequent analytical procedures.

  5. Method of evaluation of process of red blood cell sedimentation based on photometry of droplet samples.

    PubMed

    Aristov, Alexander; Nosova, Ekaterina

    2017-04-01

    The paper focuses on research aimed at creating and testing a new approach to evaluate the processes of aggregation and sedimentation of red blood cells for purpose of its use in clinical laboratory diagnostics. The proposed method is based on photometric analysis of blood sample formed as a sessile drop. The results of clinical approbation of this method are given in the paper. Analysis of the processes occurring in the sample in the form of sessile drop during the process of blood cells sedimentation is described. The results of experimental studies to evaluate the effect of the droplet sample focusing properties on light radiation transmittance are presented. It is shown that this method significantly reduces the sample volume and provides sufficiently high sensitivity to the studied processes.

  6. Method and apparatus for automated processing and aliquoting of whole blood samples for analysis in a centrifugal fast analyzer

    DOEpatents

    Burtis, Carl A.; Johnson, Wayne F.; Walker, William A.

    1988-01-01

    A rotor and disc assembly for use in a centrifugal fast analyzer. The assembly is designed to process multiple samples of whole blood followed by aliquoting of the resultant serum into precisely measured samples for subsequent chemical analysis. The assembly requires minimal operator involvement with no mechanical pipetting. The system comprises (1) a whole blood sample disc, (2) a serum sample disc, (3) a sample preparation rotor, and (4) an analytical rotor. The blood sample disc and serum sample disc are designed with a plurality of precision bore capillary tubes arranged in a spoked array. Samples of blood are loaded into the blood sample disc in capillary tubes filled by capillary action and centrifugally discharged into cavities of the sample preparation rotor where separation of serum and solids is accomplished. The serum is loaded into the capillaries of the serum sample disc by capillary action and subsequently centrifugally expelled into cuvettes of the analytical rotor for analysis by conventional methods.

  7. Method and apparatus for automated processing and aliquoting of whole blood samples for analysis in a centrifugal fast analyzer

    DOEpatents

    Burtis, C.A.; Johnson, W.F.; Walker, W.A.

    1985-08-05

    A rotor and disc assembly for use in a centrifugal fast analyzer. The assembly is designed to process multiple samples of whole blood followed by aliquoting of the resultant serum into precisely measured samples for subsequent chemical analysis. The assembly requires minimal operator involvement with no mechanical pipetting. The system comprises: (1) a whole blood sample disc; (2) a serum sample disc; (3) a sample preparation rotor; and (4) an analytical rotor. The blood sample disc and serum sample disc are designed with a plurality of precision bore capillary tubes arranged in a spoked array. Samples of blood are loaded into the blood sample disc by capillary action and centrifugally discharged into cavities of the sample preparation rotor where separation of serum and solids is accomplished. The serum is loaded into the capillaries of the serum sample disc by capillary action and subsequently centrifugally expelled into cuvettes of the analyticaly rotor for conventional methods. 5 figs.

  8. Targeting prohibited substances in doping control blood samples by means of chromatographic-mass spectrometric methods.

    PubMed

    Thevis, Mario; Thomas, Andreas; Schänzer, Wilhelm

    2013-12-01

    Urine samples have been the predominant matrix for doping controls for several decades. However, owing to the complementary information provided by blood (as well as serum or plasma and dried blood spots (DBS)), the benefits of its analysis have resulted in continuously increasing appreciation by anti-doping authorities. On the one hand, blood samples allow for the detection of various different methods of blood doping and the abuse of erythropoiesis-stimulating agents (ESAs) via the Athlete Biological Passport; on the other hand, targeted and non-targeted drug detection by means of chromatographic-mass spectrometric methods represents an important tool to increase doping control frequencies out-of-competition and to determine drug concentrations particularly in in-competition scenarios. Moreover, blood analysis seldom requires in-depth knowledge of drug metabolism, and the intact substance rather than potentially unknown or assumed metabolic products can be targeted. In this review, the recent developments in human sports drug testing concerning mass spectrometry-based techniques for qualitative and quantitative analyses of therapeutics and emerging drug candidates are summarized and reviewed. The analytical methods include both low and high molecular mass compounds (e.g., anabolic agents, stimulants, metabolic modulators, peptide hormones, and small interfering RNA (siRNA)) determined from serum, plasma, and DBS using state-of-the-art instrumentation such as liquid chromatography (LC)-high resolution/high accuracy (tandem) mass spectrometry (LC-HRMS), LC-low resolution tandem mass spectrometry (LC-MS/MS), and gas chromatography-mass spectrometry (GC-MS).

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

    PubMed

    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.

  10. Utility of the microculture method for Leishmania detection in non-invasive samples obtained from a blood bank.

    PubMed

    Ates, Sezen Canim; Bagirova, Malahat; Allahverdiyev, Adil M; Kocazeybek, Bekir; Kosan, Erdogan

    2013-10-01

    In recent years, the role of donor blood has taken an important place in epidemiology of Leishmaniasis. According to the WHO, the numbers of patients considered as symptomatic are only 5-20% of individuals with asymptomatic leishmaniasis. In this study for detection of Leishmania infection in donor blood samples, 343 samples from the Capa Red Crescent Blood Center were obtained and primarily analyzed by microscopic and serological methods. Subsequently, the traditional culture (NNN), Immuno-chromatographic test (ICT) and Polymerase Chain Reaction (PCR) methods were applied to 21 samples which of them were found positive with at least one method. Buffy coat (BC) samples from 343 blood donors were analyzed: 15 (4.3%) were positive by a microculture method (MCM); and 4 (1.1%) by smear. The sera of these 343 samples included 9 (2.6%) determined positive by ELISA and 7 (2%) positive by IFAT. Thus, 21 of (6.1%) the 343 subjects studied by smear, MCM, IFAT and ELISA techniques were identified as positive for leishmaniasis at least one of the techniques and the sensitivity assessed. According to our data, the sensitivity of the methods are identified as MCM (71%), smear (19%), IFAT (33%), ELISA (42%), NNN (4%), PCR (14%) and ICT (4%). Thus, with this study for the first time, the sensitivity of a MCM was examined in blood donors by comparing MCM with the methods used in the diagnosis of leishmaniasis. As a result, MCM was found the most sensitive method for detection of Leishmania parasites in samples obtained from a blood bank. In addition, the presence of Leishmania parasites was detected in donor bloods in Istanbul, a non-endemic region of Turkey, and these results is a vital importance for the health of blood recipients.

  11. Validation and Clinical Evaluation of a Novel Method To Measure Miltefosine in Leishmaniasis Patients Using Dried Blood Spot Sample Collection.

    PubMed

    Kip, A E; Rosing, H; Hillebrand, M J X; Blesson, S; Mengesha, B; Diro, E; Hailu, A; Schellens, J H M; Beijnen, J H; Dorlo, T P C

    2016-04-01

    To facilitate future pharmacokinetic studies of combination treatments against leishmaniasis in remote regions in which the disease is endemic, a simple cheap sampling method is required for miltefosine quantification. The aims of this study were to validate a liquid chromatography-tandem mass spectrometry method to quantify miltefosine in dried blood spot (DBS) samples and to validate its use with Ethiopian patients with visceral leishmaniasis (VL). Since hematocrit (Ht) levels are typically severely decreased in VL patients, returning to normal during treatment, the method was evaluated over a range of clinically relevant Ht values. Miltefosine was extracted from DBS samples using a simple method of pretreatment with methanol, resulting in >97% recovery. The method was validated over a calibration range of 10 to 2,000 ng/ml, and accuracy and precision were within ±11.2% and ≤7.0% (≤19.1% at the lower limit of quantification), respectively. The method was accurate and precise for blood spot volumes between 10 and 30 μl and for Ht levels of 20 to 35%, although a linear effect of Ht levels on miltefosine quantification was observed in the bioanalytical validation. DBS samples were stable for at least 162 days at 37°C. Clinical validation of the method using paired DBS and plasma samples from 16 VL patients showed a median observed DBS/plasma miltefosine concentration ratio of 0.99, with good correlation (Pearson'sr= 0.946). Correcting for patient-specific Ht levels did not further improve the concordance between the sampling methods. This successfully validated method to quantify miltefosine in DBS samples was demonstrated to be a valid and practical alternative to venous blood sampling that can be applied in future miltefosine pharmacokinetic studies with leishmaniasis patients, without Ht correction.

  12. Validation and Clinical Evaluation of a Novel Method To Measure Miltefosine in Leishmaniasis Patients Using Dried Blood Spot Sample Collection

    PubMed Central

    Rosing, H.; Hillebrand, M. J. X.; Blesson, S.; Mengesha, B.; Diro, E.; Hailu, A.; Schellens, J. H. M.; Beijnen, J. H.

    2016-01-01

    To facilitate future pharmacokinetic studies of combination treatments against leishmaniasis in remote regions in which the disease is endemic, a simple cheap sampling method is required for miltefosine quantification. The aims of this study were to validate a liquid chromatography-tandem mass spectrometry method to quantify miltefosine in dried blood spot (DBS) samples and to validate its use with Ethiopian patients with visceral leishmaniasis (VL). Since hematocrit (Ht) levels are typically severely decreased in VL patients, returning to normal during treatment, the method was evaluated over a range of clinically relevant Ht values. Miltefosine was extracted from DBS samples using a simple method of pretreatment with methanol, resulting in >97% recovery. The method was validated over a calibration range of 10 to 2,000 ng/ml, and accuracy and precision were within ±11.2% and ≤7.0% (≤19.1% at the lower limit of quantification), respectively. The method was accurate and precise for blood spot volumes between 10 and 30 μl and for Ht levels of 20 to 35%, although a linear effect of Ht levels on miltefosine quantification was observed in the bioanalytical validation. DBS samples were stable for at least 162 days at 37°C. Clinical validation of the method using paired DBS and plasma samples from 16 VL patients showed a median observed DBS/plasma miltefosine concentration ratio of 0.99, with good correlation (Pearson's r = 0.946). Correcting for patient-specific Ht levels did not further improve the concordance between the sampling methods. This successfully validated method to quantify miltefosine in DBS samples was demonstrated to be a valid and practical alternative to venous blood sampling that can be applied in future miltefosine pharmacokinetic studies with leishmaniasis patients, without Ht correction. PMID:26787691

  13. Comparison of blood plasma sample preparation methods for combined LC-MS lipidomics and metabolomics.

    PubMed

    Patterson, Rainey E; Ducrocq, Antoine J; McDougall, Danielle J; Garrett, Timothy J; Yost, Richard A

    2015-10-01

    The goal of this research was to find the most comprehensive lipid extraction of blood plasma, while also providing adequate aqueous preparation for metabolite analysis. Comparisons have been made previously of the Folch, Bligh-Dyer, and Matyash lipid extractions; furthermore, this paper provides an additional comparison of a phospholipid removal plate for analysis. This plate was used for lipid extraction rather than its intended use in lipid removal for polar analysis, and it proves to be robust for targeted lipid analysis. Folch and Matyash provided reproducible recovery over a range of lipid classes, however the Matyash aqueous layer compared well to a typical methanol preparation for polar metabolite analysis. Thus, the Matyash method is the best choice for an untargeted biphasic extraction for metabolomics and lipidomics in blood plasma.

  14. An effective method to purify Plasmodium falciparum DNA directly from clinical blood samples for whole genome high-throughput sequencing.

    PubMed

    Auburn, Sarah; Campino, Susana; Clark, Taane G; Djimde, Abdoulaye A; Zongo, Issaka; Pinches, Robert; Manske, Magnus; Mangano, Valentina; Alcock, Daniel; Anastasi, Elisa; Maslen, Gareth; Macinnis, Bronwyn; Rockett, Kirk; Modiano, David; Newbold, Christopher I; Doumbo, Ogobara K; Ouédraogo, Jean Bosco; Kwiatkowski, Dominic P

    2011-01-01

    Highly parallel sequencing technologies permit cost-effective whole genome sequencing of hundreds of Plasmodium parasites. The ability to sequence clinical Plasmodium samples, extracted directly from patient blood without a culture step, presents a unique opportunity to sample the diversity of "natural" parasite populations in high resolution clinical and epidemiological studies. A major challenge to sequencing clinical Plasmodium samples is the abundance of human DNA, which may substantially reduce the yield of Plasmodium sequence. We tested a range of human white blood cell (WBC) depletion methods on P. falciparum-infected patient samples in search of a method displaying an optimal balance of WBC-removal efficacy, cost, simplicity, and applicability to low resource settings. In the first of a two-part study, combinations of three different WBC depletion methods were tested on 43 patient blood samples in Mali. A two-step combination of Lymphoprep plus Plasmodipur best fitted our requirements, although moderate variability was observed in human DNA quantity. This approach was further assessed in a larger sample of 76 patients from Burkina Faso. WBC-removal efficacy remained high (<30% human DNA in >70% samples) and lower variation was observed in human DNA quantities. In order to assess the Plasmodium sequence yield at different human DNA proportions, 59 samples with up to 60% human DNA contamination were sequenced on the Illumina Genome Analyzer platform. An average ~40-fold coverage of the genome was observed per lane for samples with ≤ 30% human DNA. Even in low resource settings, using a simple two-step combination of Lymphoprep plus Plasmodipur, over 70% of clinical sample preparations should exhibit sufficiently low human DNA quantities to enable ~40-fold sequence coverage of the P. falciparum genome using a single lane on the Illumina Genome Analyzer platform. This approach should greatly facilitate large-scale clinical and epidemiologic studies of P

  15. A method for the determination of environmental contaminants in living marine mammals using microscale samples of blubber and blood.

    PubMed

    Newman, J W; Vedder, J M; Jarman, W M; Chang, R R

    1994-08-01

    As part of a study examining the possible effects of organochlorine compounds on juvenile northern elephant seals (Mirounga angustirostris), blubber and blood samples were taken from animals present on the Año Nuevo (California) rookery, and from animals admitted for rehabilitation at The Marine Mammal Center (Sausalito CA). Blubber samples were collected from immobilized seals. A pre-cleaned 6 mm K-medic biopsy punch was used to extract the blubber from a 1 cm incision near the hip, near the dorsal mid point. Blood samples were taken from the extradural vein; two mL of serum was analyzed for organochlorine compounds. Blubber samples (approximately 0.1g) were ground with Na2SO4 and extracted with 20 mL hexane:methylene chloride (1:1). Sera samples were extracted using commercially available disposable C18 columns. The extracts were separated on a micro-Florisil column, and analyzed by HRGC-ECD. Lipid determination in the serum was obtained by colorimetric analysis with 20 microliters samples. Results from the analysis of replicates and standard reference materials showed good recoveries, precision, and accuracy for both the blubber and blood methods.

  16. A flow cytometric method for enumeration of lymphocyte sub-populations in sample containing lysis-resistant red blood cells.

    PubMed

    Kasinrerk, Watchara

    2003-05-01

    Determination of lymphocyte sub-populations is usually carried out by flow cytometry using two-color immunophenotyping reagent. By this technique, however, the combination of FSC and SSC with CD45-FITC/CD14-PE is unable to identify the lymphocyte population in a sample containing lysis-resistant red blood cells (RBC). The actual values of lymphocyte sub-populations, therefore, cannot be determined in these RBC contaminated samples. To overcome this problem, we describe here the use of 7-aminoactinomycin D (7-AAD) to exclude lysis-resistant RBC from white blood cells (WBC). By adding 7-AAD, lymphocytes of samples containing RBC could be identified by using FL3/SSC, therefore, the actual number of lymphocyte sub-populations of the stained cells was obtained. We have proved that 7-AAD can be used to exclude contaminated RBC and has no effect on the measurement of lymphocyte sub-populations by using two-color immunophenotyping reagent. In routine blood samples that contain lysis-resistant RBC, 7-AAD markedly increased the purity of lymphocytes in the lymphocyte gate to >95% and the lymphocyte sub-populations therefore could be correctly determined. The described method is inexpensive, simple and gives successful analysis of lymphocyte sub-populations in a sample containing lysis-resistant RBC.

  17. A dried blood spots technique based LC-MS/MS method for the analysis of posaconazole in human whole blood samples.

    PubMed

    Reddy, Todime M; Tama, Cristina I; Hayes, Roger N

    2011-11-15

    A rugged and robust liquid chromatographic tandem mass spectrometric (LC-MS/MS) method utilizing dried blood spots (DBS) was developed and validated for the analysis of posaconazole in human whole blood. Posaconazole fortified blood samples were spotted (15 μL) onto Ahlstrom Alh-226 DBS cards and dried for at least 2h. Punched spots were then extracted by using a mixture of acetonitrile and water containing stable labeled internal standard (IS). Posaconazole and its IS were separated from endogenous matrix components on a Kinetex™ C18 column under gradient conditions with a mobile phase A consisting of 0.1% formic acid and a mobile phase B consisting of 0.1% formic acid in acetonitrile/methanol (70/30, v/v). The analyte and IS were detected using a Sciex API 4000 triple quadrupole LC-MS/MS system equipped with a TurboIonSpray™ source operated in the positive ion mode. The assay was linear over the concentration range of 5-5000 ng/mL. The inter-run accuracy and precision of the assay were -1.8% to 0.8% and 4.0% to 10.4%, respectively. Additional assessments unique to DBS were investigated including sample spot homogeneity, spot volume, and hematocrit. Blood spot homogeneity was maintained and accurate and precise quantitation results were obtained when using a blood spot volume of between 15 and 35 μL. Human blood samples with hematocrit values ranging between 25% and 41% gave acceptable quantitation results. The validation results indicate that the method is accurate, precise, sensitive, selective and reproducible.

  18. International Study to Evaluate PCR Methods for Detection of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients

    PubMed Central

    Schijman, Alejandro G.; Bisio, Margarita; Orellana, Liliana; Sued, Mariela; Duffy, Tomás; Mejia Jaramillo, Ana M.; Cura, Carolina; Auter, Frederic; Veron, Vincent; Qvarnstrom, Yvonne; Deborggraeve, Stijn; Hijar, Gisely; Zulantay, Inés; Lucero, Raúl Horacio; Velazquez, Elsa; Tellez, Tatiana; Sanchez Leon, Zunilda; Galvão, Lucia; Nolder, Debbie; Monje Rumi, María; Levi, José E.; Ramirez, Juan D.; Zorrilla, Pilar; Flores, María; Jercic, Maria I.; Crisante, Gladys; Añez, Néstor; De Castro, Ana M.; Gonzalez, Clara I.; Acosta Viana, Karla; Yachelini, Pedro; Torrico, Faustino; Robello, Carlos; Diosque, Patricio; Triana Chavez, Omar; Aznar, Christine; Russomando, Graciela; Büscher, Philippe; Assal, Azzedine; Guhl, Felipe; Sosa Estani, Sergio; DaSilva, Alexandre; Britto, Constança; Luquetti, Alejandro; Ladzins, Janis

    2011-01-01

    Background A century after its discovery, Chagas disease still represents a major neglected tropical threat. Accurate diagnostics tools as well as surrogate markers of parasitological response to treatment are research priorities in the field. The purpose of this study was to evaluate the performance of PCR methods in detection of Trypanosoma cruzi DNA by an external quality evaluation. Methodology/Findings An international collaborative study was launched by expert PCR laboratories from 16 countries. Currently used strategies were challenged against serial dilutions of purified DNA from stocks representing T. cruzi discrete typing units (DTU) I, IV and VI (set A), human blood spiked with parasite cells (set B) and Guanidine Hidrochloride-EDTA blood samples from 32 seropositive and 10 seronegative patients from Southern Cone countries (set C). Forty eight PCR tests were reported for set A and 44 for sets B and C; 28 targeted minicircle DNA (kDNA), 13 satellite DNA (Sat-DNA) and the remainder low copy number sequences. In set A, commercial master mixes and Sat-DNA Real Time PCR showed better specificity, but kDNA-PCR was more sensitive to detect DTU I DNA. In set B, commercial DNA extraction kits presented better specificity than solvent extraction protocols. Sat-DNA PCR tests had higher specificity, with sensitivities of 0.05–0.5 parasites/mL whereas specific kDNA tests detected 5.10−3 par/mL. Sixteen specific and coherent methods had a Good Performance in both sets A and B (10 fg/µl of DNA from all stocks, 5 par/mL spiked blood). The median values of sensitivities, specificities and accuracies obtained in testing the Set C samples with the 16 tests determined to be good performing by analyzing Sets A and B samples varied considerably. Out of them, four methods depicted the best performing parameters in all three sets of samples, detecting at least 10 fg/µl for each DNA stock, 0.5 par/mL and a sensitivity between 83.3–94.4%, specificity of 85–95

  19. Detection of genetically modified corn (Bt176) in spiked cow blood samples by polymerase chain reaction and immunoassay methods.

    PubMed

    Petit, Laetitia; Baraige, Fabienne; Bertheau, Yves; Brunschwig, Philippe; Diolez, Annick; Duhem, Koenraad; Duplan, Marie-Noëlle; Fach, Patrick; Kobilinsky, André; Lamart, Stephen; Schattner, Alexandra; Martin, Patrice

    2005-01-01

    The fate of DNA and protein transgenic sequences in products derived from animals fed transgenic crops has recently raised public interest. Sensitive molecular tests targeting the Bt176 genetic construct and the transgenic Cry1Ab protein were developed to determine whether plant sequences, especially transgenic sequences, are present in animal products. A protocol for total DNA extraction and purification from cow whole blood samples was first drawn up and assessed by spiking with known amounts of DNA from Bt176 maize. The limit of detection for transgenic sequences (35S promoter and Bt176-specific junction sequence) was determined by both the polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA) and the 5'-nuclease PCR assay. Four additional PCR systems were built to substantiate the results. The first detects a mono-copy maize-specific sequence (ADH promoter). Two others target multi-copy sequences from plant nucleus (26S rRNA gene) and chloroplast (psaB gene). The last one, used as a positive control, targets a mono-copy animal sequence (alpha(s1)-casein gene). Both methods detected a minimum spiking at 25 copies of Bt176 maize/mL in 10 mL whole blood samples. The sandwich ELISA kit used detected down to 1 ng transgenic Cry1Ab protein/mL spiked whole blood.

  20. Sensitivity testing of trypanosome detection by PCR from whole blood samples using manual and automated DNA extraction methods.

    PubMed

    Dunlop, J; Thompson, C K; Godfrey, S S; Thompson, R C A

    2014-11-01

    Automated extraction of DNA for testing of laboratory samples is an attractive alternative to labour-intensive manual methods when higher throughput is required. However, it is important to maintain the maximum detection sensitivity possible to reduce the occurrence of type II errors (false negatives; failure to detect the target when it is present), especially in the biomedical field, where PCR is used for diagnosis. We used blood infected with known concentrations of Trypanosoma copemani to test the impact of analysis techniques on trypanosome detection sensitivity by PCR. We compared combinations of a manual and an automated DNA extraction method and two different PCR primer sets to investigate the impact of each on detection levels. Both extraction techniques and specificity of primer sets had a significant impact on detection sensitivity. Samples extracted using the same DNA extraction technique performed substantially differently for each of the separate primer sets. Type I errors (false positives; detection of the target when it is not present), produced by contaminants, were avoided with both extraction methods. This study highlights the importance of testing laboratory techniques with known samples to optimise accuracy of test results.

  1. Blood plasma sample preparation method for the assessment of thyroid hormone-disrupting potency in effect-directed analysis.

    PubMed

    Simon, Eszter; Bytingsvik, Jenny; Jonker, Willem; Leonards, Pim E G; de Boer, Jacob; Jenssen, Bjørn M; Lie, Elisabeth; Aars, Jon; Hamers, Timo; Lamoree, Marja H

    2011-09-15

    A sample preparation method combining solid-phase extraction (SPE) and liquid-liquid extraction (LLE) was developed to be used in Effect-Directed Analysis (EDA) of blood plasma. Until now such a method was not available. It can be used for extraction of a broad range of thyroid hormone (TH)-disruptors from plasma with high recoveries. Validation of the method using spiked cow plasma showed good recoveries for hydroxylated polybrominated diphenyl ethers (OH-PBDEs; 93.8 ± 19.5%), hydroxylated polychlorinated biphenyls (OH-PCBs; 93.8 ± 15.5%), other halogenated phenols (OHPs; 107 ± 8.1%), and for short-chain (<8 C-atoms) perfluoroalkyl substances (PFASs; 85.2 ± 24.6%). In the same extracts, the potency of the compound classes spiked to the cow plasma to competitively bind to transthyretin (TTR) was recovered by 84.9 ± 8.8%. Furthermore, the SPE-LLE method efficiently removed endogenous THs from the extracts, thereby eliminating their possible contribution to the binding assay response. The SPE-LLE method was applied to polar bear plasma samples to investigate its applicability in future EDA studies focusing on TH-disrupting compounds in this top predator species that is exposed to relatively high levels of bioaccumulating pollutants. A first screening revealed TTR-binding potency in the polar bear plasma extracts, which could be explained for 60-85% by the presence of OH-PCBs.

  2. Detection of progesterone in whole blood samples.

    PubMed

    Ehrentreich-Förster, Eva; Scheller, Frieder W; Bier, Frank F

    2003-04-01

    The progesterone concentration in blood samples can be utilised as a marker for the diagnosis of early pregnancy, endocrinopathy and virilism. Here, we describe a method for progesterone detection and measurement in whole blood samples by a surface sensitive biosensor used in conjunction with an integrated optical grating coupler. This device determines refractive index changes near the biosensor's surface. Hence, biological species bound to a surface layer can be measured in real-time without any label. For the measurements, we have modified the indirect competitive immunoassay principle. The concentration of the progesterone antibody was kept at 1 microg/ml. Progesterone concentration was determined in buffer solution and whole blood in a range between 0.005 and 10 ng/ml. The detection limit was determined to be 3 pM. The relative standard deviation was calculated to be 3.5%.

  3. Validation of a method to quantify chromium, cadmium, manganese, nickel and lead in human whole blood, urine, saliva and hair samples by electrothermal atomic absorption spectrometry.

    PubMed

    Olmedo, P; Pla, A; Hernández, A F; López-Guarnido, O; Rodrigo, L; Gil, F

    2010-02-05

    For biological monitoring of heavy metal exposure in occupational toxicology, usually whole blood and urine samples are the most widely used and accepted matrix to assess internal xenobiotic exposure. Hair samples and saliva are also of interest in occupational and environmental health surveys but procedures for the determination of metals in saliva and hair are very scarce and to our knowledge there is no validation of a method to quantify Cr, Cd, Mn, Ni and Pb in four different human biological materials (whole blood, urine, saliva and axilary hair) by electrothermal atomization atomic absorption spectrometry (ETAAS). In the present study, quantification methods for the determination of Cr, Cd, Mn, Ni and Pb in whole blood, urine, saliva and axilary hair were validated according to the EU common standards. Pyrolisis and atomization temperatures have been determined. The main parameters evaluated were: detection and quantification limits, linearity range, repeatability, reproducibility, recovery and uncertainty. Accuracy of the methods was tested with the whole blood, urine and hair certified reference materials and recoveries of the spiked samples were acceptable ranged from 96.3 to 107.8%.

  4. Higher detectability method for the analysis of nucleosides, putative tumor biomarkers, in blood serum samples by CE-UV with reversed EOF.

    PubMed

    Buzatto, Adriana Zardini; Guedes, Sumaya Ferreira; de Oliveira Silva, Mariana; Gallafrio, Jéssica Mirela; Simionato, Ana Valéria Colnaghi

    2015-12-01

    The development and validation of methodologies for the analysis of biological samples is of outcome importance in order to obtain trustworthy results. This work reports a novel CE-UV method for the assessment of nucleosides, putative tumor biomarkers, in blood serum. The separation of seven nucleosides within c.a. 20 min has been achieved with: BGE 30 mmol/L borate at pH 9.90, 50 mmol/L CTAB, and 10% methanol; V = -10 kV; T = 20°C; and capillary dimensions of 56 cm × 50 μm. The sample plug was concentrated by a modified large volume sample stacking strategy that provided better detectability. Validation showed that the method is suitable for bioanalytical purposes and initial applications in serum samples from healthy subjects are also presented. Finally, statistical methods were applied to verify the effect of characteristics such as age, smoking habits, and alcohol consumption on nucleoside concentrations in blood serum. Univariate statistical analysis tests emphasized the need for age matching, which was confirmed by PCA-DA and PLS-DA. Cancer history in the nearby family may also interfere in nucleoside levels in blood serum, since adenosine concentrations were statistically higher for volunteers who declared having diseased relatives.

  5. Guide to capillary heelstick blood sampling in infants.

    PubMed

    Folk, Laura A

    2007-08-01

    Capillary blood sampling is an essential method of blood collection performed by nurses of all skill levels to obtain samples for routine laboratory tests in neonates. Accuracy of results depends on proper heelstick and sample collection technique. Recent advances including development of devices designed specifically for heelstick capillary blood sampling and research into expanded safe heel capillary sampling sites are discussed. A step-by-step guide to capillary blood sampling is outlined along with evidence-based practice incorporating neonatal-appropriate disinfection and nonpharmacological analgesia that contribute to improved infant safety and comfort during and after the procedure.

  6. A novel fully validated LC-MS/MS method for quantification of pyridoxal-5'-phosphate concentrations in samples of human whole blood.

    PubMed

    Ghassabian, Sussan; Griffiths, Lyn; Smith, Maree T

    2015-09-01

    Quantification of pyridoxal-5'-phosphate (PLP) in biological samples is challenging due to the presence of endogenous PLP in matrices used for preparation of calibrators and quality control samples (QCs). Hence, we have developed an LC-MS/MS method for accurate and precise measurement of the concentrations of PLP in samples (20μL) of human whole blood that addresses this issue by using a surrogate matrix and minimizing the matrix effect. We used a surrogate matrix comprising 2% bovine serum albumin (BSA) in phosphate buffer saline (PBS) for making calibrators, QCs and the concentrations were adjusted to include the endogenous PLP concentrations in the surrogate matrix according to the method of standard addition. PLP was separated from the other components of the sample matrix using protein precipitation with trichloroacetic acid 10% w/v. After centrifugation, supernatant were injected directly into the LC-MS/MS system. Calibration curves were linear and recovery was >92%. QCs were accurate, precise, stable for four freeze-thaw cycles, and following storage at room temperature for 17h or at -80°C for 3 months. There was no significant matrix effect using 9 different individual human blood samples. Our novel LC-MS/MS method has satisfied all of the criteria specified in the 2012 EMEA guideline on bioanalytical method validation.

  7. Gas chromatographic method using electron-capture detection for the determination of musk xylene in human blood samples. Biological monitoring of the general population.

    PubMed

    Angerer, J; Käfferlein, H U

    1997-05-23

    Musk xylene (2,4,6-trinitro-1,3-dimethyl-5-tert.-butylbenzene, MX), a synthetic musk often used in different fragrances and soaps to substitute the natural musk, is a potential contaminant of humans. In this publication, a specific and sensitive detection method for the determination of musk xylene in human blood samples is described. The clean-up of the blood samples includes an extraction step followed by a solid-phase adsorption to separate MX from other plasma components. Separation and detection was carried out by capillary gas chromatography and an electron capture detector (GC-ECD). The results were verified using qualitative capillary gas chromatography and a mass selective detector with electron impact ionisation (GC-EI-MS). epsilon-Hexachlorocyclohexane (epsilon-HCH) is used as internal standard. The reliability of the GC-ECD method has been proved. The relative standard deviations of the within-series imprecision were 12.7% for samples with a concentration of 0.5 microg/l and 2.1% for samples with a concentration of 5.0 microg/l, whereas the relative standard deviations for the between-day imprecision were 14.9% (0.5 microg/l samples) and 3.4% (5.0 microg/l samples). The losses during sample treatment were between 10.1% and 17.8%. No interfering peaks were observed. The absolute detection limit was 0.1 microg/l plasma. A total of 72 human blood samples were analysed to determine the MX concentrations within the general population. In 66 of the 72 human blood samples, the MX concentrations ranged from 0.10 to 1.12 microg/l plasma for the described method. In six samples no MX was detected. The median concentration was 0.24+/-0.23 microg MX/l plasma. The 95 percentile was 0.79 microg/l. No correlation could be found between MX concentrations and smoking habit, broca index, age, sex as well as fish consumption habits. Nevertheless, the results demonstrate the exposure of the general population to MX.

  8. Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients

    PubMed Central

    Ramírez, Juan Carlos; Cura, Carolina Inés; Moreira, Otacilio da Cruz; Lages-Silva, Eliane; Juiz, Natalia; Velázquez, Elsa; Ramírez, Juan David; Alberti, Anahí; Pavia, Paula; Flores-Chávez, María Delmans; Muñoz-Calderón, Arturo; Pérez-Morales, Deyanira; Santalla, José; Guedes, Paulo Marcos da Matta; Peneau, Julie; Marcet, Paula; Padilla, Carlos; Cruz-Robles, David; Valencia, Edward; Crisante, Gladys Elena; Greif, Gonzalo; Zulantay, Inés; Costales, Jaime Alfredo; Alvarez-Martínez, Miriam; Martínez, Norma Edith; Villarroel, Rodrigo; Villarroel, Sandro; Sánchez, Zunilda; Bisio, Margarita; Parrado, Rudy; Galvão, Lúcia Maria da Cunha; da Câmara, Antonia Cláudia Jácome; Espinoza, Bertha; de Noya, Belkisyole Alarcón; Puerta, Concepción; Riarte, Adelina; Diosque, Patricio; Sosa-Estani, Sergio; Guhl, Felipe; Ribeiro, Isabela; Aznar, Christine; Britto, Constança; Yadón, Zaida Estela; Schijman, Alejandro G.

    2015-01-01

    An international study was performed by 26 experienced PCR laboratories from 14 countries to assess the performance of duplex quantitative real-time PCR (qPCR) strategies on the basis of TaqMan probes for detection and quantification of parasitic loads in peripheral blood samples from Chagas disease patients. Two methods were studied: Satellite DNA (SatDNA) qPCR and kinetoplastid DNA (kDNA) qPCR. Both methods included an internal amplification control. Reportable range, analytical sensitivity, limits of detection and quantification, and precision were estimated according to international guidelines. In addition, inclusivity and exclusivity were estimated with DNA from stocks representing the different Trypanosoma cruzi discrete typing units and Trypanosoma rangeli and Leishmania spp. Both methods were challenged against 156 blood samples provided by the participant laboratories, including samples from acute and chronic patients with varied clinical findings, infected by oral route or vectorial transmission. kDNA qPCR showed better analytical sensitivity than SatDNA qPCR with limits of detection of 0.23 and 0.70 parasite equivalents/mL, respectively. Analyses of clinical samples revealed a high concordance in terms of sensitivity and parasitic loads determined by both SatDNA and kDNA qPCRs. This effort is a major step toward international validation of qPCR methods for the quantification of T. cruzi DNA in human blood samples, aiming to provide an accurate surrogate biomarker for diagnosis and treatment monitoring for patients with Chagas disease. PMID:26320872

  9. Analytical Validation of Quantitative Real-Time PCR Methods for Quantification of Trypanosoma cruzi DNA in Blood Samples from Chagas Disease Patients.

    PubMed

    Ramírez, Juan Carlos; Cura, Carolina Inés; da Cruz Moreira, Otacilio; Lages-Silva, Eliane; Juiz, Natalia; Velázquez, Elsa; Ramírez, Juan David; Alberti, Anahí; Pavia, Paula; Flores-Chávez, María Delmans; Muñoz-Calderón, Arturo; Pérez-Morales, Deyanira; Santalla, José; Marcos da Matta Guedes, Paulo; Peneau, Julie; Marcet, Paula; Padilla, Carlos; Cruz-Robles, David; Valencia, Edward; Crisante, Gladys Elena; Greif, Gonzalo; Zulantay, Inés; Costales, Jaime Alfredo; Alvarez-Martínez, Miriam; Martínez, Norma Edith; Villarroel, Rodrigo; Villarroel, Sandro; Sánchez, Zunilda; Bisio, Margarita; Parrado, Rudy; Maria da Cunha Galvão, Lúcia; Jácome da Câmara, Antonia Cláudia; Espinoza, Bertha; Alarcón de Noya, Belkisyole; Puerta, Concepción; Riarte, Adelina; Diosque, Patricio; Sosa-Estani, Sergio; Guhl, Felipe; Ribeiro, Isabela; Aznar, Christine; Britto, Constança; Yadón, Zaida Estela; Schijman, Alejandro G

    2015-09-01

    An international study was performed by 26 experienced PCR laboratories from 14 countries to assess the performance of duplex quantitative real-time PCR (qPCR) strategies on the basis of TaqMan probes for detection and quantification of parasitic loads in peripheral blood samples from Chagas disease patients. Two methods were studied: Satellite DNA (SatDNA) qPCR and kinetoplastid DNA (kDNA) qPCR. Both methods included an internal amplification control. Reportable range, analytical sensitivity, limits of detection and quantification, and precision were estimated according to international guidelines. In addition, inclusivity and exclusivity were estimated with DNA from stocks representing the different Trypanosoma cruzi discrete typing units and Trypanosoma rangeli and Leishmania spp. Both methods were challenged against 156 blood samples provided by the participant laboratories, including samples from acute and chronic patients with varied clinical findings, infected by oral route or vectorial transmission. kDNA qPCR showed better analytical sensitivity than SatDNA qPCR with limits of detection of 0.23 and 0.70 parasite equivalents/mL, respectively. Analyses of clinical samples revealed a high concordance in terms of sensitivity and parasitic loads determined by both SatDNA and kDNA qPCRs. This effort is a major step toward international validation of qPCR methods for the quantification of T. cruzi DNA in human blood samples, aiming to provide an accurate surrogate biomarker for diagnosis and treatment monitoring for patients with Chagas disease.

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

    NASA Technical Reports Server (NTRS)

    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.

  11. Comparison of different blood sample processing methods for sensitive detection of low level chimerism by RHD real-time PCR assay.

    PubMed

    Javadi, Ahmad; Verduin, Esther P; Brand, Anneke; Schonewille, Henk

    2013-01-01

    The rhesus D blood group, which is expressed on the red blood cells (RBC) of 85% of the Caucasian population, is one of the most immunogenic RBC antigens, inducing D antibody formation in up to 20-80% of D-negative transfusion recipients and about 10% of pregnancies at risk. Pregnancy-induced D-antibodies can persist for many years, but the mechanisms underlying this persistence are unclear. The LOTUS study, a long-term follow-up study of mothers from severely affected children with hemolytic disease of the fetus and newborn investigates, among other endpoints, whether persistent feto-maternal chimerism is associated with long-term maternal anti-D persistence. We questioned which blood sample processing method should be used to detect low levels of RHD chimerism with the highest sensitivity and specificity using qPCR. After optimization of primer and probe concentrations for singleplex RHD exon 5 and 7 qPCR, sensitivity, specificity and efficiency of RHD and DYS1 qPCR were investigated in artificial chimeric samples. Sensitivity of DYS1 was one log higher (0.0001%) in enriched mononuclear cell fractions as compared with whole blood. Comparable linear sensitivity (0.007%) and mean efficiency (84-99%) for RHD qPCR were observed in all samples regardless whether whole blood or pre- or post-mixing of cellular fractions had been used. We conclude that RHD chimerism using singleplex exon 5 and 7 qPCR is linearly detectable down to 1.0 GE, without an advantage of fraction enrichment.

  12. A single-step extraction method for the determination of nicotine and cotinine in Jordanian smokers' blood and urine samples by RP-HPLC and GC-MS.

    PubMed

    Massadeh, Adnan M; Gharaibeh, Ahmad A; Omari, Khaled W

    2009-02-01

    A simple, rapid, reliable, and low cost one-step extraction method is developed and validated for the determination of nicotine and cotinine in human plasma and urine in smokers using reversed-phase high-performance liquid chromatography (RP-HPLC) and gas chromatography-mass spectrometry (GC-MS). The run times are 16 and 10 min for HPLC and GC-MS, respectively. The method is validated over a wide linear range of 1-5000 ng/mL with correlation coefficients being consistently greater than 0.9985. The criteria considered for validation are: limit of quantitation, linearity, accuracy, precision, recovery, specificity, and selectivity. This study is aimed to estimate the nicotine and cotinine in Jordanian smokers' blood and urine samples; to study the relationship between the concentration of nicotine in urine and plasma samples; and to investigate the effect of pH on the extraction of nicotine and cotinine in urine samples. In the presented study, one hundred blood and urine samples are collected from eighty smokers and twenty nonsmokers. Samples are taken from the same volunteer at the same time after each volunteer fills in a questionnaire. Results of nicotine concentrations in smokers' plasma are in the range of 181-3702 ng/mL with an average of 1263.1 ng/mL, whereas nicotine in urine samples is in the range of 1364-1972 ng/mL, with an average of 1618 ng/mL. Cotinine concentrations in smokers' plasma are in the range of 21-4420 ng/mL with an average of 379.4 ng/mL, whereas cotinine in urine is in the range of 6-3946 ng/mL with an average of 865 ng/mL. Statistical analysis indicates highly significant differences in nicotine and cotinine concentrations in smoker samples compared with nonsmoker samples (p<0.05).

  13. Analysis of a novel field dilution method for testing samples that exceed the analytic range of point-of-care blood lead analyzers.

    PubMed

    Neri, Antonio James; Roy, Joannie; Jarrett, Jeffery; Pan, Yi; Dooyema, Carrie; Caldwell, Kathleen; Umar-Tsafe, Nasir Tsafe; Olubiyo, Ruth; Brown, Mary Jean

    2014-01-01

    Investigators developed and evaluated a dilution method for the LeadCare II analyzer (LCII) for blood lead levels >65 μg/dL, the analyzer's maximum reporting value. Venous blood samples from lead-poisoned children were initially analyzed in the field using the dilution method. Split samples were analyzed at the US Centers for Disease Control and Prevention (CDC) laboratory using both the dilution method and inductively coupled plasma-mass spectrometry (ICP-MS). The concordance correlation coefficient of CDC LCII vs. ICP-MS values (N = 211) was 0.976 (95 % confidence interval (CI) 0.970-0.981); of Field LCII vs. ICP-MS (N = 68) was 0.910 (95% CI 0.861-0.942), and CDC LCII vs. Field LCII (N = 53) was 0.721 (95% CI 0.565-0.827). Sixty percent of CDC and 54% of Field LCII values were within ±10% of the ICP-MS value. Results from the dilution method approximated ICP-MS values and were useful for field-based decision-making. Specific recommendations for additional evaluation are provided.

  14. Sampling system and method

    DOEpatents

    Decker, David L.; Lyles, Brad F.; Purcell, Richard G.; Hershey, Ronald Lee

    2013-04-16

    The present disclosure provides an apparatus and method for coupling conduit segments together. A first pump obtains a sample and transmits it through a first conduit to a reservoir accessible by a second pump. The second pump further conducts the sample from the reservoir through a second conduit.

  15. The effects of fin rot disease and sampling method on blood chemistry and hematocrit measurements of winter flounder, Pseudopleuronectes americanus from New Haven Harbor (1987--1990).

    PubMed

    Ziskowski, J; Mercaldo-Allen, R; Pereira, J J; Kuropat, C; Goldberg, R

    2008-04-01

    Winter flounder from New Haven, Connecticut were evaluated for fin rot disease. Blood samples collected from healthy and diseased fish were used to measure bilirubin, calcium, hematocrit, inorganic phosphorus, osmolality, and total protein. Blood measurements were significantly affected by the presence of fin rot disease and by sampling mode (bled immediately or after 18 h). A reduction in blood chemistry values was associated with fin rot disease. Logistic regression modeling was used to identify explanatory variables contributing to the fin rot outcome in winter flounder. Blood constituent levels were higher in fish bled immediately versus 18 h post-capture, especially among fish without fin rot, suggesting that a waiting period is necessary for blood values to stabilize following initial sampling stress. This study presents evidence that winter flounder blood chemistry and hematocrit measurements are affected by fin rot disease.

  16. Percutaneous umbilical cord blood sampling - slideshow

    MedlinePlus

    ... htm Percutaneous umbilical cord blood sampling - series—Normal anatomy To use the sharing features on this page, ... Bethesda, MD 20894 U.S. Department of Health and Human Services National Institutes of Health Page last updated: ...

  17. Comparison of Eleven Methods for Genomic DNA Extraction Suitable for Large-Scale Whole-Genome Genotyping and Long-Term DNA Banking Using Blood Samples

    PubMed Central

    Psifidi, Androniki; Dovas, Chrysostomos I.; Bramis, Georgios; Lazou, Thomai; Russel, Claire L.; Arsenos, Georgios; Banos, Georgios

    2015-01-01

    Over the recent years, next generation sequencing and microarray technologies have revolutionized scientific research with their applications to high-throughput analysis of biological systems. Isolation of high quantities of pure, intact, double stranded, highly concentrated, not contaminated genomic DNA is prerequisite for successful and reliable large scale genotyping analysis. High quantities of pure DNA are also required for the creation of DNA-banks. In the present study, eleven different DNA extraction procedures, including phenol-chloroform, silica and magnetic beads based extractions, were examined to ascertain their relative effectiveness for extracting DNA from ovine blood samples. The quality and quantity of the differentially extracted DNA was subsequently assessed by spectrophotometric measurements, Qubit measurements, real-time PCR amplifications and gel electrophoresis. Processing time, intensity of labor and cost for each method were also evaluated. Results revealed significant differences among the eleven procedures and only four of the methods yielded satisfactory outputs. These four methods, comprising three modified silica based commercial kits (Modified Blood, Modified Tissue, Modified Dx kits) and an in-house developed magnetic beads based protocol, were most appropriate for extracting high quality and quantity DNA suitable for large-scale microarray genotyping and also for long-term DNA storage as demonstrated by their successful application to 600 individuals. PMID:25635817

  18. Sampling system and method

    DOEpatents

    Decker, David L.; Lyles, Brad F.; Purcell, Richard G.; Hershey, Ronald Lee

    2017-03-07

    In one embodiment, the present disclosure provides an apparatus and method for supporting a tubing bundle during installation or removal. The apparatus includes a clamp for securing the tubing bundle to an external wireline. In various examples, the clamp is external to the tubing bundle or integral with the tubing bundle. According to one method, a tubing bundle and wireline are deployed together and the tubing bundle periodically secured to the wireline using a clamp. In another embodiment, the present disclosure provides an apparatus and method for coupling conduit segments together. A first pump obtains a sample and transmits it through a first conduit to a reservoir accessible by a second pump. The second pump further conducts the sample from the reservoir through a second conduit. In a specific example, one or more clamps are used to connect the first and/or second conduits to an external wireline.

  19. Establishing and evaluating bar-code technology in blood sampling system: a model based on human centered human-centered design method.

    PubMed

    Chou, Shin-Shang; Yan, Hsiu-Fang; Huang, Hsiu-Ya; Tseng, Kuan-Jui; Kuo, Shu-Chen

    2012-01-01

    This study intended to use a human-centered design study method to develop a bar-code technology in blood sampling process. By using the multilevel analysis to gather the information, the bar-code technology has been constructed to identify the patient's identification, simplify the work process, and prevent medical error rates. A Technology Acceptance Model questionnaire was developed to assess the effectiveness of system and the data of patient's identification and sample errors were collected daily. The average scores of 8 items users' perceived ease of use was 25.21(3.72), 9 items users' perceived usefulness was 28.53(5.00), and 14 items task-technology fit was 52.24(7.09), the rate of patient identification error and samples with order cancelled were down to zero, however, new errors were generated after the new system deployed; which were the position of barcode stickers on the sample tubes. Overall, more than half of nurses (62.5%) were willing to use the new system.

  20. Evaluation of Mutual Drug-Drug Interaction within Geneva Cocktail for Cytochrome P450 Phenotyping using Innovative Dried Blood Sampling Method.

    PubMed

    Bosilkovska, Marija; Samer, Caroline; Déglon, Julien; Thomas, Aurélien; Walder, Bernhard; Desmeules, Jules; Daali, Youssef

    2016-09-01

    Cytochrome P450 (CYP) activity can be assessed using a 'cocktail' phenotyping approach. Recently, we have developed a cocktail (Geneva cocktail) which combines the use of low-dose probes with a low-invasiveness dried blood spots (DBS) sampling technique and a single analytical method for the phenotyping of six major CYP isoforms. We have previously demonstrated that modulation of CYP activity after pre-treatment with CYP inhibitors/inducer could be reliably predicted using Geneva cocktail. To further validate this cocktail, in this study, we have verified whether probe drugs contained in the latter cause mutual drug-drug interactions. In a randomized, four-way, Latin-square crossover study, 30 healthy volunteers received low-dose caffeine, flurbiprofen, omeprazole, dextromethorphan and midazolam (a previously validated combination with no mutual drug-drug interactions); fexofenadine alone; bupropion alone; or all seven drugs simultaneously (Geneva cocktail). Pharmacokinetic profiles of the probe drugs and their metabolites were determined in DBS samples using both conventional micropipette sampling and new microfluidic device allowing for self-sampling. The 90% confidence intervals for the geometric mean ratios of AUC metabolite/AUC probe for CYP probes administered alone or within Geneva cocktail fell within the 0.8-1.25 bioequivalence range indicating the absence of pharmacokinetic interaction. The same result was observed for the chosen phenotyping indices, that is metabolic ratios at 2 hr (CYP1A2, CYP3A) or 3 hr (CYP2B6, CYP2C9, CYP2C19, CYP2D6) post-cocktail administration. DBS sampling could successfully be performed using a new microfluidic device. In conclusion, Geneva cocktail combined with an innovative DBS sampling device can be used routinely as a test for simultaneous CYP phenotyping.

  1. A new supramolecular based liquid solid microextraction method for preconcentration and determination of trace bismuth in human blood serum and hair samples by electrothermal atomic absorption spectrometry.

    PubMed

    Kahe, Hadi; Chamsaz, Mahmoud

    2016-11-01

    A simple and reliable supramolecule-aggregated liquid solid microextraction method is described for preconcentration and determination of trace amounts of bismuth in water as well as human blood serum and hair samples. Catanionic microstructures of cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulfate (SDS) surfactants, dissolved in deionized water/propanol, are used as a green solvent to extract bismuth (III)-diethyldithiocarbamate complexes by dispersive microextraction methodology. The extracted solid phase is easily removed and dissolved in 50 μL propanol for subsequent measurement by electrothermal atomic absorption spectrometry (ET-AAS). The procedure benefits the merits of supramolecule aggregates' properties and dispersive microextraction technique using water as the main component of disperser solvent, leading to direct interaction with analyte. Phase separation behavior of extraction solvent and different parameters influencing the extraction efficiency of bismuth ion such as salt concentration, pH, centrifugation time, amount of chelating agent, SDS:CTAB mole ratio, and solvent amounts were thoroughly optimized. Under the optimal experimental conditions, the calibration curve was linear in the range of 0.3-6 μg L(-1) Bi (III) with a limit of detection (LOD) of 0.16 μg L(-1) (S/N = 3). The relative standard deviations (RSD) of determination were obtained to be 5.1 and 6.2 % for 1 and 3 μg L(-1) of Bi (III), respectively. The developed method was successfully applied as a sensitive and accurate technique for determination of bismuth ion in human blood serum, hair samples, and a certified reference material.

  2. [Blood sampling using "dried blood spot": a clinical biology revolution underway?].

    PubMed

    Hirtz, Christophe; Lehmann, Sylvain

    2015-01-01

    Blood testing using the dried blood spot (DBS) is used since the 1960s in clinical analysis, mainly within the framework of the neonatal screening (Guthrie test). Since then numerous analytes such as nucleic acids, small molecules or lipids, were successfully measured on the DBS. While this pre-analytical method represents an interesting alternative to classic blood sampling, its use in routine is still limited. We review here the different clinical applications of the blood sampling on DBS and estimate its future place, supported by the new methods of analysis as the LC-MS mass spectrometry.

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

    PubMed

    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.

  4. Method for measuring lead concentrations in blood

    DOEpatents

    Nogar, Nicholas S.

    2001-01-01

    Method for measuring lead concentrations in blood. The present invention includes the use of resonant laser ablation to analyze .ltoreq.1 .mu.L (or equivalent mass) samples of blood for lead content. A typical finger prick, for example, yields about 10 .mu.L. Solid samples may also readily be analyzed by resonant laser ablation. The sample is placed on a lead-free, electrically conducting substrate and irradiated with a single, focused laser beam which simultaneously vaporizes, atomizes, and resonantly ionizes an analyte of interest in a sample. The ions are then sorted, collected and detected using a mass spectrometer.

  5. Malaria PCR detection in Cambodian low-transmission settings: dried blood spots versus venous blood samples.

    PubMed

    Canier, Lydie; Khim, Nimol; Kim, Saorin; Eam, Rotha; Khean, Chanra; Loch, Kaknika; Ken, Malen; Pannus, Pieter; Bosman, Philippe; Stassijns, Jorgen; Nackers, Fabienne; Alipon, SweetC; Char, Meng Chuor; Chea, Nguon; Etienne, William; De Smet, Martin; Kindermans, Jean-Marie; Ménard, Didier

    2015-03-01

    In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.

  6. Improved Sampling Method Reduces Isokinetic Sampling Errors.

    ERIC Educational Resources Information Center

    Karels, Gale G.

    The particulate sampling system currently in use by the Bay Area Air Pollution Control District, San Francisco, California is described in this presentation for the 12th Conference on Methods in Air Pollution and Industrial Hygiene Studies, University of Southern California, April, 1971. The method represents a practical, inexpensive tool that can…

  7. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 8 2014-04-01 2014-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  8. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 8 2013-04-01 2013-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  9. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 8 2011-04-01 2011-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  10. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 8 2012-04-01 2012-04-01 false Arterial blood sampling kit. 868.1100 Section 868...) MEDICAL DEVICES ANESTHESIOLOGY DEVICES Diagnostic Devices § 868.1100 Arterial blood sampling kit. (a) Identification. An arterial blood sampling kit is a device, in kit form, used to obtain arterial blood...

  11. 21 CFR 868.1100 - Arterial blood sampling kit.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

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

  12. Determination of chlorinated insecticides in blood samples of agricultural workers.

    PubMed

    Rosell, M G; Obiols, J; Berenguer, M J; Guardino, X; López, F; Brosa, J

    1993-11-26

    Lindane, aldrin and p,p'-DDT were determined in blood samples from 71 farmers by means of an analytical method which combines a direct whole-blood extraction with n-hexane and gas chromatography (GC)-electron-capture detection (ECD), using a capillary column, applied to the organic extract. This technique allowed the determination of pesticides at levels varying from 0.1 to 180 micrograms per l of blood, the detection limit for every pesticide being 0.1 microgram/l. GC-mass spectrometry was used to confirm the identity of each pesticide. The advantage of capillary column GC-ECD for pesticide determination is its sensitivity and high resolution, which makes it possible to separate pesticides from a complex n-hexane extract obtained in a very simple pretreatment of the blood sample, which is itself a very complex matrix.

  13. Sampling system and method

    SciTech Connect

    Decker, David L; Lyles, Brad F; Purcell, Richard G; Hershey, Ronald Lee

    2014-05-20

    An apparatus and method for supporting a tubing bundle during installation or removal. The apparatus includes a clamp for securing the tubing bundle to an external wireline. The method includes deploying the tubing bundle and wireline together, The tubing bundle is periodically secured to the wireline using a clamp.

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

    NASA Technical Reports Server (NTRS)

    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.

  15. Blood viscometer applying electromagnetically spinning method.

    PubMed

    Fukunaga, Kazuyoshi; Onuki, Masaya; Ohtsuka, Yoshinori; Hirano, Taichi; Sakai, Keiji; Ohgoe, Yasuharu; Katoh, Ayako; Yaguchi, Toshiyuki; Funakubo, Akio; Fukui, Yasuhiro

    2013-09-01

    Viscosity is an important parameter which affects hemodynamics during extracorporeal circulation and long-term cardiac support. In this study, we have aimed to develop a novel viscometer with which we can easily measure blood viscosity by applying the electromagnetically spinning (EMS) method. In the EMS method, we can rotate an aluminum ball 2 mm in diameter indirectly in a test tube with 0.3 ml sample of a liquid by utilizing the moment caused by the Lorentz force as well as separate the test tube from the viscometer body. First, we calibrated the EMS viscometer by means of liquid samples with known viscosities and computational fluid dynamics. Then, when we measured the viscosity of 9.4 mPa s silicone oil in order to evaluate the performance of the EMS viscometer, the mean viscosity was found to be 9.55 ± 0.10 mPa s at available shear rates from 10 to 240 s(-1). Finally, we measured the viscosity of bovine blood. We prepared four blood samples whose hematocrit levels were adjusted to 23, 45, 50, and 70% and a plasma sample without hemocyte components. As a result, the measurements of blood viscosities showed obedience to Casson's equation. We found that the viscosity was approximately constant in Newtonian silicone oil, whereas the viscosity decreased with increasing shear rate in non-Newtonian bovine blood. These results suggest that the EMS viscometer will be useful to measure blood viscosity at the clinical site.

  16. Manual versus automated blood sampling: impact of repeated blood sampling on stress parameters and behavior in male NMRI mice

    PubMed Central

    Kalliokoski, Otto; Sørensen, Dorte B; Hau, Jann; Abelson, Klas S P

    2014-01-01

    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 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 measured, and expressed less anxious behavior. We conclude that repeated blood sampling by automated blood sampling and from the tail vein is less stressful than cheek blood sampling. The choice between automated blood sampling and tail blood sampling should be based on the study requirements, the resources of the laboratory and skills of the staff. PMID:24958546

  17. Design of Er:YAG laser blood-sampling device

    NASA Astrophysics Data System (ADS)

    Wu, Zhi-chao; Jin, Guang-yong; Tan, Xue-chun; Ling, Ming; Liang, Zhu

    2009-07-01

    Laser blood-sampling device is one of the foremost tasks in medicine domain. It has a lot of merits such as un-touching, avoiding infection, indolence, and fast healing etc. The Er:YAG laser with wavelength of 2.94μm which is just close to the absorbency peak of water can be strongly absorbed by water molecular, so it has very wide application value in clinical medicine. In the paper, based on the mutual action characters of the laser with 2.94μm wave length on biological tissues, such as high absorption, acting on surface, the design of a new type of laser blood-sampling device is introduced. According to the needs of practice, the main component of the blood-sampling device is the laser, which includes optical resonator, optical collector, pumping source, optical guidance and focusing system. All of them are designed in the paper, and the reflection index of output coupling mirror of laser is optimized, the laser threshold is reduced, and pumping efficiency is improved. Moreover, thermal effect of Er:YAG solid-state laser is analyzed and a reasonable cooling method is designed. As a result, an excellent laser blood- sampling is obtained, the maximum output power is about 1J, the optical to optical conversion efficiency is 1.2%. For the better production-grade, the cuprum-based conduction is adopt to eliminate heat, the precision modulation and fixing of the optical resonance is achieved by the special adjusting structure that not only improve the stability and reliability, but also reduce the size of laser bloodsampling device. The size is 110×190×320mm, the weight is about 5.8kg, and the laser blood- sampling efficiency is 100%.

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

    NASA Astrophysics Data System (ADS)

    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. Long-Term Blood Alcohol Stability in Forensic Antemortem Whole Blood Samples.

    PubMed

    Tiscione, Nicholas B; Vacha, Ruth E; Alford, Ilene; Yeatman, Dustin Tate; Shan, Xiaoqin

    2015-01-01

    The effect of long-term room temperature storage on the stability of ethanol in whole blood specimens was investigated. One hundred and seventeen preserved whole blood case samples (110 of 117 with two tubes of blood in each case) were used for this study. One tube from each case was initially tested for blood alcohol concentration (BAC) for criminal driving under the influence proceedings. Cases positive for ethanol ranged in BAC from 0.023 to 0.281 g/dL. The second tube, if present, remained sealed. All blood samples were then stored at room temperature. After 5.4-10.3 years, the opened tubes were reanalyzed for BAC by the same laboratory that performed the initial testing using the same method and same instrumentation. After the same storage period, the unopened tubes were sent to a different laboratory, using a different method and different instrumentation, and reanalyzed for BAC after a total of 5.6-10.5 years of room temperature storage. Seven samples initially negative for alcohol remained negative. All samples initially positive for ethanol demonstrated a decrease in BAC over time with a statistically significant difference in loss observed based on blood sample volume and whether or not the tube had been previously opened. The decrease in BAC ranged from 0.005 to 0.234 g/dL. Tubes that were not previously opened and were more than half full demonstrated better BAC stability with 89% of these tubes demonstrating a loss of BAC between 0.01 and 0.05 g/dL.

  20. Bacterial and fungal DNA extraction from blood samples: manual protocols.

    PubMed

    Lorenz, Michael G; Mühl, Helge; Disqué, Claudia

    2015-01-01

    A critical point of molecular diagnosis of systemic infections is the method employed for the extraction of microbial DNA from blood. A DNA isolation method has to be able to fulfill several fundamental requirements for optimal performance of diagnostic assays. First of all, low- and high-molecular-weight substances of the blood inhibitory to downstream analytical reactions like PCR amplification have to be removed. This includes human DNA which is a known source of false-positive results and factor decreasing the analytical sensitivity of PCR assays by unspecific primer binding. At the same time, even extremely low amounts of microbial DNA need to be supplied to molecular diagnostic assays in order to detect low pathogen loads in the blood. Further, considering the variety of microbial etiologies of sepsis, a method should be capable of lysing Gram-positive, Gram-negative, and fungal organisms. Last, extraction buffers, reagents, and consumables have to be free of microbial DNA which leads to false-positive results. Here, we describe manual methods which allow the extraction of microbial DNA from small- and large-volume blood samples for the direct molecular analysis of pathogen.

  1. Development of blood extraction system designed by female mosquito's blood sampling mechanism for bio-MEMS

    NASA Astrophysics Data System (ADS)

    Tsuchiya, Kazuyoshi; Nakanishi, Naoyuki; Nakamachi, Eiji

    2005-02-01

    A compact and wearable wristwatch type Bio-MEMS such as a health monitoring system (HMS) to detect blood sugar level for diabetic patient, was newly developed. The HMS consists of (1) a indentation unit with a microneedle to generate the skin penetration force using a shape memory alloy(SMA) actuator, (2) a pumping unit using a bimorph PZT piezoelectric actuator to extract the blood and (3) a gold (Au) electrode as a biosensor immobilized GOx and attached to the gate electrode of MOSFET to detect the amount of Glucose in extracted blood. GOx was immobilized on a self assembled spacer combined with an Au electrode by the cross-link method using BSA as an additional bonding material. The device can extract blood in a few microliter through a painless microneedle with the negative pressure by deflection of the bimorph PZT piezoelectric actuator produced in the blood chamber, by the similar way the female mosquito extracts human blood with muscle motion to flex or relax. The performances of the liquid sampling ability of the pumping unit through a microneedle (3.8mm length, 100μm internal diameter) using the bimorph PZT piezoelectric microactuator were measured. The blood extraction micro device could extract human blood at the speed of 2μl/min, and it is enough volume to measure a glucose level, compared to the amount of commercial based glucose level monitor. The electrode embedded in the blood extraction device chamber could detect electrons generated by the hydrolysis of hydrogen peroxide produced by the reaction between GOx and glucose in a few microliter extracted blood, using the constant electric current measurement system of the MOSFET type hybrid biosensor. The output voltage for the glucose diluted in the chamber was increased lineally with increase of the glucose concentration.

  2. A technique for improved blood sampling during sleep studies.

    PubMed

    Ona, E; Dimsdale, J E; Ancoli-Israel, S; Dillon, E; Watkins, L; Coy, T V; Clausen, J

    1994-11-01

    Research protocols often require that blood samples be drawn during sleep. This study compares the efficacy of obtaining nocturnal blood samples using a standard heparinized intravenous setup versus the same intravenous setup used in conjunction with a small chemical heating pad. The chemical heating pad significantly improved the number of blood samples obtained and the maintenance of intravenous patency. The use of a chemical heating pad is an economical way to resolve the frustration of lost blood samples while maintaining a reasonable environment to monitor sleep.

  3. Characterization of differences between blood sample matrices in untargeted metabolomics.

    PubMed

    Denery, Judith R; Nunes, Ashlee A K; Dickerson, Tobin J

    2011-02-01

    Large-scale proteomic and metabolomic technologies are increasingly gaining attention for their use in the diagnosis of human disease. In order to ensure the statistical power of relevant markers, such analyses must incorporate a large number of representative samples. While in a best-case scenario these samples are collected through a study design that is specifically tailored for the desired analysis, often studies must rely upon the analysis of large numbers of previously banked samples that may or may not have complete and accurate documentation of their associated collection and storage methods. In this study, several human blood matrices were analyzed and compared for the quality of metabolomic output. The sample types that were tested include plasma prepared with a variety of anticoagulants and serum collected by venipuncture and capillary blood collection protocols. Analysis with liquid chromatography-mass spectrometry (LC-MS) revealed only subtle differences between the various plasma preparation methods. Differences between the serum and plasma samples appear to be largely peptide/protein-based and are consistent with the biological distinction of the two matrices. Interestingly, the small molecule lysophosphatidylinositol was found to be in higher abundance in plasma, as a possible consequence of the effect of the intrinsic clotting cascade on adjacent metabolic pathways. Comparison of the small-molecule profiles of the capillary- and venipuncture-collected samples revealed 23 statistically significant compound differences between these sample types. Most of these features can be attributed to surfactants and detergents used to pretreat the skin in order to maintain the sterility of sample collection. However, several have identical mass and molecular formulas as endogenous human metabolites and could be erroneously attributed to actual metabolic perturbations. Understanding the extent of these matrix effects is important for control of systematic bias

  4. An HPLC method with diode array detector for the simultaneous quantification of chloroquine and desethylchloroquine in plasma and whole blood samples from Plasmodium vivax patients in Vietnam, using quinine as an internal standard

    PubMed Central

    Pham Nguyen, Phuong; Nguyen Duc Khanh, Tho; Nguyen Thanh Thuy, Nhien; Nguyen Thuy Nha, Ca; Pouplin, Thomas; Farrar, Jeremy; Thwaites, Guy E.; Tran Tinh, Hien

    2016-01-01

    Abstract A sensitive, simple method for quantification of chloroquine (CQ) and desethylchloroquine (MCQ) in whole blood and plasma from Plasmodium vivax patients has been developed using HPLC with diode array detection (DAD). Solid‐phase extraction on Isolute‐96‐CBA was employed to process 100 μL of plasma/whole blood samples. CQ, MCQ and quinine were separated using a mobile phase of phosphate buffer 25 mm, pH 2.60–acetonitrile (88:12, v/v) with 2 mm sodium perchlorate on a Zorbax SB‐CN 150 × 4.6 mm, 5 μm column at a flow rate of 1.2 mL/min, at ambient temperature in 10 min, with the DAD wavelength of 343 nm. The method was linear over the range of 10–5000 ng/mL for both CQ and MCQ in plasma and whole blood. The limit of detection was 4 ng/mL and limit of quantification was 10 ng/mL in both plasma and blood for CQ and MCQ. The intra‐, inter‐ and total assay precision were <10% for CQ and MCQ in plasma and whole blood. In plasma, the accuracies varied between 101 and 103%, whereas in whole blood, the accuracies ranged from 97.0 to 102% for CQ and MCQ. The method is an ideal technique with simple facilities and instruments, bringing about good separation in comparison with previous methods. © 2016 The Authors Biomedical Chromatography Published by John Wiley & Sons Ltd PMID:26578224

  5. Improvement of the Eakins and Brown method for measuring 59Fe and 55Fe in blood and other iron-containing materials by liquid scintillation counting and sample preparation using microwave digestion and ion-exchange column purification of iron.

    PubMed

    Viteri, F E; Kohaut, B A

    1997-01-01

    The simultaneous measurement of 59Fe and 55Fe in whole blood by liquid scintillation counting by the Eakins and Brown (EB) method is extensively used in iron absorption studies. The EB method requires many steps which increase the chances of error and decrease its sensitivity. We describe two modifications to the above method consisting of microwave digestion and column purification of iron. This "New Method" (NM) is simpler and more precise, and sensitive than the EB method. Counting efficiencies with the NM are similar for 59Fe (75%) as with the EB method but are better for 55Fe (29% for NM vs 22%), and cross counting from 59Fe into the 55Fe window is lower with the NM (3.7-4.5%) than with the EB method (10-12%). For the NM, recoveries of radioactive blood samples, in relation to processed standards ranged from 100 to 103% for 59Fe and 101 to 113% for 55Fe. For the EB method, recoveries ranged from 94 to 99% for 59Fe and from 88 to 93% for 55Fe. Even with very low counts, average intrarun CV with the NM was lower than 5.4% for either isotope, while it was as high as 10.0% for 55Fe with the EB method.

  6. Sampling problems in the micro determination of blood lead.

    PubMed

    Juselius, R E; Lupovich, P; Moriarty, R

    1975-01-01

    Sampling tube and fingertip contamination were found to present potential problems in the collection of samples for micro blood lead analyses. Large differences between micro screening and macro confirming lead levels were frequently observed when the time between collection of the two samples was 1-2 weeks. The magnitude of these differences decreased as macro blood lead concentration increased and were apparently a result of episodic lead ingestion in the population.

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

    PubMed

    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.

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

    USGS Publications Warehouse

    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.

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

    PubMed Central

    2014-01-01

    Background 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. Methods 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. Results 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. Conclusions 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. PMID:24406140

  10. Elaborating transition interface sampling methods

    SciTech Connect

    Erp, Titus S. van . E-mail: bolhuis@science.uva.nl

    2005-05-01

    We review two recently developed efficient methods for calculating rate constants of processes dominated by rare events in high-dimensional complex systems. The first is transition interface sampling (TIS), based on the measurement of effective fluxes through hypersurfaces in phase space. TIS improves efficiency with respect to standard transition path sampling (TPS) rate constant techniques, because it allows a variable path length and is less sensitive to recrossings. The second method is the partial path version of TIS. Developed for diffusive processes, it exploits the loss of long time correlation. We discuss the relation between the new techniques and the standard reactive flux methods in detail. Path sampling algorithms can suffer from ergodicity problems, and we introduce several new techniques to alleviate these problems, notably path swapping, stochastic configurational bias Monte Carlo shooting moves and order-parameter free path sampling. In addition, we give algorithms to calculate other interesting properties from path ensembles besides rate constants, such as activation energies and reaction mechanisms.

  11. Kinetics of ethanol degradation in forensic blood samples.

    PubMed

    Ferrari, L A; Triszcz, J M; Giannuzzi, L

    2006-09-12

    To determine ethanol in human post-mortem blood samples is problematic, largely due to the inappropriate and variable methods of preserving and storing, which can cause decomposition and loss of alcohol concentration. In this study, four crucial parameters of sample conservation were studied: temperature (T), percentage of air chamber in container (%CA), ethanol concentration in blood and post-mortem time. Blood samples from post-mortem cases were stored under different conditions (ethanol levels were known in all cases); factorial design variables: (%CA) 0, 5, 20, 35, 65%; storage temperature: 25, 4 and -10 degrees C; in a total of 15 experiments. No preserving agent was used in samples. Quantification of ethanol in blood was carried out by gas chromatography with head-space FID detector. Initial ethanol concentration ranged from 0.50 to 4.30 g/L. The kinetics of degradation observed was pseudo-first-order. The parameter that characterised the kinetics of ethanol degradation (k(0)) ranged from (4 x 10(-4) and 5.0 x 10(-1) day(-1)), depending on storage conditions. A strong dependence between ethanol degradation and the content of the air chamber was observed and this dependence was found to be stronger than that between degradation and temperature; there was an experimental relation between (k(0)) and (%CA). Activation energy for different conditions, i.e. 0, 5, 20, 35 and 65 (%CA), were calculated and contour plots were made. A mathematical equation relating air chamber, temperature and ethanol concentration at a certain time was determined. This equation allowed estimation of initial concentrations of ethanol with minimal error. A good correlation between experimental data and data calculated with the equation was obtained (r(2) = 0.9998). The best storage conditions were: 0% CA and storage at -10 degrees C, obtaining an ethanol degradation of 0.01% after 15 days. However, 33% of ethanol degradation was obtained with 35% CA at 25 degrees C after 15 days. This

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

    ERIC Educational Resources Information Center

    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…

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

    NASA Technical Reports Server (NTRS)

    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.

  14. Comparison of the MagNA pure LC automated system and the RiboPure-Blood RNA manual method for RNA extraction from multiple myeloma bone marrow samples conserved in an RNA stabilizer.

    PubMed

    Garcia-Effron, G; Gamarra, S; Crooke, A; Martínez-Sánchez, P; Lahuerta, J; Martínez-López, J

    2007-04-01

    A total of 62 frozen bone marrow specimens conserved in RNA later (Ambion) were processed using two different extraction methods, the MagNA Pure LC system (MAG; Roche) and the manual RiboPure-Blood RNA method (RIBO; Ambion); Beta glucoronidase RNA (GUS) was amplified by LightCycler PCR to evaluate the quality of both extraction procedures. Less than 1000 GUS copies/ml was detected in 26 of 62 specimens (41.94%) processed by MAG and in five of 62 specimens (8.06%) processed by RIBO. Moreover, RNA recovery from the 62 specimens by MAG is, on average, 2.91 cycle threshold-fold higher than RIBO (P = 0.0008). Furthermore, we compared the extraction times and reagent costs of both methods. In conclusion, RNA extraction using MAG is faster to process 32 samples and less expensive than RIBO but it is not sensitive enough to be employed for research purpose in our laboratory.

  15. Stability of γ-Hydroxybutyrate in Blood Samples from Impaired Drivers after Storage at 4°C and Comparison of GC-FID-GBL and LC-MS-MS Methods of Analysis.

    PubMed

    Jones, Alan Wayne; Gladh, Sven-Åke; Windberg, Charlotte Norup; Johansen, Sys Stybe

    2015-05-01

    The stability of γ-hydroxybutyrate (GHB) was determined in 50 blood samples from impaired drivers after storage at 4 °: C for up to 12 months. GHB was determined in whole blood by gas chromatography-flame ionization detector (GC-FID) after conversion into γ-butyrolactone (GBL) and results were compared with LC-MS-MS. Both analytical methods showed a linear response (R(2) > 0.99) to GHB concentrations from 2 to 250 mg/kg. The mean decrease in concentration after storage was 4.8 mg/kg, with extreme changes of +13 mg/kg or -29 mg/kg. Results by the GC-FID-GBL method (y-variate) and the LC-MS-MS method (x-variate) were highly correlated (R(2) = 0.974). The regression equation was y = 0.85x + 2.2 and residual standard deviation (SD) was 7.8 mg/kg. The y-intercept (2.2 mg/kg) was not significantly different from zero (P > 0.05), although the slope of the regression line (0.85) differed from unity (P < 0.001), indicating a proportional bias of 15%. The LC-MS-MS method tended to give higher results than the GC-FID-GBL method. The mean difference (bias) was 12 mg/kg (P < 0.001). The SD of individual differences was 11.3 mg/kg and 95% limits of agreement were -11 to +33 mg/kg. The results of this study show that concentrations of GHB in whole blood are stable during storage at 4 °: C for up to 6 months.

  16. Palpatory Method of Measuring Diastolic Blood Pressure

    PubMed Central

    Sahu, Dinesh; Bhaskaran, M

    2010-01-01

    Background: Most common method for measuring blood pressure is palpatory but only systolic pressure can be measured with this method. In this study we are describing palpatory method of measuring diastolic blood pressure as well. Patients & Methods: We have studied in 200 patients and compared systolic as well as diastolic blood pressures with two methods, auscutatory and palpatory. Systolic and diastolic blood pressure were measured by one of the authors with new palpatory method and noted down. Then an independent observer, who was blinded to the palpatory method's values, measured blood pressure by auscultatory method and noted down. The values were compared in term of range and percentage. Results: The difference were analysed and found that 102 (51%) patients had systolic and diastolic blood pressure measured by palpatory method, within ± 2 mmHg of auscutatory method, 37 (19%) patients had within ± 4 mmHg, 52 (25%) patients had same readings as with auscutatory method, and in 9 (0.5%) patients it could not be measured. Conclusion: The palpatory method would be very useful where frequent blood pressure measurement are being done manually like in wards, in busy OPD, patient on treadmill and also whenever stethoscope is not available. The blood pressure can be measured in noisy environment too. PMID:21547184

  17. An evaluation of blood smears made by a new method using a spinner and diluted blood.

    PubMed

    Nourbakhsh, M; Atwood, J G; Raccio, J; Seligson, D

    1978-12-01

    Blood smears were prepared with the use of a spinner, which rotated with a fixed velocity for a fixed time. All blood samples used for spun smears were diluted with a fixed ratio of buffered isotonic saline solution. Distribution of cells in these smears was found to be random. The average number of cells per unit area was substantially uniform from place to place on the same slide and on multiple slides made with the smae sample. The distribution of leukocytes by type was also iniform. For different blood samples, the average number of cells per unit area in the smears correlated well with the measured cell concentrations per unit volume in the samples for leukocytes, erythrocytes and platelets. Leukocyte differential counts on replicate spun smears using the same bloods also agreed to within the sampling error. They similarly agreed with differential counts on pulled smears made from undiluted samples of the same bloods. With few exceptions, erythrocytic morphology on the spun smears was comparable to that on the good areas of pulled smears made with undiluted samples of the same bloods. Nearly all the spun smears were suitable for both viual and fully automated hematologic examination for leukocytes, erythrocytes, and platelets. This was true over nearly the whole area of each spun slide. In these ways this spinner method makes smears whose consistently high quality is little affected by either the properties of the blood sample or the skill of maker.

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

    PubMed

    Batterman, Stuart A; Chernyak, Sergey; Su, Feng-Chiao

    2016-01-01

    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 (R (2) > 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

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

    PubMed

    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.

  20. Accuracy of cyclosporin measurements made in capillary blood samples obtained by skin puncture.

    PubMed

    Merton, G; Jones, K; Lee, M; Johnston, A; Holt, D W

    2000-10-01

    International consensus guidelines suggest that cyclosporin should be measured in whole blood. In some instances it may be advantageous to collect capillary blood, by a finger or ear prick method. However, drug concentrations in skin-puncture blood may not necessarily correlate with those measured in venous blood. This study compared cyclosporin concentrations in blood collected from the fingertip or earlobe with blood collected by standard venipuncture. Patient preference for each of the blood collection methods was also assessed. Specimens were obtained from organ transplant patients receiving cyclosporin, using each of the three methods: venipuncture, finger prick, and earlobe prick. The samples were assayed using a specific radioimmunoassay and the results were compared. In the 102 sets of samples collected, the mean difference (+/- standard deviation) in cyclosporin concentration between finger prick and venipuncture and ear prick and venipuncture was 2.6% (+/- 9.5%) and 2.7% (+/- 12.1%), respectively, while the comparable median (IQR) differences were 1.9% (-3.4% to +6.6%) and -1.1% (-2.8% to +7.2%), respectively. A high degree of correlation was observed between finger prick and venipuncture or ear prick and venipuncture or ear prick and finger prick (r2 > 0.86). Of the three methods of blood collection, finger prick was the patients' preferred method (P < 0.01). These data suggest that capillary blood collected by skin puncture is suitable for use in cyclosporin blood monitoring and acceptable to patients.

  1. Comparison of five blood-typing methods for the feline AB blood group system.

    PubMed

    Seth, Mayank; Jackson, Karen V; Giger, Urs

    2011-02-01

    Objective-To compare the ease of use and accuracy of 5 feline AB blood-typing methods: card agglutination (CARD), immunochromatographic cartridge (CHROM), gel-based (GEL), and conventional slide (SLIDE) and tube (TUBE) agglutination assays. Sample Population-490 anticoagulated blood samples from sick and healthy cats submitted to the Transfusion or Clinical Laboratory at the Veterinary Hospital of the University of Pennsylvania. Procedures-Sample selection was purposely biased toward those from anemic, type B, or type AB cats or those with autoagglutination. All blood samples were tested by use of GEL, SLIDE, and TUBE methods. Fifty-eight samples were also tested by use of CARD and CHROM methods. The presence of alloantibodies in all cats expressing the B antigen as detected by use of any method was also assessed. Results-Compared with the historical gold-standard TUBE method, good to excellent agreement was achieved with the other typing tests: CARD, 53 of 58 (91% agreement); CHROM, 55 of 58 (95%); GEL, 487 of 490 (99%); and SLIDE, 482 of 487 (99%; 3 samples were excluded because of autoagglutination). Four of the samples with discordant test results originated from cats with FeLV-related anemia. Conclusions and Clinical Relevance-Current laboratory and in-clinic methods provide simple and accurate typing for the feline AB blood group system with few discrepancies. Retyping after in-clinic typing with the GEL or TUBE laboratory methods is recommended to confirm any type B or AB cats.

  2. Viscosity measurements on very small capillary blood samples.

    PubMed

    Eugster, M; Häusler, K; Reinhart, W H

    2007-01-01

    Viscosity measurements on very small capillary blood samples could be of considerable clinical interest. We have developed an oscillating viscometer for very small volumes, which consists of a glass capillary containing 7 mul of blood, which is part of an oscillating torsional resonator. The damping of the sinusoidal oscillations depends on the density and viscosity of the fluid, which allows blood viscosity measurements. The instrument was first evaluated in comparison with a standard blood viscometer (Contraves LS 30). Blood from healthy volunteers anticoagulated with EDTA was adjusted to hematocrit levels of 20, 30, 40, 50, and 60%, respectively. A strong correlation was found between hematocrit and oscillating viscosity (y=0.17x-2.05, r=0.969, p<0.0001) and between oscillating and conventional high shear viscosity (y=1.11x-0.62, r=0.971, p<0.0001). Blood viscosity measured in venous or capillary blood of normal subjects was similar (p=0.63). Bedside viscosity measurements on capillary blood drawn from a finger prick during routine blood glucose measurements in patients with diabetes mellitus showed lower blood viscosity than controls (3.62+/-0.87 vs 4.79+/-0.59 mPa.s, p=0.0007), which is in contrast to earlier publications, and may be explained by the lower hematocrit in our diabetic patients (34.7+/-6.0% vs. 43.1+/-1.9%, p<0.0001). Blood viscosity was independent of the actual glucose level (range 3-17 mmol/l). Capillary blood anticoagulated with EDTA was drawn by heel prick from 23 newborns. Blood viscosity was higher (5.66 +/-2.47 mPa.s) than in adult controls (see above), which could be explained by the dependence on the higher hematocrit (46.4 +/-8.6%). We conclude that viscosity measurements can be made on very small samples such as capillary blood from diabetic patients or newborn babies with this new oscillating viscometer. It remains to be determined if such new informations have clinical implications.

  3. Polybrominated diphenyl ethers in maternal and fetal blood samples.

    PubMed Central

    Mazdai, Anita; Dodder, Nathan G; Abernathy, Mary Pell; Hites, Ronald A; Bigsby, Robert M

    2003-01-01

    Polybrominated diphenyl ethers (PBDEs) are widely used as flame retardants in consumer goods, such as plastics, electronics, textiles, and construction material. PBDEs have been found in human milk, fat, and blood samples. Rodent studies indicate that PBDEs may be detrimental to neurodevelopment, possibly by lowering thyroid hormone concentrations in blood. In the present study, we determined concentrations of PBDEs and thyroid hormones in human fetal and maternal serum. Patients presenting in labor to Indiana University and Wishard Memorial County hospitals in Indianapolis, who were older than 18 years, were recruited to participate. Twelve paired samples of maternal and cord blood were obtained and analyzed using gas chromatographic mass spectrometry; thyroid hormone concentrations were determined by radioimmunoassay. Six congeners of PBDE were measured in maternal and fetal serum samples. The concentrations of total PBDEs found in maternal sera ranged from 15 to 580 ng/g lipid, and the concentrations found in fetal samples ranged from 14 to 460 ng/g lipid. Individual fetal blood concentrations did not differ from the corresponding maternal concentrations, indicating that measurement of maternal PBDE blood levels is useful in predicting fetal exposure; similarly, other reports have shown a high correlation between PBDE in mother's milk and fetal exposure. In accord with reports on other biologic samples, the tetrabrominated PBDE congener BDE-47 accounted for 53-64% of total PBDEs in the serum. The concentrations of PBDEs found in maternal and fetal serum samples were 20-106-fold higher than the levels reported previously in a similar population of Swedish mothers and infants. In this small sample, there was no apparent correlation between serum PBDEs and thyroid hormone concentrations. Our study shows that human fetuses in the United States may be exposed to relatively high levels of PBDEs. Further investigation is required to determine if these levels are

  4. Long-term preservation of blood samples for diagnosis of Trypanosoma cruzi infection.

    PubMed

    Pérez, A C; Cura, E; Subías, E; Lansetti, J C; Segura, E L

    1990-03-01

    Feasability and suitability for field research of a whole-blood preservation method was evaluated through the screening of anti-Trypanosoma cruzi antibodies in 1209 samples under different conditions. Antibody reactivity of paired samples from preserved capillary blood (CBP) and sera from venous blood (VBS) were studied by specific techniques. Over 96% concordance was found on indoor studies carried out with samples without storage or after 15 or 30 days preservation of CBP at 37 degrees C and VBS at -20 degrees C. Outdoor studies performed at field conditions, achieved a 92.1% concordance.

  5. Duplex sampling apparatus and method

    DOEpatents

    Brown, Paul E.; Lloyd, Robert

    1992-01-01

    An improved apparatus is provided for sampling a gaseous mixture and for measuring mixture components. The apparatus includes two sampling containers connected in series serving as a duplex sampling apparatus. The apparatus is adapted to independently determine the amounts of condensable and noncondensable gases in admixture from a single sample. More specifically, a first container includes a first port capable of selectively connecting to and disconnecting from a sample source and a second port capable of selectively connecting to and disconnecting from a second container. A second container also includes a first port capable of selectively connecting to and disconnecting from the second port of the first container and a second port capable of either selectively connecting to and disconnecting from a differential pressure source. By cooling a mixture sample in the first container, the condensable vapors form a liquid, leaving noncondensable gases either as free gases or dissolved in the liquid. The condensed liquid is heated to drive out dissolved noncondensable gases, and all the noncondensable gases are transferred to the second container. Then the first and second containers are separated from one another in order to separately determine the amount of noncondensable gases and the amount of condensable gases in the sample.

  6. Blood Samples of Peripheral Venous Catheter or The Usual Way: Do Infusion Fluid Alters the Biochemical Test Results?

    PubMed Central

    Taghizadeganzadeh, Mahboobeh; Yazdankhahfard, Mohammadreza; Farzaneh, Mohammadreza; Mirzaei, Kamran

    2016-01-01

    Background: Most blood tests require venous blood samples. Puncturing the vein also causes pain, infection, or damage to the blood, and lymph flow, or long-term healing. This study aimed to determine and compare the biochemical laboratory value of the blood samples that were provided through: peripheral vein infusion (PVI) receiving continuous intravenous fluid; and the usual method of blood sampling. Methods: This is an interventional, quasi-experimental, and controlled study. The selected study sample included 60 patients, who were hospitalized during 2014, in the Internal Medicine, part of Martyrs of Persian Gulf, teaching hospital at Bushehr. Three blood samples were taken from each patient that were provided through PVI line (5 ml blood collected at beginning of IVC and then another 5 cc), and another case was prepared by common blood sampling (control). All the samples were analyzed in terms of sodium, potassium, urea and creatinine using SPSS Ver.19 software, by paired t-test and Pearson’s correlation coefficients. Results: There was a statistically significant difference between the amount of sodium and potassium in the first blood samples taken from the intravenous infusion line and vein puncture. However, no significant differences were found among the biochemical amount in the second blood samples taken from the intravenous infusion line and vein puncture. Conclusions: We can use blood samples taken from peripheral intravenous infusion lines after 5cc discarding from the first part of the sample for measuring the value of sodium, potassium, urea and creatinine. PMID:26925892

  7. Detection methods for autologous blood doping.

    PubMed

    Segura, J; Monfort, N; Ventura, R

    2012-11-01

    The use of blood doping is forbidden by the World Anti-Doping Agency. Several practices, such as blood transfusions are used to increase oxygen delivery to muscles and all of them are highly pursued. In this regard, the development of accurate methodologies for detecting these prohibited practices is one of the current aims of the anti-doping control laboratories. Flow cytometry methods are able to detect allogeneic blood transfusions but there is no official methodology available to detect autologous blood transfusions. This paper reviews protocols, including the Athlete Biological Passport, that use indirect markers to detect misuse of blood transfusions, especially autologous blood transfusions. The methods of total haemoglobin mass measurements and the detection of metabolites of blood bags plasticizers in urine are reviewed. The latter seems to be an important step forward because it is a fast screening method and it is based on urine, a fluid widely available for doping control. Other innovative approaches to blood transfusion detection are also mentioned. A combination of the reported methodologies and the implementation of the Athlete Biological Passport is becoming a promising approach.

  8. Investigation of prothrombin time in human whole-blood samples with a quartz crystal biosensor.

    PubMed

    Müller, Lothar; Sinn, Stefan; Drechsel, Hartmut; Ziegler, Christiane; Wendel, Hans-Peter; Northoff, Hinnak; Gehring, Frank K

    2010-01-15

    Monitoring of blood coagulation and fibrinolysis is an important issue in treatment of patients with cardiovascular problems and in surgery when blood gets into contact with artificial surfaces. In this work a new method for measuring the coagulation time (prothrombin time, PT) of human whole-blood samples based on a quartz crystal microbalance (QCM) biosensor is presented. The 10 MHz sensors used in this work respond with a frequency shift to changes in viscosity during blood clot formation. For driving and for readout of the quartz, both a network analyzer and an oscillator circuit were utilized. The sensor surfaces were specifically coated with a thin polyethylene layer. We found that both frequency analysis methods are suitable to measure exact prothrombin times in a very good conformity with a mechanical coagulometer as a reference. The anticoagulant effect of heparin on the prothrombin time was exemplarily shown as well as the reverse effect of the heparin antagonist polybrene. The change of the viscoelastic properties during blood coagulation, reflected by the ratio of frequency and dissipation shifts, is discussed for different dilutions of the whole-blood samples. In conclusion, QCM is a distinguished biosensor technique to determine prothrombin time and to monitor heparin therapy in whole-blood samples. Due to the excellent potential of miniaturization and the availability of direct digital signals, the method is predestinated for incorporation and integration into other devices and is thus opening the field of application for inline coagulation diagnostic in extracorporeal blood circuits.

  9. Method for Reducing Pumping Damage to Blood

    NASA Technical Reports Server (NTRS)

    Bozeman, Richard J., Jr. (Inventor); Akkerman, James W. (Inventor); Aber, Gregory S. (Inventor); VanDamm, George Arthur (Inventor); Bacak, James W. (Inventor); Svejkovsky, Robert J. (Inventor); Benkowski, Robert J. (Inventor)

    1997-01-01

    Methods are provided for minimizing damage to blood in a blood pump wherein the blood pump comprises a plurality of pump components that may affect blood damage such as clearance between pump blades and housing, number of impeller blades, rounded or flat blade edges, variations in entrance angles of blades, impeller length, and the like. The process comprises selecting a plurality of pump components believed to affect blood damage such as those listed herein before. Construction variations for each of the plurality of pump components are then selected. The pump components and variations are preferably listed in a matrix for easy visual comparison of test results. Blood is circulated through a pump configuration to test each variation of each pump component. After each test, total blood damage is determined for the blood pump. Preferably each pump component variation is tested at least three times to provide statistical results and check consistency of results. The least hemolytic variation for each pump component is preferably selected as an optimized component. If no statistical difference as to blood damage is produced for a variation of a pump component, then the variation that provides preferred hydrodynamic performance is selected. To compare the variation of pump components such as impeller and stator blade geometries, the preferred embodiment of the invention uses a stereolithography technique for realizing complex shapes within a short time period.

  10. Apparatus and method for handheld sampling

    DOEpatents

    Staab, Torsten A.

    2005-09-20

    The present invention includes an apparatus, and corresponding method, for taking a sample. The apparatus is built around a frame designed to be held in at least one hand. A sample media is used to secure the sample. A sample media adapter for securing the sample media is operated by a trigger mechanism connectively attached within the frame to the sample media adapter.

  11. Soil sampling kit and a method of sampling therewith

    DOEpatents

    Thompson, C.V.

    1991-02-05

    A soil sampling device and a sample containment device for containing a soil sample is disclosed. In addition, a method for taking a soil sample using the soil sampling device and soil sample containment device to minimize the loss of any volatile organic compounds contained in the soil sample prior to analysis is disclosed. The soil sampling device comprises two close fitting, longitudinal tubular members of suitable length, the inner tube having the outward end closed. With the inner closed tube withdrawn a selected distance, the outer tube can be inserted into the ground or other similar soft material to withdraw a sample of material for examination. The inner closed end tube controls the volume of the sample taken and also serves to eject the sample. The soil sample containment device has a sealing member which is adapted to attach to an analytical apparatus which analyzes the volatile organic compounds contained in the sample. The soil sampling device in combination with the soil sample containment device allows an operator to obtain a soil sample containing volatile organic compounds and minimizing the loss of the volatile organic compounds prior to analysis of the soil sample for the volatile organic compounds. 11 figures.

  12. Soil sampling kit and a method of sampling therewith

    DOEpatents

    Thompson, Cyril V.

    1991-01-01

    A soil sampling device and a sample containment device for containing a soil sample is disclosed. In addition, a method for taking a soil sample using the soil sampling device and soil sample containment device to minimize the loss of any volatile organic compounds contained in the soil sample prior to analysis is disclosed. The soil sampling device comprises two close fitting, longitudinal tubular members of suitable length, the inner tube having the outward end closed. With the inner closed tube withdrawn a selected distance, the outer tube can be inserted into the ground or other similar soft material to withdraw a sample of material for examination. The inner closed end tube controls the volume of the sample taken and also serves to eject the sample. The soil sample containment device has a sealing member which is adapted to attach to an analytical apparatus which analyzes the volatile organic compounds contained in the sample. The soil sampling device in combination with the soil sample containment device allow an operator to obtain a soil sample containing volatile organic compounds and minimizing the loss of the volatile organic compounds prior to analysis of the soil sample for the volatile organic compounds.

  13. A study of blood alcohol stability in forensic antemortem blood samples.

    PubMed

    Shan, Xiaoqin; Tiscione, Nicholas B; Alford, Ilene; Yeatman, Dustin Tate

    2011-09-10

    The effect of long-term storage on alcohol stability in preserved forensic antemortem blood samples was investigated. Thirty-two whole blood case samples (each with two tubes of blood) were used for this study. One tube from each case was analyzed for blood alcohol concentration (BAC) for court proceedings of driving under the influence (DUI), and all blood samples were then stored under refrigeration. After the storage time (ranging from 13 to 39 months) both tubes of blood for each case were reanalyzed for BAC and the results were compared to the original analysis. Seven samples originally negative for alcohol analysis remained negative. The comparative data for 25 samples demonstrated various losses in BAC in both tubes. A significant loss with a mean of 0.015g/dL, was observed in previously opened tubes compared to a mean loss of 0.010g/dL in unopened tubes. In order to determine the effect of other storage conditions, the same blood samples were then stored at room temperature for 6 months followed by 38°C for 7 and 28 days and analyzed for BAC at the end of each storage time period. The seven alcohol negative cases remained negative when stored at room temperature or at 38°C. Six months of storage at room temperature decreased BAC further for both tubes of the alcohol positive cases with a mean loss of 0.014g/dL. Further storage at 38°C for 7 days did not cause any significant change in BAC. Storage at 38°C for 28 days caused some loss in BAC which was determined to be significant by statistical analysis.

  14. Vesicle-associated microRNAs are released from blood cells on incubation of blood samples.

    PubMed

    Köberle, Verena; Kakoschky, Bianca; Ibrahim, Ahmed Atef; Schmithals, Christian; Peveling-Oberhag, Jan; Zeuzem, Stefan; Kronenberger, Bernd; Waidmann, Oliver; Pleli, Thomas; Piiper, Albrecht

    2016-03-01

    MicroRNAs (miRNAs) circulating extracellularly in the blood are currently intensively studied as novel disease markers. However, the preanalytical factors influencing the levels of the extracellular miRNAs are still incompletely explored. In particular, it is unknown, whether the incubation of blood samples as occurring in clinical routine can lead to a release of miRNAs from blood cells and thus alter the extracellular miRNA levels before the preparation of serum or plasma from the blood cells. Using a set of marker miRNAs and quantitative RT-PCR, we found that the levels of extracellular miRNA-1, miRNA-16, and miRNA-21 were increased in EDTA and serum collection tubes incubated for 1-3 hours at room temperature and declined thereafter; the levels of the liver-specific miRNA-122 declined monophasically. These events occurred in the absence of significant hemolysis. When the blood was supplemented with Ribonuclease A inhibitor, the levels of miRNA-1, miRNA-16, and miRNA-21 increased substantially during the initial 3 hours of incubation and those of miRNA-122 remained unchanged, indicating that the release of blood cell-derived miRNAs occurred during the initial 3 hours of incubation of the blood tubes, but not at later time points. Separation of 5-hour preincubated blood into vesicle and nonvesicle fractions revealed a selective increase in the portion of vesicle-associated miRNAs. Together, these data indicate that the release of vesicle-associated miRNAs from blood cells can occur in blood samples within the time elapsing in normal clinical practice until their processing without significant hemolysis. This becomes particularly visible on the inhibition of miRNA degradation by Ribonuclease A inhibitor.

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

    PubMed Central

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

    2010-01-01

    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. PMID:20234395

  16. Foetal blood sampling. Practical approach to management of foetal distress.

    PubMed

    Coltart, T M; Trickey, N R; Beard, R W

    1969-02-08

    The practical application of foetal blood sampling in the routine management of patients in labour has been reviewed in a six-month survey, during which time 1,668 patients were delivered at Queen Charlotte's Hospital.Foetal acidaemia (pH 7.25 or less) occurred in 45 of the 295 patients who showed clinical signs of foetal distress. Foetal tachycardia was the presenting sign in 33 of these 45 patients, underlining the importance of this physical sign. Foetal acidaemia in association with clinical foetal distress occurred twice as often in patients who had complications of pregnancy and who were therefore regarded as obstetrically "at risk" as it did in patients who were obstetrically "normal" No cases of acidaemia were detected in any of the foetal blood samples performed routinely on "at-risk" patients in the absence of clinical foetal distress.

  17. Blood proteins analysis by Raman spectroscopy method

    NASA Astrophysics Data System (ADS)

    Artemyev, D. N.; Bratchenko, I. A.; Khristoforova, Yu. A.; Lykina, A. A.; Myakinin, O. O.; Kuzmina, T. P.; Davydkin, I. L.; Zakharov, V. P.

    2016-04-01

    This work is devoted to study the possibility of plasma proteins (albumin, globulins) concentration measurement using Raman spectroscopy setup. The blood plasma and whole blood were studied in this research. The obtained Raman spectra showed significant variation of intensities of certain spectral bands 940, 1005, 1330, 1450 and 1650 cm-1 for different protein fractions. Partial least squares regression analysis was used for determination of correlation coefficients. We have shown that the proposed method represents the structure and biochemical composition of major blood proteins.

  18. Molecular recognition of HER-1 in whole-blood samples.

    PubMed

    Moldoveanu, Iuliana; Stanciu Gavan, Camelia; Stefan-van Staden, Raluca-Ioana

    2014-11-01

    Multimode sensing was proposed for molecular screening and recognition of HER-1 in whole blood. The tools used for molecular recognition were platforms based on nanostructured materials such as the complex of Mn(III) with meso-tetra (4-carboxyphenyl) porphyrin, and maltodextrin (dextrose equivalence between 4 and 7), immobilized in diamond paste, graphite paste or C60 fullerene paste. The identification of HER-1 in whole-blood samples, at molecular level, is performed using stochastic mode and is followed by the quantification of it using stochastic and differential pulse voltammetry modes. HER-1 can be identified in the concentration range between 280 fg/ml and 4.86 ng/ml using stochastic mode, this making possible the early detection of cancers such as gastrointestinal, pancreatic and lung cancers. The recovery tests performed using whole-blood samples proved that the platforms can be used for identification and quantification of HER-1 with high sensitivity and reliability in such samples, these making them good molecular screening tools.

  19. A Clinical Evaluation of Routine Blood Sampling Practices in Patients With Diabetes

    PubMed Central

    Pineau, Mitchel; Pynes, Mary Kate; Katz, Laurence B.; Ginsberg, Barry

    2014-01-01

    Background: There is a perception that patients with diabetes struggle to produce sufficient blood to fill glucose test strips, including strips with 1-µL fill requirements. The purpose of this study was to determine the volume of blood expressed when these patients perform routine fingersticks using their own lancing device and sampling technique and to evaluate the relationship between blood volume and pain. Methods: Sixty-four patients (type 1 or type 2 diabetes) performed 8 fingersticks using their own lancing device and preferred depth setting and lancing technique. Eight different commercially available lancing systems were used (8 patients/system). Blood volume and perceived pain were recorded after each fingerstick. Results: The mean blood volume across all patients was 3.1 µL (512 fingersticks), with 97% of patients expressing a mean of ≥1.0 µL of blood. There was no correlation between pain response and the volume of blood expressed. Nearly all patients agreed that they could easily and comfortably obtain a 1-µL blood sample, and most patients actually preferred a larger drop size to ease sampling and avoid wasting strips. Conclusion: These results provide evidence across 8 lancing systems that challenge the current perceptions that patients with diabetes struggle to produce sufficient blood samples to fill most test strips, including those with 1-µL fill requirements, and that obtaining larger volumes of blood is more painful. These results are consistent with the previous literature suggesting that patients derive no real benefits from very low strip volumes and generally prefer a blood drop size that enables them to confidently fill their test strip. PMID:24876439

  20. An easy method for the determination of active concentrations of cholinesterase reactivators in blood samples: Application to the efficacy assessment of non quaternary reactivators compared to HI-6 and pralidoxime in VX-poisoned mice.

    PubMed

    Calas, André-Guilhem; Dias, José; Rousseau, Catherine; Arboléas, Mélanie; Touvrey-Loiodice, Mélanie; Mercey, Guillaume; Jean, Ludovic; Renard, Pierre-Yves; Nachon, Florian

    2017-04-01

    Organophosphorus nerve agents, like VX, are highly toxic due to their strong inhibition potency against acetylcholinesterase (AChE). AChE inhibited by VX can be reactivated using powerful nucleophilic molecules, most commonly oximes, which are one major component of the emergency treatment in case of nerve agent intoxication. We present here a comparative in vivo study on Swiss mice of four reactivators: HI-6, pralidoxime and two uncharged derivatives of 3-hydroxy-2-pyridinaldoxime that should more easily cross the blood-brain barrier and display a significant central nervous system activity. The reactivability kinetic profile of the oximes is established following intraperitoneal injection in healthy mice, using an original and fast enzymatic method based on the reactivation potential of oxime-containing plasma samples. HI-6 displays the highest reactivation potential whatever the conditions, followed by pralidoxime and the two non quaternary reactivators at the dose of 50 mg/kg bw. But these three last reactivators display equivalent reactivation potential at the same dose of 100 μmol/kg bw. Maximal reactivation potential closely correlates to surviving test results of VX intoxicated mice.

  1. Sample taking during orthopedic surgery: sensitivity and specificity using the BACTEC blood culture system.

    PubMed

    Podleska, L E; Lendemans, S; Schmid, E; Hussmann, B; Nast-Kolb, D; Taeger, G

    2012-02-01

    The use of blood culture systems for sterile body fluids other than blood has proven to be superior to routine culture methods. This study was conducted in order to assess the performance of the BACTEC blood culture system compared to swab/tissue sample collection for the detection of infection from intraoperative samples taken during surgical procedures. Sensitivity was determined by taking samples (BACTEC and swab/tissue samples) from patients with clinically evident infection (Infection group). Specificity was tested by taking the same sample sets from patients who had aseptic operations with no history of infection (Control group). The sensitivity was found to be much higher for the BACTEC group (50 isolates from 56 samples, sensitivity: 89%) compared to the swab/tissue samples (29 isolates out of 56 samples, sensitivity: 52%). The specificity was lower in the BACTEC group (32 isolates out of 44 samples, specificity: 27%) compared to the swab/tissue samples (1 isolate out of 44 samples, specificity: 98%). We conclude that BACTEC is useful for intraoperative sample collection in cases of low-grade infection. However, it is less specific and there is always the possibility for contamination. Therefore, it is advisable to use this technique in combination with regular tissue samples.

  2. Nanoliter viscometer for analyzing blood plasma and other liquid samples.

    PubMed

    Srivastava, Nimisha; Davenport, Robertson D; Burns, Mark A

    2005-01-15

    We have developed a microfabricated nanoliter capillary viscometer that quickly, easily, and inexpensively measures the viscosity of liquids. The measurement of viscosity is based on capillary pressure-driven flow inside microfluidic channels (depth approximately 30 microm and width approximately 300 microm). Accurate and precise viscosity measurements can be made in less than 100 s while using only 600 nL of liquid sample. The silicon-glass hybrid device (18 mm by 15 mm) contains on-chip components that measure the driving capillary pressure difference and the relevant geometrical parameters; these components make the nanoliter viscometer completely self-calibrating, robust, and easy to use. Several different microfabricated viscometers were tested using solutions with viscosities ranging from 1 to 5 cP, a range relevant to biological fluids (urine, blood, blood plasma, etc.). Blood plasma samples collected from patients with the symptoms of hyperviscosity syndrome were tested on the nanoliter capillary viscometer to an accuracy of 3%. Such self-calibrating nanoliter viscometers may have widespread applications in chemical, biological, and medical laboratories as well as in personal health care.

  3. Effects of sample storage time, temperature and syringe type on blood gas tensions in samples with high oxygen partial pressures.

    PubMed Central

    Pretto, J. J.; Rochford, P. D.

    1994-01-01

    BACKGROUND--Although plastic arterial sampling syringes are now commonly used, the effects of sample storage time and temperature on blood gas tensions are poorly described for samples with a high oxygen partial pressure (PaO2) taken with these high density polypropylene syringes. METHODS--Two ml samples of tonometered whole blood (PaO2 86.7 kPa, PaCO2 4.27 kPa) were placed in glass syringes and in three brands of plastic blood gas syringes. The syringes were placed either at room temperature or in iced water and blood gas analysis was performed at baseline and after 5, 10, 20, 40, 60, 90, and 120 minutes. RESULTS--In the first 10 minutes measured PaO2 in plastic syringes at room temperature fell by an average of 1.21 kPa/min; placing the sample on ice reduced the rate of PaO2 decline to 0.19 kPa/min. The rate of fall of PaO2 in glass at room temperature was 0.49 kPa/min. The changes in PaCO2 were less dramatic and at room temperature averaged increases of 0.47 kPa for plastic syringes and 0.71 kPa for glass syringes over the entire two hour period. These changes in gas tension for plastic syringes would lead to an overestimation of pulmonary shunt measured by the 100% oxygen technique of 0.6% for each minute left at room temperature before analysis. CONCLUSIONS--Glass syringes are superior to plastic syringes in preserving samples with a high PaO2, and prompt and adequate cooling of such samples is essential for accurate blood gas analysis. PMID:8016801

  4. Hematological Assessment in Pet Guinea Pigs (Cavia porcellus): Blood Sample Collection and Blood Cell Identification.

    PubMed

    Zimmerman, Kurt; Moore, David M; Smith, Stephen A

    2015-09-01

    Pet guinea pigs are presented to veterinary clinics for routine care and treatment of clinical diseases. In addition to obtaining clinical history and exam findings, diagnostic testing may be required, including hematological assessments. This article describes common blood collection methods, including venipuncture sites, the volume of blood that can be safely collected, and handling of the blood. Hematological parameters for normal guinea pigs are provided for comparison with in-house or commercial test results. A description of the morphology of guinea pig leukocytes is provided to assist in performing a differential count.

  5. Hematological assessment in pet guinea pigs (Cavia porcellus): blood sample collection and blood cell identification.

    PubMed

    Zimmerman, Kurt; Moore, David M; Smith, Stephen A

    2015-01-01

    Pet guinea pigs are presented to veterinary clinics for routine care and treatment of clinical diseases. In addition to obtaining clinical history and exam findings, diagnostic testing may be required, including hematological assessments. This article describes common blood collection methods, including venipuncture sites, the volume of blood that can be safely collected, and handling of the blood. Hematological parameters for normal guinea pigs are provided for comparison with in-house or commercial test results. A description of the morphology of guinea pig leukocytes is provided to assist in performing a differential count.

  6. Limited blood sampling for pharmacokinetic dose tailoring of FVIII in the prophylactic treatment of haemophilia A.

    PubMed

    Björkman, S

    2010-07-01

    The aim of this study was to evaluate the use of limited blood sampling and Bayesian analysis to estimate the pharmacokinetics (PK) and tailor the dose of factor VIII (FVIII) in an individual patient. In a Bayesian analysis, PK parameters are estimated from only a few plasma concentration measurements, using a previously established PK model. First the necessary model was created using intense blood sampling FVIII data from 10 patients. Then FVIII data from another 21 patients were used for 'clinical' evaluation. Three scenarios were created retrospectively by reduction of the original 7-sample data set; blood sampling at 4, 24 and 48 h, at 8 and 30 h and at 24 h after the infusion. PK parameters were estimated for each individual using Bayesian analysis and compared with those obtained using conventional methods from the full data. The accuracy of predictions of FVIII levels during prophylactic treatment 5-17 months later and implications for dose tailoring were also investigated. Blood sampling at 4, 24 and 48 h was found to give practically the same PK information as a full, conventional (7-10-sample) study. Even a single 24-h FVIII level provided adequate data for initial dose tailoring and gave predictions of FVIII levels 5-17 months later that were not appreciably worse than predictions based on the full PK analysis. By contrast, dose tailoring based on body weight failed completely. In conclusion, PK-based dose tailoring of FVIII can be performed using limited blood sampling during prophylactic treatment.

  7. Experience of fetal scalp blood sampling during labor.

    PubMed

    Liljeström, Lena; Wikström, Anna-Karin; Skalkidou, Alkistis; Akerud, Helena; Jonsson, Maria

    2014-01-01

    Fetal scalp blood sampling (FBS) is often claimed to be painful for women in labor and difficult for obstetricians to perform. Our aim was to assess women's experience of pain during FBS and obstetricians' experience of difficulty in performing the test. At a tertiary center in Sweden, a questionnaire with answers on a 10-point scale was completed by 51 women and the obstetricians performing the test. Women's experience of pain had a median of 3.5. FBS was well tolerated in women who had epidural analgesia but might be associated with pain in women without. Higher maternal body mass index and less cervical dilation were associated with higher pain ratings. Obstetricians did not generally experience scalp sampling as difficult to perform (median score 3.0). However, the sampling procedure can be more complicated in situations with higher maternal body mass index, less cervical dilation, and a higher station of the fetal head.

  8. Hematologic Assessment in Pet Rats, Mice, Hamsters, and Gerbils: Blood Sample Collection and Blood Cell Identification.

    PubMed

    Lindstrom, Nicole M; Moore, David M; Zimmerman, Kurt; Smith, Stephen A

    2015-09-01

    Hamsters, gerbils, rats, and mice are presented to veterinary clinics and hospitals for prophylactic care and treatment of clinical signs of disease. Physical examination, history, and husbandry practice information can be supplemented greatly by assessment of hematologic parameters. As a resource for veterinarians and their technicians, this article describes the methods for collection of blood, identification of blood cells, and interpretation of the hemogram in mice, rats, gerbils, and hamsters.

  9. Hematologic assessment in pet rats, mice, hamsters, and gerbils: blood sample collection and blood cell identification.

    PubMed

    Lindstrom, Nicole M; Moore, David M; Zimmerman, Kurt; Smith, Stephen A

    2015-01-01

    Hamsters, gerbils, rats, and mice are presented to veterinary clinics and hospitals for prophylactic care and treatment of clinical signs of disease. Physical examination, history, and husbandry practice information can be supplemented greatly by assessment of hematologic parameters. As a resource for veterinarians and their technicians, this article describes the methods for collection of blood, identification of blood cells, and interpretation of the hemogram in mice, rats, gerbils, and hamsters.

  10. Donating blood for research: a potential method for enhancing customer satisfaction of permanently deferred blood donors

    PubMed Central

    Waller, Daniel; Thijsen, Amanda; Garradd, Allira; Hayman, Jane; Smith, Geoff

    2017-01-01

    Background Each year, a large number of individuals in Australia are deferred from donating blood. A deferral may have a negative impact on donor satisfaction and subsequent word-of-mouth communication. The Australian Red Cross Blood Service (the Blood Service) is, therefore, investigating options for managing service interactions with deferred donors to maintain positive relationships. While public research institutes in Australia have established independent research donor registries, other countries provide programmes allowing deferred donors to donate blood for research via blood collection agencies. This study examined attitudes towards donating blood for research use in a sample of permanently deferred Australian donors. Materials and methods Donors permanently deferred because of a risk of variant Creutzfeldt-Jakob disease (n=449) completed a postal survey that examined attitudes towards research donation. Results The majority of participants were interested in donating blood for research (96%), and joining a registry of research donors (93%). Participants preferred to donate for transfusion or clinical research, and were willing to travel large distances. Results indicated that positive attitudes towards the Blood Service would be extended if the opportunity to donate blood was provided. These findings indicate a desire for continued engagement with the Blood Service despite deferral. Discussion Donating blood for research is a potential way of maintaining positive relationships with permanently deferred donors which also benefits the health research community. Through maintaining positive relationships with deferred donors, positive word-of-mouth activity can be stimulated. Further work is needed to determine the feasibility of implementing research donation through the Blood Service in Australia. PMID:26674813

  11. Blood Density Is Nearly Equal to Water Density: A Validation Study of the Gravimetric Method of Measuring Intraoperative Blood Loss.

    PubMed

    Vitello, Dominic J; Ripper, Richard M; Fettiplace, Michael R; Weinberg, Guy L; Vitello, Joseph M

    2015-01-01

    Purpose. The gravimetric method of weighing surgical sponges is used to quantify intraoperative blood loss. The dry mass minus the wet mass of the gauze equals the volume of blood lost. This method assumes that the density of blood is equivalent to water (1 gm/mL). This study's purpose was to validate the assumption that the density of blood is equivalent to water and to correlate density with hematocrit. Methods. 50 µL of whole blood was weighed from eighteen rats. A distilled water control was weighed for each blood sample. The averages of the blood and water were compared utilizing a Student's unpaired, one-tailed t-test. The masses of the blood samples and the hematocrits were compared using a linear regression. Results. The average mass of the eighteen blood samples was 0.0489 g and that of the distilled water controls was 0.0492 g. The t-test showed P = 0.2269 and R (2) = 0.03154. The hematocrit values ranged from 24% to 48%. The linear regression R (2) value was 0.1767. Conclusions. The R (2) value comparing the blood and distilled water masses suggests high correlation between the two populations. Linear regression showed the hematocrit was not proportional to the mass of the blood. The study confirmed that the measured density of blood is similar to water.

  12. Blood Density Is Nearly Equal to Water Density: A Validation Study of the Gravimetric Method of Measuring Intraoperative Blood Loss

    PubMed Central

    Vitello, Dominic J.; Ripper, Richard M.; Fettiplace, Michael R.; Weinberg, Guy L.; Vitello, Joseph M.

    2015-01-01

    Purpose. The gravimetric method of weighing surgical sponges is used to quantify intraoperative blood loss. The dry mass minus the wet mass of the gauze equals the volume of blood lost. This method assumes that the density of blood is equivalent to water (1 gm/mL). This study's purpose was to validate the assumption that the density of blood is equivalent to water and to correlate density with hematocrit. Methods. 50 µL of whole blood was weighed from eighteen rats. A distilled water control was weighed for each blood sample. The averages of the blood and water were compared utilizing a Student's unpaired, one-tailed t-test. The masses of the blood samples and the hematocrits were compared using a linear regression. Results. The average mass of the eighteen blood samples was 0.0489 g and that of the distilled water controls was 0.0492 g. The t-test showed P = 0.2269 and R2 = 0.03154. The hematocrit values ranged from 24% to 48%. The linear regression R2 value was 0.1767. Conclusions. The R2 value comparing the blood and distilled water masses suggests high correlation between the two populations. Linear regression showed the hematocrit was not proportional to the mass of the blood. The study confirmed that the measured density of blood is similar to water. PMID:26464949

  13. Cancer diagnosis by discrimination between normal and malignant human blood samples using attenuated total reflectance-Fourier transform infrared spectroscopy.

    PubMed

    Khanmohammadi, M; Ansari, M A; Garmarudi, A Bagheri; Hassanzadeh, G; Garoosi, G

    2007-09-01

    FTIR spectroscopy is a common technique for cancer diagnosis. Applied tissue samples are heterogeneous and may be damaged in preparation procedures. Easier sampling, more available samples and also easier process with assured results would be interesting. Whole blood samples include all of these qualifications and our hypothesis was the bio-molecular changes in blood which manifest themselves in different optical signatures, detectable by FTIR spectroscopy. Noncancerous blood samples were differentiated from cancerous ones using ATR-FTIR spectroscopy and LDA classification method. Procedure was 100 percent and 90 percent accurate in prediction of cancerous or noncancerous situation for 33 known and 10 unknown samples, respectively.

  14. Rapid and reliable determination of the halogenating peroxidase activity in blood samples.

    PubMed

    Flemmig, Jörg; Schwarz, Pauline; Bäcker, Ingo; Leichsenring, Anna; Lange, Franziska; Arnhold, Jürgen

    2014-12-15

    By combining easy and fast leukocyte enrichment with aminophenyl-fluorescein (APF) staining we developed a method to quickly and specifically address the halogenating activity of the immunological relevant blood heme peroxidases myeloperoxidase and eosinophil peroxidase, respectively. For leukocyte enrichment a two-fold hypotonic lysis procedure of the blood with Millipore water was chosen which represents a cheap, fast and reliable method to diminish the amount of erythrocytes in the samples. This procedure is shown to be suitable both to human and murine blood micro-samples, making it also applicable to small animal experiments with recurring blood sampling. As all types of leukocytes are kept in the sample during the preparation, they can be analysed separately after discrimination during the flow cytometry analysis. This also holds for all heme peroxidase-containing cells, namely neutrophils, eosinophils and monocytes. Moreover additional parameters (e.g. antibody staining) can be combined with the heme peroxidase activity determination to gain additional information about the different immune cell types. Based on previous results we applied APF for specifically addressing the halogenating activity of leukocyte peroxidases in blood samples. This dye is selectively oxidized by the MPO and EPO halogenation products hypochlorous and hypobromous acid. This approach may provide a suitable tool to gain more insights into the immune-physiological role of the halogenating activity of heme peroxidases.

  15. [Quantitation of cerebral blood flow and partition coefficient using 123I-IMP dynamic SPECT with single arterial blood sampling].

    PubMed

    Mizumura, S; Kumita, S; Kumazaki, T

    1996-03-01

    A method base on the two-compartment model was developed to measure quantitative cerebral blood flow (CBF) and partition coefficient (lambda) of IMP from dynamic SPECT and single arterial blood sampling. In this method, the linear differential equation of two-compartment model, Yokoi proposed, was employed and quantitative CBF and lambda values were measured with the standard input function calibrated by single arterial sampling. The input function was derived from the standard input function scaled by a factor determined by the single arterial blood sample. This new technique was applied to 5 normal volunteers (Ages ranged from 25 to 29 yr., average 26 yr.). The optimal time to calibrate the standard input function in the individual study and optimal the period of the upper limit time to which input function is integrated from IMP administration for analysis of the equation were determined to minimize the difference between integration of the calibrated standard input function and of the individual input function. Minimization of the difference yields an optimal calibration time (4 to 10 min after IMP administration) and the period of the upper limit time (8 to 60 min after acquisition start). Comparison of CBF and lambda values obtained by the graphical method using the calibrated standard data and individual input function were performed. It should be noted that CBF values were in good agreement between the two methods, respectively (r = 0.92, P<0.01; r = 0.72, p = 0.01). This method is easy to estimate CBF and lambda by only single arterial blood sampling and IMP dynamic SPECT, and useful for routine studies.

  16. [Investigation of uninterpretative HLA typing in 311 umbilical cord blood samples].

    PubMed

    Hong, Jing-Xin; Liang, Xiao-Lan; Han, Jun-Ling; Li, Qian; Qiu, Lu-Gui

    2009-10-01

    The aim of this study was to investigate the factors which affect HLA typing in 311 umbilical cord blood (UCB) samples. The HLA low resolution typing of UCB samples with misinterpreted HLA types from 311 UCB samples analyzed by PCR-SSO and PCR-SSP was performed. 7 samples difficult to determine their HLA genotype were sequenced directly and the reason leading to misinterpret HLA typing was analyzed. The results indicated that 99.4% of misinterpreted samples resulted from the restriction of HLA typing method itself and 0.6% of misinterpreted samples were suspected to be contaminated with maternal blood in UCB. It is concluded that HLA typing is mainly affected by the shortcomings of oligonucleotide probe design for PCR-SSO and lack of allele specific primers of PCR-SSP.

  17. Using blood samples to estimate persistent organic pollutants and metals in green sea turtles (Chelonia mydas).

    PubMed

    van de Merwe, Jason P; Hodge, Mary; Olszowy, Henry A; Whittier, Joan M; Lee, Shing Y

    2010-04-01

    Persistent organic pollutants (POPs) and heavy metals have been reported in a number of green turtle (Chelonia mydas) populations worldwide. However, due to ethical considerations, these studies have generally been on tissues from deceased and stranded animals. The purpose of this study was to investigate the use of blood samples to estimate the tissue contamination of live C. mydas populations. This study analysed 125 POP compounds and eight heavy metals in the blood, liver, kidney and muscle of 16 C. mydas from the Sea World Sea Turtle Rehabilitation Program, Gold Coast, Australia. Strong correlations were observed between blood and tissue concentrations for a number of POPs and metals. Furthermore, these correlations were observed over large ranges of turtle size, sex and condition. These results indicate that blood samples are a reliable non-lethal method for predicting chemical contamination in C. mydas.

  18. Toxoplasma polymerase chain reaction on experimental blood samples.

    PubMed

    Joss, A W; Chatterton, J M; Evans, R; Ho-Yen, D O

    1993-01-01

    A two-stage polymerase chain reaction (PCR) assay employing oligonucleotide primers from the B1 gene of Toxoplasma gondii was developed and assessed for sensitivity and specificity. It was able to detect T. gondii DNA from as little as one parasite/sample in mock-infected rat or mouse leucocyte preparations. Parasitaemia was also identified in animals at five stages between 16 and 66 h after infection with the virulent RH strain, and at 12 stages between 2 and 38 days after infection with the cyst-forming Beverley strain. In the latter case, PCR was more sensitive than animal culture. No cross-reactions were observed in samples containing various opportunist pathogens which may also be found in the blood of immunocompromised patients.

  19. Subrandom methods for multidimensional nonuniform sampling

    NASA Astrophysics Data System (ADS)

    Worley, Bradley

    2016-08-01

    Methods of nonuniform sampling that utilize pseudorandom number sequences to select points from a weighted Nyquist grid are commonplace in biomolecular NMR studies, due to the beneficial incoherence introduced by pseudorandom sampling. However, these methods require the specification of a non-arbitrary seed number in order to initialize a pseudorandom number generator. Because the performance of pseudorandom sampling schedules can substantially vary based on seed number, this can complicate the task of routine data collection. Approaches such as jittered sampling and stochastic gap sampling are effective at reducing random seed dependence of nonuniform sampling schedules, but still require the specification of a seed number. This work formalizes the use of subrandom number sequences in nonuniform sampling as a means of seed-independent sampling, and compares the performance of three subrandom methods to their pseudorandom counterparts using commonly applied schedule performance metrics. Reconstruction results using experimental datasets are also provided to validate claims made using these performance metrics.

  20. Method and apparatus for data sampling

    DOEpatents

    Odell, D.M.C.

    1994-04-19

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

  1. Method and apparatus for data sampling

    DOEpatents

    Odell, Daniel M. C.

    1994-01-01

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

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

    PubMed

    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.

  3. Liquid chromatography coupled with multi-channel electrochemical detection for the determination of daidzin in rat blood sampled by an automated blood sampling system.

    PubMed

    Tian, Feifei; Zhu, Yongxin; Long, Hong; Cregor, Meloney; Xie, Fuming; Kissinger, Candice B; Kissinger, Peter T

    2002-05-25

    Daidzin, a soy-derived biologically active natural product, has been reported to inhibit mitochondrial aldehyde dehydrogenase and suppress ethanol intake. This paper describes a method for the determination of daidzin in rat blood. After administration of daidzin, blood samples were periodically collected from awake, freely moving animals by a Culex automated blood sampler. Daidzin was extracted from 50 microl of diluted blood (blood and saline at a ratio of 1:1) with ethyl acetate. Chromatographic separation was achieved within 12 min using a microbore C(18) (100 x 1.0 mm) 3 microm column with a mobile phase containing 20 mM sodium acetate, 0.25 mM EDTA, pH 4.3, 4% methanol and 11% acetonitrile at a flow-rate of 90 microl/min. Detection was attained using a four-channel electrochemical detector with glassy carbon electrodes using oxidation potentials of +1100, 950, 850, 750 mV vs. Ag/AgCl. The limit of detection for daidzin in rat plasma was 5 ng/ml at a signal-to-noise ratio of 3:1. The extraction recovery of daidzin from rat plasma was over 74%. Linearity was obtained for the range of 25-1000 ng/ml. The intra- and inter-assay precisions were in the ranges of 2.7-6.6 and 1.9-3.7%, respectively. This method is suitable to routine in vivo monitoring of daidzin in rat plasma.

  4. Mixed Methods Sampling: A Typology with Examples

    ERIC Educational Resources Information Center

    Teddlie, Charles; Yu, Fen

    2007-01-01

    This article presents a discussion of mixed methods (MM) sampling techniques. MM sampling involves combining well-established qualitative and quantitative techniques in creative ways to answer research questions posed by MM research designs. Several issues germane to MM sampling are presented including the differences between probability and…

  5. Multivariate analysis methods for spectroscopic blood analysis

    NASA Astrophysics Data System (ADS)

    Wood, Michael F. G.; Rohani, Arash; Ghazalah, Rashid; Vitkin, I. Alex; Pawluczyk, Romuald

    2012-01-01

    Blood tests are an essential tool in clinical medicine with the ability diagnosis or monitor various diseases and conditions; however, the complexities of these measurements currently restrict them to a laboratory setting. P&P Optica has developed and currently produces patented high performance spectrometers and is developing a spectrometer-based system for rapid reagent-free blood analysis. An important aspect of this analysis is the need to extract the analyte specific information from the measured signal such that the analyte concentrations can be determined. To this end, advanced chemometric methods are currently being investigated and have been tested using simulated spectra. A blood plasma model was used to generate Raman, near infrared, and optical rotatory dispersion spectra with glucose as the target analyte. The potential of combined chemometric techniques, where multiple spectroscopy modalities are used in a single regression model to improve the prediction ability was investigated using unfold partial least squares and multiblock partial least squares. Results show improvement in the predictions of glucose levels using the combined methods and demonstrate potential for multiblock chemometrics in spectroscopic blood analysis.

  6. Small and cheap: accurate differential blood count with minimal sample volume by laser scanning cytometry (LSC)

    NASA Astrophysics Data System (ADS)

    Mittag, Anja; Lenz, Dominik; Smith, Paul J.; Pach, Susanne; Tarnok, Attila

    2005-04-01

    Aim: In patients, e.g. with congenital heart diseases, a differential blood count is needed for diagnosis. To this end by standard automatic analyzers 500 μl of blood is required from the patients. In case of newborns and infants this is a substantial volume, especially after operations associated with blood loss. Therefore, aim of this study was to develop a method to determine a differential blood picture with a substantially reduced specimen volume. Methods: To generate a differential blood picture 10 μl EDTA blood were mixed with 10 μl of a DRAQ5 solution (500μM, Biostatus) and 10 μl of an antibody mixture (CD45-FITC, CD14-PE, diluted with PBS). 20 μl of this cell suspension was filled into a Neubauer counting chamber. Due to the defined volume of the chamber it is possible to determine the cell count per volume. The trigger for leukocyte counting was set on DRAQ5 signal in order to be able to distinguish nucleated white blood cells from erythrocytes. Different leukocyte subsets could be distinguished due to the used fluorescence labeled antibodies. For erythrocyte counting cell suspension was diluted another 150 times. 20 μl of this dilution was analyzed in a microchamber by LSC with trigger set on forward scatter signal. Results: This method allows a substantial decrease of blood sample volume for generation of a differential blood picture (10 μl instead of 500μl). There was a high correlation between our method and the results of routine laboratory (r2=0.96, p<0.0001 n=40). For all parameters intra-assay variance was less than 7 %. Conclusions: In patients with low blood volume such as neonates and in critically ill infants every effort has to be taken to reduce the blood volume needed for diagnostics. With this method only 2% of standard sample volume is needed to generate a differential blood picture. Costs are below that of routine laboratory. We suggest this method to be established in paediatric cardiology for routine diagnostics and for

  7. Microwaving Blood as a Non-Destructive Technique for Haemoglobin Measurements on Microlitre Samples

    PubMed Central

    Basey-Fisher, Toby H.; Guerra, Nadia; Triulzi, Chiara; Gregory, Andrew; Hanham, Stephen M.; Stevens, Molly M.; Maier, Stefan A.; Klein, Norbert

    2016-01-01

    The non-destructive ex vivo determination of haemoglobin (Hgb) concentration offers the capability to conduct multiple red blood cell haematological measurements on a single sample, an advantage that current optical techniques are unable to offer. Here, a microwave method and device for the accurate and non-destructive determination of Hgb concentration in microlitre blood samples are described. Using broadband microwave spectroscopy, a relationship is established between the dielectric properties of murine blood and Hgb concentration that is utilized to create a technique for the determination of Hgb concentration. Subsequently, a microwave dielectric resonator-microfluidic system is implemented in the analysis of 52 murine samples with microlitre volumes and Hgb concentrations ranging from 0 to 17 g dL−1. Using the characterized relationship, independent and minimally invasive Hgb measurements are made on nine healthy mice as well as seven with mutations in the Adenomatous polyposis coli (APC) gene that leads to colorectal cancer and consequently anaemia. PMID:24002989

  8. ALARA ASSESSMENT OF SETTLER SLUDGE SAMPLING METHODS

    SciTech Connect

    NELSEN LA

    2009-01-30

    The purpose of this assessment is to compare underwater and above water settler sludge sampling methods to determine if the added cost for underwater sampling for the sole purpose of worker dose reductions is justified. Initial planning for sludge sampling included container, settler and knock-out-pot (KOP) sampling. Due to the significantly higher dose consequence of KOP sludge, a decision was made to sample KOP underwater to achieve worker dose reductions. Additionally, initial plans were to utilize the underwater sampling apparatus for settler sludge. Since there are no longer plans to sample KOP sludge, the decision for underwater sampling for settler sludge needs to be revisited. The present sampling plan calls for spending an estimated $2,500,000 to design and construct a new underwater sampling system (per A21 C-PL-001 RevOE). This evaluation will compare and contrast the present method of above water sampling to the underwater method that is planned by the Sludge Treatment Project (STP) and determine if settler samples can be taken using the existing sampling cart (with potentially minor modifications) while maintaining doses to workers As Low As Reasonably Achievable (ALARA) and eliminate the need for costly redesigns, testing and personnel retraining.

  9. Microwave Blood Thawing: Biochemical Analysis of Small Samples of Thawed Red Blood Cells.

    DTIC Science & Technology

    1984-01-01

    lactate + NAD+ ( Lehninger , 1977) The large increase in pyruvate observed at 6 hours post-wash was most likely due to the large lactate concentrations at...Storage of Blood. London: Academic Press. Lehninger , A.L. 1977. Biochemistry. New York: Worth Publishers, Inc. Lewis, G.P. 1965. Method using o-tolidine

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

    PubMed Central

    Sun, Xiaocun; Flatland, Bente

    2016-01-01

    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. Discussion Canine

  11. Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

    PubMed Central

    Boardman, Anna K.; Campbell, Jennifer; Wirz, Holger; Sharon, Andre; Sauer-Budge, Alexis F.

    2015-01-01

    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 (MSSA)to 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. PMID:25675242

  12. Comparison of manual and automated nucleic acid extraction from whole-blood samples.

    PubMed

    Riemann, Kathrin; Adamzik, Michael; Frauenrath, Stefan; Egensperger, Rupert; Schmid, Kurt W; Brockmeyer, Norbert H; Siffert, Winfried

    2007-01-01

    Nucleic acid extraction and purification from whole blood is a routine application in many laboratories. Automation of this procedure promises standardized sample treatment, a low error rate, and avoidance of contamination. The performance of the BioRobot M48 (Qiagen) and the manual QIAmp DNA Blood Mini Kit (Qiagen) was compared for the extraction of DNA from whole blood. The concentration and purity of the extracted DNAs were determined by spectrophotometry. Analytical sensitivity was assessed by common PCR and genotyping techniques. The quantity and quality of the generated DNAs were slightly higher using the manual extraction method. The results of downstream applications were comparable to each other. Amplification of high-molecular-weight PCR fragments, genotyping by restriction digest, and pyrosequencing were successful for all samples. No cross-contamination could be detected. While automated DNA extraction requires significantly less hands-on time, it is slightly more expensive than the manual extraction method.

  13. Fast multi-spectral imaging technique for detection of circulating endothelial cells in human blood samples

    NASA Astrophysics Data System (ADS)

    Berezhnyy, Ihor V.; Berezhna, Svitlana Y.

    2012-08-01

    The appearance of non-blood cells circulating in human peripheral bloodstream indicates an abnormal condition. One important category of these cells is circulating endothelial cells (CECs) shed by compromised blood vessels. Clinical applications that measure the blood level of CECs are hindered due to a lack of standardized instruments. The major challenge in detecting circulating non-blood cells is their extreme scarcity; 1 in 106 to 107. Described here is a new method for detection of rare cells in blood samples deposited on the adhesive microscopic slides and immunostained with distinct fluorescent markers. The key novelty of the proposed approach is an intelligent search principle and a dual-mode scanner to implement this principle. To begin, a fast scanning that uses a single beam is performed in the spectral channel where only rare cells produce florescence. Once a target cell is registered, the scanner switches on the imaging mode, auto-focuses and then records images in multiple spectral channels at the selected area. The instrument runs in repetitive cycles until the entire slide is scanned. The technology has been validated via detection of human umbilical vein endothelial cells spiked into human blood samples. In addition, the operational principle can be adapted for detection of other types of rare cells in blood.

  14. Uniform sampling table method and its applications: establishment of a uniform sampling method.

    PubMed

    Chen, Yibin; Chen, Jiaxi; Wang, Wei

    2013-01-01

    A novel uniform sampling method is proposed in this paper. According to the requirements of uniform sampling, we propose the properties that must be met by analyzing the distribution of samples. Based on this, the proposed uniform sampling method is demonstrated and evaluated strictly by mathematical means such as inference. The uniform sampling tables with respect to Cn(t2) and Cn(t3) are established. Furthermore, a one-dimension uniform sampling method and a multidimension method are proposed. The proposed novel uniform sampling method, which is guided by uniform design theory, enjoys the advantages of simplified use and good representativeness of the whole sample.

  15. Evaluation of pesticide residues in human blood samples from Punjab (India)

    PubMed Central

    Bedi, Jasbir Singh; Gill, J. P. S.; Kaur, P.; Sharma, A.; Aulakh, R. S.

    2015-01-01

    Aim: The present study was undertaken to estimate the current status of residues of organochlorine pesticides (OCPs), organophosphates (OPs) and synthetic pyrethroids (SPs) pesticides in human blood. Materials and Methods: Human blood samples were analyzed by gas chromatography and confirmed by gas chromatography-mass spectrometry in selective ion monitoring mode. Results: The gas chromatographic analysis of human blood samples collected from Punjab revealed the presence of p,p’-dichlorodiphenyl dichloroethylene (DDE), p,p’ dichlorodiphenyl dichloroethane (DDD), o,p’ DDE and β-endosulfan at mean levels of 15.26, 2.71, 5.62 and 4.02 ng/ml, respectively. p,p’ DDE residue was observed in 18.0% blood samples, and it contributes 55% of the total pesticide burden in human blood. The difference of total dichlorordiphenyl trichloroethane (DDT) between different age groups of humans was found to be statistically significant (p<0.05). The difference of DDT and endosulfan between dietary habits, gender and spraying of pesticides was found statistically non-significant, however endosulfan residues were observed only in pesticide sprayer’s population. Conclusion: Occurrence of p,p’ DDE, p,p’ DDD, o,p’ DDE in human blood indicated restricted use of DDT. However, presence of endosulfan residues in occupationally exposed population is a matter of public health concern. PMID:27046999

  16. Sampling and storage conditions influencing the measurement of parathyroid hormone in blood samples: a systematic review.

    PubMed

    Hanon, Elodie A; Sturgeon, Catharine M; Lamb, Edmund J

    2013-10-01

    Parathyroid hormone (PTH) is relatively unstable: optimisation of pre-analytical conditions, including specimen type, sampling time and storage conditions, is essential. We have undertaken a systematic review of these pre-analytical conditions. An electronic search of the PubMed, Embase, Cochrane, Centre for Research and Dissemination and Bandolier databases was undertaken. Of 5511 papers identified, 96 underwent full text review, of which 83 were finally included. At room temperature PTH was stable in ethylenediaminetetraacetic acid (EDTA) preserved whole blood for at least 24 h and in EDTA plasma for at least 48 h after venepuncture. Losses were observed in clotted blood samples after 3 h and in serum after 2 h. At 4°C PTH was more stable in EDTA plasma (at least 72 h) than serum (at least 24 h). Central venous PTH concentrations were higher than peripheral venous concentrations. In the northern hemisphere, PTH concentrations were higher in winter than summer. PTH has a circadian rhythm characterised by a nocturnal acrophase and mid-morning nadir. Data related to frozen storage of PTH (-20°C and -80°C) were limited and contradictory. We recommend that blood samples for PTH measurement should be taken into tubes containing EDTA, ideally between 10:00 and 16:00, and plasma separated within 24 h of venepuncture. Plasma samples should be stored at 4°C and analysed within 72 h of venepuncture. Particular regard must be paid to the venepuncture site when interpreting PTH concentration. Further research is required to clarify the suitability of freezing samples prior to PTH measurement.

  17. Microfluidic, marker-free isolation of circulating tumor cells from blood samples

    PubMed Central

    Karabacak, Nezihi Murat; Spuhler, Philipp S; Fachin, Fabio; Lim, Eugene J; Pai, Vincent; Ozkumur, Emre; Martel, Joseph M; Kojic, Nikola; Smith, Kyle; Chen, Pin-i; Yang, Jennifer; Hwang, Henry; Morgan, Bailey; Trautwein, Julie; Barber, Thomas A; Stott, Shannon L; Maheswaran, Shyamala; Kapur, Ravi; Haber, Daniel A; Toner, Mehmet

    2014-01-01

    The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen–independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 107 cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2–5 d. PMID:24577360

  18. Microfluidic, marker-free isolation of circulating tumor cells from blood samples.

    PubMed

    Karabacak, Nezihi Murat; Spuhler, Philipp S; Fachin, Fabio; Lim, Eugene J; Pai, Vincent; Ozkumur, Emre; Martel, Joseph M; Kojic, Nikola; Smith, Kyle; Chen, Pin-i; Yang, Jennifer; Hwang, Henry; Morgan, Bailey; Trautwein, Julie; Barber, Thomas A; Stott, Shannon L; Maheswaran, Shyamala; Kapur, Ravi; Haber, Daniel A; Toner, Mehmet

    2014-03-01

    The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-iChip technology. The CTC-iChip uses deterministic lateral displacement, inertial focusing and magnetophoresis to sort up to 10⁷ cells/s. By using two-stage magnetophoresis and depletion antibodies against leukocytes, we achieve 3.8-log depletion of white blood cells and a 97% yield of rare cells with a sample processing rate of 8 ml of whole blood/h. The CTC-iChip is compatible with standard cytopathological and RNA-based characterization methods. This protocol describes device production, assembly, blood sample preparation, system setup and the CTC isolation process. Sorting 8 ml of blood sample requires 2 h including setup time, and chip production requires 2-5 d.

  19. A LITERATURE REVIEW OF WIPE SAMPLING METHODS ...

    EPA Pesticide Factsheets

    Wipe sampling is an important technique for the estimation of contaminant deposition in buildings, homes, or outdoor surfaces as a source of possible human exposure. Numerousmethods of wipe sampling exist, and each method has its own specification for the type of wipe, wetting solvent, and determinative step to be used, depending upon the contaminant of concern. The objective of this report is to concisely summarize the findings of a literature review that was conducted to identify the state-of-the-art wipe sampling techniques for a target list of compounds. This report describes the methods used to perform the literature review; a brief review of wipe sampling techniques in general; an analysis of physical and chemical properties of each target analyte; an analysis of wipe sampling techniques for the target analyte list; and asummary of the wipe sampling techniques for the target analyte list, including existing data gaps. In general, no overwhelming consensus can be drawn from the current literature on how to collect a wipe sample for the chemical warfare agents, organophosphate pesticides, and other toxic industrial chemicals of interest to this study. Different methods, media, and wetting solvents have been recommended and used by various groups and different studies. For many of the compounds of interest, no specific wipe sampling methodology has been established for their collection. Before a wipe sampling method (or methods) can be established for the co

  20. Spectral feature characterization methods for blood stain detection in crime scene backgrounds

    NASA Astrophysics Data System (ADS)

    Yang, Jie; Mathew, Jobin J.; Dube, Roger R.; Messinger, David W.

    2016-05-01

    Blood stains are one of the most important types of evidence for forensic investigation. They contain valuable DNA information, and the pattern of the stains can suggest specifics about the nature of the violence that transpired at the scene. Blood spectral signatures containing unique reflectance or absorption features are important both for forensic on-site investigation and laboratory testing. They can be used for target detection and identification applied to crime scene hyperspectral imagery, and also be utilized to analyze the spectral variation of blood on various backgrounds. Non-blood stains often mislead the detection and can generate false alarms at a real crime scene, especially for dark and red backgrounds. This paper measured the reflectance of liquid blood and 9 kinds of non-blood samples in the range of 350 nm - 2500 nm in various crime scene backgrounds, such as pure samples contained in petri dish with various thicknesses, mixed samples with different colors and materials of fabrics, and mixed samples with wood, all of which are examined to provide sub-visual evidence for detecting and recognizing blood from non-blood samples in a realistic crime scene. The spectral difference between blood and non-blood samples are examined and spectral features such as "peaks" and "depths" of reflectance are selected. Two blood stain detection methods are proposed in this paper. The first method uses index to denote the ratio of "depth" minus "peak" over"depth" add"peak" within a wavelength range of the reflectance spectrum. The second method uses relative band depth of the selected wavelength ranges of the reflectance spectrum. Results show that the index method is able to discriminate blood from non-blood samples in most tested crime scene backgrounds, but is not able to detect it from black felt. Whereas the relative band depth method is able to discriminate blood from non-blood samples on all of the tested background material types and colors.

  1. Venepuncture is preferable to heel lance for blood sampling in term neonates

    PubMed Central

    Ogawa, S; Ogihara, T; Fujiwara, E; Ito, K; Nakano, M; Nakayama, S; Hachiya, T; Fujimoto, N; Abe, H; Ban, S; Ikeda, E; Tamai, H

    2005-01-01

    Background: The analgesic effect of oral sucrose in newborn infants undergoing painful procedures is generally accepted. For blood sampling, some studies have shown that venepuncture (VP) is less painful than heel lance (HL). Objective: To determine the least painful and most effective method among blood sampling by VP or HL with or without sucrose. Design: Randomised, double blind, placebo controlled trial. Subjects: A total of 100 healthy, full term newborn infants being screened for inborn errors of metabolism were randomly allocated to one of four experimental groups (25 infants in each). Intervention and outcome measure: Seven specially trained nurses took turns to carry out blood sampling two minutes after administration of oral sucrose or water. Neonatal pain was assessed by the neonatal facial coding system (NFCS), as well as by crying. Results: Without sucrose, the NFCS score was higher in the HL group than the VP group during blood sampling (median 58 v 23, p<0.001). Oral sucrose significantly reduced the score of the HL group (58 v 47, p<0.01) and also tended to reduce the score of the VP group (23 v 2, p<0.1). However, the HL with sucrose group still had a higher score than the VP without sucrose group (47 v 23, p<0.01). Crying and the total procedure time showed the same trends as the NFCS score. Conclusions: VP is less painful and more effective than HL for blood sampling in newborn infants. Although oral sucrose may have an additive analgesic effect, it is not necessarily required if VP is used for blood sampling. PMID:15871991

  2. Dynamic Method for Identifying Collected Sample Mass

    NASA Technical Reports Server (NTRS)

    Carson, John

    2008-01-01

    G-Sample is designed for sample collection missions to identify the presence and quantity of sample material gathered by spacecraft equipped with end effectors. The software method uses a maximum-likelihood estimator to identify the collected sample's mass based on onboard force-sensor measurements, thruster firings, and a dynamics model of the spacecraft. This makes sample mass identification a computation rather than a process requiring additional hardware. Simulation examples of G-Sample are provided for spacecraft model configurations with a sample collection device mounted on the end of an extended boom. In the absence of thrust knowledge errors, the results indicate that G-Sample can identify the amount of collected sample mass to within 10 grams (with 95-percent confidence) by using a force sensor with a noise and quantization floor of 50 micrometers. These results hold even in the presence of realistic parametric uncertainty in actual spacecraft inertia, center-of-mass offset, and first flexibility modes. Thrust profile knowledge is shown to be a dominant sensitivity for G-Sample, entering in a nearly one-to-one relationship with the final mass estimation error. This means thrust profiles should be well characterized with onboard accelerometers prior to sample collection. An overall sample-mass estimation error budget has been developed to approximate the effect of model uncertainty, sensor noise, data rate, and thrust profile error on the expected estimate of collected sample mass.

  3. Direct RNA-based detection of CTX-M β-lactamases in human blood samples.

    PubMed

    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.

  4. Innovative methods for inorganic sample preparation

    SciTech Connect

    Essling, A.M.; Huff, E.A.; Graczyk, D.G.

    1992-04-01

    Procedures and guidelines are given for the dissolution of a variety of selected materials using fusion, microwave, and Parr bomb techniques. These materials include germanium glass, corium-concrete mixtures, and zeolites. Emphasis is placed on sample-preparation approaches that produce a single master solution suitable for complete multielement characterization of the sample. In addition, data are presented on the soil microwave digestion method approved by the Environmental Protection Agency (EPA). Advantages and disadvantages of each sample-preparation technique are summarized.

  5. Application of the Triage panel for drugs of abuse to forensic blood samples.

    PubMed

    Moriya, F; Hashimoto, Y

    1996-04-01

    A simple and rapid screening procedure with Triage has been developed to detect 7 classes of drugs of abuse, phencyclidine (PCP), benzodiazepines (BZO), cocaine metabolite (COC), amphetamines (AMP), cannabinoids (THC), opiates (OPI), and barbiturates (BAR), in hemolyzed blood. A clear supernatant was obtained by mixing the blood with sulfosalicylic acid. The supernatant was neutralized with ammonium acetate and then screened using Triage. The lower limits of detection of the Triage screening method for PCP, diazepam, benzolyecgonine, methamphetamine, morphine, 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid (THC-COOH), phenobarbital, and secobarbital were 50 ng/mL, 900 ng/mL, 1,000 ng/mL, 600 ng/mL, 900 ng/mL, and 900 ng/mL, respectively. The sensitivity of Triage for THC-COOH in deproteinized blood samples was much lower than that in urine samples. No false positive reactions were observed for the 6 classes of the drugs of abuse with the exception of AMP when the blood was decomposed. Phenethylamine, a putrefactive amine, gave positive results for AMP at concentrations over 5,000 ng/mL. The method was applied to 9 hemolyzed blood samples and 3 turbid urine samples from 12 forensic autopsy cases suspected of drug misuse. Among these, 5 were positive for AMP, 1 for OPI, and 4 for BAR. The presence of methamphetamine is only one of the 5, codeine in 1, and phenobarbital in 4 was confirmed by gas chromatography. All 4 samples which were false positive for AMP contained phenethylamine at relatively high concentrations because of moderate to heavy putrefaction. This method, although disadvantageous to test for AMP and THC, is helpful for the forensic toxicologist because any kind of bloody fluid can be tested rapidly with Triage to detect toxic levels of PCP, BZO, COC, OPI, and BAR.

  6. Method and apparatus for sampling atmospheric mercury

    DOEpatents

    Trujillo, Patricio E.; Campbell, Evan E.; Eutsler, Bernard C.

    1976-01-20

    A method of simultaneously sampling particulate mercury, organic mercurial vapors, and metallic mercury vapor in the working and occupational environment and determining the amount of mercury derived from each such source in the sampled air. A known volume of air is passed through a sampling tube containing a filter for particulate mercury collection, a first adsorber for the selective adsorption of organic mercurial vapors, and a second adsorber for the adsorption of metallic mercury vapor. Carbon black molecular sieves are particularly useful as the selective adsorber for organic mercurial vapors. The amount of mercury adsorbed or collected in each section of the sampling tube is readily quantitatively determined by flameless atomic absorption spectrophotometry.

  7. The rank product method with two samples.

    PubMed

    Koziol, James A

    2010-11-05

    Breitling et al. (2004) introduced a statistical technique, the rank product method, for detecting differentially regulated genes in replicated microarray experiments. The technique has achieved widespread acceptance and is now used more broadly, in such diverse fields as RNAi analysis, proteomics, and machine learning. In this note, we extend the rank product method to the two sample setting, provide distribution theory attending the rank product method in this setting, and give numerical details for implementing the method.

  8. Blood storage device and method for oxygen removal

    DOEpatents

    Bitensky, Mark W.; Yoshida, Tatsuro

    2000-01-01

    The present invention relates to a storage device and method for the long-term storage of blood and, more particularly, to a blood storage device and method capable of removing oxygen from the stored blood and thereby prolonging the storage life of the deoxygenated blood.

  9. Method development for detecting the novel cyanide antidote dimethyl trisulfide from blood and brain, and its interaction with blood.

    PubMed

    Kiss, Lóránd; Holmes, Secondra; Chou, Ching-En; Dong, Xinmei; Ross, James; Brown, Denise; Mendenhall, Brooke; Coronado, Valerie; De Silva, Deepthika; Rockwood, Gary A; Petrikovics, Ilona; Thompson, David E

    2017-02-15

    The antidotal potency of dimethyl trisulfide (DMTS) against cyanide poisoning was discovered and investigated in our previous studies. Based on our results it has better efficacy than the Cyanokit and the Nithiodote therapies that are presently used against cyanide intoxication in the US. Because of their absence in the literature, the goal of this work was to develop analytical methods for determining DMTS from blood and brain that could be employed in future pharmacokinetic studies. An HPLC-UV method for detection of DMTS from blood, a GC-MS method for detection of DMTS from brain, and associated validation experiments are described here. These analytical methods were developed using in vitro spiking of brain and blood, and are suitable for determining the in vivo DMTS concentrations in blood and brain in future pharmacokinetic and distribution studies. An important phenomenon was observed in the process of developing these methods. Specifically, recoveries from fresh blood spiked with DMTS were found to be significantly lower than recoveries from aged blood spiked in the same manner with DMTS. This decreased DMTS recovery from fresh blood is important, both because of the role it may play in the antidotal action of DMTS in the presence of cyanide, and because it adds the requirement of sample stabilization to the method development process. Mitigation procedures for stabilizing DMTS samples in blood are reported.

  10. Determination of ketorolac in blood and plasma samples by high-performance liquid chromatography.

    PubMed

    Flores-Murrieta, F J; Granados-Soto, V; Hong, E

    1994-10-01

    A new method for determination of ketorolac in blood or plasma samples by reversed-phase high-performance liquid chromatography has been developed. The method includes a double extraction with diethyl ether and detection by absorbance at 313 nm. Quantitation was performed by height ratios of ketorolac and the internal standard (sodium tolmetin). Detection limit of the method was 3 ng/ml using 1 ml of plasma and 10 ng/ml using 0.2 ml of blood. The method is linear in the range of concentrations typically obtained after therapeutic doses of the drug, has the advantages of using low volume of body fluid and the internal standard used is commercially available. Those characteristics allow us to conclude that this method is suitable for pharmacokinetic or drug monitoring studies.

  11. Screening for lead poisoning in urban school children of southern India using capillary and venous blood samples.

    PubMed

    D'Souza, Herman S; Menezes, Geraldine; Venkatesh, T

    2002-01-01

    Our study aimed at comparing lead and zinc protoporphyrin (ZPP) levels in capillary and venous blood samples in a small population and to employ an easier method of sample collection for a major screening program in school children in major Indian cities. An awareness program on lead and its effects was conducted in two different schools. A total of thirty urban school children from South India, with an age group between 4-12 years consented for dual blood sampling and reported for the study. Venous and capillary blood samples were obtained simultaneously. Blood lead and zinc protoporphyrin (ZPP) levels were estimated using ESA Lead Analyzer and Haematofluorometer respectively. A significant correlation between capillary and venous ZPP (r=0.98) and lead (r=0.99) was observed. Rank sum test showed that there is no statistically significant difference between capillary and venous ZPP (P=0.891) and lead (P=0.672) values. This pilot study recommends that screening for lead may be done using capillary blood samples since significant correlation is observed between capillary and venous blood measurements. Obtaining samples using this mode is a non-invasive, less expensive, quick and easy method in children. Appropriately performed capillary sampling may be considered as an acceptable alternative to venipuncture for screening of blood for lead poisoning both in children and adults.

  12. The Use of Dried Blood Spot Sampling in the National Social Life, Health, and Aging Project

    PubMed Central

    McDade, Thomas W.

    2009-01-01

    Objectives This paper describes the methods used for and issues associated with collection and analysis of dried blood spot (DBS) samples for the National Social Life, Health, and Aging Project and provides the basic distributions of the resulting analytes. Methods DBSs from capillary finger sticks were collected by nonmedically trained interviewers from 2,044 individuals, aged 57–85 years. The quality and quantity of DBS samples were evaluated to allow for analysis of interviewer performance. Levels of C-reactive protein, antibodies to the Epstein–Barr virus, hemoglobin, and glycosylated hemoglobin were assayed using various analytic methods. Results Cooperation rate for DBS collection was 84.5%, with 99% of the cards yielding enough sample for at least one analysis. The distribution, mean, and standard deviation of the analytes obtained from DBSs are also presented in this paper. Conclusions The high cooperation rate and quality of the spots collected suggest that the collection of DBSs in population-based research is a feasible and viable alternative to venous blood draws. The relative ease of sample collection, transport, and storage are significant benefits. Care should be taken, however, when comparing results from analysis of DBS samples with those obtained from serum or plasma samples. PMID:19244547

  13. Sample preparation methods for determination of drugs of abuse in hair samples: A review.

    PubMed

    Vogliardi, Susanna; Tucci, Marianna; Stocchero, Giulia; Ferrara, Santo Davide; Favretto, Donata

    2015-02-01

    Hair analysis has assumed increasing importance in the determination of substances of abuse, both in clinical and forensic toxicology investigations. Hair analysis offers particular advantages over other biological matrices (blood and urine), including a larger window of detection, ease of collection and sample stability. In the present work, an overview of sample preparation techniques for the determination of substances of abuse in hair is provided, specifically regarding the principal steps in hair sample treatment-decontamination, extraction and purification. For this purpose, a survey of publications found in the MEDLINE database from 2000 to date was conducted. The most widely consumed substances of abuse and psychotropic drugs were considered. Trends in simplification of hair sample preparation, washing procedures and cleanup methods are discussed. Alternative sample extraction techniques, such as head-space solid phase microextraction (HS-SPDE), supercritical fluid extraction (SFE) and molecularly imprinted polymers (MIP) are also reported.

  14. Differences between blood donors and a population sample: implications for case–control studies

    PubMed Central

    Golding, Jean; Northstone, Kate; Miller, Laura L; Davey Smith, George; Pembrey, Marcus

    2013-01-01

    Background Selecting appropriate controls for studies of genetic variation in case series is important. The two major candidates involve the use of blood donors or a random sample of the population. Methods We compare and contrast the two different populations of controls for studies of genetic variation using data from parents enrolled in the Avon Longitudinal Study of Parents and Children (ALSPAC). In addition we compute different biases using a series of hypothetical assumptions. Results The study subjects who had been blood donors differed markedly from the general population in social, health-related, anthropometric, and personality-related variables. Using theoretical examples, we show that blood donors are a poor control group for non-genetic studies of diseases related to environmentally, behaviourally, or socially patterned exposures. However, we show that if blood donors are used as controls in genetic studies, these factors are unlikely to make a major difference in detecting true associations with relatively rare disorders (cumulative incidence through life of <10%). Nevertheless, for more common disorders, the reduction in accuracy resulting from the inclusion in any control population of individuals who have or will develop the disease in question can create a greater bias than can socially patterned factors. Conclusions Information about the medical history of a control and the parents of the control (as a proxy for whether the control will develop the disease) is more important with regard to the choice of controls than whether the controls are a random population sample or blood donors. PMID:23825379

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

    PubMed Central

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

    2017-01-01

    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 the laboratory protocol. Results 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 (<2+ reaction grade), extra 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. Conclusions 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. PMID:28028997

  16. Fluidics platform and method for sample preparation and analysis

    SciTech Connect

    Benner, W. Henry; Dzenitis, John M.; Bennet, William J.; Baker, Brian R.

    2014-08-19

    Herein provided are fluidics platform and method for sample preparation and analysis. The fluidics platform is capable of analyzing DNA from blood samples using amplification assays such as polymerase-chain-reaction assays and loop-mediated-isothermal-amplification assays. The fluidics platform can also be used for other types of assays and analyzes. In some embodiments, a sample in a sealed tube can be inserted directly. The following isolation, detection, and analyzes can be performed without a user's intervention. The disclosed platform may also comprises a sample preparation system with a magnetic actuator, a heater, and an air-drying mechanism, and fluid manipulation processes for extraction, washing, elution, assay assembly, assay detection, and cleaning after reactions and between samples.

  17. Neutron activation analysis of certified samples by the absolute method

    NASA Astrophysics Data System (ADS)

    Kadem, F.; Belouadah, N.; Idiri, Z.

    2015-07-01

    The nuclear reactions analysis technique is mainly based on the relative method or the use of activation cross sections. In order to validate nuclear data for the calculated cross section evaluated from systematic studies, we used the neutron activation analysis technique (NAA) to determine the various constituent concentrations of certified samples for animal blood, milk and hay. In this analysis, the absolute method is used. The neutron activation technique involves irradiating the sample and subsequently performing a measurement of the activity of the sample. The fundamental equation of the activation connects several physical parameters including the cross section that is essential for the quantitative determination of the different elements composing the sample without resorting to the use of standard sample. Called the absolute method, it allows a measurement as accurate as the relative method. The results obtained by the absolute method showed that the values are as precise as the relative method requiring the use of standard sample for each element to be quantified.

  18. Diagnosis of Carrion’s Disease by Direct Blood PCR in Thin Blood Smear Negative Samples

    PubMed Central

    Tinco Valdez, Carmen; Pons, Maria J.; del Valle, Luis J.; Oré, Verónica Casabona; Michelena, Denisse Champin; Mayra, Jorge Bazán; Gavidea, Víctor Zavaleta; Vargas, Martha; Ruiz, Joaquim

    2014-01-01

    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. PMID:24651298

  19. A comparative evaluation of four DNA extraction protocols from whole blood sample.

    PubMed

    Ghaheri, M; Kahrizi, D; Yari, K; Babaie, A; Suthar, R S; Kazemi, E

    2016-03-31

    All organisms have Deoxyribonucleic acid (DNA) within their cells. DNA is a complex molecule that contains all of the information necessary to build and maintain an organism. DNA extraction is one of the most basic and essential techniques in the study of DNA that allow huge advances in molecular biology, biotechnology and bioinformatics laboratories. Whole blood samples are one of the main sources used to obtain DNA and there are many different protocols available in this issue. In current research, compared four DNA extraction protocols from blood samples; include modified phenol-chloroform protocol, two salting-out and enzyme free method and from commercial kit. The extracted DNAs by these protocols were analyzed according to their time demands, quality and quantity, toxicity and functionality in PCR method. Also the quality and quantity of the extracted DNA were surveyed by gel electrophoresis and Nanodrop spectrophotometry methods. It was observed that there are not significantly differences between these methods about DNA Purity (A260/A280), but the DNA yield (ng DNA/μl) of phenol/chloroform method was higher than other methods. In addition, phenol/chloroform was the most toxic method and it takes more time than other methods. Roche diagnostics GmbH kit was the most expensive among the four methods but the least extraction time was required and it was the safest method.

  20. Raman Spectroscopy: A New Proposal for the Detection of Leukemia Using Blood Samples

    SciTech Connect

    Martinez-Espinosa, J. C.; Gonzalez-Solis, J. L.; Miranda-Beltran, M. L.; Soria-Fregoso, C.; Medina-Valtierra, J.; Sanchez-Gomez, R.

    2008-08-11

    The use of Raman spectroscopy to analyze blood biochemistry and hence distinguish between normal and abnormal blood was investigated. The blood samples were obtained from 6 patients who were clinically diagnosed with leukemia and 6 healthy volunteer. The imprint was put under the microscope and several points were chosen for Raman measurement. All spectra were collected at confocal Raman micro-spectroscopy (Renishaw) with NIR 830 nm laser. It is shown that the serum samples from patients with leukemia and from the control group can be discriminated when the multivariate statistical methods of principal component analysis (PCA) and linear discriminated analysis (LDA) is applied to their Raman spectra. The ratios of some band intensities were analyzed and some band ratios were significant and corresponded to proteins, phospholipids, and polysaccharides. In addition, currently the degree of damage to the bone marrow is estimated through biopsies and therefore it is a very procedure painful. The preliminary results suggest that Raman spectroscopy could be a new technique to study the bone marrow using just blood samples.

  1. Quantitative analysis of trace chromium in blood samples. Combination of the advanced oxidation process with catalytic adsorptive stripping voltammetry.

    PubMed

    Yong, Li; Armstrong, Kristie C; Dansby-Sparks, Royce N; Carrington, Nathan A; Chambers, James Q; Xue, Zi-Ling

    2006-11-01

    A new method for pretreating blood samples for trace Cr analysis is described. The advanced oxidation process (AOP with H2O2 and 5.5-W UV irradiation for 60 min) is used to remove biological/organic species for subsequent analysis. Prior to the AOP pretreatment, acid (HNO3) is used at pH 3.0 to inhibit the enzyme catalase in the blood samples. Catalytic adsorptive stripping voltammetry at a bismuth film electrode gives a Cr concentration of 6.0 +/- 0.3 ppb in the blood samples. This concentration was confirmed by dry-ashing the blood samples and subsequent analysis by atomic absorption spectroscopy. This current method may be used to monitor chromium, a trace metal in humans, and the efficacy and safety of chromium supplements as adjuvant therapy for diabetes.

  2. Quantification of rifapentine, a potent antituberculosis drug, from dried blood spot samples using liquid chromatographic-tandem mass spectrometric analysis.

    PubMed

    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.

  3. Quantification of Rifapentine, a Potent Antituberculosis Drug, from Dried Blood Spot Samples Using Liquid Chromatographic-Tandem Mass Spectrometric Analysis

    PubMed Central

    Parsons, Teresa L.; Marzinke, Mark A.; Hoang, Thuy; Bliven-Sizemore, Erin; Weiner, Marc; Mac Kenzie, William R.; Dorman, Susan E.

    2014-01-01

    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. PMID:25182637

  4. Comparisons of polybrominated diphenyl ethers levels in paired South Korean cord blood, maternal blood, and breast milk samples.

    PubMed

    Kim, Tae Hyung; Bang, Du Yeon; Lim, Hyun Jung; Won, A Jin; Ahn, Mee Young; Patra, Nabanita; Chung, Ki Kyung; Kwack, Seung Jun; Park, Kui Lea; Han, Soon Young; Choi, Wahn Soo; Han, Jung Yeol; Lee, Byung Mu; Oh, Jeong-Eun; Yoon, Jeong-Hyun; Lee, Jaewon; Kim, Hyung Sik

    2012-03-01

    Polybrominated diphenyl ethers (PBDEs), commonly used flame retardants, have been reported as potential endocrine disruptor and neurodevelopmental toxicants, thus giving rise to the public health concern. The goal of this study was to investigate the relationship between umbilical cord blood, maternal blood, and breast milk concentrations of PBDEs in South Korean. We assessed PBDE levels in paired samples of umbilical cord blood, maternal blood, and breast milk. The levels of seven PBDE congeners were measured in 21 paired samples collected from the Cheil Woman's Hospital (Seoul, Korea) in 2008. We also measured thyroid hormones levels in maternal and cord blood to assess the association between PBDEs exposure and thyroid hormone levels. However, there was no correlation between serum thyroxin (T4) and total PBDEs concentrations. The total PBDEs concentrations in the umbilical cord blood, maternal blood, and breast milk were 10.7±5.1 ng g(-1) lipid, 7.7±4.2 ng g(-1) lipid, and 3.0±1.8 ng g(-1) lipid, respectively. The ranges of total PBDE concentrations observed were 2.28-30.94 ng g(-1) lipid in umbilical cord blood, 1.8-17.66 ng g(-1) lipid in maternal blood, and 1.08-8.66 ng g(-1) lipid in breast milk. BDE-47 (45-73% of total PBDEs) was observed to be present dominantly in all samples, followed by BDE-153. A strong correlation was found for major BDE-congeners between breast milk and cord blood or maternal blood and cord blood samples. The measurement of PBDEs concentrations in maternal blood or breast milk may help to determine the concentration of PBDEs in infant.

  5. A new solid phase microextraction method using organic ligand in micropipette tip syringe system packed with modified carbon cloth for preconcentration of cadmium in drinking water and blood samples of kidney failure patients.

    PubMed

    Panhwar, Abdul Haleem; Kazi, Tasneem Gul; Afridi, Hassan Imran; Arain, Salma Aslam; Naeemullah; Brahman, Kapil Dev; Arain, Mariam Shahzadi

    2015-03-05

    A simple and efficient miniaturized solid phase microextraction (M-SPμE) in a syringe system was developed for preconcentration of cadmium (Cd) in environmental and biological samples, followed by flame atomic absorption technique. The syringe system contains the micropipette tip packed with activated carbon cloth, coated with modified magnetic nanoparticles of iron oxide Triton X114 (ACC-NPs). Scanning electron microscopy and energy dispersive spectroscopy used for characterization of the size, morphology and elemental composition of ACC-NPs. The sample solution treated with a complexing reagent 8-hydroxyqunilone (8-HQ), and drawn into the syringe, filled with ACC-MNPs and dispensed manually for 2-10 drawing/discharging cycles. The analyte retained on ACC-NPs in micropipette tip-syringe system were then eluted with different volume of 1.5molL(-1) HCl by 1-5 drawing/discharging cycles. The syringe system directly couple with FAAS for analysis. The influence of different variables on the extraction efficiency of Cd, including adsorbent dosage, pH, sample volume, eluent volume and drawing/discharging cycles of syringe system were optimized. At optimized extraction conditions, the method showed good linearity in the range of 5-250μgL(-1), with a limit of detection 0.15μgL(-1). Repeatability of the extraction (%RSD) was <5%, n=5. The validity and accuracy of the method was checked by the certified reference materials. The proposed method was successfully applied for the determination of Cd in different drinking water and biological samples of kidney failure patients and healthy controls.

  6. A new solid phase microextraction method using organic ligand in micropipette tip syringe system packed with modified carbon cloth for preconcentration of cadmium in drinking water and blood samples of kidney failure patients

    NASA Astrophysics Data System (ADS)

    Panhwar, Abdul Haleem; Kazi, Tasneem Gul; Afridi, Hassan Imran; Arain, Salma Aslam; Naeemullah; Brahman, Kapil Dev; Arain, Mariam Shahzadi

    2015-03-01

    A simple and efficient miniaturized solid phase microextraction (M-SPμE) in a syringe system was developed for preconcentration of cadmium (Cd) in environmental and biological samples, followed by flame atomic absorption technique. The syringe system contains the micropipette tip packed with activated carbon cloth, coated with modified magnetic nanoparticles of iron oxide Triton X114 (ACC-NPs). Scanning electron microscopy and energy dispersive spectroscopy used for characterization of the size, morphology and elemental composition of ACC-NPs. The sample solution treated with a complexing reagent 8-hydroxyqunilone (8-HQ), and drawn into the syringe, filled with ACC-MNPs and dispensed manually for 2-10 drawing/discharging cycles. The analyte retained on ACC-NPs in micropipette tip-syringe system were then eluted with different volume of 1.5 mol L-1 HCl by 1-5 drawing/discharging cycles. The syringe system directly couple with FAAS for analysis. The influence of different variables on the extraction efficiency of Cd, including adsorbent dosage, pH, sample volume, eluent volume and drawing/discharging cycles of syringe system were optimized. At optimized extraction conditions, the method showed good linearity in the range of 5-250 μg L-1, with a limit of detection 0.15 μg L-1. Repeatability of the extraction (%RSD) was <5%, n = 5. The validity and accuracy of the method was checked by the certified reference materials. The proposed method was successfully applied for the determination of Cd in different drinking water and biological samples of kidney failure patients and healthy controls.

  7. Direct identification of bacteria in blood culture samples using an electronic nose.

    PubMed

    Trincavelli, Marco; Coradeschi, Silvia; Loutfi, Amy; Söderquist, Bo; Thunberg, Per

    2010-12-01

    In this paper, we introduce a method for identification of bacteria in human blood culture samples using an electronic nose. The method uses features, which capture the static (steady state) and dynamic (transient) properties of the signal from the gas sensor array and proposes a means to ensemble results from consecutive samples. The underlying mechanism for ensembling is based on an estimation of posterior probability, which is extracted from a support vector machine classifier. A large dataset representing ten different bacteria cultures has been used to validate the presented methods. The results detail the performance of the proposed algorithm and show that through ensembling decisions on consecutive samples, significant reliability in classification accuracy can be achieved.

  8. Bridging the Gap between Sample Collection and Laboratory Analysis: Using Dried Blood Spots to Identify Human Exposure to Chemical Agents.

    PubMed

    Hamelin, Elizabeth I; Blake, Thomas A; Perez, Jonas W; Crow, Brian S; Shaner, Rebecca L; Coleman, Rebecca M; Johnson, Rudolph C

    2016-05-13

    Public health response to large scale chemical emergencies presents logistical challenges for sample collection, transport, and analysis. Diagnostic methods used to identify and determine exposure to chemical warfare agents, toxins, and poisons traditionally involve blood collection by phlebotomists, cold transport of biomedical samples, and costly sample preparation techniques. Use of dried blood spots, which consist of dried blood on an FDA-approved substrate, can increase analyte stability, decrease infection hazard for those handling samples, greatly reduce the cost of shipping/storing samples by removing the need for refrigeration and cold chain transportation, and be self-prepared by potentially exposed individuals using a simple finger prick and blood spot compatible paper. Our laboratory has developed clinical assays to detect human exposures to nerve agents through the analysis of specific protein adducts and metabolites, for which a simple extraction from a dried blood spot is sufficient for removing matrix interferents and attaining sensitivities on par with traditional sampling methods. The use of dried blood spots can bridge the gap between the laboratory and the field allowing for large scale sample collection with minimal impact on hospital resources while maintaining sensitivity, specificity, traceability, and quality requirements for both clinical and forensic applications.

  9. Bridging the gap between sample collection and laboratory analysis: using dried blood spots to identify human exposure to chemical agents

    NASA Astrophysics Data System (ADS)

    Hamelin, Elizabeth I.; Blake, Thomas A.; Perez, Jonas W.; Crow, Brian S.; Shaner, Rebecca L.; Coleman, Rebecca M.; Johnson, Rudolph C.

    2016-05-01

    Public health response to large scale chemical emergencies presents logistical challenges for sample collection, transport, and analysis. Diagnostic methods used to identify and determine exposure to chemical warfare agents, toxins, and poisons traditionally involve blood collection by phlebotomists, cold transport of biomedical samples, and costly sample preparation techniques. Use of dried blood spots, which consist of dried blood on an FDA-approved substrate, can increase analyte stability, decrease infection hazard for those handling samples, greatly reduce the cost of shipping/storing samples by removing the need for refrigeration and cold chain transportation, and be self-prepared by potentially exposed individuals using a simple finger prick and blood spot compatible paper. Our laboratory has developed clinical assays to detect human exposures to nerve agents through the analysis of specific protein adducts and metabolites, for which a simple extraction from a dried blood spot is sufficient for removing matrix interferents and attaining sensitivities on par with traditional sampling methods. The use of dried blood spots can bridge the gap between the laboratory and the field allowing for large scale sample collection with minimal impact on hospital resources while maintaining sensitivity, specificity, traceability, and quality requirements for both clinical and forensic applications.

  10. Cost Evaluation of Dried Blood Spot Home Sampling as Compared to Conventional Sampling for Therapeutic Drug Monitoring in Children

    PubMed Central

    Martial, Lisa C.; Aarnoutse, Rob E.; Schreuder, Michiel F.; Henriet, Stefanie S.; Brüggemann, Roger J. M.; Joore, Manuela A.

    2016-01-01

    Dried blood spot (DBS) sampling for the purpose of therapeutic drug monitoring can be an attractive alternative for conventional blood sampling, especially in children. This study aimed to compare all costs involved in conventional sampling versus DBS home sampling in two pediatric populations: renal transplant patients and hemato-oncology patients. Total costs were computed from a societal perspective by adding up healthcare cost, patient related costs and costs related to loss of productivity of the caregiver. Switching to DBS home sampling was associated with a cost reduction of 43% for hemato-oncology patients (€277 to €158) and 61% for nephrology patients (€259 to €102) from a societal perspective (total costs) per blood draw. From a healthcare perspective, costs reduced with 7% for hemato-oncology patients and with 21% for nephrology patients. Total savings depend on the number of hospital visits that can be avoided by using home sampling instead of conventional sampling. PMID:27941974

  11. Cost Evaluation of Dried Blood Spot Home Sampling as Compared to Conventional Sampling for Therapeutic Drug Monitoring in Children.

    PubMed

    Martial, Lisa C; Aarnoutse, Rob E; Schreuder, Michiel F; Henriet, Stefanie S; Brüggemann, Roger J M; Joore, Manuela A

    2016-01-01

    Dried blood spot (DBS) sampling for the purpose of therapeutic drug monitoring can be an attractive alternative for conventional blood sampling, especially in children. This study aimed to compare all costs involved in conventional sampling versus DBS home sampling in two pediatric populations: renal transplant patients and hemato-oncology patients. Total costs were computed from a societal perspective by adding up healthcare cost, patient related costs and costs related to loss of productivity of the caregiver. Switching to DBS home sampling was associated with a cost reduction of 43% for hemato-oncology patients (€277 to €158) and 61% for nephrology patients (€259 to €102) from a societal perspective (total costs) per blood draw. From a healthcare perspective, costs reduced with 7% for hemato-oncology patients and with 21% for nephrology patients. Total savings depend on the number of hospital visits that can be avoided by using home sampling instead of conventional sampling.

  12. Actinide recovery method -- Large soil samples

    SciTech Connect

    Maxwell , S.L. III

    2000-04-25

    There is a need to measure actinides in environmental samples with lower and lower detection limits, requiring larger sample sizes. This analysis is adversely affected by sample-matrix interferences, which make analyzing soil samples above five-grams very difficult. A new Actinide-Recovery Method has been developed by the Savannah River Site Central Laboratory to preconcentrate actinides from large-soil samples. Diphonix Resin (Eichrom Industries), a 1994 R and D 100 winner, is used to preconcentrate the actinides from large soil samples, which are bound powerfully to the resin's diphosphonic acid groups. A rapid microwave-digestion technique is used to remove the actinides from the Diphonix Resin, which effectively eliminates interfering matrix components from the soil matrix. The microwave-digestion technique is more effective and less tedious than catalyzed hydrogen peroxide digestions of the resin or digestion of diphosphonic stripping agents such as HEDPA. After resin digestion, the actinides are recovered in a small volume of nitric acid which can be loaded onto small extraction chromatography columns, such as TEVA Resin, U-TEVA Resin or TRU Resin (Eichrom Industries). Small, selective extraction columns do not generate large volumes of liquid waste and provide consistent tracer recoveries after soil matrix elimination.

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

    PubMed

    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.

  14. Microcontroller-based system for collecting anaerobic blood samples from a running greyhound.

    PubMed

    Schmalzried, R T; Toll, P W; Devore, J J; Fedde, M R

    1992-04-01

    Many physiological variables change rapidly in the blood during sprint exercise. To characterize the dynamics and extent of these changes, blood samples must be obtained during exercise. We describe herein a portable, microcontroller-based system used to automatically obtain repeated, anaerobic, arterial blood samples from greyhounds before, during, and following a race on a track. In addition, the system also records the blood temperature in the pulmonary artery each time a blood sample is taken. The system has been tested for more than 2 years and has proven to be reliable and effective.

  15. A micro blood sampling system for catheterized neonates and pediatrics in intensive care unit.

    PubMed

    Jung, Wooseok; Ahn, Chong H

    2013-04-01

    A new micro blood sampling system has been designed, fabricated, and characterized to reduce iatrogenic blood loss from the catheterized neonates and pediatrics in intensive care unit by providing micro-volume of blood to analytical biomedical microdevices which can do point-of-care testing for their critical care. The system can not only save enormous iatrogenic blood loss through 1 to 10 μL of blood sampling and re-infusion of 1 to 5 mL of discard blood but also reduce the infection risk through the closed structure while satisfying the key criteria of the blood sampler. The sampled blood preserved its quality without rupturing of red blood cells verified by blood potassium concentrations of 3.86 ± 0.07 mM on the sampled blood which is similar to 3.81 ± 0.04 mM measured from the blood which did not go through the system. The sampling volume among the sampling channels showed consistency with the relative standard deviation of 1.41 %. In addition to the micro blood sampling capability, the sampling system showed negligible sample cross-contamination. The analyte-free samples collected after aspirating 7,500 times higher signal sample showed the same output signal as blank. The system was also demonstrated not to cause air-embolism by having no bubble generation during flushing procedure and the system was verified as leak-free since there was no fluid leakage under 30 times higher pressure than central venous pressure for 24 h.

  16. Direct Trace Element Analysis of Liquid Blood Samples by In-Air Ion Beam Analytical Techniques (PIXE-PIGE).

    PubMed

    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.

  17. Determination of proflavine in rat whole blood without sample pretreatment by laser desorption postionization mass spectrometry.

    PubMed

    Chen, Jiaxin; Hu, Yongjun; Lu, Qiao; Wang, Pengchao; Zhan, Huaqi

    2017-02-10

    A novel pretreatment-free method involving laser desorption postionization (LDPI) coupled with time-of-flight mass spectrometry (MS) was developed for the monitoring of proflavine level in rat whole blood. It comprises a protocol for dosing via intravenous administration and collection of whole blood, followed by direct LDPI-MS analysis without any sample pretreatment. An intense ion signal at m/z 209 was observed from whole blood without any interference signals, except some background signals below m/z 100. The calibration curve was established with use of 9-phenylacridine as the internal standard for proflavine determination from the plotting of the peak ratios of proflavine to the internal standard, with a correlation coefficient (R (2)) greater than 0.99. The limit of detection was estimated to be 0.48 pmol/mm(2) and the quantification range was 0.5-16.5 μg/mL for proflavine. In addition, only a minimal matrix effect was observed, as expected from considerations of the desorption and ionization mechanism. Interday and intraday accuracy and precision were calculated to be within 13% and 82-114%, respectively. Estimated concentrations of proflavine residue in whole blood were also successfully obtained at selected time points after dosing. The proposed method is simple, low cost, and sensitive, and should be seen as a complementary method for monitoring drug levels in blood. Graphical Abstract Monitoring proflavine levels in rat whole blood at different time points using laser desorption postionization mass spectrometry (LDPI-MS).

  18. Use of an extracorporeal arterio-venous shunt and capillary tubes for frequent micro-blood sampling in rats.

    PubMed

    Wiegmann, T B; Artzer, D

    1983-02-01

    Because of the importance of frequent sampling during kinetic analysis, we have developed a method that permits frequent, reliable, and rapid access to the blood stream in rats. The method comprises the establishment of an extracorporeal arterio-venous shunt between femoral blood vessels. Quantitative samples of whole blood and plasma are obtained with capillary micro hematocrit tubes. The small sample volume permits repetitive analysis without major changes in hematocrit. There is good agreement between these measures and ordinary pipetting techniques. Kinetic analysis showed good resolution with close correspondence between actual measurements and the final curve of best fit. This method could be employed advantageously in small conscious animals, when frequent small samples are required by experimental design.

  19. Constrained sampling method for analytic continuation.

    PubMed

    Sandvik, Anders W

    2016-12-01

    A method for analytic continuation of imaginary-time correlation functions (here obtained in quantum Monte Carlo simulations) to real-frequency spectral functions is proposed. Stochastically sampling a spectrum parametrized by a large number of δ functions, treated as a statistical-mechanics problem, it avoids distortions caused by (as demonstrated here) configurational entropy in previous sampling methods. The key development is the suppression of entropy by constraining the spectral weight to within identifiable optimal bounds and imposing a set number of peaks. As a test case, the dynamic structure factor of the S=1/2 Heisenberg chain is computed. Very good agreement is found with Bethe ansatz results in the ground state (including a sharp edge) and with exact diagonalization of small systems at elevated temperatures.

  20. Constrained sampling method for analytic continuation

    NASA Astrophysics Data System (ADS)

    Sandvik, Anders W.

    2016-12-01

    A method for analytic continuation of imaginary-time correlation functions (here obtained in quantum Monte Carlo simulations) to real-frequency spectral functions is proposed. Stochastically sampling a spectrum parametrized by a large number of δ functions, treated as a statistical-mechanics problem, it avoids distortions caused by (as demonstrated here) configurational entropy in previous sampling methods. The key development is the suppression of entropy by constraining the spectral weight to within identifiable optimal bounds and imposing a set number of peaks. As a test case, the dynamic structure factor of the S =1 /2 Heisenberg chain is computed. Very good agreement is found with Bethe ansatz results in the ground state (including a sharp edge) and with exact diagonalization of small systems at elevated temperatures.

  1. Actinide Recovery Method for Large Soil Samples

    SciTech Connect

    Maxwell, S.L. III; Nichols, S.

    1998-11-01

    A new Actinide Recovery Method has been developed by the Savannah River Site Central Laboratory to preconcentrate actinides in very large soil samples. Diphonix Resin(r) is used eliminate soil matrix interferences and preconcentrate actinides after soil leaching or soil fusion. A rapid microwave digestion technique is used to remove the actinides from the Diphonix Resin(r). After the resin digestion, the actinides are recovered in a small volume of nitric acid which can be easily loaded onto small extraction-chromatography columns, such as TEVA Resin(r), U-TEVA Resin(r) or TRU Resin(r) (Eichrom Industries). This method enables the application of small, selective extraction-columns to recover actinides from very large soil samples with high selectivity, consistent tracer recoveries and minimal liquid waste.

  2. Methods for Sampling of Airborne Viruses

    PubMed Central

    Verreault, Daniel; Moineau, Sylvain; Duchaine, Caroline

    2008-01-01

    Summary: To better understand the underlying mechanisms of aerovirology, accurate sampling of airborne viruses is fundamental. The sampling instruments commonly used in aerobiology have also been used to recover viruses suspended in the air. We reviewed over 100 papers to evaluate the methods currently used for viral aerosol sampling. Differentiating infections caused by direct contact from those caused by airborne dissemination can be a very demanding task given the wide variety of sources of viral aerosols. While epidemiological data can help to determine the source of the contamination, direct data obtained from air samples can provide very useful information for risk assessment purposes. Many types of samplers have been used over the years, including liquid impingers, solid impactors, filters, electrostatic precipitators, and many others. The efficiencies of these samplers depend on a variety of environmental and methodological factors that can affect the integrity of the virus structure. The aerodynamic size distribution of the aerosol also has a direct effect on sampler efficiency. Viral aerosols can be studied under controlled laboratory conditions, using biological or nonbiological tracers and surrogate viruses, which are also discussed in this review. Lastly, general recommendations are made regarding future studies on the sampling of airborne viruses. PMID:18772283

  3. SOIL AND SEDIMENT SAMPLING METHODS | Science ...

    EPA Pesticide Factsheets

    The EPA Office of Solid Waste and Emergency Response's (OSWER) Office of Superfund Remediation and Technology Innovation (OSRTI) needs innovative methods and techniques to solve new and difficult sampling and analytical problems found at the numerous Superfund sites throughout the United States. Inadequate site characterization and a lack of knowledge of surface and subsurface contaminant distributions hinders EPA's ability to make the best decisions on remediation options and to conduct the most effective cleanup efforts. To assist OSWER, NERL conducts research to improve their capability to more accurately, precisely, and efficiently characterize Superfund, RCRA, LUST, oil spills, and brownfield sites and to improve their risk-based decision making capabilities, research is being conducted on improving soil and sediment sampling techniques and improving the sampling and handling of volatile organic compound (VOC) contaminated soils, among the many research programs and tasks being performed at ESD-LV.Under this task, improved sampling approaches and devices will be developed for characterizing the concentration of VOCs in soils. Current approaches and devices used today can lose up to 99% of the VOCs present in the sample due inherent weaknesses in the device and improper/inadequate collection techniques. This error generally causes decision makers to markedly underestimate the soil VOC concentrations and, therefore, to greatly underestimate the ecological

  4. New method for determination of ten pesticides in human blood.

    PubMed

    García-Repetto, R; Giménez, M P; Repetto, M

    2001-01-01

    An analytical method was developed for precise identification and quantitation of 10 pesticides in human blood. The pesticides studied, which have appeared frequently in actual cases, were endosulfan, lindane, parathion, ethyl-azinphos, diazinon, malathion, alachlor, tetradifon, fenthion and dicofol (o-p' and p-p' isomers). The current method replaces an earlier method which involved liquid-liquid extraction with a mixture of n-hexane-benzene (1 + 1). The extraction is performed by solid-phase extraction, with C18 cartridges and 2 internal standards, perthane and triphenylphosphate. Eluates were analyzed by gas chromatography (GC) with nitrogen-phosphorus and electrochemical detectors. Results were confirmed by GC-mass spectrometry in the electron impact mode. Blood blank samples spiked with 2 standard mixtures and an internal standard were used for quantitation. Mean recoveries ranged from 71.83 to 97.10%. Detection and quantitation limits are reported for each pesticide. Examples are provided to show the application of the present method to actual samples.

  5. Sample to answer visualization pipeline for low-cost point-of-care blood cell counting

    NASA Astrophysics Data System (ADS)

    Smith, Suzanne; Naidoo, Thegaran; Davies, Emlyn; Fourie, Louis; Nxumalo, Zandile; Swart, Hein; Marais, Philip; Land, Kevin; Roux, Pieter

    2015-03-01

    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 solutions [1], and can be particularly beneficial for blood cell count tests, which are often the starting point in the process of diagnosing a patient [2]. The initial focus of this work is on total white and red blood cell counts, using a microfluidic cartridge [3] for sample processing. Analysis of the processed samples has been implemented by means of two main optical visualization systems developed in-house: 1) a fluidic operation analysis system using high speed video data to determine volumes, mixing efficiency and flow rates, and 2) a microscopy analysis system to investigate homogeneity and concentration of blood cells. Fluidic parameters were derived from the optical flow [4] as well as color-based segmentation of the different fluids using a hue-saturation-value (HSV) color space. Cell count estimates were obtained using automated microscopy analysis and were compared to a widely accepted manual method for cell counting using a hemocytometer [5]. The results using the first iteration microfluidic device [3] showed that the most simple - and thus low-cost - approach for microfluidic component implementation was not adequate as compared to techniques based on manual cell counting principles. An improved microfluidic design has been developed to incorporate enhanced mixing and metering components, which together with this work provides the foundation on which to successfully implement automated, rapid and low-cost blood cell counting tests.

  6. Flow cytometric detection method for DNA samples

    DOEpatents

    Nasarabadi,Shanavaz; Langlois, Richard G.; Venkateswaran, Kodumudi S.

    2011-07-05

    Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM.TM. on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA.TM., on the 5' end.

  7. Flow cytometric detection method for DNA samples

    DOEpatents

    Nasarabadi, Shanavaz; Langlois, Richard G.; Venkateswaran, Kodumudi S.

    2006-08-01

    Disclosed herein are two methods for rapid multiplex analysis to determine the presence and identity of target DNA sequences within a DNA sample. Both methods use reporting DNA sequences, e.g., modified conventional Taqman.RTM. probes, to combine multiplex PCR amplification with microsphere-based hybridization using flow cytometry means of detection. Real-time PCR detection can also be incorporated. The first method uses a cyanine dye, such as, Cy3.TM., as the reporter linked to the 5' end of a reporting DNA sequence. The second method positions a reporter dye, e.g., FAM, on the 3' end of the reporting DNA sequence and a quencher dye, e.g., TAMRA, on the 5' end.

  8. Measurement of hematological, clinical chemistry, and infection parameters from hirudinized blood collected in universal blood sampling tubes.

    PubMed

    Menssen, H D; Brandt, N; Leben, R; Müller, F; Thiel, E; Melber, K

    2001-08-01

    Hirudin, the anticoagulatory polypeptide of the leech Hirudo medicinalis, strongly inhibits thrombus formation by specifically interacting with thrombin. For diagnostic purposes, hirudin should be superior to other anticlotting compounds because it only minimally alters the mineral, protein, and cellular blood constituents. To test this hypothesis, hirudinized and routinely processed venous blood from 80 healthy volunteers and patients was subjected to a variety of automated blood tests. A strong correlation was found between the results of automated complete blood counts obtained from K(2)-ethylenediaminetetraacetic acid (EDTA) anticoagulated and hirudinized blood (1000 antithrombin units [ATU] hirudin/ml). In addition, clinical chemistry and serological infection parameters (asparlat amintransferase [ASAT], lactate dehydrogenase [LDH], sodium, and so on, and antibodies against hepatitis B and C and human immunodeficiency virus [HIV]1/2, respectively) correlated well when measured in serum as compared with hirudinized plasma. Contrary to single clotting factors, global coagulation parameters (activated partial thromboplastin time [aPTT], prothrombin time [PT]) could not be measured in hirudinized blood. Recombinant hirudin neither interfered with immunophenotyping of mononuclear cells using FACScan analysis, nor did it alter the detection of Wilms' tumor gene expression by RT-PCR technology even at high doses (5000 ATU hirudin). Thus, a hirudin-containing blood sampling tube can be designed as a universal blood sampling tube (UBT) for testing the majority of diagnostic blood parameters.

  9. Blood doping: potential of blood and urine sampling to detect autologous transfusion.

    PubMed

    Segura, J; Lundby, C

    2014-05-01

    The collection of blood, its storage as red blood cell (RBC) concentrates and its reinjection is prohibited; until now, the practice cannot be reliably detected. A recent innovation-the haematological module of the athlete's biological passport-can provide authorities with indications towards autologous blood transfusion. In situations where a given athlete has been exposed to altitude, heat stress, sickness, etc, additional evidence may be needed to establish beyond any reasonable doubt that a blood transfusion may actually have occurred. Additional evidence may be obtained from at least three different approaches using parameters related to blood and urine matrices.Genomics applied to mRNA or miRNA is one of the most promising analytical tools. Proteomics of changes associated with RBC membranes may reveal the presence of cells stored for some time, as can an abnormal pattern of size distribution of aged cells. In urine, high concentrations of metabolites of plasticisers originating from the blood storing bags strongly suggest a recent blood transfusion. We emphasise the usefulness of simultaneously obtaining and then analysing blood and urine for complementary evidence of autologous blood transfusion ('blood doping').

  10. Validation of a minimally invasive blood-sampling technique for the analysis of hormones in domestic rabbits, Oryctolagus cuniculus (Lagomorpha).

    PubMed

    Voigt, Christian C; Fassbender, Mirja; Dehnhard, Martin; Wibbelt, Gudrun; Jewgenow, Katarina; Hofer, Heribert; Schaub, Günter A

    2004-01-01

    Previous studies in small mammals showed that blood-sucking bugs (Reduviidae, Heteroptera) can be used to obtain blood from veins difficult to access by human experimenters. In the present study, we validated the use of reduviid bugs for endocrinological studies in endotherms using domestic rabbits as a model organism. Two processes could alter the hormone concentrations in the blood ingested by the bug: (1) Mixing of ingested blood with saliva, gut fluid, or hemolymph and (2) digestive processes. We compared concentrations of progesterone, testosterone, and hydrocortisone in blood samples that were acquired from domestic rabbits (Oryctolagus cuniculus) by bugs (Dipetalogaster maxima) with hormone concentrations in blood obtained from the same individual rabbits with a conventional method, i.e., syringe. We found no significant differences in hormone concentrations between the two methods. Thus, the mixing effect is negligible immediately after the blood meal. In addition, we also could not find significant changes in concentrations of progesterone and hydrocortisone for up to 8h after the blood meal. Whereas levels of hydrocortisone remained unchanged for even 24h, progesterone levels significantly increased between eight and 24h. Thus, the bugs' excretory apparatus did not fractionate between water and hormones. Thirdly, we hypothesized that reduviid bugs impose less stress on the rabbits than the conventional method. We showed that deviations in hydrocortisone concentrations between the two blood sampling routines were lower when the bug method was used first and higher when the conventional method was used first. Thus, bugs imposed less stress on the study animals than the conventional method. Overall, we conclude that reduviid bugs present a minimally invasive method for obtaining blood from endotherm animals for endocrinological studies.

  11. Whole blood is the sample matrix of choice for monitoring systemic triclocarban levels.

    PubMed

    Schebb, Nils Helge; Ahn, Ki Chang; Dong, Hua; Gee, Shirley J; Hammock, Bruce D

    2012-05-01

    The antibacterial triclocarban (TCC) concentrates in the cellular fraction of blood. Consequently, plasma levels are at least two-fold lower than the TCC amount present in blood. Utilizing whole blood sampling, a low but significant absorption of TCC from soap during showering is demonstrated for a small group of human subjects.

  12. Hemoglobin Variant Analysis via Direct Surface Sampling of Dried Blood Spots Coupled with High-Resolution Mass Spectrometry

    PubMed Central

    2011-01-01

    Hemoglobinopathies are the most common inherited disorders. Newborn blood screening for clinically significant hemoglobin variants, including sickle (HbS), HbC, and HbD, has been adopted in many countries as it is widely acknowledged that early detection improves the outcome. We present a method for determination of Hb variants by direct surface sampling of dried blood spots by use of an Advion Triversa Nanomate automated electrospray system coupled to a high-resolution mass spectrometer. The method involves no sample preparation. It is possible to unambiguously identify homozygous and heterozygous HbS, HbC, and HbD variants in <10 min without the need for additional confirmation. The method allows for repeated analysis of a single blood spot over a prolonged time period and is tolerant of blood spot storage conditions. PMID:21341716

  13. Active tracking of rejected dried blood samples in a large program in Nigeria

    PubMed Central

    Inalegwu, Auchi; Phillips, Sunny; Datir, Rawlings; Chime, Christopher; Ozumba, Petronilla; Peters, Samuel; Ogbanufe, Obinna; Mensah, Charles; Abimiku, Alash’Le; Dakum, Patrick; Ndembi, Nicaise

    2016-01-01

    AIM: To study the impact of rejection at different levels of health care by retrospectively reviewing records of dried blood spot samples received at the molecular laboratory for human immunodeficiency virus (HIV) early infant diagnosis (EID) between January 2008 and December 2012. METHODS: The specimen rejection rate, reasons for rejection and the impact of rejection at different levels of health care was examined. The extracted data were cleaned and checked for consistency and then de-duplicated using the unique patient and clinic identifiers. The cleaned data were ciphered and exported to SPSS version 19 (SPSS 2010 IBM Corp, New York, United States) for statistical analyses. RESULTS: Sample rejection rate of 2.4% (n = 786/32552) and repeat rate of 8.8% (n = 69/786) were established. The mean age of infants presenting for first HIV molecular test among accepted valid samples was 17.83 wk (95%CI: 17.65-18.01) vs 20.30 wk (95%CI: 16.53-24.06) for repeated samples. HIV infection rate was 9.8% vs 15.9% for accepted and repeated samples. Compared to tertiary healthcare clinics, secondary and primary clinics had two-fold and three-fold higher likelihood of sample rejection, respectively (P < 0.05). We observed a significant increase in sample rejection rate with increasing number of EID clinics (r = 0.893, P = 0.041). The major reasons for rejection were improper sample collection (26.3%), improper labeling (16.4%) and insufficient blood (14.8%). CONCLUSION: Programs should monitor pre-analytical variables and incorporate continuous quality improvement interventions to reduce errors associated with sample rejection and improve patient retention. PMID:27175352

  14. Determination of DDT and related compounds in blood samples from agricultural workers.

    PubMed

    Guardino, X; Serra, C; Obiols, J; Rosell, M G; Berenguer, M J; López, F; Brosa, J

    1996-01-05

    An analytical method combining a solid-phase (C18) clean-up and GC-electron-capture detection using a capillary column, was implemented to determine p,p'-DDT and its metabolites (p,p'-DDD and p,p'-DDE), as well as other organochlorine pesticides in whole blood samples from 30 farmers and 24 non-occupationally exposed workers. The average concentrations for the quantified pesticides, p,p'-DDT, p,p'-DDD and p,p'-DDE, were 0.9, 1.5 and 8.0 micrograms/l whole blood for exposed workers and 0.3, 0.5 and 3.3 micrograms/l for unexposed workers, respectively. GC-MS was used to confirm the identity of the pesticides found. Solid-phase extraction and the protocol used give a cleaner analytical matrix, not only improving sensitivity and resolution, but also allowing analyses with smaller blood samples as compared to other methods.

  15. New HPLC method for separation of blood plasma phospholipids.

    PubMed

    Suchocka, Zofia; Gronostajska, Dorota; Suchocki, Piotr; Pachecka, Jan

    2003-08-08

    The aim of the present work was to develop a new HPLC method for separation of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and lysophosphatidylcholine (LPC) from small-volume samples of blood plasma. Human plasma glycerophospholipids were separated by liquid-liquid extraction method followed by solid phase extraction (SPE) on aminopropyl columns. Reversed-phase Sephasil C8 column (10 cm x 2.1 mm, I.D. 5 microm) and micropreparative chromatograph "SMART" were used for separation of PC, PE, LPC and PI from SPE phospholipids extract. Binary-step gradient of eluent A: acetonitrile-methanol (130:5, v/v) and B (0.01% trifluoroacetic acid) provided good, fast and reproducible resolution of investigated phospholipids classes in 12 min at 30 degrees C. Eluted phospholipids were detected at wavelengths lambda=235 and 254 nm. This method made it possible to determine quantitatively: 5 microg ml(-1) PC, 1 microg ml(-1) LPC, 4 microg ml(-1) PE and 3 microg ml(-1) PI in blood plasma samples.

  16. Slurry sampling in serum blood for mercury determination by CV-AFS.

    PubMed

    Aranda, Pedro R; Gil, Raúl A; Moyano, Susana; De Vito, Irma; Martinez, Luis D

    2009-01-30

    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 microg 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 microg L(-1)Hg.

  17. Well purge and sample apparatus and method

    DOEpatents

    Schalla, Ronald; Smith, Ronald M.; Hall, Stephen H.; Smart, John E.; Gustafson, Gregg S.

    1995-01-01

    The present invention specifically permits purging and/or sampling of a well but only removing, at most, about 25% of the fluid volume compared to conventional methods and, at a minimum, removing none of the fluid volume from the well. The invention is an isolation assembly with a packer, pump and exhaust, that is inserted into the well. The isolation assembly is designed so that only a volume of fluid between the outside diameter of the isolation assembly and the inside diameter of the well over a fluid column height from the bottom of the well to the top of the active portion (lower annulus) is removed. The packer is positioned above the active portion thereby sealing the well and preventing any mixing or contamination of inlet fluid with fluid above the packer. Ports in the wall of the isolation assembly permit purging and sampling of the lower annulus along the height of the active portion.

  18. Well purge and sample apparatus and method

    DOEpatents

    Schalla, R.; Smith, R.M.; Hall, S.H.; Smart, J.E.; Gustafson, G.S.

    1995-10-24

    The present invention specifically permits purging and/or sampling of a well but only removing, at most, about 25% of the fluid volume compared to conventional methods and, at a minimum, removing none of the fluid volume from the well. The invention is an isolation assembly with a packer, pump and exhaust, that is inserted into the well. The isolation assembly is designed so that only a volume of fluid between the outside diameter of the isolation assembly and the inside diameter of the well over a fluid column height from the bottom of the well to the top of the active portion (lower annulus) is removed. The packer is positioned above the active portion thereby sealing the well and preventing any mixing or contamination of inlet fluid with fluid above the packer. Ports in the wall of the isolation assembly permit purging and sampling of the lower annulus along the height of the active portion. 8 figs.

  19. Blood monitoring systems and methods thereof

    NASA Technical Reports Server (NTRS)

    Mir, Jose (Inventor); Zander, Dennis (Inventor)

    2012-01-01

    A blood monitoring system is capable of monitoring the blood of a subject in vivo. The blood monitoring system comprises: 1) an array of movable microneedle micromachined within associated wells; 2) array of motion actuators able to move each needle in and out of their associated wells; 3) array of microvalves associated with each microneedle able to control the flow of air around the microneedle; 4) an array of chemical sensors inserted into patient by movable microneedles; 5) an array of inductors able to measure chemical concentration in the vicinity of inserted chemical sensors; 6) conducting vias that provide timed actuating signal signals from a control system to each motion actuator; 7) conducting vias that transmit signal produced by array of chemical sensors to the control system for processing, although the blood monitoring system can comprise other numbers and types of elements in other configurations.

  20. Method and device for supporting blood vessels during anastomosis

    DOEpatents

    Doss, J.D.

    1985-05-20

    A device and method for preventing first and second severed blood vessels from collapsing during attachment to each other. The device comprises a dissolvable non-toxic stent that is sufficiently rigid to prevent the blood vessels from collapsing during anastomosis. The stent can be hollow or have passages to permit blood flow before it dissolves. A single stent can be inserted with an end in each of the two blood vessels or separate stents can be inserted into each blood vessel. The stent may include a therapeutically effective amount of a drug which is slowly released into the blood stream as the stent dissolves. 12 figs.

  1. Dried Blood Spot sampling in psychiatry: Perspectives for improving therapeutic drug monitoring.

    PubMed

    Martial, Lisa C; Aarnoutse, Rob E; Mulder, Martina; Schellekens, Arnt; Brüggemann, Roger J M; Burger, David M; Schene, Aart H; Batalla, Albert

    2017-03-01

    Assessment of drug concentrations is indicated to guide dosing of a selected number of drugs used in psychiatry. Conventionally this is done by vena puncture. Novel sampling strategies such as dried blood spot (DBS) sampling have been developed for various drugs, including antipsychotics, antidepressants and mood-stabilizers. DBS sampling is typically performed by means of a finger prick. This method allows for remote sampling, which means that patients are not required to travel to a health care facility. The number of DBS assays for drugs used in psychiatry has increased over the last decade and includes antidepressants (tricyclic and serotonin and/or norepinephrine reuptake inhibitors), mood stabilizers and first- and second-generation antipsychotics. Available assays often comply with analytical validation criteria but are seldom used in routine clinical care. Little attention has been paid to the clinical validation and implementation processes of home sampling. Ideally, not only medicines but also clinical chemistry parameters should be measured within the same sample. This article reflects on the position of DBS remote sampling in psychiatry and provides insight in the requisites of making such a sampling tool successful.

  2. An indirect spectrophotometric method for the determination of silicon in serum, whole blood and erythrocytes.

    PubMed

    Tamada, Tomoko

    2003-09-01

    An indirect method for the determination of silicon in blood samples has been developed. The proposed method overcame interference from a large amount of salts and phosphate in blood samples, and enabled us to determine the silicon contents in serum and whole blood by the same operation. After blood samples were digested by microwave heating, silicon, present as silicate in the sample solution, was reacted with molybdate to form a silicomolybdate complex. The complex was then separated from unreacted molybdate by a cation-exchange resin column. The molybdate liberated from the complex was spectrophotometrically determined in place of silicon. Since the method is not affected the composition of matrices between serum and whole blood, it could achieve good precision and accuracy, and could also estimate the silicon contents in erythrocytes from those in serum and whole blood. The sensitivity of the method was almost equal to that of the conventional silicomolybdenum blue method, and the calibration curve was linear up to 50 micromol l(-1) of silicon with a detection limit of 1.1 micromol l(-1) in whole blood. The mean concentrations of silicon in five healthy subjects were 11 micromol l(-1) for serum, 28 micromol l(-1) for whole blood and 50 micromol l(-1) for erythrocytes. Thus, the obtained distribution ratio between serum and erythrocytes was in the range of 0.15-0.39, and was found to be included in a narrow range.

  3. A convenient method to measure blood-plasma concentration ratio using routine plasma collection in in vivo pharmacokinetic studies.

    PubMed

    Berezhkovskiy, Leonid M; Zhang, Xiaolin; Cheong, Jonathan

    2011-12-01

    A practical time-saving method of determination of equilibrium blood-plasma concentration ratio is described. The method is based on the analysis of compound plasma concentrations in regular blood sample and the blood sample diluted with blank plasma. Since only plasma concentrations are analyzed, the method can be conveniently applied in routine pharmacokinetic studies with minimal additional work for obtaining blood-plasma ratio. The method can also be easily used in in vitro experiment. The results obtained by suggested method are in good agreement with that obtained by common in vitro measurements of blood-plasma ratio.

  4. A round robin approach to the analysis of bisphenol a (BPA) in human blood samples

    PubMed Central

    2014-01-01

    Background Human exposure to bisphenol A (BPA) is ubiquitous, yet there are concerns about whether BPA can be measured in human blood. This Round Robin was designed to address this concern through three goals: 1) to identify collection materials, reagents and detection apparatuses that do not contribute BPA to serum; 2) to identify sensitive and precise methods to accurately measure unconjugated BPA (uBPA) and BPA-glucuronide (BPA-G), a metabolite, in serum; and 3) to evaluate whether inadvertent hydrolysis of BPA-G occurs during sample handling and processing. Methods Four laboratories participated in this Round Robin. Laboratories screened materials to identify BPA contamination in collection and analysis materials. Serum was spiked with concentrations of uBPA and/or BPA-G ranging from 0.09-19.5 (uBPA) and 0.5-32 (BPA-G) ng/mL. Additional samples were preserved unspiked as ‘environmental’ samples. Blinded samples were provided to laboratories that used LC/MSMS to simultaneously quantify uBPA and BPA-G. To determine whether inadvertent hydrolysis of BPA metabolites occurred, samples spiked with only BPA-G were analyzed for the presence of uBPA. Finally, three laboratories compared direct and indirect methods of quantifying BPA-G. Results We identified collection materials and reagents that did not introduce BPA contamination. In the blinded spiked sample analysis, all laboratories were able to distinguish low from high values of uBPA and BPA-G, for the whole spiked sample range and for those samples spiked with the three lowest concentrations (0.5-3.1 ng/ml). By completion of the Round Robin, three laboratories had verified methods for the analysis of uBPA and two verified for the analysis of BPA-G (verification determined by: 4 of 5 samples within 20% of spiked concentrations). In the analysis of BPA-G only spiked samples, all laboratories reported BPA-G was the majority of BPA detected (92.2 – 100%). Finally, laboratories were more likely to be verified

  5. Field evaluation of a VOST sampling method

    SciTech Connect

    Jackson, M.D.; Johnson, L.D.; Fuerst, R.G.; McGaughey, J.F.; Bursey, J.T.; Merrill, R.G.

    1994-12-31

    The VOST (SW-846 Method 0030) specifies the use of Tenax{reg_sign} and a particular petroleum-based charcoal (SKC Lot 104, or its equivalent), that is no longer commercially available. In field evaluation studies of VOST methodology, a replacement petroleum-based charcoal has been used: candidate replacement sorbents for charcoal were studied, and Anasorb{reg_sign} 747, a carbon-based sorbent, was selected for field testing. The sampling train was modified to use only Anasorb{reg_sign} in the back tube and Tenax{reg_sign} in the two front tubes to avoid analytical difficulties associated with the analysis of the sequential bed back tube used in the standard VOST train. The standard (SW-846 Method 0030) and the modified VOST methods were evaluated at a chemical manufacturing facility using a quadruple probe system with quadruple trains. In this field test, known concentrations of the halogenated volatile organic compounds, that are listed in the Clean Air Act Amendments of 1990, Title 3, were introduced into the VOST train and the modified VOST train, using the same certified gas cylinder as a source of test compounds. Statistical tests of the comparability of methods were performed on a compound-by-compound basis. For most compounds, the VOST and modified VOST methods were found to be statistically equivalent.

  6. Validation of a non-invasive blood-sampling technique for doubly-labelled water experiments.

    PubMed

    Voigt, Christian C; Helversen, Otto Von; Michener, Robert H; Kunz, Thomas H

    2003-04-01

    Two techniques for bleeding small mammals have been used in doubly-labeled water (DLW) studies, including vena puncture and the use of starved nymphal stages of hematophagous reduviid bugs (Reduviidae, Hemiptera). In this study, we tested the validity of using reduviid bugs in doubly-labeled water experiments. We found that the isotope enrichment in initial blood samples collected with bugs was significantly lower compared to isotope enrichment in blood samples obtained using vena puncture. We therefore used the desiccation method for estimating total body water (TBW) in DLW experiments because TBW calculated using the isotope dilution method was overestimated when blood samples were collected using reduviid bugs. In our validation experiment with nectar-feeding bats (Glossophaga soricina), we compared estimates of daily energy expenditure (DEE) using DLW with those derived from the energy balance method. We considered Speakman's equation (controlling for 25% fractionated water loss) as the most appropriate for our study animal and calculated DEE accordingly. On average, DEE estimated with DLW was not significantly different from the mean value obtained with the energy balance method (mean deviation 1.2%). We conclude that although bug hemolymph or intestinal liquids most likely contaminate the samples, estimates of DEE are still valid because the DLW method does not depend on absolute isotope enrichments but on the rate of isotope decrease over time. However, dilution of blood with intestinal liquids or hemolymph from a bug may lead to larger variation in DEE estimates. We also tested how the relative error of DLW estimates changed with varying assumptions about fractionation. We used three additional equations for calculating DEE in DLW experiments. The basic equation for DLW experiments published by Lifson and McClintock (LM-6) assumes no fractionation, resulted in an overestimate of DEE by 10%. Nagy's equation (N-2) controls for changes in body mass but not for

  7. [Comparison of the determination of cyclosporin-A in blood samples collected on filter paper and by the ordinary technique].

    PubMed

    Azevedo, L S; Manrique, R; Sabbaga, E

    1995-01-01

    Monitoring cyclosporin-A (CsA) blood levels is of utmost importance for the rational use of this drug. Although many centers perform transplants, in Brazil there are few laboratories able to measure CsA blood levels. Therefore making blood samples reach the laboratory emerged as a problem. Collection of blood on filter paper has been a technique used for a long time in special cases. PURPOSE--To confirm the usefulness of measuring CsA blood levels in blood samples collected on filter paper and in the usual way. METHOD--We studied twenty renal cadaver kidney recipients who were receiving CsA, azathioprine and prednisone. Ninety five blood samples were collected and divided into two aliquots. One of them was sent routinely to one laboratory to perform whole blood CsA measurements. From the other aliquot, 20 microliters were pipetted on filter paper. When dried they were mailed to the other laboratory, where, after elution, CsA was measured. In both cases radioimmunoassay with polyclonal antibody was used. RESULTS--Linear correlation between both measurements revealed r = 0.81 with no statistical difference. CONCLUSION--The technique showed to be useful in clinical practice. In countries with continental size, as Brazil, it may be very helpful.

  8. Comparison of Submental Blood Collection with the Retroorbital and Submandibular Methods in Mice (Mus musculus)

    PubMed Central

    Regan, Rainy D; Fenyk-Melody, Judy E; Tran, Sam M; Chen, Guang; Stocking, Kim L

    2016-01-01

    Nonterminal blood sample collection of sufficient volume and quality for research is complicated in mice due to their small size and anatomy. Large (>100 μL) nonterminal volumes of unhemolyzed or unclotted blood currently are typically collected from the retroorbital sinus or submandibular plexus. We developed a third method—submental blood collection—which is similar in execution to the submandibular method but with minor changes in animal restraint and collection location. Compared with other techniques, submental collection is easier to perform due to the direct visibility of the target vessels, which are located in a sparsely furred region. Compared with the submandibular method, the submental method did not differ regarding weight change and clotting score but significantly decreased hemolysis and increased the overall number of high-quality samples. The submental method was performed with smaller lancets for the majority of the bleeds, yet resulted in fewer repeat collection attempts, fewer insufficient samples, and less extraneous blood loss and was qualitatively less traumatic. Compared with the retroorbital technique, the submental method was similar regarding weight change but decreased hemolysis, clotting, and the number of overall high-quality samples; however the retroorbital method resulted in significantly fewer incidents of insufficient sample collection. Extraneous blood loss was roughly equivalent between the submental and retroorbital methods. We conclude that the submental method is an acceptable venipuncture technique for obtaining large, nonterminal volumes of blood from mice. PMID:27657712

  9. Sensitivity of PCR Assays for Murine Gammaretroviruses and Mouse Contamination in Human Blood Samples

    PubMed Central

    Lee, Li Ling; Lin, Lin; Bell, David S.; Levine, Susan; Hanson, Maureen R.

    2012-01-01

    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. PMID:22629404

  10. Simplified [18F]FDG Image-Derived Input Function Using the Left Ventricle, Liver, and One Venous Blood Sample

    PubMed Central

    Tantawy, Mohammed Noor; Peterson, Todd E.

    2014-01-01

    A relatively simple, almost entirely noninvasive imaging-based method is presented for deriving arterial blood input functions for quantitative [18F]2-fluoro-2-deoxy-D-glucose (FDG) positron emission tomographic (PET) studies in rodents. It requires only one venous blood sample at the end of the scan. MicroPET images and arterial blood time-activity curves (TACs) were downloaded from the Mouse Quantitation Program database at the University of California, Los Angeles. Three-dimensional regions of interest were drawn around the blood-pool region of the left ventricle and within the liver to derive their respective TACs. To construct the “hybrid” image-derived input function (IDIF), the initial part of the left ventricle TAC, containing the peak concentration of [18F]FDG in the arterial blood, was corrected for spillout (ie, partial-volume effect yielding a recovery coefficient < 1) and then joined to the liver TAC (normalized to the 60-minute arterial blood sample) immediately after it peaks. To validate our method, the [18F]FDG influx constant (Ki) was estimated using a two-tissue compartment model and compared to estimates of Ki obtained using measured arterial blood TACs. No significant difference in the Ki estimates was obtained with the arterial blood input function and our hybrid IDIF. We conclude that the normalized hybrid IDIF can be used in practice to obtain reliable Ki estimates. PMID:20236605

  11. Using Dried Blood Spot Sampling to Improve Data Quality and Reduce Animal Use in Mouse Pharmacokinetic Studies

    PubMed Central

    Wickremsinhe, Enaksha R; Perkins, Everett J

    2015-01-01

    Traditional pharmacokinetic analysis in nonclinical studies is based on the concentration of a test compound in plasma and requires approximately 100 to 200 µL blood collected per time point. However, the total blood volume of mice limits the number of samples that can be collected from an individual animal—often to a single collection per mouse—thus necessitating dosing multiple mice to generate a pharmacokinetic profile in a sparse-sampling design. Compared with traditional methods, dried blood spot (DBS) analysis requires smaller volumes of blood (15 to 20 µL), thus supporting serial blood sampling and the generation of a complete pharmacokinetic profile from a single mouse. Here we compare plasma-derived data with DBS-derived data, explain how to adopt DBS sampling to support discovery mouse studies, and describe how to generate pharmacokinetic and pharmacodynamic data from a single mouse. Executing novel study designs that use DBS enhances the ability to identify and streamline better drug candidates during drug discovery. Implementing DBS sampling can reduce the number of mice needed in a drug discovery program. In addition, the simplicity of DBS sampling and the smaller numbers of mice needed translate to decreased study costs. Overall, DBS sampling is consistent with 3Rs principles by achieving reductions in the number of animals used, decreased restraint-associated stress, improved data quality, direct comparison of interanimal variability, and the generation of multiple endpoints from a single study. PMID:25836959

  12. A novel method for blood-typing using nitrocellulose.

    PubMed

    Afshari, Parastoo; Abolfathi, Nabiollah

    2016-12-07

    Blood wicking in its steady-state form, i.e. the uniform distribution of blood cells in plasma, is completely different from that in its coagulated form on a porous surface like paper. The hydrophilic property of the cellulose leads to a significant wicking of the blood cells on paper fibers after rinsing with isotonic solution. The difference in the wicking length of the blood cells in steady state and that in the coagulated form could be considered as a criterion to recognize the blood type in a paper-based kit. However, owing to the molecular structure of the nitrocellulose, a better process occurs while separating the coagulated blood from the steady-state form of cells. Therefore, it is possible to use the nitrocellulose for the blood-typing kit which leads to a simpler way to diagnose a blood type. Two series of experiments were performed on nitrocellulose membrane. First, antibody solutions and blood samples were sequentially absorbed on nitrocellulose strips, allowed to interact, rinsed with an isotonic solution and distilled water, and image processing performed on a digital picture of the remaining blood cells. The efficiency of the agglutinated blood cell fixation was quantified by red color intensity. Then, it was demonstrated that there is no considerable difference in fixation of agglutinated blood cells with rinsing using isotonic and nonisotonic solutions. This fact can be a considerable advantage over paper since it can eliminate the probable mistake from using unisotonic solution for rinsing. Second, owing to the nonwicking property of the blood cells on the hydrophobic nitrocellulose fibers, we employed another diagnostic criterion and investigated nitrocellulose blood-typing prototypes. The nitrocellulose blood-typing kit provides more simple, sensitive and trustworthy assay for rapid blood typing in situations with no access to laboratory facilities.

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

    NASA Astrophysics Data System (ADS)

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

    2005-08-01

    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.

  14. Comparison of a new cobinamide-based method to a standard laboratory method for measuring cyanide in human blood.

    PubMed

    Swezey, Robert; Shinn, Walter; Green, Carol; Drover, David R; Hammer, Gregory B; Schulman, Scott R; Zajicek, Anne; Jett, David A; Boss, Gerry R

    2013-01-01

    Most hospital laboratories do not measure blood cyanide concentrations, and samples must be sent to reference laboratories. A simple method is needed for measuring cyanide in hospitals. The authors previously developed a method to quantify cyanide based on the high binding affinity of the vitamin B12 analog, cobinamide, for cyanide and a major spectral change observed for cyanide-bound cobinamide. This method is now validated in human blood, and the findings include a mean inter-assay accuracy of 99.1%, precision of 8.75% and a lower limit of quantification of 3.27 µM cyanide. The method was applied to blood samples from children treated with sodium nitroprusside and it yielded measurable results in 88 of 172 samples (51%), whereas the reference laboratory yielded results in only 19 samples (11%). In all 19 samples, the cobinamide-based method also yielded measurable results. The two methods showed reasonable agreement when analyzed by linear regression, but not when analyzed by a standard error of the estimate or paired t-test. Differences in results between the two methods may be because samples were assayed at different times on different sample types. The cobinamide-based method is applicable to human blood, and can be used in hospital laboratories and emergency rooms.

  15. Blood Sampling Seasonality as an Important Preanalytical Factor for Assessment of Vitamin D Status

    PubMed Central

    Bonelli, Patrizia; Buonocore, Ruggero; Aloe, Rosalia

    2016-01-01

    Summary 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. Methods 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. Results 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. Conclusions 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. PMID:28356869

  16. System and Method for Isolation of Samples

    NASA Technical Reports Server (NTRS)

    Zhang, Ye (Inventor); Wu, Honglu (Inventor)

    2014-01-01

    Systems and methods for isolating samples are provided. The system comprises a first membrane and a second membrane disposed within an enclosure. First and second reservoirs can also be disposed within the enclosure and adapted to contain one or more reagents therein. A first valve can be disposed within the enclosure and in fluid communication with the first reservoir, the second reservoir, or both. The first valve can also be in fluid communication with the first or second membranes or both. The first valve can be adapted to selectively regulate the flow of the reagents from the first reservoir, through at least one of the first and second membranes, and into the second reservoir.

  17. A micro-rheological method for determination of blood type.

    PubMed

    Makulska, Sylwia; Jakiela, Slawomir; Garstecki, Piotr

    2013-07-21

    The measurement of time and distance can be used for determining agglutination in small (nL) samples of liquid. We demonstrate the use of this new scheme of detection in typing and subtyping blood in a simple microfluidic system that monitors the speed of flow of microdroplets. The system (i) accepts small samples of liquids deposited directly onto the chip, (ii) forms droplets on demand from these samples, (iii) merges the droplets, and (iv) measures their speed in a microchannel. A sequence of measurements on different combinations of blood and antibodies can thus be used to determine blood type with the estimated probability of mistyping being less than 1 in a million tests. In addition, in the agglutinated samples, red blood cells concentrate at the rear of the droplets yielding an additional vista for detection and suggesting a possible mechanism for separations.

  18. A general method to determine sampling windows for nonlinear mixed effects models with an application to population pharmacokinetic studies.

    PubMed

    Foo, Lee Kien; McGree, James; Duffull, Stephen

    2012-01-01

    Optimal design methods have been proposed to determine the best sampling times when sparse blood sampling is required in clinical pharmacokinetic studies. However, the optimal blood sampling time points may not be feasible in clinical practice. Sampling windows, a time interval for blood sample collection, have been proposed to provide flexibility in blood sampling times while preserving efficient parameter estimation. Because of the complexity of the population pharmacokinetic models, which are generally nonlinear mixed effects models, there is no analytical solution available to determine sampling windows. We propose a method for determination of sampling windows based on MCMC sampling techniques. The proposed method attains a stationary distribution rapidly and provides time-sensitive windows around the optimal design points. The proposed method is applicable to determine sampling windows for any nonlinear mixed effects model although our work focuses on an application to population pharmacokinetic models.

  19. DNA methylome profiling using neonatal dried blood spot samples: a proof-of-principle study.

    PubMed

    Hollegaard, Mads Vilhelm; Grauholm, Jonas; Nørgaard-Pedersen, Bent; Hougaard, David Michael

    2013-04-01

    DNA methylation is the most common DNA modification and perhaps the best described epigenetic modification. It is believed to be important for genomic imprinting and gene regulation and has been associated with the development of diseases such as schizophrenia and some types of cancer. Neonatal dried blood spot samples, commonly known as Guthrie cards, are routinely collected worldwide to screen newborns for diseases. Some countries, including Denmark, have been storing the excess neonatal dried blood spot samples in biobanks for decades. Representing a high percentage of the population under a certain age, the neonatal dried blood spot samples are a potential alternative to collecting new samples to study diseases. As such, neonatal dried blood spot samples have previously been used for DNA genotyping studies with excellent results. However, the amount of material available for research is often limited, challenging researchers to generate the most data from a limited quantity of material. In this proof-of-principle study, we address whether two 3.2mm disks punched from a neonatal dried blood spot sample contain enough DNA for genome-wide methylome profiling, measuring 27,578 loci at the same time. We selected two subjects and carried out the following with each: 1) collected an adult whole-blood sample as reference, 2) spotted a fraction of the whole-blood sample onto a similar type of filter paper as used in the newborn screening and stored it for 3years to serve as a dried blood spot reference, and 3) identified the archived neonatal dried blood spot samples, stored for 26-28years, in the Danish Newborn Screening Biobank as a representative of the archived samples. For comparison, we used two different kits for DNA extraction. The DNA, extracted using the Extract-N-Amp Blood PCR kit, was analyzed, and no statistically significant differences were observed (P<0.001) when we compared the methylation profile of the reference whole-blood samples to the dried blood

  20. Luminol chemiluminescence biosensor for glycated hemoglobin (HbA1c) in human blood samples.

    PubMed

    Ahn, Kwang-Soo; Lee, JungHoon; Park, Jong-Myeon; Choi, Han Nim; Lee, Won-Yong

    2016-01-15

    Luminol chemiluminescence (CL) biosensor based on boronic acid modified gold substrate has been developed for the determination of glycated hemoglobin (HbA1c) in human blood samples. In order to selectively capture HbA1c in sample, carboxy-EG6-undecanethiol was self-assembled on a gold thin-film substrate, followed by covalent coupling of 3-aminophenyl boronic acid (3-APBA). The captured HbA1c containing four iron heme groups plays as a catalyst for luminol CL reaction in the presence of hydrogen peroxide, and thus the luminol CL response is linearly proportional to the amount of HbA1c captured on the biosensor surface. The present biosensor showed linear dynamic range of HbA1c from 2.5% to 17.0%, which well covers the clinically important concentration range. In addition, the present biosensor exhibited negligible response to interfering species such as hemoglobin, fructose, and sorbitol. The present HbA1c biosensor was applied to the determination of HbA1c in human blood samples and the results were well agreed with that obtained with a conventional method.

  1. Chromatographic resolution, characterisation and quantification of VX enantiomers in hemolysed swine blood samples.

    PubMed

    Reiter, Georg; Mikler, John; Hill, Ira; Weatherby, Kendal; Thiermann, Horst; Worek, Franz

    2008-09-15

    The present study was initiated to develop a sensitive and highly selective method for the analysis of the enantiomers of the nerve agent VX (O-ethyl S-[2(diisopropylamino)ethyl] methylphosphonothioate) in blood samples for toxicokinetic and therapeutic research. To achieve this goal, analytical and semi-preparative enantioseparation of VX were carried out with gas and liquid chromatography. The GC chiral stationary phase was HYDRODEX-beta-TBDAc (beta cyclodextrin), on which VX was baseline-resolved. On the chiral HPLC phase CHIRALCEL OD-H the enantiomers of VX were isolated with enantiomeric excess >99.99%. They were characterised by specific optical rotation (+/-25.8 deg ml dm(-1)g(-1) at 20 degrees C and 589 nm) and by determination of cholinesterase inhibition rate constants. For the quantitative chiral detection of VX the enantioresolution was realized on the HPLC chiral phase CHIRAL AGP. A specific procedure was developed to isolate VX from swine blood samples thereby stabilising its enantiomers. The limit of detection was 200 fg per enantiomer on column. The absolute recovery of the overall sample preparation procedure was 75%. After an intravenous and percutaneous administration of a supralethal dose of VX in anesthetised swine (+)-VX and (-)-VX could be quantified up to 720 min.

  2. Characterization at the individual cell level and in whole blood samples of shear stress preventing red blood cells aggregation.

    PubMed

    Lee, K; Kinnunen, M; Danilina, A V; Ustinov, V D; Shin, S; Meglinski, I; Priezzhev, A V

    2016-05-03

    The aggregation of red blood cells (RBC) is an intrinsic feature of blood that has a strong impact on its microcirculation. For a number of years it has been attracting a great attention in basic research and clinical studies. Here, we study a relationship between the RBC aggregation parameters measured at the individual cell level and in a whole blood sample. The home made optical tweezers were used to measure the aggregating and disaggregating forces for a pair of interacting RBCs, at the individual cell level, in order to evaluate the corresponding shear stresses. The RheoScan aggregometer was used for the measurements of critical shear stress (CSS) in whole blood samples. The correlation between CSS and the shear stress required to stop an RBC pair from aggregating was found. The shear stress required to disaggregate a pair of RBCs using the double channel optical tweezers appeared to be about 10 times higher than CSS. The correlation between shear stresses required to prevent RBCs from aggregation at the individual cell level and in whole blood samples was estimated and assessed quantitatively. The experimental approach developed has a high potential for advancing hemorheological studies.

  3. Automatic disease screening method using image processing for dried blood microfluidic drop stain pattern recognition.

    PubMed

    Sikarwar, Basant S; Roy, Mukesh; Ranjan, Priya; Goyal, Ayush

    2016-07-01

    This paper examines programmed automatic recognition of infection from samples of dried stains of micro-scale drops of patient blood. This technique has the upside of being low-cost and less-intrusive and not requiring puncturing the patient with a needle for drawing blood, which is especially critical for infants and the matured. It also does not require expensive pathological blood test laboratory equipment. The method is shown in this work to be successful for ailment identification in patients suffering from tuberculosis and anaemia. Illness affects the physical properties of blood, which thus influence the samples of dried micro-scale blood drop stains. For instance, if a patient has a severe drop in platelet count, which is often the case of dengue or malaria patients, the blood's physical property of viscosity drops substantially, i.e. the blood is thinner. Thus, the blood micro-scale drop stain samples can be utilised for diagnosing maladies. This paper presents programmed automatic examination of the dried micro-scale drop blood stain designs utilising an algorithm based on pattern recognition. The samples of micro-scale blood drop stains of ordinary non-infected people are clearly recognisable as well as the samples of micro-scale blood drop stains of sick people, due to key distinguishing features. As a contextual analysis, the micro-scale blood drop stains of patients infected with tuberculosis have been contrasted with the micro-scale blood drop stains of typical normal healthy people. The paper dives into the fundamental flow mechanics behind how the samples of the dried micro-scale blood drop stain is shaped. What has been found is a thick ring like feature in the dried micro-scale blood drop stains of non-ailing people and thin shape like lines in the dried micro-scale blood drop stains of patients with anaemia or tuberculosis disease. The ring like feature at the periphery is caused by an outward stream conveying suspended particles to the edge

  4. Comparison of S. stercoralis Serology Performed on Dried Blood Spots and on Conventional Serum Samples

    PubMed Central

    Formenti, Fabio; Buonfrate, Dora; Prandi, Rosanna; Marquez, Monica; Caicedo, Cintia; Rizzi, Eleonora; Guevara, Angel G.; Vicuña, Yosselin; Huerlo, Francisco R.; Perandin, Francesca; Bisoffi, Zeno; Anselmi, Mariella

    2016-01-01

    Background: Dried blood spots (DBS) are used for epidemiological surveys on infectious diseases in settings where limited resources are available. In fact, DBS can help to overcome logistic difficulties for the collection, transport and storage of biological specimens. Objective: To evaluate the accuracy of Strongyloides stercoralis serology performed on DBS. Methods: A survey was proposed to children attending a school in the village of Borbon, Ecuador, and to their parents/guardians. Each participant gave consent to the collection of both serum and DBS specimens. DBS absorbed on filter papers were analyzed with a commercially available ELISA test for S. stercoralis antibodies, as well as with standard serology. The agreement between the two methods was assessed through the Cohen’s kappa coefficient. Results: The study sample was composed of 174 children and 61 adults, for a total of 235 serum and 235 DBS samples. The serology was positive in 31/235 (13%) serum samples, and in 27/235 (11%) DBS: 4 samples resulted discordant (positive at standard serology). Cohen’s kappa coefficient was 0.921 (95% CI 0.845 – 0.998), indicating a high rate of concordance. Conclusion: DBS are suitable for in field-surveys requiring serological testing for S. stercoralis. PMID:27877170

  5. Cytokine assays: an assessment of the preparation and treatment of blood and tissue samples.

    PubMed

    Keustermans, Genoveva C E; Hoeks, Sanne B E; Meerding, Jenny M; Prakken, Berent J; de Jager, Wilco

    2013-05-15

    Cytokines are key components of the innate and adaptive immune system. As pivotal players in the progression or regression of a pathological process, these molecules provide a window through which diseases can be monitored and can thus act as biomarkers. In order to measure cytokine levels, a plethora of protocols can be applied. These methods include bioassays, protein microarrays, high-performance liquid chromatography (HPLC), sandwich enzyme-linked immunosorbent assay (ELISA), Meso Scale Discovery (MSD) electrochemiluminescence and bead based multiplex immunoassays (MIA). Due to the interaction and activity of cytokines, multiplex immunoassays are at the forefront of cytokine analysis by allowing multiple cytokines to be measured in parallel. However, even with optimized protocols, sample standardization needs to occur before these proteins can optimally act as biomarkers. This review describes various factors influencing the levels of cytokines measured in plasma, serum, dried blood spots and tissue biopsies, focusing on sample collection and handling, long term storage and the repetitive use of samples. By analyzing how each of these factors influences protein levels, it is concluded that samples should be stored at low temperatures in order to maintain cytokine stability. In addition, within a study, sample manipulations should be kept the same, with measurement protocols being chosen for their compatibility with the research in question. By having a clear understanding of what factors influence cytokine levels and how to overcome these technical issues, minimally confounded data can be obtained and cytokines can achieve optimal biomarker activity.

  6. Evaluation of current derivative spectrophotometric methodology for the determination of percent carboxyhemoglobin saturation in postmortem blood samples.

    PubMed

    Perrigo, B J; Joynt, B P

    1989-01-01

    Carbon monoxide intoxication continues to be a commonly encountered cause of death in most areas of Canada. The forensic nature of the samples in these cases presents special problems that are not normally encountered in clinical determinations. A study was undertaken to assess various methods of determining the percent carboxyhemoglobin saturation in blood, more specifically, those using derivative spectrophotometric measurements in the Soret region of the UV spectrum. At the same time, other studies were carried out: the effects of storage time on the carboxyhemoglobin levels; evaluation of sample containers; comparison of percent carboxyhemoglobin saturation in blood samples taken ante-mortem and post-mortem. Blood for the study was obtained from laboratory animals that were exposed to carbon monoxide before death.

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

    MedlinePlus

    National Cancer Institute How You Can Help Medical Research U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES National Institutes of Health Donating Your Blood, Tissue, and Other Samples You have the choice to ...

  8. The history of the microsphere method for measuring blood flows with special reference to myocardial blood flow: a personal memoir.

    PubMed

    Hoffman, Julien I E

    2017-04-01

    We use many types of equipment and technologies to make our measurements but give little thought to how they developed. Evolution was once described as a series of recoils from blind alleys, and this is exemplified by the gradual development of the microsphere method of measuring blood flows. The microsphere method is one of the most frequently used methods for measuring blood flow to organs and portions of organs. The method can measure myocardial blood flow with reasonable accuracy (within 10%) down to samples weighing >50 mg but probably will not do so for samples weighing 1-10 mg. Microspheres with diameters from 10 to 15 μm provide the best compromise between accurate flow measurement and retention in tissue. Radioactive labels have been almst entirely replaced by fluorescent labels, but colored microspheres and neutron-activated labels are also used.NEW & NOTEWORTHY The contributions of the various individuals who developed the microsphere method of measuring regional blood flows and how these advances took place are brought to light in this paper.

  9. Noninvasive method of estimating human newborn regional cerebral blood flow

    SciTech Connect

    Younkin, D.P.; Reivich, M.; Jaggi, J.; Obrist, W.; Delivoria-Papadopoulos, M.

    1982-12-01

    A noninvasive method of estimating regional cerebral blood flow (rCBF) in premature and full-term babies has been developed. Based on a modification of the /sup 133/Xe inhalation rCBF technique, this method uses eight extracranial NaI scintillation detectors and an i.v. bolus injection of /sup 133/Xe (approximately 0.5 mCi/kg). Arterial xenon concentration was estimated with an external chest detector. Cerebral blood flow was measured in 15 healthy, neurologically normal premature infants. Using Obrist's method of two-compartment analysis, normal values were calculated for flow in both compartments, relative weight and fractional flow in the first compartment (gray matter), initial slope of gray matter blood flow, mean cerebral blood flow, and initial slope index of mean cerebral blood flow. The application of this technique to newborns, its relative advantages, and its potential uses are discussed.

  10. Performance characteristics of the HemoCue B-Glucose analyzer using whole-blood samples.

    PubMed

    Voss, E M; Cembrowski, G S

    1993-07-01

    We evaluated the HemoCue B-Glucose (HemoCue Inc, Mission Viejo, Calif) analyzer for accuracy, precision, linearity, and recovery. One hundred eighteen capillary whole-blood samples were analyzed in duplicate on the HemoCue B-Glucose and the YSI 2300 STAT Glucose/L-Lactate (Yellow Springs [Ohio] Instruments) analyzers; corresponding plasma glucose levels were measured in duplicate on the Roche Cobas MIRA (Roche Diagnostic Systems, Nutley, NJ) analyzer. Plasma glucose levels were converted to whole-blood equivalent glucose levels by using a factor of 1.11. The following regression equations were obtained: HemoCue = 1.02 (YSI) + 0.19, Sy/x = 0.52, r2 = .984; and HemoCue = 0.98 (whole-blood equivalent glucose levels) + 0.26, Sy/x = 0.55, r2 = .982. Within-run coefficients of variation were 4.0%, 3.5%, 2.2%, and 1.0% at glucose concentrations of 3.9, 5.4, 8.7, and 17.1 mmol/L (71, 97, 156, and 308 mg/dL), respectively. Between-run imprecision and total imprecision using lyopholized materials with three lot numbers of cuvettes were 4.2% and 5.6% at 2.1 mmol/L (37 mg/dL) and 2.4% and 2.7% at 5.2 mmol/L (95 mg/dL), respectively. The HemoCue B-Glucose analyzer displayed linearity between 0 and 22.2 mmol/L (0 and 400 mg/dL), and the percent recovery averaged 98.7% +/- 4.5% (mean +/- SD). The HemoCue B-Glucose analyzer is a rapid, simple, and reliable method for determinations of whole-blood glucose levels.

  11. Evaluation of Sampling Methods for Bacillus Spore ...

    EPA Pesticide Factsheets

    Journal Article Following a wide area release of biological materials, mapping the extent of contamination is essential for orderly response and decontamination operations. HVAC filters process large volumes of air and therefore collect highly representative particulate samples in buildings. HVAC filter extraction may have great utility in rapidly estimating the extent of building contamination following a large-scale incident. However, until now, no studies have been conducted comparing the two most appropriate sampling approaches for HVAC filter materials: direct extraction and vacuum-based sampling.

  12. Enzymatic method for determining ketone body ratio in arterial blood.

    PubMed

    Uno, S; Takehiro, O; Tabata, R; Ozawa, K

    1995-12-01

    We have developed a new, sensitive, and rapid method for measuring the ketone body concentration in arterial blood and determining the arterial blood ketone body ratio. The procedure involves the sequential use of the enzymes 3-hydroxybutyrate dehydrogenase (3-HBDH; EC 1.1.1.30) and NADH oxidase, followed by a color-generating reaction with the hydrogen peroxide produced by the oxidase reaction. The amount of oxidized chromogen produced is proportional to the 3-hydroxybutyrate (3-HBA) concentration. The acetoacetate (AcAc) concentration is obtained after complete conversion of the AcAc to 3-HBA, in the presence of 3-HBDH. The total 3-HBA concentration is measured and then subtracted from the total ketone body concentration to give the AcAc concentration. This procedure may be applied to plasma samples and the absorbance change measured with an automated chemistry analyzer. Ketone body concentration may be determined over the range 0 to 400 mumol/L. The analysis takes approximately 12 min and requires only 30 microL of plasma.

  13. Dried Blood Spot Proteomics: Surface Extraction of Endogenous Proteins Coupled with Automated Sample Preparation and Mass Spectrometry Analysis

    NASA Astrophysics Data System (ADS)

    Martin, Nicholas J.; Bunch, Josephine; Cooper, Helen J.

    2013-08-01

    Dried blood spots offer many advantages as a sample format including ease and safety of transport and handling. To date, the majority of mass spectrometry analyses of dried blood spots have focused on small molecules or hemoglobin. However, dried blood spots are a potentially rich source of protein biomarkers, an area that has been overlooked. To address this issue, we have applied an untargeted bottom-up proteomics approach to the analysis of dried blood spots. We present an automated and integrated method for extraction of endogenous proteins from the surface of dried blood spots and sample preparation via trypsin digestion by use of the Advion Biosciences Triversa Nanomate robotic platform. Liquid chromatography tandem mass spectrometry of the resulting digests enabled identification of 120 proteins from a single dried blood spot. The proteins identified cross a concentration range of four orders of magnitude. The method is evaluated and the results discussed in terms of the proteins identified and their potential use as biomarkers in screening programs.

  14. Postmortem measurement of caffeine in bone marrow: influence of sample location and correlation with blood concentration.

    PubMed

    Cartiser, N; Bévalot, F; Chatenay, C; Le Meur, C; Gaillard, Y; Malicier, D; Guitton, J; Fanton, L

    2011-07-15

    Bone marrow (BM) analysis is of forensic interest in postmortem toxicological investigation in case of limited, unavailable or unusable blood samples. However, it remains difficult to determine whether a drug BM concentration is therapeutic or represents overdose, due to the lack of studies on this alternative matrix. Given the variations in BM composition in the body, sample location was suggested to be a relevant factor in assessing BM concentration. The aim of the present study was to compare postmortem caffeine concentrations in various BM sample locations and secondly to consider the correlation between BM and blood concentrations. Six BM samples (right and left side: proximal and medial femur and 5th rib) and a blood sample were collected from 21 forensic autopsies. Gas chromatography coupled to tandem mass spectrometry was performed. Blood caffeine concentrations ranged from 60 to 7591ng/mL. Femoral and rib BM concentrations ranged from 51 to 6171ng/g and 66 to 7280ng/g, respectively. Blood concentrations were always higher than BM concentrations. As a good correlation was demonstrated between blood and rib BM and between blood and the average of the four femoral BM concentrations, blood caffeine concentrations could be correctly extrapolated from BM concentrations. BM caffeine concentration was found to depend on sample location. Rib BM caffeine concentrations appeared to be systematically greater than averaged femur values and concentrations were much more variable between the 4 femur BM samples than between the 2 ribs. From a practical point of view, for caffeine analysis, rib BM appeared more relevant than femoral BM, which requires multisampling to overcome the concentration variability problem.

  15. Heat stabilization of blood spot samples for determination of metabolically unstable drug compounds

    PubMed Central

    Blessborn, Daniel; Sköld, Karl; Zeeberg, David; Kaewkhao, Karnrawee; Sköld, Olof; Ahnoff, Martin

    2014-01-01

    Background Sample stability is critical for accurate analysis of drug compounds in biosamples. The use of additives to eradicate the enzymatic activity causing loss of these analytes has its limitations. Results A novel technique for sample stabilization by rapid, high-temperature heating was used. The stability of six commercial drugs in blood and blood spots was investigated under various conditions with or without heat stabilization at 95°C. Oseltamivir, cefotaxime and ribavirin were successfully stabilized by heating whereas significant losses were seen in unheated samples. Amodiaquine was stable with and without heating. Artemether and dihydroartemisinin were found to be very heat sensitive and began to decompose even at 60°C. Conclusion Heat stabilization is a viable technique to maintain analytes in blood spot samples, without the use of chemical additives, by stopping the enzymatic activity that causes sample degradation. PMID:23256470

  16. Use of dried blood samples for monitoring hepatitis B virus infection

    PubMed Central

    Lira, Rosalia; Maldonado-Rodriguez, Angelica; Rojas-Montes, Othon; Ruiz-Tachiquin, Martha; Torres-Ibarra, Rocio; Cano-Dominguez, Carlos; Valdez-Salazar, Hilda; Gomez-Delgado, Alejandro; Muñoz, Onofre; Alvarez-Muñoz, Ma-Teresa

    2009-01-01

    Background Hepatitis B virus (HBV) infection is a problem in several regions of the world with limited resources. Blood samples dried on filter paper (DBS) have been successfully used to diagnose and monitor several infectious diseases. In Mexico there is an urgent need for an affordable and easy sampling method for viral load (VL) testing and monitoring of chronic HBV infection. The purpose of this work was to validate the utility of DBS samples for monitoring HBV infection in patients from Mexico City. Methods Matched samples of plasma and DBS on filter paper from 47 HBV infected patients from the Instituto Mexicano del Seguro Social (IMSS), were included. To evaluate the DNA stability and purity from DBS stored at different temperature conditions, samples from ten patients were stored at 4 degree, 25 degree, and 37 degree C for 7 days. After DBS elution and DNA extraction, the purity of these samples was determined measuring the O.D. rate 260/280. The DBS utility for molecular studies was assessed with PCR assays to amplify a 322 bp fragment from the "a" determinant region of the HBV "S" gene. The VL from all samples was determined to evaluate the correlation between plasma and DBS matched samples. Results The quality of the DNA from DBS specimen is not adversely affected by storage at 4 degree, 25 degree and 37 degree C for up 7 days. Statistical ANOVA analyses did not show any significant difference. The same amplification efficiency was observed between DNA templates from samples stored at different temperatures. The Pearson correlation between the VL from DBS and plasma matched samples was 0.93 (p = 0.01). The SD was 1.48 for DBS vs.1.32 for Plasma, and an average of log10 copies/mL of 5.32 vs. 5.53. ANOVA analysis did not show any statistically significant difference between the analyzed groups (p = 0.92). Conclusion The results provide strong evidence that the isolation and quantification of DNA-HBV from DBS is a viable alternative for patient monitoring

  17. Sulfatide Analysis by Mass Spectrometry for Screening of Metachromatic Leukodystrophy in Dried Blood and Urine Samples

    PubMed Central

    Spacil, Zdenek; Kumar, Arun Babu; Liao, Hsuan-Chieh; Auray-Blais, Christiane; Stark, Samantha; Suhr, Teryn R.; Scott, C. Ronald; Turecek, Frantisek; Gelb, Michael H.

    2016-01-01

    BACKGROUND Metachromatic leukodystrophy (MLD) is an autosomal recessive disorder caused by deficiency in arylsulfatase A activity, leading to accumulation of sulfatide substrates. Diagnostic and monitoring procedures include demonstration of reduced arylsulfatase A activity in peripheral blood leukocytes or detection of sulfatides in urine. However, the development of a screening test is challenging because of instability of the enzyme in dried blood spots (DBS), the widespread occurrence of pseudodeficiency alleles, and the lack of available urine samples from newborn screening programs. METHODS We measured individual sulfatide profiles in DBS and dried urine spots (DUS) from MLD patients with LC-MS/MS to identify markers with the discriminatory power to differentiate affected individuals from controls. We also developed a method for converting all sulfatide molecular species into a single species, allowing quantification in positive-ion mode upon derivatization. RESULTS In DBS from MLD patients, we found up to 23.2-fold and 5.1-fold differences in total sulfatide concentrations for early- and late-onset MLD, respectively, compared with controls and pseudodeficiencies. Corresponding DUS revealed up to 164-fold and 78-fold differences for early- and late-onset MLD patient samples compared with controls. The use of sulfatides converted to a single species simplified the analysis and increased detection sensitivity in positive-ion mode, providing a second option for sulfatide analysis. CONCLUSIONS This study of sulfatides in DBS and DUS suggests the feasibility of the mass spectrometry method for newborn screening of MLD and sets the stage for a larger-scale newborn screening pilot study. PMID:26585924

  18. Diagnosis of basal cell carcinoma by infrared spectroscopy of whole blood samples applying soft independent modeling class analogy.

    PubMed

    Khanmohammadi, Mohammadreza; Nasiri, Razieh; Ghasemi, Keyvan; Samani, Simin; Bagheri Garmarudi, Amir

    2007-12-01

    Attenuated total reflectance-Fourier transform infrared (ATR-FTIR) spectroscopy was applied to discriminate the blood samples obtained from healthy people and those with basal cell carcinoma, demonstrating high accuracy while soft independent modeling class analogy (SIMCA) chemometric technique is benefited. It was aimed to classify the normal case and cancer case blood samples through the use of ATR-FTIR spectroscopy as a rapid method while the sample preparation is so easy in comparison with the common pathologic methods. A total of 72 blood samples, including 32 cancer and 40 normal cases, were analyzed in 1,800-900 cm(-1) spectral region. Results showed 97.6% of accuracy being compared with the current clinical methods. Research results were exemplified with comparable data of other classification methods such as principal component analysis (PCA) and Cluster analysis. The residual errors in prediction (REP) of calibration model for normal and cancerous groups in SIMCA method were 0.00362 and 0.00343, respectively.

  19. System and method for extracting a sample from a surface

    DOEpatents

    Van Berkel, Gary; Covey, Thomas

    2015-06-23

    A system and method is disclosed for extracting a sample from a sample surface. A sample is provided and a sample surface receives the sample which is deposited on the sample surface. A hydrophobic material is applied to the sample surface, and one or more devices are configured to dispense a liquid on the sample, the liquid dissolving the sample to form a dissolved sample material, and the one or more devices are configured to extract the dissolved sample material from the sample surface.

  20. In search of blood--detection of minute particles using spectroscopic methods.

    PubMed

    De Wael, K; Lepot, L; Gason, F; Gilbert, B

    2008-08-25

    An examination protocol for rapid detection of remnants of blood particles on garments of suspects in bloody murder cases is described. Microparticles of blood are sampled along with fibres and hairs using the tape lifting method. The tapings are searched with a low power microscope for red particles with morphology similar to blood. Presumed blood traces are further examined using microspectrophotometry on the cut out piece of taping. The typical visible spectrum of haemoglobin is characteristic for blood. Alternatively Raman spectroscopy can be used to measure the characteristic vibrational spectrum of haemoglobin. At a later stage, these particles may be removed from the piece of taping in order to extract the blood and attempt to obtain a genetic profile.

  1. Development of a Modular Assay for Detailed Immunophenotyping of Peripheral Human Whole Blood Samples by Multicolor Flow Cytometry

    PubMed Central

    Rühle, Paul F.; Fietkau, Rainer; Gaipl, Udo S.; Frey, Benjamin

    2016-01-01

    The monitoring of immune cells gained great significance in prognosis and prediction of therapy responses. For analyzing blood samples, the multicolor flow cytometry has become the method of choice as it combines high specificity on single cell level with multiple parameters and high throughput. Here, we present a modular assay for the detailed immunophenotyping of blood (DIoB) that was optimized for an easy and direct application in whole blood samples. The DIoB assay characterizes 34 immune cell subsets that circulate the peripheral blood including all major immune cells such as T cells, B cells, natural killer (NK) cells, monocytes, dendritic cells (DCs), neutrophils, eosinophils, and basophils. In addition, it evaluates their functional state and a few non-leukocytes that also have been associated with the outcome of cancer therapy. This DIoB assay allows a longitudinal and close-meshed monitoring of a detailed immune status in patients requiring only 2.0 mL of peripheral blood and it is not restricted to peripheral blood mononuclear cells. It is currently applied for the immune monitoring of patients with glioblastoma multiforme (IMMO-GLIO-01 trial, NCT02022384), pancreatic cancer (CONKO-007 trial, NCT01827553), and head and neck cancer (DIREKHT trial, NCT02528955) and might pave the way for immune biomarker identification for prediction and prognosis of therapy outcome. PMID:27529227

  2. Exploration and Sampling Methods for Borrow Areas

    DTIC Science & Technology

    1990-12-01

    The principal geomorphic features associated with stratified deposits are kames, eskers , and proglacial outwash material. 60. Kames are moundlike...deposits without obtaining core samples or geophysical data. 61. Eskers are sinuous ridgelike features that form in channels, tun- nels, and crevasses in...the ice by meltwater transport and deposition of gla- cial material. Eskers are variable in dimensions but, at their largest, can be up to 100 m high

  3. Lead and cadmium levels in blood samples from the general population of Sweden

    SciTech Connect

    Elinder, C.G.; Friberg, L.; Lind, B.; Jawaid, M.

    1983-02-01

    Lead and cadmium was determined in whole blood samples obtained from 473 nonoccupationally exposed adult persons in Sweden in 1980. Analyses were performed using atomic absorption spectrophotometry equipped with an electrothermal atomization unit. Blood lead concentrations were shown to be significantly influenced by sex, smoking habits, and alcohol consumption. Current male smokers had a median blood lead level of 92 ..mu..g Pb/liter, as compared to 77 ..mu..g Pb/liter for nonsmokers. For females the corresponding values were 69 ..mu..g Pb/liter and 57 ..mu..g Pb/liter for current smokers and nonsmokers, respectively. Highly significant correlations were found between stated alcohol consumption and blood lead in most of the different sex and smoking categories. People living in apartments close to streets with heavy traffic in Stockholm had slightly, but not significantly, higher blood lead levels when compared to people living in areas of this city with low traffic density. Blood cadmium levels were very strongly affected by smoking habits. A significant correlation existed between the number of cigarettes consumed daily and blood cadmium concentration. The median blood cadmium level for nonsmoking males was 0.2 ..mu..g Cd/liter (less than or equal to0.2, detection limit) and for females 0.3 ..mu..g Cd/liter. About 90% of the current male and female smokers had cadmium concentrations in blood of 0.6 ..mu..g Cd/liter or more.

  4. Blood viscosity measurement: an integral method using Doppler ultrasonic profiles

    NASA Astrophysics Data System (ADS)

    Flaud, P.; Bensalah, A.

    2005-12-01

    The aim of this work is to present a new indirect and noninvasive method for the measurement of the Newtonian blood viscosity. Based on an integral form of the axial Navier-Stokes equation, this method is particularly suited for in vivo investigations using ultrasonic arterial blood velocity profiles. Its main advantage is that it is applicable to periodic as well as non periodic flows. Moreover it does not require classical filtering methods enhancing signal to noise ratio of the physiological signals. This method only requires the knowledge of the velocimetric data measured inside a spatially and temporally optimized zone of the Doppler velocity profiles. The results obtained using numerical simulation as well as in vitro or in vivo experiments prove the effectiveness of the method. It is then well adapted to the clinical environment as a systematic quasi on-line method for the measurement of the blood viscosity.

  5. A spectral and morphologic method for white blood cell classification

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Chang, Li; Zhou, Mei; Li, Qingli; Liu, Hongying; Guo, Fangmin

    2016-10-01

    The identification of white blood cells is important as it provides an assay for diagnosis of various diseases. To overcome the complexity and inaccuracy of traditional methods based on light microscopy, we proposed a spectral and morphologic method based on hyperspectral blood images. We applied mathematical morphology-based methods to extract spatial information and supervised method is employed for spectral analysis. Experimental results show that white blood cells could be segmented and classified into five types with an overall accuracy of more than 90%. Moreover, the experiments including spectral features reached higher accuracy than the spatial-only cases, with a maximum improvement of nearly 20%. By combing both spatial and spectral features, the proposed method provides higher classification accuracy than traditional methods.

  6. Enhanced Stability of Blood Matrices Using a Dried Sample Spot Assay to Measure Human Butyrylcholinesterase Activity and Nerve Agent Adducts

    PubMed Central

    Perez, Jonas W.; Pantazides, Brooke G.; Watson, Caroline M.; Thomas, Jerry D.; Blake, Thomas A.; Johnson, Rudolph C.

    2015-01-01

    Dried matrix spots are safer to handle and easier to store than wet blood products, but factors such as intra-spot variability and unknown sample volumes have limited their appeal as a sampling format for quantitative analyses. In this work, we introduce a dried spot activity assay for quantifying butyrylcholinesterase (BChE) specific activity which is BChE activity normalized to the total protein content in a sample spot. The method was demonstrated with blood, serum, and plasma spotted on specimen collection devices (cards) which were extracted to measure total protein and BChE activity using a modified Ellman assay. Activity recovered from dried spots was ∼80% of the initial spotted activity for blood and >90% for plasma and serum. Measuring total protein in the sample and calculating specific activity substantially improved quantification and reduced intra-spot variability. Analyte stability of nerve agent adducts was also evaluated, and the results obtained via BChE-specific activity measurements were confirmed by quantification of BChE adducts using a previously established LC-MS/MS method. The spotted samples were up to 10-times more resistant to degradation compared to unspotted control samples when measuring BChE inhibition by the nerve agents sarin and VX. Using this method, both BChE activity and adducts can be accurately measured from a dried sample spot. This use of a dried sample spot with normalization to total protein is robust, demonstrates decreased intra-spot variability without the need to control for initial sample volume, and enhances analyte stability. PMID:25955132

  7. Enhanced Detection of Rift Valley Fever Virus using Molecular Assays on Whole Blood Samples

    PubMed Central

    Grolla, Allen; Mehedi, Masfique; Lindsay, Robbin; Bosio, Catharine; Duse, Adriano; Feldmann, Heinz

    2012-01-01

    Background Rift Valley fever (RVF) is an emerging arthropod-borne zoonoses of global agricultural and public health importance. In December 2006, an RVF outbreak was recognized in Kenya which led to the deployment of international response laboratory teams to the area. Objectives A field laboratory was operated in Malindi, Kenya to provide safe sample handling and molecular testing for RVF virus (RVFV) as well as selected other pathogens for differential diagnosis. Study Design Safe sample handling was carried out using a negative pressure flexible film isolator (glovebox) and commercial reagents to inactivate clinical specimens and purify nucleic acid. Whole blood was routinely used for diagnostic testing although paired plasma samples were also tested in select cases. Subsequently, human macrophages were tested in vitro for their susceptibility to RVFV. Results The field laboratory received samples from 33 individuals and a definite laboratory diagnosis was provided in 16 of these cases. Using molecular diagnostic techniques, RVFV was more consistently detected in whole blood than in plasma samples most likely due to association of RVFV with blood cells. Subsequent in vitro studies identified macrophages as a target cell for RVFV replication. Conclusions RVFV appears to replicate in blood cells such as macrophages. Thus, the sensitivity of molecular diagnostic testing is improved if whole blood is used as the clinical specimen rather than plasma or serum. PMID:22632901

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

    PubMed Central

    Seemann, Tine Lindberg; Nybo, Mads

    2016-01-01

    Introduction An observational study was conducted using a structured observation scheme to assess compliance with the local phlebotomy guideline, to identify necessary focus items, and to investigate whether adherence to the phlebotomy guideline improved. Materials and methods The questionnaire from the EFLM Working Group for the Preanalytical Phase was adapted to local procedures. A pilot study of three months duration was conducted. Based on this, corrective actions were implemented and a follow-up study was conducted. All phlebotomists at the Department of Clinical Biochemistry and Pharmacology were observed. Three blood collections by each phlebotomist were observed at each session conducted at the phlebotomy ward and the hospital wards, respectively. Error frequencies were calculated for the phlebotomy ward and the hospital wards and for the two study phases. Results A total of 126 blood drawings by 39 phlebotomists were observed in the pilot study, while 84 blood drawings by 34 phlebotomists were observed in the follow-up study. In the pilot study, the three major error items were hand hygiene (42% error), mixing of samples (22%), and order of draw (21%). Minor significant differences were found between the two settings. After focus on the major aspects, the follow-up study showed significant improvement for all three items at both settings (P < 0.01, P < 0.01, and P = 0.01, respectively). Conclusion Continuous quality control of the phlebotomy procedure revealed a number of items not conducted in compliance with the local phlebotomy guideline. It supported significant improvements in the adherence to the recommended phlebotomy procedures and facilitated documentation of the phlebotomy quality. PMID:27812302

  9. The Optimization of Molecular Detection of Clinical Isolates of Brucella in Blood Cultures by eryD Transcriptase Gene for Confirmation of Culture-Negative Samples

    PubMed Central

    Tabibnejad, Mahsa; Alikhani, Mohammad Yousef; Arjomandzadegan, Mohammad; Hashemi, Seyed Hamid; Naseri, Zahra

    2016-01-01

    Background Brucellosis is a zoonosis disease which is widespread across the world. Objectives The aim of the present study is the evaluation of culture-negative blood samples. Materials and Methods A total of 100 patients with suspected brucellosis were included in this experimental study and given positive serological tests. Diagnosis was performed on patients with clinical symptoms of the disease, followed by the detection of a titer that was equal to or more than 1:160 (in endemic areas) by the standard tube agglutination method. Blood samples were cultured by a BACTEC 9050 system, and subsequently by Brucella agar. At the same time, DNA from all blood samples was extracted by Qiagen Kit Company (Qia Amp Mini Kit). A molecular assay of blood samples was carried out by detection of eryD transcriptase and bcsp 31 genes in specific double PCR reactions. The specificity of the primers was evaluated by DNA from pure and approved Brucella colonies found in the blood samples, by DNA from other bacteria, and by ordinary PCR. DNA extraction from the pure colonies was carried out by both Qiagen Kit and Chelex 100 methods; the two were compared. Results 39 cases (39%) had positive results when tested by the BACTEC system, and 61 cases (61%) became negative. 23 culture-positive blood samples were randomly selected for PCR reactions; all showed 491 bp for the eryD gene and 223 bp for the bcsp 31 gene. Interestingly, out of 14 culture-negative blood samples, 13 cases showed positive bonds in PCR. The specificity of the PCR method was equal to 100%. DNA extraction from pure cultures was done by both Chelex 100 and Qiagen Kit; these showed the same results for all samples. Conclusions The results prove that the presented double PCR method could be used to detect positive cases from culture-negative blood samples. The Chelex 100 method is simpler and safer than the use of Qiagen Kit for DNA extraction. PMID:27330831

  10. A method for sampling waste corn

    USGS Publications Warehouse

    Frederick, R.B.; Klaas, E.E.; Baldassarre, G.A.; Reinecke, K.J.

    1984-01-01

    Corn had become one of the most important wildlife food in the United States. It is eaten by a wide variety of animals, including white-tailed deer (Odocoileus virginianus ), raccoon (Procyon lotor ), ring-necked pheasant (Phasianus colchicus , wild turkey (Meleagris gallopavo ), and many species of aquatic birds. Damage to unharvested crops had been documented, but many birds and mammals eat waste grain after harvest and do not conflict with agriculture. A good method for measuring waste-corn availability can be essential to studies concerning food density and food and feeding habits of field-feeding wildlife. Previous methods were developed primarily for approximating losses due to harvest machinery. In this paper, a method is described for estimating the amount of waste corn potentially available to wildlife. Detection of temporal changes in food availability and differences caused by agricultural operations (e.g., recently harvested stubble fields vs. plowed fields) are discussed.

  11. Rapid Detection of Candida albicans by Polymerase Spiral Reaction Assay in Clinical Blood Samples

    PubMed Central

    Jiang, Xiaoqun; Dong, Derong; Bian, Lihong; Zou, Dayang; He, Xiaoming; Ao, Da; Yang, Zhan; Huang, Simo; Liu, Ningwei; Liu, Wei; Huang, Liuyu

    2016-01-01

    Candida albicans is the most common human yeast pathogen which causes mucosal infections and invasive fungal diseases. Early detection of this pathogen is needed to guide preventative and therapeutic treatment. The aim of this study was to establish a polymerase spiral reaction (PSR) assay that rapidly and accurately detects C. albicans and to assess the clinical applicability of PSR-based diagnostic testing. Internal transcribed spacer 2 (ITS2), a region between 5.8S and 28S fungal ribosomal DNA, was used as the target sequence. Four primers were designed for amplification of ITS2 with the PSR method, which was evaluated using real time turbidity monitoring and visual detection using a pH indicator. Fourteen non-C. albicans yeast strains were negative for detection, which indicated the specificity of PSR assay was 100%. A 10-fold serial dilution of C. albicans genomic DNA was subjected to PSR and conventional polimerase chain reaction (PCR) to compare their sensitivities. The detection limit of PSR was 6.9 pg/μl within 1 h, 10-fold higher than that of PCR (69.0 pg/μl). Blood samples (n = 122) were collected from intensive care unit and hematological patients with proven or suspected C. albicans infection at two hospitals in Beijing, China. Both PSR assay and the culture method were used to analyze the samples. Of the 122 clinical samples, 34 were identified as positive by PSR. The result was consistent with those obtained by the culture method. In conclusion, a novel and effective C. albicans detection assay was developed that has a great potential for clinical screening and point-of-care testing. PMID:27379048

  12. Detection of canine distemper virus in blood samples by reverse transcription loop-mediated isothermal amplification.

    PubMed

    Cho, H S; Park, N Y

    2005-11-01

    Reverse transcription loop-mediated isothermal amplification (RT-LAMP) was used to detect canine distemper virus (CDV) genomic RNA. A set of four primers, two outer and two inner, were designed from CDV genomic RNA targeting the nucleocapsid protein gene. The optimal reaction time and temperature for LAMP were determined to be 60 min at 65 degrees C. The relative sensitivity and specificity of RT-LAMP was found to be 100% and 93.3%, respectively, based on 50 canine blood samples and using RT-PCR as the gold standard. The detection limit of the RT-LAMP method was 100 times lower than with RT-PCR (10-1TCID50 ml(-1) versus 10TCID50 ml(-1)). In addition to the advantage resulting from the visual detection of the end-product, the LAMP method is fast, requiring only 1 h to complete the assay. The LAMP method is a viable alternative to RT-PCR for diagnosing CDV infection in dogs. The LAMP method might be useful as an on site diagnostic assay for detecting CDV.

  13. Blood grouping based on PCR methods and agarose gel electrophoresis.

    PubMed

    Sell, Ana Maria; Visentainer, Jeane Eliete Laguila

    2015-01-01

    The study of erythrocyte antigens continues to be an intense field of research, particularly after the development of molecular testing methods. More than 300 specificities have been described by the International Society for Blood Transfusion as belonging to 33 blood group systems. The polymerase chain reaction (PCR) is a central tool for red blood cells (RBC) genotyping. PCR and agarose gel electrophoresis are low cost, easy, and versatile in vitro methods for amplifying defined target DNA (RBC polymorphic region). Multiplex-PCR, AS-PCR (Specific Allele Polymerase Chain Reaction), and RFLP-PCR (Restriction Fragment Length Polymorphism-Polymerase Chain Reaction) techniques are usually to identify RBC polymorphisms. Furthermore, it is an easy methodology to implement. This chapter describes the PCR methodology and agarose gel electrophoresis to identify the polymorphisms of the Kell, Duffy, Kidd, and MNS blood group systems.

  14. Evaluation of the Secretor Status of ABO Blood Group Antigens in Saliva among Southern Rajasthan Population Using Absorption Inhibition Method

    PubMed Central

    Khajuria, Nidhi; Mamta; Ramesh, Gayathri

    2016-01-01

    Introduction The ABO blood group system was the significant element for forensic serological examination of blood and body fluids in the past before the wide adaptation of DNA typing. A significant proportion of individuals (80%) are secretors, meaning that antigens present in the blood are also found in other body fluids such as saliva. Absorption inhibition is one such method that works by reducing strength of an antiserum based on type and amount of antigen present in the stains. Aim To check the efficacy of identifying the blood group antigens in saliva and to know the secretor status using absorption inhibition method among southern Rajasthan population. Materials and Methods Blood and saliva samples were collected from 80 individuals comprising 20 individuals in each blood group. The absorption inhibition method was used to determine the blood group antigens in the saliva and then the results were correlated with the blood group of the collected blood sample. The compiled data was statistically analysed using chi-square test. Results Blood groups A & O revealed 100% secretor status for both males and females. While blood groups B and AB revealed 95% secretor status. Conclusion Secretor status evaluation of the ABO blood group antigen in saliva using absorption inhibition method can be a useful tool in forensic examination. PMID:27042574

  15. Modified electrokinetic sample injection method in chromatography and electrophoresis analysis

    DOEpatents

    Davidson, J. Courtney; Balch, Joseph W.

    2001-01-01

    A sample injection method for horizontal configured multiple chromatography or electrophoresis units, each containing a number of separation/analysis channels, that enables efficient introduction of analyte samples. This method for loading when taken in conjunction with horizontal microchannels allows much reduced sample volumes and a means of sample stacking to greatly reduce the concentration of the sample. This reduction in the amount of sample can lead to great cost savings in sample preparation, particularly in massively parallel applications such as DNA sequencing. The essence of this method is in preparation of the input of the separation channel, the physical sample introduction, and subsequent removal of excess material. By this method, sample volumes of 100 nanoliter to 2 microliters have been used successfully, compared to the typical 5 microliters of sample required by the prior separation/analysis method.

  16. Genotyping of celiac disease-related-risk haplotypes using a closed-tube polymerase chain reaction analysis of dried blood and saliva disk samples.

    PubMed

    Ollikka, Pia; Raussi, Hanna-Mari; Laitala, Ville; Jaakkola, Lassi; Hovinen, Jari; Hemmilä, Ilkka; Ylikoski, Alice

    2009-03-01

    Expansion of molecular diagnostics more widely into clinical routines requires simplified methods allowing automation. We developed a homogeneous, multilabel polymerase chain reaction (PCR) method based on time-resolved fluorometry, and studied the use of dried disk samples in PCR. Celiac disease-related HLA-DQA1*05, HLA-DQB1*02, and HLA-DQB1*0302 genotyping was used to verify the method with blood and saliva samples dried on S&S 903 and IsoCode sample collection papers. Three sample preparation procedures, including manufacturer's manual elution, an automated elution, and direct use of disk samples, were compared using dried disk samples. The three procedures gave successful amplification and correct genotyping results. Owing to the simplicity of the direct use of disk samples in PCR, this method was chosen for the subsequent homogeneous analysis of blood (n=194) and saliva (n=30) disk samples on S&S 903 paper. The results revealed that, in addition to DNA samples (n=29), both blood and saliva disk samples were successfully amplified and genotyped using the homogeneous PCR assays for HLA-DQA1 and HLA-DQB1. The homogeneous PCR assays developed provide a useful tool to genotype celiac disease-related HLA-DQA1*05, HLA-DQB1*02, and HLA-DQB1*0302 alleles. Furthermore, the method provides a direct way to perform a closed-tube PCR analysis of dried blood and saliva disk samples enabling simple automation.

  17. Surface Sampling Methods for Bacillus anthracis Spore Contamination

    PubMed Central

    Hein, Misty J.; Taylor, Lauralynn; Curwin, Brian D.; Kinnes, Gregory M.; Seitz, Teresa A.; Popovic, Tanja; Holmes, Harvey T.; Kellum, Molly E.; McAllister, Sigrid K.; Whaley, David N.; Tupin, Edward A.; Walker, Timothy; Freed, Jennifer A.; Small, Dorothy S.; Klusaritz, Brian; Bridges, John H.

    2002-01-01

    During an investigation conducted December 17–20, 2001, we collected environmental samples from a U.S. postal facility in Washington, D.C., known to be extensively contaminated with Bacillus anthracis spores. Because methods for collecting and analyzing B. anthracis spores have not yet been validated, our objective was to compare the relative effectiveness of sampling methods used for collecting spores from contaminated surfaces. Comparison of wipe, wet and dry swab, and HEPA vacuum sock samples on nonporous surfaces indicated good agreement between results with HEPA vacuum and wipe samples. However, results from HEPA vacuum sock and wipe samples agreed poorly with the swab samples. Dry swabs failed to detect spores >75% of the time they were detected by wipe and HEPA vacuum samples. Wipe samples collected after HEPA vacuum samples and HEPA vacuum samples after wipe samples indicated that neither method completely removed spores from the sampled surfaces. PMID:12396930

  18. Identifying the potential of changes to blood sample logistics using simulation.

    PubMed

    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.

  19. Comparison of genomic DNA extraction techniques from whole blood samples: a time, cost and quality evaluation study.

    PubMed

    Chacon-Cortes, Diego; Haupt, Larisa M; Lea, Rod A; Griffiths, Lyn R

    2012-05-01

    Genomic DNA obtained from patient whole blood samples is a key element for genomic research. Advantages and disadvantages, in terms of time-efficiency, cost-effectiveness and laboratory requirements, of procedures available to isolate nucleic acids need to be considered before choosing any particular method. These characteristics have not been fully evaluated for some laboratory techniques, such as the salting out method for DNA extraction, which has been excluded from comparison in different studies published to date. We compared three different protocols (a traditional salting out method, a modified salting out method and a commercially available kit method) to determine the most cost-effective and time-efficient method to extract DNA. We extracted genomic DNA from whole blood samples obtained from breast cancer patient volunteers and compared the results of the product obtained in terms of quantity (concentration of DNA extracted and DNA obtained per ml of blood used) and quality (260/280 ratio and polymerase chain reaction product amplification) of the obtained yield. On average, all three methods showed no statistically significant differences between the final result, but when we accounted for time and cost derived for each method, they showed very significant differences. The modified salting out method resulted in a seven- and twofold reduction in cost compared to the commercial kit and traditional salting out method, respectively and reduced time from 3 days to 1 hour compared to the traditional salting out method. This highlights a modified salting out method as a suitable choice to be used in laboratories and research centres, particularly when dealing with a large number of samples.

  20. 7 CFR 58.245 - Method of sample analysis.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Method of sample analysis. 58.245 Section 58.245... Procedures § 58.245 Method of sample analysis. Samples shall be tested according to the applicable methods of laboratory analysis contained in either DA Instruction 918-RL as issued by the USDA, Agricultural...

  1. 7 CFR 58.812 - Methods of sample analysis.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 Agriculture 3 2013-01-01 2013-01-01 false Methods of sample analysis. 58.812 Section 58.812... Procedures § 58.812 Methods of sample analysis. Samples shall be tested according to the applicable methods of laboratory analysis contained in either DA Instruction 918-RL, as issued by the USDA,...

  2. 7 CFR 58.812 - Methods of sample analysis.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Methods of sample analysis. 58.812 Section 58.812... Procedures § 58.812 Methods of sample analysis. Samples shall be tested according to the applicable methods of laboratory analysis contained in either DA Instruction 918-RL, as issued by the USDA,...

  3. 7 CFR 58.245 - Method of sample analysis.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Method of sample analysis. 58.245 Section 58.245... Procedures § 58.245 Method of sample analysis. Samples shall be tested according to the applicable methods of laboratory analysis contained in either DA Instruction 918-RL as issued by the USDA, Agricultural...

  4. Microfluidic DNA sample preparation method and device

    DOEpatents

    Krulevitch, Peter A.; Miles, Robin R.; Wang, Xiao-Bo; Mariella, Raymond P.; Gascoyne, Peter R. C.; Balch, Joseph W.

    2002-01-01

    Manipulation of DNA molecules in solution has become an essential aspect of genetic analyses used for biomedical assays, the identification of hazardous bacterial agents, and in decoding the human genome. Currently, most of the steps involved in preparing a DNA sample for analysis are performed manually and are time, labor, and equipment intensive. These steps include extraction of the DNA from spores or cells, separation of the DNA from other particles and molecules in the solution (e.g. dust, smoke, cell/spore debris, and proteins), and separation of the DNA itself into strands of specific lengths. Dielectrophoresis (DEP), a phenomenon whereby polarizable particles move in response to a gradient in electric field, can be used to manipulate and separate DNA in an automated fashion, considerably reducing the time and expense involved in DNA analyses, as well as allowing for the miniaturization of DNA analysis instruments. These applications include direct transport of DNA, trapping of DNA to allow for its separation from other particles or molecules in the solution, and the separation of DNA into strands of varying lengths.

  5. Biomarkers for Monitoring Pre-Analytical Quality Variation of mRNA in Blood Samples

    PubMed Central

    Zhang, Hui; Korenková, Vlasta; Sjöback, Robert; Švec, David; Björkman, Jens; Kruhøffer, Mogens; Verderio, Paolo; Pizzamiglio, Sara; Ciniselli, Chiara Maura; Wyrich, Ralf; Oelmueller, Uwe; Kubista, Mikael; Lindahl, Torbjørn; Lönneborg, Anders; Rian, Edith

    2014-01-01

    There is an increasing need for proper quality control tools in the pre-analytical phase of the molecular diagnostic workflow. The aim of the present study was to identify biomarkers for monitoring pre-analytical mRNA quality variations in two different types of blood collection tubes, K2EDTA (EDTA) tubes and PAXgene Blood RNA Tubes (PAXgene tubes). These tubes are extensively used both in the diagnostic setting as well as for research biobank samples. Blood specimens collected in the two different blood collection tubes were stored for varying times at different temperatures, and microarray analysis was performed on resultant extracted RNA. A large set of potential mRNA quality biomarkers for monitoring post-phlebotomy gene expression changes and mRNA degradation in blood was identified. qPCR assays for the potential biomarkers and a set of relevant reference genes were generated and used to pre-validate a sub-set of the selected biomarkers. The assay precision of the potential qPCR based biomarkers was determined, and a final validation of the selected quality biomarkers using the developed qPCR assays and blood samples from 60 healthy additional subjects was performed. In total, four mRNA quality biomarkers (USP32, LMNA, FOSB, TNRFSF10C) were successfully validated. We suggest here the use of these blood mRNA quality biomarkers for validating an experimental pre-analytical workflow. These biomarkers were further evaluated in the 2nd ring trial of the SPIDIA-RNA Program which demonstrated that these biomarkers can be used as quality control tools for mRNA analyses from blood samples. PMID:25369468

  6. Detection of malaria infection in blood transfusion: a comparative study among real-time PCR, rapid diagnostic test and microscopy: sensitivity of Malaria detection methods in blood transfusion.

    PubMed

    Hassanpour, Gholamreza; Mohebali, Mehdi; Raeisi, Ahmad; Abolghasemi, Hassan; Zeraati, Hojjat; Alipour, Mohsen; Azizi, Ebrahim; Keshavarz, Hossein

    2011-06-01

    The transmission of malaria by blood transfusion was one of the first transfusion-transmitted infections recorded in the world. Transfusion-transmitted malaria may lead to serious problems because infection with Plasmodium falciparum may cause rapidly fatal death. This study aimed to compare real-time polymerase chain reaction (real-time PCR) with rapid diagnostic test (RDT) and light microscopy for the detection of Plasmodium spp. in blood transfusion, both in endemic and non-endemic areas of malaria disease in Iran. Two sets of 50 blood samples were randomly collected. One set was taken from blood samples donated in blood bank of Bandar Abbas, a city located in a malarious-endemic area, and the other set from Tehran, a non-endemic one. Light microscopic examination on both thin and thick smears, RDTs, and real-time PCR were performed on the blood samples and the results were compared. Thin and thick light microscopic examinations of all samples as well as RDT results were negative for Plasmodium spp. Two blood samples from endemic area were positive only with real-time PCR. It seems that real-time PCR as a highly sensitive method can be helpful for the confirmation of malaria infection in different units of blood transfusion organization especially in malaria-endemic areas where the majority of donors may be potentially infected with malaria parasites.

  7. Preterm Cord Blood Contains a Higher Proportion of Immature Hematopoietic Progenitors Compared to Term Samples

    PubMed Central

    Podestà, Marina; Bruschettini, Matteo; Cossu, Claudia; Sabatini, Federica; Dagnino, Monica; Romantsik, Olga; Spaggiari, Grazia Maria; Ramenghi, Luca Antonio; Frassoni, Francesco

    2015-01-01

    Background Cord blood contains high number of hematopoietic cells that after birth disappear. In this paper we have studied the functional properties of the umbilical cord blood progenitor cells collected from term and preterm neonates to establish whether quantitative and/or qualitative differences exist between the two groups. Methods and Results Our results indicate that the percentage of total CD34+ cells was significantly higher in preterm infants compared to full term: 0.61% (range 0.15–4.8) vs 0.3% (0.032–2.23) p = 0.0001 and in neonates <32 weeks of gestational age (GA) compared to those ≥32 wks GA: 0.95% (range 0.18–4.8) and 0.36% (0.15–3.2) respectively p = 0.0025. The majority of CD34+ cells co-expressed CD71 antigen (p<0.05 preterm vs term) and grew in vitro large BFU-E, mostly in the second generation. The subpopulations CD34+CD38- and CD34+CD45- resulted more represented in preterm samples compared to term, conversely, Side Population (SP) did not show any difference between the two group. The absolute number of preterm colonies (CFCs/10microL) resulted higher compared to term (p = 0.004) and these progenitors were able to grow until the third generation maintaining an higher proportion of CD34+ cells (p = 0.0017). The number of colony also inversely correlated with the gestational age (Pearson r = -0.3001 p<0.0168). Conclusions We found no differences in the isolation and expansion capacity of Endothelial Colony Forming Cells (ECFCs) from cord blood of term and preterm neonates: both groups grew in vitro large number of endothelial cells until the third generation and showed a transitional phenotype between mesenchymal stem cells and endothelial progenitors (CD73, CD31, CD34 and CD144)The presence, in the cord blood of preterm babies, of high number of immature hematopoietic progenitors and endothelial/mesenchymal stem cells with high proliferative potential makes this tissue an important source of cells for developing new cells therapies

  8. A DNA-based method for detecting homologous blood doping.

    PubMed

    Manokhina, Irina; Rupert, James L

    2013-12-01

    Homologous (or allogeneic) blood doping, in which blood is transferred from a donor into a recipient athlete, is the easiest, cheapest, and fastest way to increase red cell mass (hematocrit) and therefore the oxygen-carrying capacity of the blood. Although thought to have been rendered obsolete as a doping strategy by the increased use of rhEPO to increased hematocrits, there is evidence that athletes are still using this potentially dangerous method to improve endurance performance. Current testing for homologous blood doping is based on identification of mixed populations of red blood cells by flow cytometry. This paper proposes that homologous blood doping could also be tested for by high-resolution qPCR-based genotyping and demonstrates that assays could be developed that would detect second populations of cells even if the "donor" blood was depleted of 99% of the DNA-containing leukocytes. Issues of test specificity and sensitivity are discussed as well as some of the ethical considerations that would have to be addressed if athletes' genotypes were to be used by the anti-doping authorities to prevent, or detect, the use of prohibited ergogenic practices.

  9. Estimation of glomerular filtration rate from low-dose injection of iohexol and a single blood sample

    SciTech Connect

    Thomsen, H.S.; Hvid-Jacobsen, K. )

    1991-04-01

    Clearance of a small dose of iohexol (7 g I) was compared with the glomerular filtration rate (GFR) marker {sup 51}Cr EDTA in 11 healthy volunteers. The two tracers were injected simultaneously. The plasma concentration of iohexol was measured with x-ray fluorescence technique. Glomerular filtration rate was determined using blood samples drawn three and four hours after injection. An excellent correlation (0.92 less than r less than 0.97) between iohexol clearance and {sup 51}Cr EDTA clearance was found. Glomerular filtration rate can be reliably determined with a low dose of iohexol and a single blood sample obtained three hours after the injection in persons with normal serum creatinine. This new method is a good alternative to the methods using radiopharmaceuticals; it causes no radioactive burden to the patients, increases patient comfort, reduces costs, and requires no special license.

  10. Direct and long-term detection of gene doping in conventional blood samples.

    PubMed

    Beiter, T; Zimmermann, M; Fragasso, A; Hudemann, J; Niess, A M; Bitzer, M; Lauer, U M; Simon, P

    2011-03-01

    The misuse of somatic gene therapy for the purpose of enhancing athletic performance is perceived as a coming threat to the world of sports and categorized as 'gene doping'. This article describes a direct detection approach for gene doping that gives a clear yes-or-no answer based on the presence or absence of transgenic DNA in peripheral blood samples. By exploiting a priming strategy to specifically amplify intronless DNA sequences, we developed PCR protocols allowing the detection of very small amounts of transgenic DNA in genomic DNA samples to screen for six prime candidate genes. Our detection strategy was verified in a mouse model, giving positive signals from minute amounts (20 μl) of blood samples for up to 56 days following intramuscular adeno-associated virus-mediated gene transfer, one of the most likely candidate vector systems to be misused for gene doping. To make our detection strategy amenable for routine testing, we implemented a robust sample preparation and processing protocol that allows cost-efficient analysis of small human blood volumes (200 μl) with high specificity and reproducibility. The practicability and reliability of our detection strategy was validated by a screening approach including 327 blood samples taken from professional and recreational athletes under field conditions.

  11. Quantification of Cell-Free DNA in Red Blood Cell Units in Different Whole Blood Processing Methods

    PubMed Central

    Bhagirath, Vinai C.; Heddle, Nancy M.; Liu, Yang; Eikelboom, John W.; Liaw, Patricia C.

    2016-01-01

    Background. Whole blood donations in Canada are processed by either the red cell filtration (RCF) or whole blood filtration (WBF) methods, where leukoreduction is potentially delayed in WBF. Fresh WBF red blood cells (RBCs) have been associated with increased in-hospital mortality after transfusion. Cell-free DNA (cfDNA) is released by neutrophils prior to leukoreduction, degraded during RBC storage, and is associated with adverse patient outcomes. We explored cfDNA levels in RBCs prepared by RCF and WBF and different storage durations. Methods. Equal numbers of fresh (stored ≤14 days) and older RBCs were sampled. cfDNA was quantified by spectrophotometry and PicoGreen. Separate regression models determined the association with processing method and storage duration and their interaction on cfDNA. Results. cfDNA in 120 RBC units (73 RCF, 47 WBF) were measured. Using PicoGreen, WBF units overall had higher cfDNA than RCF units (p = 0.0010); fresh WBF units had higher cfDNA than fresh RCF units (p = 0.0093). Using spectrophotometry, fresh RBC units overall had higher cfDNA than older units (p = 0.0031); fresh WBF RBCs had higher cfDNA than older RCF RBCs (p = 0.024). Conclusion. Higher cfDNA in fresh WBF was observed compared to older RCF blood. Further study is required for association with patient outcomes. PMID:27774338

  12. Rapid Diagnosis of Staphylococcal Catheter-Related Bacteraemia in Direct Blood Samples by Real-Time PCR

    PubMed Central

    Soto, Marcelo; Sampietro-Colom, Laura; Soriano, Alex; Alvarez-Martínez, Míriam José; Almela, Manel; Marco, Francesc; Arjona, Ruth; Cobos-Trigueros, Nazaret; Morata, Laura; Mensa, José; Martínez, José Antonio; Mira, Aurea

    2016-01-01

    Catheter-related bacteremia (CRB) is an important cause of morbidity and mortality among hospitalized patients, being staphylococci the main etiologic agents. The objective of this study was to assess the use of a PCR-based assay for detection of staphylococci directly from blood obtained through the catheter to diagnose CRB caused by these microorganisms and to perform a cost-effectiveness analysis. A total of 92 patients with suspected CRB were included in the study. Samples were obtained through the catheter. Paired blood cultures were processed by standard culture methods and 4 ml blood samples were processed by GeneXpert-MRSA assay for the detection of methicillin-susceptible (MSSA) or methicillin-resistant (MRSA) Staphylococcus aureus, and methicillin-resistant coagulase-negative staphylococci (MR-CoNS). Sixteen CRB caused by staphylococci were diagnosed among 92 suspected patients. GeneXpert detected 14 out of 16 cases (87.5%), including 4 MSSA and 10 MR-CoNS in approximately 1 hour after specimen receipt. The sensitivity and specificity of GeneXpert were 87.5% (CI 95%: 60.4–97.8) and 92.1% (CI 95%: 83–96.7), respectively, compared with standard culture methods. The sensitivity of GeneXpert for S. aureus was 100%. Regarding a cost-effectiveness analysis, the incremental cost of using GeneXpert was of 31.1€ per patient while the incremental cost-effectiveness ratio of GeneXpert compared with blood culture alones was about 180€ per life year gained. In conclusion, GeneXpert can be used directly with blood samples obtained through infected catheters to detect S. aureus and MR-CoNS in approximately 1h after sampling. In addition, it is cost-effective especially in areas with high prevalence of staphylococcal CRB. PMID:27571200

  13. Persistent organic pollutants in blood samples of Southern Giant Petrels (Macronectes giganteus) from the South Shetland Islands, Antarctica.

    PubMed

    Colabuono, Fernanda I; Vander Pol, Stacy S; Huncik, Kevin M; Taniguchi, Satie; Petry, Maria V; Kucklick, John R; Montone, Rosalinda C

    2016-09-01

    Seabirds play an important role as top consumers in the food web and can be used as biomonitors of exposure to pollutants. Contamination studies involving non-destructive sampling methods are of considerable importance, allowing better evaluation of the levels of pollutants and their toxic effects. In the present study, organohalogen contaminants were analyzed in 113 blood samples from Southern Giant Petrel (Macronectes giganteus) adults and chicks collected in the austral summer of 2011/2012 and 2012/2013 from colonies on Elephant and Livingston Islands, South Shetland, Antarctica. Polychlorinated biphenyls (PCBs), hexachlorobenzene (HCB), pentachlorobenzene (PeCB), mirex, dichlorodiphenyltrichloroetane and derivatives (DDTs) and chlordanes were detected in all birds, whereas polybrominated diphenyl ethers (PBDEs) were not detected in any blood samples. No significant differences were found in organochlorine levels between sampling events. Adults exhibited significantly higher levels than chicks, except for PeCB. PCBs, HCB, mirex and DDTs were statistically similar in males and females from Elephant Island. Females on Livingston Island exhibited higher HCB values than males, but no sex differences were found regarding other organochlorines. The similarity in organochlorine levels between sexes in birds with very marked sexual segregation in feeding habits during the breeding season may indicate that significant amounts of contaminants are acquired during migration to lower latitudes, when the diets of males and females are similar. Birds sampled on Livingston Island exhibited significantly lower levels of PCBs, HCB, DDTs, mirex and chlordanes in comparison to those on Elephant Island, which could be the result of distinct foraging patterns between the two colonies. Organochlorine levels were similar between years in birds captured in two consecutive breeding seasons. Blood samples from Southern Giant Petrels adults and chicks proved to be useful for the comparison

  14. 1-Hydroxypyrene Levels in Blood Samples of Rats After Exposure to Generator Fumes

    PubMed Central

    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

  15. Method for sampling sub-micron particles

    DOEpatents

    Gay, Don D.; McMillan, William G.

    1985-01-01

    Apparatus and method steps for collecting sub-micron sized particles include a collection chamber and cryogenic cooling. The cooling is accomplished by coil tubing carrying nitrogen in liquid form, with the liquid nitrogen changing to the gas phase before exiting from the collection chamber in the tubing. Standard filters are used to filter out particles of diameter greater than or equal to 0.3 microns; however the present invention is used to trap particles of less than 0.3 micron in diameter. A blower draws air to said collection chamber through a filter which filters particles with diameters greater than or equal to 0.3 micron. The air is then cryogenically cooled so that moisture and sub-micron sized particles in the air condense into ice on the coil. The coil is then heated so that the ice melts, and the liquid is then drawn off and passed through a Buchner funnel where the liquid is passed through a Nuclepore membrane. A vacuum draws the liquid through the Nuclepore membrane, with the Nuclepore membrane trapping sub-micron sized particles therein. The Nuclepore membrane is then covered on its top and bottom surfaces with sheets of Mylar.RTM. and the assembly is then crushed into a pellet. This effectively traps the sub-micron sized particles for later analysis.

  16. Method and apparatus for non-invasive monitoring of blood glucose

    DOEpatents

    Thomas, Graham H.; Watson, Roger M.; Noell, J. Oakey

    1992-06-09

    A new and improved method and apparatus are provided for non-invasive monitoring of changes in blood glucose concentration in a tissue specimen and particularly in an individual. The method uses acoustic velocity measurements for monitoring the effect of glucose concentration upon the density and adiabatic compressibility of the serum. In a preferred embodiment, the acoustic velocity measurements are made through the earlobe of a subject by means of an acoustic probe or monitor which includes a transducer for transmitting and receiving ultrasonic energy pulses to and from the blood flowing in the subject's earlobe and a reflector for facilitating reflection of the acoustic pulses from the blood. The probe is designed in such a way that when properly affixed to an ear, the transducer is positioned flush against the anterior portion of an earlobe while the reflector is positioned flush against the interior portion of the earlobe. A microthermocouple is provided on the probe for monitoring the internal temperature of the blood being sampled. An electrical system, essentially comprising a frequency generator, a time intervalometer and an oscilloscope, is linked to the glucose monitoring probe. The electrical system analyzes selected ones of the pulses reflected from the blood sample in order to determine therefrom the acoustic velocity of the blood which, in turn, provides a representation of the blood glucose concentration levels at the time of the acoustic velocity measurements.

  17. Methods for blood flow measurements using ultrasound contrast agents

    NASA Astrophysics Data System (ADS)

    Fowlkes, J. Brian

    2003-10-01

    Blood flow measurements using ultrasound contrast agents are being investigated for myocardial perfusion and more recently in other organ systems. The methods are based largely on the relative increase in echogenicity due to the concentration of bubbles present in the ultrasound beam. In the simplest form, regional differences in blood volume can be inferred but the possibility exists to extract perfusion from the transit of contrast agent through tissue. Perfusion measurements rely on determining the flux of blood through a tissue volume and as such require knowledge of the fractional blood volume (FBV), i.e., ml blood/g tissue and the rate of exchange, commonly measured as the mean transit time (MTT). This presentation will discuss methods of determining each of these values and their combination to estimate tissue perfusion. Underlying principles of indicator-dilution theory will be provided in the context of ultrasound contrast agents. Current methods for determining MTT will include imaging of the intravenous bolus, in-plane contrast disruption with interval and real-time contrast recovery imaging, and control of contrast agent flow using arterial disruption (contrast interruption). The advantages and limitations of the methods will be examined along with current applications. [Work supported in part by NIH.

  18. A new algorithm for quadratic sample entropy optimization for very short biomedical signals: application to blood pressure records.

    PubMed

    Cirugeda-Roldán, E M; Cuesta-Frau, D; Miró-Martínez, P; Oltra-Crespo, S; Vigil-Medina, L; Varela-Entrecanales, M

    2014-05-01

    This paper describes a new method to optimize the computation of the quadratic sample entropy (QSE) metric. The objective is to enhance its segmentation capability between pathological and healthy subjects for short and unevenly sampled biomedical records, like those obtained using ambulatory blood pressure monitoring (ABPM). In ABPM, blood pressure is measured every 20-30 min during 24h while patients undergo normal daily activities. ABPM is indicated for a number of applications such as white-coat, suspected, borderline, or masked hypertension. Hypertension is a very important clinical issue that can lead to serious health implications, and therefore its identification and characterization is of paramount importance. Nonlinear processing of signals by means of entropy calculation algorithms has been used in many medical applications to distinguish among signal classes. However, most of these methods do not perform well if the records are not long enough and/or not uniformly sampled. That is the case for ABPM records. These signals are extremely short and scattered with outliers or missing/resampled data. This is why ABPM Blood pressure signal screening using nonlinear methods is a quite unexplored field. We propose an additional stage for the computation of QSE independently of its parameter r and the input signal length. This enabled us to apply a segmentation process to ABPM records successfully. The experimental dataset consisted of 61 blood pressure data records of control and pathological subjects with only 52 samples per time series. The entropy estimation values obtained led to the segmentation of the two groups, while other standard nonlinear methods failed.

  19. Detection of the BLV provirus from nasal secretion and saliva samples using BLV-CoCoMo-qPCR-2: Comparison with blood samples from the same cattle.

    PubMed

    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 (<380 copies/1×10(5) cells) than that in the peripheral blood, prolonged direct contact between infected and healthy cattle may be considered as a risk factor for BLV transmission.

  20. A one-step extraction procedure for the screening of cocaine, amphetamines and cannabinoids in postmortem blood samples.

    PubMed

    Pelição, Fabrício Souza; Peres, Mariana Dadalto; Pissinate, Jauber Fornaciari; De Martinis, Bruno Spinosa

    2014-01-01

    A gas chromatography-mass spectrometric (GC-MS) method was developed and validated for the simultaneous detection and quantification in postmortem whole blood samples of cocaine (COC), amphetamines (AMPs) and cannabis; the main drugs involved in cases of impaired driving in Brazil. The analytes were extracted by solid-phase extraction by means of Bond-Elute Certify cartridges, derivatized with N-methyl-N-(trimethylsilyl)trifluoroacetamide at 80°C for 30 min and analyzed by GC-MS. Linearity ranged from 10 to 500 ng/mL, except for ecgonine methyl ester, for which linearity ranged from 10 to 100 ng/mL. Inter- and intra-day imprecision ranged from 2.8 to 18.4% and from 1.5 to 14.9%, respectively. Accuracy values lay between 86.9 and 104.4%. The limit of quantitation for all drugs was 10 ng/mL and recoveries were >74% for all analytes, except for cannabinoids, which showed poor recovery (∼30%). The developed method was applied to real samples collected from deceased victims due to traffic accidents. These samples were selected according to the results obtained in immunoassay screening on collected urine samples. Five samples were positive for the presence of COC and metabolites, four samples were positive for cannabinoids, six samples were positive for AMPs and two samples were drug negative. Some samples were positive for more than one class of drug. Results obtained from whole blood samples showed good agreement with urine screening. The developed method proved capable of quantifying all three classes of drugs of abuse proposed in this study, through a one-step extraction procedure.

  1. Methods for characterizing, classifying, and identifying unknowns in samples

    DOEpatents

    Grate, Jay W [West Richland, WA; Wise, Barry M [Manson, WA

    2002-01-01

    Disclosed is a method for taking the data generated from an array of responses from a multichannel instrument, and determining the characteristics of a chemical in the sample without the necessity of calibrating or training the instrument with known samples containing the same chemical. The characteristics determined by the method are then used to classify and identify the chemical in the sample. The method can also be used to quantify the concentration of the chemical in the sample.

  2. Methods for characterizing, classifying, and identifying unknowns in samples

    DOEpatents

    Grate, Jay W.; Wise, Barry M.

    2003-08-12

    Disclosed is a method for taking the data generated from an array of responses from a multichannel instrument, and determining the characteristics of a chemical in the sample without the necessity of calibrating or training the instrument with known samples containing the same chemical. The characteristics determined by the method are then used to classify and identify the chemical in the sample. The method can also be used to quantify the concentration of the chemical in the sample.

  3. In vitro study of thimerosal reactions in human whole blood and plasma surrogate samples.

    PubMed

    Trümpler, Stefan; Meermann, Björn; Nowak, Sascha; Buscher, Wolfgang; Karst, Uwe; Sperling, Michael

    2014-04-01

    Because of its bactericidal and fungicidal properties, thimerosal is used as a preservative in drugs and vaccines and is thus deliberately injected into the human body. In aqueous environment, it decomposes into thiosalicylic acid and the ethylmercury cation. This organomercury fragment is a potent neurotoxin and is suspected to have similar toxicity and bioavailability like the methylmercury cation. In this work, human whole blood and physiological simulation solutions were incubated with thimerosal to investigate its behaviour and binding partners in the blood stream. Inductively coupled plasma with optical emission spectrometry (ICP-OES) was used for total mercury determination in different blood fractions, while liquid chromatography (LC) coupled to electrospray ionisation time-of-flight (ESI-TOF) and inductively coupled plasma-mass spectrometry (ICP-MS) provided information on the individual mercury species in plasma surrogate samples. Analogous behaviour of methylmercury and ethylmercury species in human blood was shown and an ethylmercury-glutathione adduct was identified.

  4. Blood, sweat, and tears: embedding biological samples in social science research on children.

    PubMed

    Kall, Denise

    2008-01-01

    In the first decade of the 21st Century, calls for interdisciplinary research are commonplace. Yet, relatively few papers discuss how to complete such research successfully. In this paper, I describe the details of data collection focused on five, six and seven-year old children. The project examined the effect of environmental contaminants on children's educational outcomes. It included a primary caregiver interview, a skill test with the child, and a venous blood draw from the child to test for lead, mercury, cadmium, arsenic, nicotine, and cotinine. This paper describes key issues and the solutions I adopted. Challenges discussed here include navigating the Institutional Review Board Process, analyzing the blood, obtaining the supplies needed to draw blood, banking blood for future research, hiring a phlebotomist, and recruiting subjects. While not all details will apply directly to other research projects, this paper provides some perspective on the current realities facing social scientists who decide to collect biological samples.

  5. Extraction of human genomic DNA from whole blood using a magnetic microsphere method.

    PubMed

    Gong, Rui; Li, Shengying

    2014-01-01

    With the rapid development of molecular biology and the life sciences, magnetic extraction is a simple, automatic, and highly efficient method for separating biological molecules, performing immunoassays, and other applications. Human blood is an ideal source of human genomic DNA. Extracting genomic DNA by traditional methods is time-consuming, and phenol and chloroform are toxic reagents that endanger health. Therefore, it is necessary to find a more convenient and efficient method for obtaining human genomic DNA. In this study, we developed urea-formaldehyde resin magnetic microspheres and magnetic silica microspheres for extraction of human genomic DNA. First, a magnetic microsphere suspension was prepared and used to extract genomic DNA from fresh whole blood, frozen blood, dried blood, and trace blood. Second, DNA content and purity were measured by agarose electrophoresis and ultraviolet spectrophotometry. The human genomic DNA extracted from whole blood was then subjected to polymerase chain reaction analysis to further confirm its quality. The results of this study lay a good foundation for future research and development of a high-throughput and rapid extraction method for extracting genomic DNA from various types of blood samples.

  6. Adaptive sampling of CT data for myocardial blood flow estimation from dose-reduced dynamic CT

    NASA Astrophysics Data System (ADS)

    Modgil, Dimple; Bindschadler, Michael D.; Alessio, Adam M.; La Rivière, Patrick J.

    2015-03-01

    Quantification of myocardial blood flow (MBF) can aid in the diagnosis and treatment of coronary artery disease (CAD). However, there are no widely accepted clinical methods for estimating MBF. Dynamic CT holds the promise of providing a quick and easy method to measure MBF quantitatively, however the need for repeated scans has raised concerns about the potential for high radiation dose. In our previous work, we explored techniques to reduce the patient dose by either uniformly reducing the tube current or by uniformly reducing the number of temporal frames in the dynamic CT sequence. These dose reduction techniques result in very noisy data, which can give rise to large errors in MBF estimation. In this work, we seek to investigate whether nonuniformly varying the tube current or sampling intervals can yield more accurate MBF estimates. Specifically, we try to minimize the dose and obtain the most accurate MBF estimate through addressing the following questions: when in the time attenuation curve (TAC) should the CT data be collected and at what tube current(s). We hypothesize that increasing the sampling rate and/or tube current during the time frames when the myocardial CT number is most sensitive to the flow rate, while reducing them elsewhere, can achieve better estimation accuracy for the same dose. We perform simulations of contrast agent kinetics and CT acquisitions to evaluate the relative MBF estimation performance of several clinically viable adaptive acquisition methods. We found that adaptive temporal and tube current sequences can be performed that impart an effective dose of about 5 mSv and allow for reductions in MBF estimation RMSE on the order of 11% compared to uniform acquisition sequences with comparable or higher radiation doses.

  7. Sampling Methods in Cardiovascular Nursing Research: An Overview.

    PubMed

    Kandola, Damanpreet; Banner, Davina; O'Keefe-McCarthy, Sheila; Jassal, Debbie

    2014-01-01

    Cardiovascular nursing research covers a wide array of topics from health services to psychosocial patient experiences. The selection of specific participant samples is an important part of the research design and process. The sampling strategy employed is of utmost importance to ensure that a representative sample of participants is chosen. There are two main categories of sampling methods: probability and non-probability. Probability sampling is the random selection of elements from the population, where each element of the population has an equal and independent chance of being included in the sample. There are five main types of probability sampling including simple random sampling, systematic sampling, stratified sampling, cluster sampling, and multi-stage sampling. Non-probability sampling methods are those in which elements are chosen through non-random methods for inclusion into the research study and include convenience sampling, purposive sampling, and snowball sampling. Each approach offers distinct advantages and disadvantages and must be considered critically. In this research column, we provide an introduction to these key sampling techniques and draw on examples from the cardiovascular research. Understanding the differences in sampling techniques may aid nurses in effective appraisal of research literature and provide a reference pointfor nurses who engage in cardiovascular research.

  8. Effects of music therapy on pain responses induced by blood sampling in premature infants: A randomized cross-over trial

    PubMed Central

    Shabani, Fidan; Nayeri, Nahid Dehghan; Karimi, Roghiyeh; Zarei, Khadijeh; Chehrazi, Mohammad

    2016-01-01

    Background: Premature infants are subjected to many painful procedures during care and treatment. The aim of this study was to assess the effect of music therapy on physiological and behavioral pain responses of premature infants during and after blood sampling. Materials and Methods: This study was a cross-over clinical trial conducted on 20 infants in a hospital affiliated to Tehran University of Medical Sciences for a 5-month period in 2011. In the experimental group, Transitions music was played from 5 min before until 10 min after blood sampling. The infants’ facial expressions and physiological measures were recorded from 10 min before until 10 min after sampling. All steps and measurements, except music therapy, were the same for the control group. Data were analyzed using SAS and SPSS software through analysis of variance (ANOVA) and Chi-square tests. Results: There were significant differences between the experimental and control groups (P = 0.022) in terms of heart rate during needle extraction and at the first 5 min after sampling (P = 0.005). Considering the infant's sleep–wake state in the second 5 min before sampling, the statistical difference was significant (P = 0.044). Difference was significant (P = 0.045) during injection of the needle, in the first 5 min after sampling (P = 0.002), and in the second 5 min after sampling (P = 0.005). There were significant difference in infants’ facial expressions of pain in the first 5 min after sampling (P = 0.001). Conclusions: Music therapy reduces the physiological and behavioral responses of pain during and after blood sampling. PMID:27563323

  9. Possible cause of lack of positive samples on homologous blood transfusion.

    PubMed

    Krotov, Grigory; Nikitina, Maria; Rodchenkov, Grigory

    2014-01-01

    Homologous blood transfusion is a prohibited method of blood manipulation that can be used to increase the number of erythrocytes circulating in the blood stream resulting in an increased oxygen transport capacity. In doping controls, homologous blood transfusions are determined by means of a procedure based on the detection of red blood cell phenotypes by flow cytometry. In the past six years, no adverse analytical findings concerning homologous blood transfusions were reported. One explanation for that phenomenon, assuming that athletes have not completely given up this kind of manipulation, would be a more careful selection of potential donors. If such a donor has the same set of minor erythrocyte antigens as the recipient, the established methodology to detect homologous transfusion would fail. We have hypothesized that any athlete can be a potential donor for teammates with the same RhD factor and AB0 blood group. Having analyzed the phenotype of erythrocytes of 535 Russian athletes in various endurance sports, several pairs of athletes with the same phenotype were observed. Based on the frequency of occurrence of red blood cell antigens, the theoretical probability of finding a donor within a team with exactly the same phenotype was calculated, and the existing number of occurrences where two individuals share the same phenotype in the same sport was in fact five times higher than the theoretical probability.

  10. Method for determining properties of red blood cells

    DOEpatents

    Gourley, Paul L.

    2001-01-01

    A method for quantifying the concentration of hemoglobin in a cell, and indicia of anemia, comprises determining the wavelength of the longitudinal mode of a liquid in a laser microcavity; determining the wavelength of the fundamental transverse mode of a red blood cell in the liquid in the laser microcavity; and determining if the cell is anemic from the difference between the wavelength of the longitudinal mode and the fundamental transverse mode. In addition to measuring hemoglobin, the invention includes a method using intracavity laser spectroscopy to measure the change in spectra as a function of time for measuring the influx of water into a red blood cell and the cell's subsequent rupture.

  11. Capillary gas chromatography with cryogenic oven temperature for headspace samples: analysis of chloroform or methylene chloride in whole blood.

    PubMed

    Watanabe, K; Seno, H; Ishii, A; Suzuki, O; Kumazawa, T

    1997-12-15

    A new and sensitive gas chromatography (GC) method for measurement of chloroform or methylene chloride in whole blood is presented. Trace levels of these analytes present in the headspace of samples were cryogenically trapped prior to on-line GC analysis. After heating of a blood sample containing chloroform and methylene chloride (internal standard, and vice versa) in a 7.0-mL vial at 55 degrees C for 20 min, 5 mL of the headspace vapor was drawn into a glass syringe. All vapor was introduced into an Rtx-Volatiles middle-bore capillary column in the splitless mode at -30 degrees C oven temperature to trap the entire analytes, and the oven temperature was programmed up to 280 degrees C for detection of the compounds and for cleaning of the column. The present conditions gave sharp peaks for both chloroform and methylene chloride and very low background noises for whole blood samples. As much as 11.5 and 20.0% of chloroform and methylene chloride, respectively, which had been added to whole blood in a vial, could be introduced into the GC column. The calibration curves showed linearity in the range of 0.05-5.0 micrograms/0.5 mL of whole blood. The detection limit was estimated to be about 2 ng/0.5 mL. The coefficients of intraday and interday variations were 1.31 and 8.90% for chloroform and 1.37 and 9.03% for methylene chloride, respectively. The data on chloroform or methylene chloride in rat blood after inhalation of each compound were also presented.

  12. Blood sampling in juvenile buff-breasted sandpipers: Movement, weight change and survival

    USGS Publications Warehouse

    Lanctot, Richard B.

    1994-01-01

    The effect of blood sampling on juvenile Buff-breasted Sandpipers (Tryngites subruficollis) was evaluated by comparing movements, mass, and survival of 10 broods (37 chicks) that were bled and eight broods (31 chicks) that were not bled. Blood was sampled from the jugular vein of chicks when they weighed 9.1 ± 0.9 g (x̄ ± SD) on or within 1 d of hatch. Chicks showed few short-term negative effects from blood sampling. Individual chicks suffered little physical injury, and five of eight chicks where injury occurred (i.e., hematomas formed) survived to fledging. Furthermore, bled broods gained mass at a comparable rate during the first 5 d post-hatch, and were resighted at similar frequencies as broods that were not bled. Bled broods moved slightly longer distances than control broods 1 d after hatch, however. This increased activity may have been stress-induced, but was only temporary; bled and control broods made similar long-term movements, and the probability of resighting was similar at fledging. With the proper precautions, it appears that Buff-breasted Sandpiper young can be safely sampled for blood at an early age without causing undue harm.

  13. Effect of Ibuprofen Dose on Platelet Aggregation and Coagulation in Blood Samples From Pigs

    DTIC Science & Technology

    2015-03-01

    responses between human and pig blood samples in trauma situations. INTRODUCTION Ibuprofen is a nonsteroidal anti-inflammatory drug (NSAID) with analgesic ... analgesic , and antipyretic effects, different profiles of actions have been observed in Aspirin and ibupro- fen. At equivalent effective doses of

  14. Novel genotype of Ehrlichia canis detected in samples of human blood bank donors in Costa Rica.

    PubMed

    Bouza-Mora, Laura; Dolz, Gaby; Solórzano-Morales, Antony; Romero-Zuñiga, Juan José; Salazar-Sánchez, Lizbeth; Labruna, Marcelo B; Aguiar, Daniel M

    2017-01-01

    This study focuses on the detection and identification of DNA and antibodies to Ehrlichia spp. in samples of blood bank donors in Costa Rica using molecular and serological techniques. Presence of Ehrlichia canis was determined in 10 (3.6%) out of 280 blood samples using polymerase chain reaction (PCR) targeting the ehrlichial dsb conserved gene. Analysis of the ehrlichial trp36 polymorphic gene in these 10 samples revealed substantial polymorphism among the E. canis genotypes, including divergent tandem repeat sequences. Nucleotide sequences of dsb and trp36 amplicons revealed a novel genotype of E. canis in blood bank donors from Costa Rica. Indirect immunofluorescence assay (IFA) detected antibodies in 35 (35%) of 100 serum samples evaluated. Thirty samples showed low endpoint titers (64-256) to E. canis, whereas five sera yielded high endpoint titers (1024-8192); these five samples were also E. canis-PCR positive. These findings represent the first report of the presence of E. canis in humans in Central America.

  15. Blood oxygen content in microliter samples using an easy-to-build galvanic oxygen cell.

    PubMed

    Grubb, B R; Mills, C D

    1981-02-01

    We have designed a simple, inexpensive, easy-to-build and operate apparatus for measuring blood oxygen content. The galvanic oxygen cell (fuel cell) requires as little as 1 microliter of blood and has a measuring time of 1-3 min. It is well suited for measuring oxygen content in fluids low in oxygen inasmuch as the sensitivity of the instrument is variable. Either air or water (at a known temperature and oxygen tension) can be used for calibration. No significant differences in blood oxygen content measured with our cell or the Van Slyke manometric method were found.

  16. Top-Down Proteomics and Direct Surface Sampling of Neonatal Dried Blood Spots: Diagnosis of Unknown Hemoglobin Variants

    NASA Astrophysics Data System (ADS)

    Edwards, Rebecca L.; Griffiths, Paul; Bunch, Josephine; Cooper, Helen J.

    2012-11-01

    We have previously shown that liquid microjunction surface sampling of dried blood spots coupled with high resolution top-down mass spectrometry may be used for screening of common hemoglobin variants HbS, HbC, and HbD. In order to test the robustness of the approach, we have applied the approach to unknown hemoglobin variants. Six neonatal dried blood spot samples that had been identified as variants, but which could not be diagnosed by current screening methods, were analyzed by direct surface sampling top-down mass spectrometry. Both collision-induced dissociation and electron transfer dissociation mass spectrometry were employed. Four of the samples were identified as β-chain variants: two were heterozygous Hb D-Iran, one was heterozygous Hb Headington, and one was heterozygous Hb J-Baltimore. The fifth sample was identified as the α-chain variant heterozygous Hb Phnom Penh. Analysis of the sixth sample suggested that it did not in fact contain a variant. Adoption of the approach in the clinic would require speed in both data collection and interpretation. To address that issue, we have compared manual data analysis with freely available data analysis software (ProsightPTM). The results demonstrate the power of top-down proteomics for hemoglobin variant analysis in newborn samples.

  17. EtG and EtS in Autopsy Blood Samples With and Without Putrefaction Using UPLC-MS-MS.

    PubMed

    Hegstad, Solfrid; Kristoffersen, Lena; Liane, Veronica H; Spigset, Olav

    2017-03-01

    Analytical challenges related to postmortem specimens are well known. The degree of putrefaction of the corpse will influence the quality of the blood samples, and both the efficiency of sample preparation and the subsequent chromatographic performance can be affected. An ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method was developed and validated for the determination of ethyl glucuronide (EtG) and ethyl sulfate (EtS) in postmortem whole blood. Sample preparation prior to UPLC-MS-MS analysis consisted of protein precipitation and filtration through a phospholipid removal plate. Chromatography was achieved using an HSS T3 column and gradient elution with formic acid in water in combination with methanol. The injection volume was 0.5 µL. Negative electrospray ionization was performed in the multiple reaction monitoring mode. Two transitions were monitored for the analytes and one for the internal standards. The between-assay relative standard deviations were in the range of 1.7-7.0% and the limits of quantification were 0.025 and 0.009 mg/L for EtG and EtS, respectively. Recovery was 51-55% and matrix effects ranged from 98% to 106% (corrected with internal standard). Blood samples from nine autopsy cases with various extents of putrefaction were analyzed. The sample preparation efficiently removed the phospholipids from the blood specimens. The samples were clean and the analytical quality of the chromatographic performance was satisfactory for both analytes irrespective of the degree of putrefaction.

  18. Satisfaction survey with DNA cards method to collect genetic samples for pharmacogenetics studies

    PubMed Central

    Vidal-Taboada, Jose M; Cucala, Mercedes; Mas Herrero, Sergio; Lafuente, Amalia; Cobos, Albert

    2006-01-01

    Background Pharmacogenetic studies are essential in understanding the interindividual variability of drug responses. DNA sample collection for genotyping is a critical step in genetic studies. A method using dried blood samples from finger-puncture, collected on DNA-cards, has been described as an alternative to the usual venepuncture technique. The purpose of this study is to evaluate the implementation of the DNA cards method in a multicentre clinical trial, and to assess the degree of investigators' satisfaction and the acceptance of the patients perceived by the investigators. Methods Blood samples were collected on DNA-cards. The quality and quantity of DNA recovered were analyzed. Investigators were questioned regarding their general interest, previous experience, safety issues, preferences and perceived patient satisfaction. Results 151 patients' blood samples were collected. Genotyping of GST polymorphisms was achieved in all samples (100%). 28 investigators completed the survey. Investigators perceived patient satisfaction as very good (60.7%) or good (39.3%), without reluctance to finger puncture. Investigators preferred this method, which was considered safer and better than the usual methods. All investigators would recommend using it in future genetic studies. Conclusion Within the clinical trial setting, the DNA-cards method was very well accepted by investigators and patients (in perception of investigators), and was preferred to conventional methods due to its ease of use and safety. PMID:16681846

  19. An HPLC-HR-MS-MS method for identification of anticoagulant rodenticides in blood.

    PubMed

    Schaff, Jason E; Montgomery, Madeline A

    2013-01-01

    This paper presents a fully validated method for the qualitative identification of bromadiolone, brodifacoum, coumachlor, coumatetralyl, difenacoum and warfarin in whole blood specimens. Samples are protein precipitated with acetonitrile, processed via solid-phase extraction and analyzed by high-performance liquid chromatography with high resolution tandem mass spectrometric detection. Limits of detection were 10 ng/mL or better for all analytes.

  20. Evaluation of Existing Methods for Human Blood mRNA Isolation and Analysis for Large Studies

    PubMed Central

    Meyer, Anke; Paroni, Federico; Günther, Kathrin; Dharmadhikari, Gitanjali; Ahrens, Wolfgang; Kelm, Sørge; Maedler, Kathrin

    2016-01-01

    Aims Prior to implementing gene expression analyses from blood to a larger cohort study, an evaluation to set up a reliable and reproducible method is mandatory but challenging due to the specific characteristics of the samples as well as their collection methods. In this pilot study we optimized a combination of blood sampling and RNA isolation methods and present reproducible gene expression results from human blood samples. Methods The established PAXgeneTM blood collection method (Qiagen) was compared with the more recent TempusTM collection and storing system. RNA from blood samples collected by both systems was extracted on columns with the corresponding Norgen and PAX RNA extraction Kits. RNA quantity and quality was compared photometrically, with Ribogreen and by Real-Time PCR analyses of various reference genes (PPIA, β-ACTIN and TUBULIN) and exemplary of SIGLEC-7. Results Combining different sampling methods and extraction kits caused strong variations in gene expression. The use of PAXgeneTM and TempusTM collection systems resulted in RNA of good quality and quantity for the respective RNA isolation system. No large inter-donor variations could be detected for both systems. However, it was not possible to extract sufficient RNA of good quality with the PAXgeneTM RNA extraction system from samples collected by TempusTM collection tubes. Comparing only the Norgen RNA extraction methods, RNA from blood collected either by the TempusTM or PAXgeneTM collection system delivered sufficient amount and quality of RNA, but the TempusTM collection delivered higher RNA concentration compared to the PAXTM collection system. The established Pre-analytix PAXgeneTM RNA extraction system together with the PAXgeneTM blood collection system showed lowest CT-values, i.e. highest RNA concentration of good quality. Expression levels of all tested genes were stable and reproducible. Conclusions This study confirms that it is not possible to mix or change sampling or

  1. Investigation of the Method to Determine Carboxyhaemoglobin in Blood,

    DTIC Science & Technology

    1985-08-01

    phic Integrator Procedure for the Determination of Carboxyhemoglo- bin Percent Saturation in Post - Mortem Samples. J. Anal. Toxicol. 4: 181, 1980. 9...5: 264, 1981. 10. Rodkey, F.L., Hill, T.A., Pitts, L.L., Robertson, R.F. Spectropho- tometric Measurement of Carboxyhemoglobin and Methemoglobin in...Simultaneous Measurement of Carboxy- and * Methemoglobin in Blood. J. Appl. Phys. 31: 154, 1971. 27. Rodkey, F.L., O’Neal, J.D. Effects of Carboxyhemoglobin on

  2. Prediction of Arterial Blood pH and Partial Pressure of Carbon dioxide from Venous Blood Samples in Patients Receiving Mechanical Ventilation.

    PubMed

    Tavakol, Kamran; Ghahramanpoori, Bahareh; Fararouei, Mohammad

    2013-07-01

    Substitution of arterial with venous blood samples to estimate blood gas status is highly preferable due to practical and safety concerns. Numerous studies support the substitution of arterial by venous blood samples, reporting strong correlations between arterial and venous values. This study further investigated the predictive ability of venous blood samples for arterial Acid-Base Balance (pH) and pressure of carbon dioxide (pCO2). Participants were 51 post-brain surgery patients receiving mechanical ventilation, who had blood samples taken simultaneously from radial artery of the wrist and elbow vein. Results showed significant associations between arterial and venous pH and pCO2. However, the variation of regression residuals was not homogenous, and the regression line did not fit properly to the data, indicating that simple linear regression is sub-optimal for prediction of arterial pH and pCO2 by venous blood sample. Although highly significant correlations were found between arterial and venous blood pH and pCO2, the results did not support the reliability of prediction of arterial blood pH and pCO2 by venous blood samples across a range of concentrations.

  3. Attic dust and human blood samples collected near a former wood treatment facility.

    PubMed

    Hensley, A R; Scott, A; Rosenfeld, P E; Clark, J J J

    2007-10-01

    A wood treatment facility operating in southern Alabama released dioxins and other hazardous substances into the surrounding community over a period of approximately 35 years. The facility used a variety of chemical insecticides including pentachlorophenol (PCP), chromated copper arsenate (CCA), and creosote (which contains polycyclic aromatic hydrocarbons (PAHs)) to treat wood. The health risks associated with the released contaminants are numerous and significant. To evaluate the historic exposure to the contaminants from the facility, blood samples and health surveys were collected from 21 current and past residents of the adjacent, isolated community and analyzed for concentrations of polychlorinated dibenzo-p-dioxins (dioxins) and polychlorinated dibenzofurans (furans). In addition, attic dust sampling was performed in 11 buildings located within a 1-mile radius of the former wood treatment facility. The average total dioxin toxic equivalent (TEQ) concentration in the residents' blood samples was 36.6 pg/g lipids. In the attic dust, the average total dioxin TEQ concentration, benzo[a]pyrene (PAH) TEQ concentration, and arsenic concentration were 145 ng/kg, 0.98 and 29.8 mg/kg, respectively. The concentrations of dioxins measured in the blood samples exceed the 90th percentile total dioxin levels found in the general US adult population. Concentrations of dioxin, arsenic, and PAHs found in the attic samples exceeded the US Environmental Protection Agency (USEPA) Region 4 soil exposure cancer risk preliminary remediation goal (PRG) values. These findings indicate a very significant potential for related health effects in these communities.

  4. Pattern recognition of monocyte chemoattractant protein-1 (MCP-1) in whole blood samples using new platforms based on nanostructured materials

    NASA Astrophysics Data System (ADS)

    Stefan-van Staden, Raluca-Ioana; Gugoasa, Livia Alexandra; Biris, Alexandru Radu

    2015-09-01

    Four stochastic microsensors based on nanostructured materials (graphene, maltodextrin (MD), and diamond) integrated in miniaturized platforms were proposed. Monocyte chemoattractant protein-1 (MCP-1) is a pro-inflammatory cytokine whose main function is to regulate cell trafficking. It is correlated with the incidence of cardiovascular diseases and obesity, and was used as the model analyte in this study. The screening of whole blood samples for MCP-1 can be done for concentrations ranging from 10-12 to 10-8 g mL-1. The method was used for both qualitative and quantitative assessments of MCP-1 in whole blood samples. The lowest quantification limits for the assay of MCP-1 (1 pg mL-1) were reached when the microsensors based on protoporphyrin IX/Graphene-Au-3 and on MD/Graphene were employed in the platform design.

  5. NEW CLINICALLY SIGNIFICANT METHOD OF DETERMINING GLYCOPROTEINS IN BLOOD SERUM

    DTIC Science & Technology

    The reaction with ammonium molybdate is a simple and at the same time a very sensitive method for determining the level of glycoproteins in the blood ... serum , which has a number of advantages over the diphehylamine reaction. The reaction with ammonium molybdate is a valuable supplementary test for

  6. Photoacoustic sample vessel and method of elevated pressure operation

    DOEpatents

    Autrey, Tom; Yonker, Clement R.

    2004-05-04

    An improved photoacoustic vessel and method of photoacoustic analysis. The photoacoustic sample vessel comprises an acoustic detector, an acoustic couplant, and an acoustic coupler having a chamber for holding the acoustic couplant and a sample. The acoustic couplant is selected from the group consisting of liquid, solid, and combinations thereof. Passing electromagnetic energy through the sample generates an acoustic signal within the sample, whereby the acoustic signal propagates through the sample to and through the acoustic couplant to the acoustic detector.

  7. Therapeutic drug monitoring of everolimus using the dried blood spot method in combination with liquid chromatography-mass spectrometry.

    PubMed

    van der Heijden, J; de Beer, Y; Hoogtanders, K; Christiaans, M; de Jong, G J; Neef, C; Stolk, L

    2009-11-01

    An assay of everolimus based on finger prick sampling and consecutive application as a blood spot on sampling paper has been developed. We explored several methods [K. Hoogtanders, J. van der Heijden, M. Christiaans, P. Edelbroek, J. van Hooff, L. Stolk, J. Pharm. Biomed. Anal. 44 (2006) 658-664; A. Allanson, M. Cotton, J. Tettey, et al., J. Pharm. Biomed. Anal. 44 (2007) 963-969] and developed a new method, namely the impregnation of sampling paper with a solution of plasma-protein, formic acid and ammonium acetate, in combination with the extraction of the blood spot by filter filtration. This kind of sample preparation provides new possibilities for blood spot sampling especially if analytes are adsorbed to the paper. The dried blood spot was analysed using the HPLC-electrospray-tandem mass spectrometry method, with 32-desmethoxyrapamycin as the internal standard. The working range of our study was 2-30 microg/l. Within this range, intra-and inter-assay variability for precision and accuracy was <15%. Everolimus blood spot samples proved stable for 3 days at 60 degrees C and for 32 days at 4 degrees C. Everolimus concentrations of one stable out-patient were compared after both blood spot sampling and conventional venous sampling on various occasions. Results indicate that this new method is promising for therapeutic drug monitoring in stable renal transplant patients.

  8. Performance comparison of publicly available retinal blood vessel segmentation methods.

    PubMed

    Vostatek, Pavel; Claridge, Ela; Uusitalo, Hannu; Hauta-Kasari, Markku; Fält, Pauli; Lensu, Lasse

    2017-01-01

    Retinal blood vessel structure is an important indicator of many retinal and systemic diseases, which has motivated the development of various image segmentation methods for the blood vessels. In this study, two supervised and three unsupervised segmentation methods with a publicly available implementation are reviewed and quantitatively compared with each other on five public databases with ground truth segmentation of the vessels. Each method is tested under consistent conditions with two types of preprocessing, and the parameters of the methods are optimized for each database. Additionally, possibility to predict the parameters of the methods by the linear regression model is tested for each database. Resolution of the input images and amount of the vessel pixels in the ground truth are used as predictors. The results show the positive influence of preprocessing on the performance of the unsupervised methods. The methods show similar performance for segmentation accuracy, with the best performance achieved by the method by Azzopardi et al. (Acc 94.0) on ARIADB, the method by Soares et al. (Acc 94.6, 94.7) on CHASEDB1 and DRIVE, and the method by Nguyen et al. (Acc 95.8, 95.5) on HRF and STARE. The method by Soares et al. performed better with regard to the area under the ROC curve. Qualitative differences between the methods are discussed. Finally, it was possible to predict the parameter settings that give performance close to the optimized performance of each method.

  9. Melting Temperature Mapping Method: A Novel Method for Rapid Identification of Unknown Pathogenic Microorganisms within Three Hours of Sample Collection.

    PubMed

    Niimi, Hideki; Ueno, Tomohiro; Hayashi, Shirou; Abe, Akihito; Tsurue, Takahiro; Mori, Masashi; Tabata, Homare; Minami, Hiroshi; Goto, Michihiko; Akiyama, Makoto; Yamamoto, Yoshihiro; Saito, Shigeru; Kitajima, Isao

    2015-07-28

    Acquiring the earliest possible identification of pathogenic microorganisms is critical for selecting the appropriate antimicrobial therapy in infected patients. We herein report the novel "melting temperature (Tm) mapping method" for rapidly identifying the dominant bacteria in a clinical sample from sterile sites. Employing only seven primer sets, more than 100 bacterial species can be identified. In particular, using the Difference Value, it is possible to identify samples suitable for Tm mapping identification. Moreover, this method can be used to rapidly diagnose the absence of bacteria in clinical samples. We tested the Tm mapping method using 200 whole blood samples obtained from patients with suspected sepsis, 85% (171/200) of which matched the culture results based on the detection level. A total of 130 samples were negative according to the Tm mapping method, 98% (128/130) of which were also negative based on the culture method. Meanwhile, 70 samples were positive according to the Tm mapping method, and of the 59 suitable for identification, 100% (59/59) exhibited a "match" or "broad match" with the culture or sequencing results. These findings were obtained within three hours of whole blood collection. The Tm mapping method is therefore useful for identifying infectious diseases requiring prompt treatment.

  10. Evaluation of PCR Approaches for Detection of Bartonella bacilliformis in Blood Samples

    PubMed Central

    Gomes, Cláudia; Martinez-Puchol, Sandra; Pons, Maria J.; Bazán, Jorge; Tinco, Carmen; del Valle, Juana; Ruiz, Joaquim

    2016-01-01

    Background The lack of an effective diagnostic tool for Carrion’s disease leads to misdiagnosis, wrong treatments and perpetuation of asymptomatic carriers living in endemic areas. Conventional PCR approaches have been reported as a diagnostic technique. However, the detection limit of these techniques is not clear as well as if its usefulness in low bacteriemia cases. The aim of this study was to evaluate the detection limit of 3 PCR approaches. Methodology/Principal Findings We determined the detection limit of 3 different PCR approaches: Bartonella-specific 16S rRNA, fla and its genes. We also evaluated the viability of dry blood spots to be used as a sample transport system. Our results show that 16S rRNA PCR is the approach with a lowest detection limit, 5 CFU/μL, and thus, the best diagnostic PCR tool studied. Dry blood spots diminish the sensitivity of the assay. Conclusions/Significance From the tested PCRs, the 16S rRNA PCR-approach is the best to be used in the direct blood detection of acute cases of Carrion’s disease. However its use in samples from dry blood spots results in easier management of transport samples in rural areas, a slight decrease in the sensitivity was observed. The usefulness to detect by PCR the presence of low-bacteriemic or asymptomatic carriers is doubtful, showing the need to search for new more sensible techniques. PMID:26959642

  11. Measured blood pressure and hypertension among young adults: a comparison between two nationally representative samples.

    PubMed

    Chyu, Laura; McDade, Thomas W; Adam, Emma K

    2011-01-01

    Measurement and distribution of systolic and diastolic blood pressure and related health risk factors in the National Longitudinal Study of Adolescent Health (Add Health) were compared with data from the National Health and Nutrition Examination Survey (NHANES) 2007-2008. Sociodemographic patterns of blood pressure, prevalence of hypertension, and measurement characteristics in Add Health were also examined. Prevalence of hypertension (20.88%) in Add Health was significantly higher than that in NHANES (4.60%). This difference was only partially explained by body mass index and waist circumference and could reflect different measurement techniques, sample composition differences, or masked hypertension.

  12. Novel method to determine instantaneous blood volume in pulsatile blood pump using electrical impedance.

    PubMed

    Sasaki, E; Nakatani, T; Taenaka, Y; Takano, H; Hirose, H

    1994-08-01

    A novel real-time volumetric method was developed for a pulsatile pump. This method, the impedance method, used electrical impedance change in the blood chamber according to volume change while pumping. This method was evaluated with two kinds of air-driven diaphragm pumps. During in vitro tests, the impedance method indicated real-time volume change, and there was excellent correlation between computed stroke volume with the impedance method and measured stroke volume with the electromagnetic flowmeter. In chronic animal tests with goats and in a clinical case, the impedance method measured pump output accurately, and it detected diaphragm motion in real-time. In addition, excellent durability was seen. Full-fill to full-empty drive was realized accurately with this method. Application of the impedance method was easy, and it did not deteriorate native antithrombogencity of the pump. The impedance method is practical and useful to estimate the pumping condition of a pulsatile blood pump, especially a diaphragm pump. This method would be useful in clinical application.

  13. Systems and methods for self-synchronized digital sampling

    NASA Technical Reports Server (NTRS)

    Samson, Jr., John R. (Inventor)

    2008-01-01

    Systems and methods for self-synchronized data sampling are provided. In one embodiment, a system for capturing synchronous data samples is provided. The system includes an analog to digital converter adapted to capture signals from one or more sensors and convert the signals into a stream of digital data samples at a sampling frequency determined by a sampling control signal; and a synchronizer coupled to the analog to digital converter and adapted to receive a rotational frequency signal from a rotating machine, wherein the synchronizer is further adapted to generate the sampling control signal, and wherein the sampling control signal is based on the rotational frequency signal.

  14. Postmortem tissue samples: an alternative to urine and blood for drug analysis in racehorses.

    PubMed

    Uboh, C E; Rudy, J A; Railing, F A; Enright, J M; Shoemaker, J M; Kahler, M C; Shellenberger, J M; Kemecsei, Z; Das, D N

    1995-09-01

    Although urine is the sample of choice for drug tests in racehorses, it is rarely obtained following the sudden death of a racehorse on the track while racing. The purpose of this study was to demonstrate the significance of postmortem tissue samples as an alternative to urine and blood samples in equine drug analysis following the sudden death of a racehorse on the track while participating in a competitive race. Postmortem tissue samples were frozen (-80 degrees C) until analyzed. A 30-40-g portion of each organ was homogenized in a 0.1 M phosphate buffer (pH 7.4), deproteinized, hydrolyzed with beta-glucuronidase, extracted, and screened by thin-layer chromatography and immunoassay. Samples that initially tested positive for drug(s) were then extracted using high-flow, solid-phase extraction cartridges. The eluates were analyzed by gas chromatography-mass spectrometry. The presence of butorphanol in horses HB355 and CD387, pentobarbital in horse HO940, and ergotamine in horses HO940 and CD387 was detected and confirmed. Thus, in the absence of urine and blood samples following sudden death, postmortem tissue samples are equally useful for forensic toxicological investigations of racehorses.

  15. Safety and blood sample volume and quality of a refined retro-orbital bleeding technique in rats using a lateral approach.

    PubMed

    Sharma, Ashish; Fish, Brian L; Moulder, John E; Medhora, Meetha; Baker, John E; Mader, Marylou; Cohen, Eric P

    2014-02-01

    The collection of blood samples from laboratory rats requires the use of bleeding techniques that provide quality samples of sufficient volume for analysis without injury to the animal. Retro-orbital bleeding (ROB) is a phlebotomy technique that can yield high-quality samples of adequate volume, but it has been criticized for its potential to cause injury. To evaluate the injury-causing potential of their refined ROB method using a lateral approach, the authors retrospectively reviewed ROB procedures carried out in their colony during an 18-month period and found that 0.6% of these procedures were associated with ocular injury. The authors also compared the quality of blood samples collected by ROB and by saphenous phlebotomy and found that ROB yielded samples of better quality. The authors conclude that, when done using a lateral approach and by an experienced technician, ROB is humane and safe and provides blood samples of adequate volume and quality for analysis.

  16. Melting Temperature Mapping Method: A Novel Method for Rapid Identification of Unknown Pathogenic Microorganisms within Three Hours of Sample Collection

    PubMed Central

    Niimi, Hideki; Ueno, Tomohiro; Hayashi, Shirou; Abe, Akihito; Tsurue, Takahiro; Mori, Masashi; Tabata, Homare; Minami, Hiroshi; Goto, Michihiko; Akiyama, Makoto; Yamamoto, Yoshihiro; Saito, Shigeru; Kitajima, Isao

    2015-01-01

    Acquiring the earliest possible identification of pathogenic microorganisms is critical for selecting the appropriate antimicrobial therapy in infected patients. We herein report the novel “melting temperature (Tm) mapping method” for rapidly identifying the dominant bacteria in a clinical sample from sterile sites. Employing only seven primer sets, more than 100 bacterial species can be identified. In particular, using the Difference Value, it is possible to identify samples suitable for Tm mapping identification. Moreover, this method can be used to rapidly diagnose the absence of bacteria in clinical samples. We tested the Tm mapping method using 200 whole blood samples obtained from patients with suspected sepsis, 85% (171/200) of which matched the culture results based on the detection level. A total of 130 samples were negative according to the Tm mapping method, 98% (128/130) of which were also negative based on the culture method. Meanwhile, 70 samples were positive according to the Tm mapping method, and of the 59 suitable for identification, 100% (59/59) exhibited a “match” or “broad match” with the culture or sequencing results. These findings were obtained within three hours of whole blood collection. The Tm mapping method is therefore useful for identifying infectious diseases requiring prompt treatment. PMID:26218169

  17. Post mortem concentrations of endogenous gamma hydroxybutyric acid (GHB) and in vitro formation in stored blood and urine samples.

    PubMed

    Busardò, Francesco Paolo; Bertol, Elisabetta; Vaiano, Fabio; Baglio, Giovanni; Montana, Angelo; Barbera, Nunziata; Zaami, Simona; Romano, Guido

    2014-10-01

    Gamma-hydroxybutyrate (GHB) is a central nervous system depressant, primarily used as a recreational drug of abuse with numerous names. It has also been involved in various instances of drug-facilitated sexual assault due to its potential incapacitating effects. The first aim of this paper is to measure the post-mortem concentration of endogenous GHB in whole blood and urine samples of 30 GHB free-users, who have been divided according to the post-mortem interval (PMI) in three groups (first group: 24-36h; second group: 37-72h; third group: 73-192h), trying to evaluate the role of PMI in affecting post mortem levels. Second, the Authors have evaluated the new formation of GHB in vitro in blood and urine samples of the three groups, which have been stored at -20°C, 4°C and 20°C over a period of one month. The concentrations were measured by GC-MS after liquid-liquid extraction according to the method validated and published by Elliot (For. Sci. Int., 2003). For urine samples, GHB concentrations were creatinine-normalized. In the first group the GHB mean concentration measured after autopsy was: 2.14mg/L (range 0.54-3.21mg/L) in blood and 3.90mg/g (range 0.60-4.81mg/g) in urine; in the second group it was: 5.13mg/L (range 1.11-9.60mg/L) in blood and 3.93mg/g (range 0.91-7.25mg/g) in urine; in the third group it was: 11.8mg/L (range 3.95-24.12mg/L) in blood and 9.83mg/g (range 3.67-21.90mg/g) in urine. The results obtained in blood and urine samples showed a statistically significant difference among groups (p<0.001) in the first analysis performed immediately after autopsy. Throughout the period of investigation up to 4 weeks, the comparison of storage temperatures within each group showed in blood and urine samples a mean difference at 20°C compared to -20°C not statistically significant at the 10% level. These findings allow us to affirm that the PMI strongly affects the post mortem production of GHB in blood and urine samples. Regarding the new formation of

  18. Sampling bee communities using pan traps: alternative methods increase sample size

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Monitoring of the status of bee populations and inventories of bee faunas require systematic sampling. Efficiency and ease of implementation has encouraged the use of pan traps to sample bees. Efforts to find an optimal standardized sampling method for pan traps have focused on pan trap color. Th...

  19. Pulsing Depolarization: New Method of In-Vivo Blood Glucometry

    NASA Astrophysics Data System (ADS)

    Romanov, Dmitri; Shvartsman, Leonid; Fine, Ilya

    2002-03-01

    We propose a new system of non-invasive measurements enabling one to monitor in-vivo the glucose content of the blood. The proposed method is based on the transmission oximetry technique [1] and on the model of red blood cell aggregation [2]. We utilize the known birefregence of glucose solutions (in this case, the blood plasma) and the absence of this optical activity on the part of the red cells. Thus, we model the blood as a birefrigent host medium filled with optically non-active inclusions of spheroidal shape, arbitrarily oriented and having one semiaxis growing with time during the cell aggregation. We trace the evolution of initially linearly polarized laser beam in this system, and calculate the ellipticity and the azimuthal angle of the resulting partially depolarized light. The azimuthal angle is obtained as time-independent quantity being a function of the hematocrit, glucose content and thickness of the host medium layer, while the ellipticity depends also on the red cell aggregation, and thus oscillates with the heartbeats. Measurements of both the azimuthal angle and the ellipticity provide us with sufficient data to determine the glucose content of the blood. [1] I. Fine, A. Weinreb, Med. and Biol. Eng. and Comput., 31, 516 (1993). [2] L.D. Shvartsman, I. Fine, SPIE Proc., 4162, 120 (2000).

  20. Stability of ethyl glucuronide in urine, post-mortem tissue and blood samples.

    PubMed

    Schloegl, Haiko; Dresen, Sebastian; Spaczynski, Karin; Stoertzel, Mylène; Wurst, Friedrich Martin; Weinmann, Wolfgang

    2006-03-01

    The stability of ethyl glucuronide (EtG) under conditions of degradation was examined in urine samples of nine volunteers and in post-mortem tissue (liver, skeletal muscle) and blood taken from seven corpses at autopsies. Analysis was performed via LC-MS/MS. EtG concentrations in urine samples ranged from 2.5 to 296.5 mg/l. When stored at 4 degrees C in airtight test tubes, EtG concentrations remained relatively constant; when stored at room temperature (RT) for 5 weeks in ventilated vials, variations of EtG concentrations ranged from a 30% decrease to an 80% increase, with an average of 37.5% increase. Liver and skeletal muscle tissue of three corpses with positive blood alcohol concentrations (BAC; ranging from 0.106 to 0.183 g%) were stored for 4 weeks and analysed periodically. EtG concentrations decreased 27.7% on average in 4 weeks storage at RT but EtG was still detectable in all samples with initial EtG concentrations higher than 1 mug/g. Blood and liver samples of four corpses with negative BACs were stored at RT after addition of 0.1 g% ethanol, and no new formation of EtG was observed.

  1. Morphine to codeine concentration ratio in blood and urine as a marker of illicit heroin use in forensic autopsy samples.

    PubMed

    Konstantinova, Svetlana V; Normann, Per T; Arnestad, Marianne; Karinen, Ritva; Christophersen, Asbjørg S; Mørland, Jørg

    2012-04-10

    A morphine to codeine ratio greater than unity (M/C>1) has been suggested as an indicator of heroin use in living individuals. The aim of this study was to examine the morphine to codeine ratio in a large population (N=2438) of forensically examined autopsy cases positive for 6-monoacetylmorphine (6-MAM) and/or morphine in blood and/or urine. Blood and urine concentrations of 6-MAM, morphine and codeine were examined using GC-MS and LC-MS/MS methods. In 6-MAM positive samples, the M/C ratio was greater than unity in 98% (N=917) of the blood samples and 96% (N=665) of the urine samples. Stratification of 6-MAM negative cases by M/C above or below unity revealed similarities in morphine and codeine concentrations in cases where M/C>1 and 6-MAM positive cases. Median blood and urine morphine concentrations were 8-10 times greater than codeine for both groups. Similarly to 6-MAM positive cases, 25-44 year-old men prevailed in the M/C>1 group. In comparison to cases where M/C ≤ 1, the M/C ratio was a hundred times higher in both 6-MAM positive and M/C>1 cases. The range of morphine concentration between the lowest and the highest quintile of codeine in M/C>1 cases was similar to that in 6-MAM positive cases. This range was much higher than for M/C ≤ 1 cases. Moreover, linear regression analyses, adjusted for age and gender, revealed a strong positive association between morphine and codeine in 6-MAM positive and M/C>1 cases. The M/C ratio appeared to be a good marker of heroin use in post-mortem cases. Both blood and urine M/C>1 can be used to separate heroin users from other cases positive for morphine and codeine.

  2. Rapid and Sensitive Salmonella Typhi Detection in Blood and Fecal Samples Using Reverse Transcription Loop-Mediated Isothermal Amplification.

    PubMed

    Fan, Fenxia; Yan, Meiying; Du, Pengcheng; Chen, Chen; Kan, Biao

    2015-09-01

    Typhoid fever caused by Salmonella enterica serovar Typhi remains a significant public health problem in developing countries. Although the main method for diagnosing typhoid fever is blood culture, the test is time consuming and not always able to detect infections. Thus, it is very difficult to distinguish typhoid from other infections in patients with nonspecific symptoms. A simple and sensitive laboratory detection method remains necessary. The purpose of this study is to establish and evaluate a rapid and sensitive reverse transcription-based loop-mediated isothermal amplification (RT-LAMP) method to detect Salmonella Typhi infection. In this study, a new specific gene marker, STY1607, was selected to develop a STY1607-RT-LAMP assay; this is the first report of specific RT-LAMP detection assay for typhoid. Human-simulated and clinical blood/stool samples were used to evaluate the performance of STY1607-RT-LAMP for RNA detection; this method was compared with STY1607-LAMP, reverse transcription real-time polymerase chain reaction (rRT-PCR), and bacterial culture methods for Salmonella Typhi detection. Using mRNA as the template, STY1607-RT-LAMP exhibited 50-fold greater sensitivity than STY1607-LAMP for DNA detection. The STY1607-RT-LAMP detection limit is 3 colony-forming units (CFU)/mL for both the pure Salmonella Typhi samples and Salmonella Typhi-simulated blood samples and was 30 CFU/g for the simulated stool samples, all of which were 10-fold more sensitive than the rRT-PCR method. RT-LAMP exhibited improved Salmonella Typhi detection sensitivity compared to culture methods and to rRT-PCR of clinical blood and stool specimens from suspected typhoid fever patients. Because it can be performed without sophisticated equipment or skilled personnel, RT-LAMP is a valuable tool for clinical laboratories in developing countries. This method can be applied in the clinical diagnosis and care of typhoid fever patients as well as for a quick public health response.

  3. Ibuprofen analysis in blood samples by palladium particles-impregnated sodium montmorillonite electrodes: Validation using high performance liquid chromatography.

    PubMed

    Loudiki, A; Boumya, W; Hammani, H; Nasrellah, H; El Bouabi, Y; Zeroual, M; Farahi, A; Lahrich, S; Hnini, K; Achak, M; Bakasse, M; El Mhammedi, M A

    2016-12-01

    The electrochemical detection of ibuprofen has been studied on Palladium-Montmorillonite (Mt) modified carbon paste electrode using differential pulse voltammetry. The optimization of the modifier preparation and the instrumental parameters was investigated. The results indicate that ibuprofen oxidation was favored in the presence of Pd-PdO particles. The quantitative determination of ibuprofen was statistically analyzed and validated using HPLC method. The detection and quantification limits, specificity and precision were found to be acceptable. Finally, the developed method was successfully applied for ibuprofen determination in human blood samples.

  4. Measurement of thyroid hormones in donkey (Equus asinus) blood and milk: validation of ELISA kits and evaluation of sample collection, handling and storage.

    PubMed

    Todini, Luca; Malfatti, Alessandro; Salimei, Elisabetta; Fantuz, Francesco

    2010-11-01

    Donkey's milk is well tolerated by human infants with cow's milk allergy and is useful in the treatment of human immune-related diseases and in the prevention of atherosclerosis. Thyroid hormones (TH) stimulate lactation and active triiodothyronine (T3) in colostrum and milk could take paracrine action supporting lactogenesis in the mother, and play physiological roles for the suckling offspring (systemic or within the gastrointestinal tract). The aims were to measure TH concentrations in donkey blood and milk, validate ELISA methods, evaluate the effects of sample collection and post-collection handling and the stability of TH in milk and blood serum and plasma samples. In milk and blood samples obtained from lactating jennies total concentrations of TH were assayed using competitive-type ELISA kits. Good validation results were obtained for both TH concentrations in blood serum and plasma and T3 in milk samples extracted with cold (-20°C) ethanol alkalinized (pH 9·0) with NH4OH. In most of the milk extract samples, thyroxine (T4) concentrations resulted below the sensitivity threshold. Intra- and inter-assay coefficients of variations of TH concentrations in different blood and milk samples were below 10%. Parallelism tests gave displacement lines parallel to those of the calibrators for both TH in blood serum and plasma and for T3 in milk extracts. Mean recovery rates were between 95% and 123%, but the concentration values approaching the highest calibrators were overestimated. Therefore, serum and plasma samples for T3 assay must be previously diluted with buffer. Both TH concentrations in blood serum and plasma and T3 in milk did not change during storage for up to 6 months at -20°C. In conclusion, the ELISA methods tested in the present study are suitable for determination of both TH concentrations in donkey blood samples, and for T3 measurement in milk, after extraction with cold alkaline ethanol.

  5. Trace-element concentrations in blood samples from welders of stainless steel or aluminium and a reference group.

    PubMed

    Ulfvarson, U; Wold, S

    1977-12-01

    The concentrations of 17 trace elements (e.g., copper, cobalt, iron, manganese, chromium, silicon and magnesium) were determined in whole blood samples of 81 persons working with different welding methods on stainless steel or aluminium and 68 nonwelders. Inorganic spark source mass spectrometry was used for the chemical analyses. The data were analyzed by the SIMCA method for pattern recognition (discriminant analysis). No differences were found between the five groups, either in the average levels of the trace elements or in the correlation structures between the trace elements. Thus no blood concentration data on the analyzed elements and collected from a single person contained any information with respect to exposure to the welding fumes investigated.

  6. Passive Samplers for Investigations of Air Quality: Method Description, Implementation, and Comparison to Alternative Sampling Methods

    EPA Science Inventory

    This Paper covers the basics of passive sampler design, compares passive samplers to conventional methods of air sampling, and discusses considerations when implementing a passive sampling program. The Paper also discusses field sampling and sample analysis considerations to ensu...

  7. DOE methods for evaluating environmental and waste management samples.

    SciTech Connect

    Goheen, S C; McCulloch, M; Thomas, B L; Riley, R G; Sklarew, D S; Mong, G M; Fadeff, S K

    1994-04-01

    DOE Methods for Evaluating Environmental and Waste Management Samples (DOE Methods) provides applicable methods in use by. the US Department of Energy (DOE) laboratories for sampling and analyzing constituents of waste and environmental samples. The development of DOE Methods is supported by the Laboratory Management Division (LMD) of the DOE. This document contains chapters and methods that are proposed for use in evaluating components of DOE environmental and waste management samples. DOE Methods is a resource intended to support sampling and analytical activities that will aid in defining the type and breadth of contamination and thus determine the extent of environmental restoration or waste management actions needed, as defined by the DOE, the US Environmental Protection Agency (EPA), or others.

  8. Coupling passive sampling with in vitro bioassays and chemical analysis to understand combined effects of bioaccumulative chemicals in blood of marine turtles.

    PubMed

    Jin, Ling; Escher, Beate I; Limpus, Colin J; Gaus, Caroline

    2015-11-01

    Conventional target analysis of biological samples such as blood limits our ability to understand mixture effects of chemicals. This study aimed to establish a rapid passive sampling technique using the polymer polydimethylsiloxane (PDMS) for exhaustive extraction of mixtures of neutral organic chemicals accumulated in blood of green turtles, in preparation for screening in in vitro bioassays. We designed a PDMS-blood partitioning system based on the partition coefficients of chemicals between PDMS and major blood components. The sampling kinetics of hydrophobic test chemicals (polychlorinated dibenzo-p-dioxins; PCDDs) from blood into PDMS were reasonably fast reaching steady state in <96 h. The geometric mean of the measured PDMS-blood partition coefficients for PCDDs, polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) was 14 L blood kg PDMS(-1) and showed little variability (95% confidence interval from 8.4 to 29) across a wide range of hydrophobicity (logKow 5.7-8.3). The mass transfer of these chemicals from 5 mL blood into 0.94 g PDMS was 62-84%, which is similar to analytical recoveries in conventional solvent extraction methods. The validated method was applied to 15 blood samples from green turtles with known concentrations of PCDD/Fs, dioxin-like PCBs, PBDEs and organochlorine pesticides. The quantified chemicals explained most of the dioxin-like activity (69-98%), but less than 0.4% of the oxidative stress response. The results demonstrate the applicability of PDMS-based passive sampling to extract bioaccumulative chemicals from blood as well as the value of in vitro bioassays for capturing the combined effects of unknown and known chemicals.

  9. Assessment of erythrocyte aggregation in whole blood samples by light backscattering: clinical applications

    NASA Astrophysics Data System (ADS)

    Priezzhev, Alexander V.; Firsov, Nikolai N.; Vyshlova, Marina G.; Lademann, Juergen; Richter, Heike; Kiesewetter, Holger; Mueller, Gerhard J.

    1999-05-01

    We report on the results of a collaborative effort made in the field of optical diagnostics of whole blood samples to study the ability of red blood cells to aggregate in a Couette chamber. We studied a possibility to quantitatively measure this ability as a function of the physiological state of blood donors. The aggregometer designed by the Russian coauthors of this paper and described in their earlier publications (see e.g. Proc SPIE 1884, 2100, 2678, 2982) was extensively used in the experiments performed in the Rheumatology Institute in Moscow and in the Charite Clinic in Berlin. The following parameters were measured: two characteristic times of RBC aggregation and the average spontaneous aggregation rate in the state of stasis, the average hydrodynamic strength of all aggregates and that of the largest aggregates. Different algorithms of the remission signal processing for the quantitative evaluation of the above parameters were compared. Reproducible alterations of the parameters from their normal values were obtained for blood samples from individuals suffering auto-immune disease and diabetes. Statistical data is reported proving high efficiency of the technique for the diagnostics of rheological disorders. Basing on these data the quantitative criteria of the heaviness of hemorheological state of the patients are proposed that are important for choosing specific therapies for which the patient is minimally resistant.

  10. Inactivation of human immunodeficiency virus type 1 in blood samples stored as high-salt lysates.

    PubMed

    Zolg, J W; Lanciotti, R S; Wendlinger, M; Meyer, W A

    1990-09-01

    Blood samples to be tested for the presence of parasite DNA by using specific DNA probes are routinely stored in our laboratory as high-salt lysates (HSL). To safeguard against the risk of accidental infection with etiological agents such as the human immunodeficiency virus type 1 (HIV-1) while manipulating large numbers of blood samples in preparation for DNA probing, we determined the residual infectivity of HIV-1 after exposure to HSL components. Both high-titer virus stocks or provirus-carrying cells, suspended either in tissue culture medium or freshly drawn blood, were completely inactivated upon contact with the HSL components. This was verified by the absence of any detectable HIV-1-specific antigen in the supernatants of long-term cultures and the absence of virus-specific DNA fragments after amplification by polymerase chain reaction with DNA from such cultures as target DNA. These results support the conclusion that the virus is in fact completely inactivated by contact with the HSL components, rendering blood specimens stored as HSL noninfectious in regard to HIV-1.

  11. Preconcentration and determination of lead and cadmium levels in blood samples of adolescent workers consuming smokeless tobacco products in Pakistan.

    PubMed

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Afridi, Hassan Imran; Brahman, Kapil Dev; Naeemullah; Khan, Sumaira; Panhwar, Abdul Haleem; Kamboh, Muhammad Afzal; Memon, Jamil R

    2015-05-01

    The present study was aimed to evaluate the cadmium (Cd) and lead (Pb) levels in the blood samples of adolescent boys, chewing different smokeless tobacco (SLT) products in Pakistan. For comparative purpose, boys of the same age group (12-15 years), not consumed any SLT products were selected as referents. To determine trace levels of Cd and Pb in blood samples, a preconcentration method, vortex-assisted liquid-liquid microextraction (VLLME) has been developed, prior to analysis by flame atomic absorption spectrometry. The hydrophobic chelates of Cd and Pb with ammonium pyrrolidinedithiocarbamate were extracted into the fine droplets of ionic liquid (IL) 1-butyl-3-methylimidazolium hexafluorophosphate, while nonionic surfactant, Triton X-114 was used as a dispersing medium. The main factors affecting the recoveries of Cd and Pb, such as concentration of APDC, centrifugation time, volume of IL and TX-114, were investigated in detail. It was also observed that adolescent boys who consumed different SLT products have 2- to 3-fold higher levels of Cd and Pb in their blood samples as compared to referent boys (p < 0.001).

  12. Surgical vascular access in the porcine model for long-term repeated blood sampling.

    PubMed

    Lombardo, Carlo; Damiano, Giuseppe; Cassata, Giovanni; Palumbo, Vincenzo Davide; Cacciabaudo, Francesco; Spinelii, Gabriele; Calvagna, Cristiano; Gioviale, Maria Concetta; Maione, Caro-lina; Lo Monte, Attilio Ignazio

    2010-09-01

    A simple technique for implanting a long-term jugular catheter in piglets under general anesthesia is described. We report our experience in 10 young female pigs with a body weight of 20-30 Kg. The surgical procedure involves implantation of a jugular central venous catheter (11Fr polyurethane) tunneled in the subcutaneous fat layer of the neck. This procedure may be performed in about 15 minutes. The maintenance of the catheter is described which allows several daily blood samples to be taken. This procedure reduces both the stress in piglets and the chance of catheter dislodgement due to the animals scratching or rubbing. Blood sampling can be easily performed with a low incidence of infection or thrombosis.

  13. Method and apparatus for imaging a sample on a device

    DOEpatents

    Trulson, Mark; Stern, David; Fiekowsky, Peter; Rava, Richard; Walton, Ian; Fodor, Stephen P. A.

    2001-01-01

    A method and apparatus for imaging a sample are provided. An electromagnetic radiation source generates excitation radiation which is sized by excitation optics to a line. The line is directed at a sample resting on a support and excites a plurality of regions on the sample. Collection optics collect response radiation reflected from the sample I and image the reflected radiation. A detector senses the reflected radiation and is positioned to permit discrimination between radiation reflected from a certain focal plane in the sample and certain other planes within the sample.

  14. Engineering Study of 500 ML Sample Bottle Transportation Methods

    SciTech Connect

    BOGER, R.M.

    1999-08-25

    This engineering study reviews and evaluates all available methods for transportation of 500-mL grab sample bottles, reviews and evaluates transportation requirements and schedules and analyzes and recommends the most cost-effective method for transporting 500-mL grab sample bottles.

  15. 40 CFR Appendix I to Part 261 - Representative Sampling Methods

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... Appendix I to Part 261 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) SOLID WASTES (CONTINUED) IDENTIFICATION AND LISTING OF HAZARDOUS WASTE Pt. 261, App. I Appendix I to Part 261—Representative Sampling Methods The methods and equipment used for sampling waste materials will vary with...

  16. GROUND WATER PURGING AND SAMPLING METHODS: HISTORY VS. HYSTERIA

    EPA Science Inventory

    It has been over 10 years since the low-flow ground water purging and sampling method was initially reported in the literature. The method grew from the recognition that well purging was necessary to collect representative samples, bailers could not achieve well purging, and high...

  17. Evaluation of Common Methods for Sampling Invertebrate Pollinator Assemblages: Net Sampling Out-Perform Pan Traps

    PubMed Central

    Popic, Tony J.; Davila, Yvonne C.; Wardle, Glenda M.

    2013-01-01

    Methods for sampling ecological assemblages strive to be efficient, repeatable, and representative. Unknowingly, common methods may be limited in terms of revealing species function and so of less value for comparative studies. The global decline in pollination services has stimulated surveys of flower-visiting invertebrates, using pan traps and net sampling. We explore the relative merits of these two methods in terms of species discovery, quantifying abundance, function, and composition, and responses of species to changing floral resources. Using a spatially-nested design we sampled across a 5000 km2 area of arid grasslands, including 432 hours of net sampling and 1296 pan trap-days, between June 2010 and July 2011. Net sampling yielded 22% more species and 30% higher abundance than pan traps, and better reflected the spatio-temporal variation of floral resources. Species composition differed significantly between methods; from 436 total species, 25% were sampled by both methods, 50% only by nets, and the remaining 25% only by pans. Apart from being less comprehensive, if pan traps do not sample flower-visitors, the link to pollination is questionable. By contrast, net sampling functionally linked species to pollination through behavioural observations of flower-visitation interaction frequency. Netted specimens are also necessary for evidence of pollen transport. Benefits of net-based sampling outweighed minor differences in overall sampling effort. As pan traps and net sampling methods are not equivalent for sampling invertebrate-flower interactions, we recommend net sampling of invertebrate pollinator assemblages, especially if datasets are intended to document declines in pollination and guide measures to retain this important ecosystem service. PMID:23799127

  18. Evaluation of common methods for sampling invertebrate pollinator assemblages: net sampling out-perform pan traps.

    PubMed

    Popic, Tony J; Davila, Yvonne C; Wardle, Glenda M

    2013-01-01

    Methods for sampling ecological assemblages strive to be efficient, repeatable, and representative. Unknowingly, common methods may be limited in terms of revealing species function and so of less value for comparative studies. The global decline in pollination services has stimulated surveys of flower-visiting invertebrates, using pan traps and net sampling. We explore the relative merits of these two methods in terms of species discovery, quantifying abundance, function, and composition, and responses of species to changing floral resources. Using a spatially-nested design we sampled across a 5000 km(2) area of arid grasslands, including 432 hours of net sampling and 1296 pan trap-days, between June 2010 and July 2011. Net sampling yielded 22% more species and 30% higher abundance than pan traps, and better reflected the spatio-temporal variation of floral resources. Species composition differed significantly between methods; from 436 total species, 25% were sampled by both methods, 50% only by nets, and the remaining 25% only by pans. Apart from being less comprehensive, if pan traps do not sample flower-visitors, the link to pollination is questionable. By contrast, net sampling functionally linked species to pollination through behavioural observations of flower-visitation interaction frequency. Netted specimens are also necessary for evidence of pollen transport. Benefits of net-based sampling outweighed minor differences in overall sampling effort. As pan traps and net sampling methods are not equivalent for sampling invertebrate-flower interactions, we recommend net sampling of invertebrate pollinator assemblages, especially if datasets are intended to document declines in pollination and guide measures to retain this important ecosystem service.

  19. Acetaminophen and Meloxicam Inhibit Platelet Aggregation and Coagulation in Blood Samples from Humans

    DTIC Science & Technology

    2014-01-01

    participant was sampled once with a total of 100-ml blood volume. Exclusion criteria included pregnancy, on- going therapeutic anticoagulation , and use...of thromboxane A2 (TxA2) from prostaglandin H2, which is generated from arachidonic acid by cyclo-oxygenase (COX-1). The antiplatelet effects of...is acetaminophen? Some practical cautions with this widely used agent . Clin Pediatr (Phila) 1973; 12:692– 696. 3 Whyte IM, Buckley NA, Reith DM

  20. Evaluating Composite Sampling Methods of Bacillus spores at Low Concentrations

    SciTech Connect

    Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.

    2016-10-13

    Restoring facility operations after the 2001 Amerithrax attacks took over three months to complete, highlighting the need to reduce remediation time. The most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite: a single cellulose sponge samples multiple coupons; 2) single medium multi-pass composite: a single cellulose sponge is used to sample multiple coupons; and 3) multi-medium post-sample composite: a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155CFU/cm2, respectively). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p-value < 0.0001) and coupon material (p-value = 0.0008). Recovery efficiency (RE) was higher overall using the post-sample composite (PSC) method compared to single medium composite from both clean and grime coated materials. RE with the PSC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, painted wall board, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but significantly lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when

  1. A Mixed Methods Sampling Methodology for a Multisite Case Study

    ERIC Educational Resources Information Center

    Sharp, Julia L.; Mobley, Catherine; Hammond, Cathy; Withington, Cairen; Drew, Sam; Stringfield, Sam; Stipanovic, Natalie

    2012-01-01

    The flexibility of mixed methods research strategies makes such approaches especially suitable for multisite case studies. Yet the utilization of mixed methods to select sites for these studies is rarely reported. The authors describe their pragmatic mixed methods approach to select a sample for their multisite mixed methods case study of a…

  2. Effect of blood sample handling post-collection on Erysipelothrix rhusiopathiae antibody titres.

    PubMed

    Neumann, Eric J; Bonistalli, Kathryn N

    2009-06-01

    A study was conducted to determine the effect of blood sample mishandling on the performance of an enzyme-linked immunosorbent assay for the detection of antibodies against Erysipelothrix rhusiopathiae. Eleven sample maltreatments (storage at -10 degrees C, storage at 4 degrees C, heat treatment of clotted blood, haemolysis, repetitive freeze-thaw cycling, and substitution of plasma in place of serum) were simulated in a laboratory environment and then run concurrently against a gold standard sample (storage at -80 degrees C). The mishandling treatment groups that simulated high levels of haemolysis had significantly lower optical density (OD) readings when compared to the gold standard. However, the magnitude of the effects was relatively small and only samples with OD values close to the cut-off changed state from positive to negative. Heat treatment had a minor, but non-significant, effect on OD values. Findings from this study suggested that immunoglobulin G antibody was stable in the face of most common sample mishandling events.

  3. The effect of sodium fluoride on the stability of cyanide in postmortem blood samples from fire victims.

    PubMed

    McAllister, J L; Roby, R J; Levine, Barry; Purser, David

    2011-06-15

    Assigning a level of significance to cyanide concentrations found in the blood of fire victims is often hampered by the fact that cyanide is inherently unstable in cadavers and in stored blood samples. A few researchers have proposed that sodium fluoride can be used to minimize the instability of cyanide in blood samples; however, controlled studies have not been performed to support validation of this hypothesis. To test the sodium fluoride hypothesis, both treated and control blood samples from 14 autopsied fire victims were tested over a 25-30 day period. A 2% concentration of sodium fluoride was added to the blood samples at the start of testing and the samples were refrigerated between testing intervals. Cyanide concentrations in the treated and control samples were measured between 9 and 11 days post treatment and between 25 and 30 days post treatment. A statistically significant difference was not present between blood cyanide concentrations in treated and control samples between 9 and 11 days. During this time period, although there were small statistically significant increases in both treated and untreated samples the fluctuations were minor. Since the treated and control samples did not exhibit instability between 9 and 11 days, it is not surprising that the sodium fluoride appeared to have no effect. However, a statistically significant difference between blood cyanide concentrations in treated and control samples was observed between 25 and 30 days. Those samples treated with sodium fluoride showed a reduction in blood cyanide variability with virtually no overall change, over a 25-30 day period when compared to control samples, while unconditioned samples showed a significant, average increase of 35%. Based on the findings of this study, it is recommended that 2% sodium fluoride be added to blood samples obtained from fire victims to reduce cyanide instability due to bacteriological activity.

  4. A rapid paper-based test for quantifying sickle hemoglobin in blood samples from patients with sickle cell disease.

    PubMed

    Piety, Nathaniel Z; Yang, Xiaoxi; Lezzar, Dalia; George, Alex; Shevkoplyas, Sergey S

    2015-06-01

    Quantification of sickle hemoglobin (HbS) in patients with sickle cell disease (SCD) undergoing hydroxyurea or chronic transfusion therapy is essential to monitoring the effectiveness of these therapies. The clinical monitoring of %HbS using conventional laboratory methods is limited by high per-test costs and long turnaround times usually associated with these methods. Here we demonstrate a simple, rapid, inexpensive paper-based assay capable of quantifying %HbS in blood samples from patients with SCD. A 20 μL droplet of whole blood and hemoglobin solubility buffer was deposited on chromatography paper. The relative color intensities of regions of the resulting blood stain, determined by automated image analysis, are used to estimate %HbS. We compared the paper-based assay with hemoglobin electrophoresis (comparison method) using blood samples from 88 subjects. The test shows high correlation (R(2)  = 0.86) and strong agreement (standard deviation of difference = 7%HbS) with conventional Hb electrophoresis measurement of %HbS, and closely approximates clinically predicted change in %HbS with transfusion therapy (mean difference 2.6%HbS, n = 5). The paper-based assay can be completed in less than 35 min and has a per-test cost less than $0.25. The assay is accurate across a wide range of HbS levels (10-97%) and hemoglobin concentrations (5.6-12.9 g/dL) and is unaffected by high levels of HbF (up to 80.6%). This study demonstrates the feasibility of the paper-based %HbS assay. The paper-based test could improve clinical care for SCD, particularly in resource-limited settings, by enabling more rapid and less expensive %HbS monitoring.

  5. Method and kit for the selective labeling of red blood cells in whole blood with TC-99M

    DOEpatents

    Srivastava, Suresh C.; Babich, John W.; Straub, Rita; Richards, Powell

    1988-01-01

    Disclosed herein are a method and kit for the preparation of .sup.99m Tc labeled red blood cells using whole blood in a closed sterile system containing stannous tin in a form such that it will enter the red blood cells and be available therein for the reduction of technetium.

  6. Method and kit for the selective labeling of red blood cells in whole blood with Tc-99m

    DOEpatents

    Srivastava, S.C.; Babich, J.W.; Straub, R.; Richards, P.

    1988-07-05

    Disclosed herein are a method and kit for the preparation of [sup 99m]Tc labeled red blood cells using whole blood in a closed sterile system containing stannous tin in a form such that it will enter the red blood cells and be available for the reduction of technetium. No Drawings

  7. Blood pressure and body mass index in an ethnically diverse sample of adolescents in Paramaribo, Suriname

    PubMed Central

    Agyemang, Charles; Oudeman, Eline; Zijlmans, Wilco; Wendte, Johannes; Stronks, Karien

    2009-01-01

    Background High blood pressure (BP) is now an important public health problem in non-industrialised countries. The limited evidence suggests ethnic inequalities in BP in adults in some non-industrialised countries. However, it is unclear whether these ethnic inequalities in BP patterns in adults reflect on adolescents. Hence, we assessed ethnic differences in BP, and the association of BP with body mass index (BMI) among adolescents aged 12–17 years in Paramaribo, Suriname. Methods Cross-sectional study with anthropometric and blood pressure measurements. A random sample of 855 adolescents (167 Hindustanis, 169 Creoles, 128 Javanese, 91 Maroons and 300 mixed-ethnicities) were studied. Ethnicity was based on self-reported ethnic origin. Results Among boys, Maroons had a lower age- and height-adjusted systolic BP than Creoles, and a lower diastolic BP than other ethnic groups. However, after further adjustment for BMI, only diastolic BP in Maroons was significantly lower than in Javanese (67.1 versus 70.9 mmHg). Creole boys had a lower diastolic BP than Hindustani (67.3 versus 70.2 mmHg) and Javanese boys after adjustment for age, height and BMI. Among girls, there were no significant differences in systolic BP between the ethnic groups. Maroon girls, however, had a lower diastolic BP (65.6 mmHg) than Hindustani (69.1 mmHg), Javanese (71.2 mmHg) and Mixed-ethnic (68.3 mmHg) girls, but only after differences in BMI had been adjusted for. Javanese had a higher diastolic BP than Creoles (71.2 versus 66.8 mmHg) and Mixed-ethnicity girls. BMI was positively associated with BP in all the ethnic groups, except for diastolic BP in Maroon girls. Conclusion The study findings indicate higher mean BP levels among Javanese and Hindustani adolescents compared with their African descent peers. These findings contrast the relatively low BP reported in Javanese and Hindustani adult populations in Suriname and underscore the need for public health measures early in life to prevent

  8. Blood

    MedlinePlus

    ... The liquid part, called plasma, is made of water, salts, and protein. Over half of your blood is plasma. The solid part of your blood contains red blood cells, white blood cells, and platelets. Red ...

  9. Array CGH Analysis of Paired Blood and Tumor Samples from Patients with Sporadic Wilms Tumor

    PubMed Central

    del Carmen Crespo, María; Vallespín, Elena; Palomares-Bralo, María; Martin-Arenas, Rubén; Rueda-Arenas, Inmaculada; Silvestre de Faria, Paulo Antonio; García-Miguel, Purificación; Lapunzina, Pablo; Regla Vargas, Fernando; Seuanez, Hector N.; Martínez-Glez, Víctor

    2015-01-01

    Wilms tumor (WT), the most common cancer of the kidney in infants and children, has a complex etiology that is still poorly understood. Identification of genomic copy number variants (CNV) in tumor genomes provides a better understanding of cancer development which may be useful for diagnosis and therapeutic targets. In paired blood and tumor DNA samples from 14 patients with sporadic WT, analyzed by aCGH, 22% of chromosome abnormalities were novel. All constitutional alterations identified in blood were segmental (in 28.6% of patients) and were also present in the paired tumor samples. Two segmental gains (2p21 and 20q13.3) and one loss (19q13.31) present in blood had not been previously described in WT. We also describe, for the first time, a small, constitutive partial gain of 3p22.1 comprising 2 exons of CTNNB1, a gene associated to WT. Among somatic alterations, novel structural chromosomal abnormalities were found, like gain of 19p13.3 and 20p12.3, and losses of 2p16.1-p15, 4q32.5-q35.1, 4q35.2-q28.1 and 19p13.3. Candidate genes included in these regions might be constitutively (SIX3, SALL4) or somatically (NEK1, PIAS4, BMP2) operational in the development and progression of WT. To our knowledge this is the first report of CNV in paired blood and tumor samples in sporadic WT. PMID:26317783

  10. [Respondent-Driven Sampling: a new sampling method to study visible and hidden populations].

    PubMed

    Mantecón, Alejandro; Juan, Montse; Calafat, Amador; Becoña, Elisardo; Román, Encarna

    2008-01-01

    The paper introduces a variant of chain-referral sampling: respondent-driven sampling (RDS). This sampling method shows that methods based on network analysis can be combined with the statistical validity of standard probability sampling methods. In this sense, RDS appears to be a mathematical improvement of snowball sampling oriented to the study of hidden populations. However, we try to prove its validity with populations that are not within a sampling frame but can nonetheless be contacted without difficulty. The basics of RDS are explained through our research on young people (aged 14 to 25) who go clubbing, consume alcohol and other drugs, and have sex. Fieldwork was carried out between May and July 2007 in three Spanish regions: Baleares, Galicia and Comunidad Valenciana. The presentation of the study shows the utility of this type of sampling when the population is accessible but there is a difficulty deriving from the lack of a sampling frame. However, the sample obtained is not a random representative one in statistical terms of the target population. It must be acknowledged that the final sample is representative of a 'pseudo-population' that approximates to the target population but is not identical to it.

  11. Modified salting-out method for DNA isolation from newborn cord blood nucleated cells.

    PubMed

    Noguera, N I; Tallano, C E; Bragós, I M; Milani, A C

    2000-01-01

    The present work describes modification of a widely used salting-out procedure to rapidly extract DNA suitable for PCR, using the ARMS method to amplify a target sequence in the beta-globin gene. The salting-out DNA extraction procedure did not completely remove or decrease the presence of inhibitors to PCR in a considerable number of cord blood samples. By introducing a simple phenol/chloroform step, before ethanol precipitation of the nucleic acid, to certain samples, we were able to eliminate or substantially reduce the presence of inhibitors to PCR without having to re-extract the samples.

  12. Method based on bioinspired sample improves autofocusing performances

    NASA Astrophysics Data System (ADS)

    Cao, Jie; Cheng, Yang; Wang, Peng; Peng, Yuxin; Zhang, Kaiyu; Wu, Leina; Xia, Wenze; Yu, Haoyong

    2016-10-01

    In order to solve the issue between fast autofocusing speed and high volume data processing, we propose a bioinspired sampling method based on a retina-like structure. We develop retina-like models and analyze the division of sampling structure. The optimal retina-like sample is obtained by analyzing two key parameters (sectors and radius of blind area) of the retina-like structure through experiments. Under the typical autofocus functions, including Vollath-4, Laplacian, Tenengrad, spatial frequency, and sum-modified-Laplacian (SML), we carry out comparative experiments of computation time based on the retina-like sample and a traditional uniform sample. The results show that the retina-like sample is suitable for those autofocus functions. Based on the autofocus function of SML, the average time of uniform sample decreases from 3.5 to 2.1 s for the retina-like sample.

  13. Identification of bacteria directly from positive blood culture samples by DNA pyrosequencing of the 16S rRNA gene.

    PubMed

    Motoshima, Maiko; Yanagihara, Katsunori; Morinaga, Yoshitomo; Matsuda, Junichi; Hasegawa, Hiroo; Kohno, Shigeru; Kamihira, Shimeru

    2012-11-01

    Rapid identification of the causative bacteria of sepsis in patients can contribute to the selection of appropriate antibiotics and improvement of patients' prognosis. Genotypic identification is an emerging technology that may provide an alternative method to, or complement, established phenotypic identification procedures. We evaluated a rapid protocol for bacterial identification based on PCR and pyrosequencing of the V1 and V3 regions of the 16S rRNA gene using DNA extracted directly from positive blood culture samples. One hundred and two positive blood culture bottles from 68 patients were randomly selected and the bacteria were identified by phenotyping and pyrosequencing. The results of pyrosequencing identification displayed 84.3 and 64.7 % concordance with the results of phenotypic identification at the genus and species levels, respectively. In the monomicrobial samples, the concordance between the results of pyrosequencing and phenotypic identification at the genus level was 87.0 %. Pyrosequencing identified one isolate in 60 % of polymicrobial samples, which were confirmed by culture analysis. Of the samples identified by pyrosequencing, 55.7 % showed consistent results in V1 and V3 targeted sequencing; other samples were identified based on the results of V1 (12.5 %) or V3 (31.8 %) sequencing alone. One isolate was erroneously identified by pyrosequencing due to high sequence similarity with another isolate. Pyrosequencing identified one isolate that was not detected by phenotypic identification. The process of pyrosequencing identification can be completed within ~4 h. The information provided by DNA-pyrosequencing for the identification of micro-organisms in positive blood culture bottles is accurate and could prove to be a rapid and useful tool in standard laboratory practice.

  14. Blood species identification using Near-Infrared diffuse transmitted spectra and PLS-DA method

    NASA Astrophysics Data System (ADS)

    Zhang, Linna; Zhang, Shengzhao; Sun, Meixiu; Wang, Zhennan; Li, Hongxiao; Li, Yingxin; Li, Gang; Lin, Ling

    2016-05-01

    Blood species identification is of great significance for blood supervision and wildlife investigations. The current methods used to identify the blood species are destructive. Near-Infrared spectroscopy method is known for its non-invasive properties. In this research, we combined Near-Infrared diffuse transmitted spectra and Partial Least Square Discrimination Analysis (PLS-DA) to identify three blood species, including macaque, human and mouse. Blind test and external test were used to assess the PLS-DA model. The model performed 100% accuracy in its identification between three blood species. This approach does not require a specific knowledge of biochemical features for each individual class but relies on a spectroscopic statistical differentiation of the whole components. This is the first time to demonstrate Near-Infrared diffuse transmitted spectra have the ability to identify the species of origin of blood samples. The results also support a good potential of absorption and scattering spectroscopy for species identification in practical applications for real-time detection.

  15. Cyanide concentrations in blood after cigarette smoking, as determined by a sensitive fluorimetric method.

    PubMed

    Lundquist, P; Rosling, H; Sörbo, B; Tibbling, L

    1987-07-01

    Using the sensitive fluorimetric method described here, we evaluated the determination of blood cyanide as a method for monitoring exposure to tobacco smoke. The mean concentration of cyanide in blood from eight nonsmokers was 0.098 (SD 0.036) mumol/L. The concentration of cyanide in blood of smokers who had refrained from smoking for at least 2 h before sampling peaked immediately after the subjects smoked a cigarette, then rapidly declined, with a half-life of about 4 min. Its rapid disappearance from blood makes cyanide an unsuitable marker of exposure to tobacco smoke. Because the ability of patients with hepatic dysfunction to detoxify cyanide has been presumed to be impaired, we monitored the concentrations of cyanide in the blood of four patients with severe hepatic insufficiency who smoked a cigarette. The rate of elimination of cyanide from blood after smoking was only slightly less in these patients than in the controls, and the difference was not statistically significant.

  16. Method for using polarization gating to measure a scattering sample

    DOEpatents

    Baba, Justin S.

    2015-08-04

    Described herein are systems, devices, and methods facilitating optical characterization of scattering samples. A polarized optical beam can be directed to pass through a sample to be tested. The optical beam exiting the sample can then be analyzed to determine its degree of polarization, from which other properties of the sample can be determined. In some cases, an apparatus can include a source of an optical beam, an input polarizer, a sample, an output polarizer, and a photodetector. In some cases, a signal from a photodetector can be processed through attenuation, variable offset, and variable gain.

  17. Dried venous blood samples for the detection and quantification of measles IgG using a commercial enzyme immunoassay.

    PubMed Central

    Riddell, Michaela A.; Byrnes, Graham B.; Leydon, Jennie A.; Kelly, Heath A.

    2003-01-01

    OBJECTIVES: To determine whether samples of dried venous blood (DVB) were an acceptable alternative to serum for detecting measles-specific IgG in a commercial enzyme immunoassay. METHODS: Paired samples of serum and DVB were collected from 98 suspected cases of measles and 1153 schoolchildren in Victoria, Australia. All samples were tested using the Dade Behring Enzygnost Anti-Measles-Virus/IgG immunoassay. DVB samples were eluted using either the sample buffer provided with the kit or 5% dry milk powder in phosphate-buffered saline-Tween 20. FINDINGS: DVB samples eluted by sample buffer showed significantly better linear correlation to the serum samples than did DVB samples eluted in 5% dry milk in phosphate-buffered saline-Tween 20. To improve the comparability of serum and DVB samples an adjustment factor of 1.28 was applied to the optical density (OD) values of DVB. This adjustment also enabled quantification of the titre of measles IgG in mIU/ml directly from the OD value using the alpha calculation as specified by the kit protocol. For DVB samples stored for less than six months at 4 degrees C, the assay showed an overall sensitivity of 98.4% and a specificity of 97.2% compared with the results of serum testing. CONCLUSION: These results illustrate the potential for DVB samples to be widely used with the Dade Behring enzyme immunoassay system for determining the immunity of the individual and the population to the measles virus. PMID:14758429

  18. Sampling and sample preparation methods for determining concentrations of mycotoxins in foods and feeds.

    PubMed

    2012-01-01

    Sample variation is often the largest error in determining concentrations of mycotoxins in food commodities. The worldwide safety evaluation of mycotoxins requires sampling plans that give acceptably accurate values for the levels of contamination in specific batches or lots of a commodity. Mycotoxin concentrations show a skewed or uneven distribution in foods and feeds, especially in whole kernels (or nuts), so it is extremely difficult to collect a sample that accurately represents the mean batch concentration. Sample variance studies and sampling plans have been published for select mycotoxins such as aflatoxin, fumonisin, and deoxynivalenol, emphasizing the importance of sample selection, sample size, and the number of incremental samples. For meaningful data to be generated from surveillance studies, representative samples should be collected from carefully selected populations (batches or lots) of food that, in turn, should be representative of clearly defined locations (e.g. a country, a region within a country). Although sampling variability is unavoidable, it is essential that the precision of the sampling plan be clearly defined and be considered acceptable by those responsible for interpreting and reporting the surveillance data. The factors influencing variability are detailed here, with reference to both major mycotoxins and major commodities. Sampling of large bag stacks, bulk shipments, and domestic supplies are all discussed. Sampling plans currently accepted in international trade are outlined. Acceptance sampling plans and the variabilities that affect operating characteristic curves of such plans are also detailed. The constraints and issues related to the sampling of harvested crops within subsistence farming areas are also discussed in this chapter, as are the essential rules of sample labelling and storage. The chapter concludes with a short section on sample preparation methods.

  19. [Recent advances in sample preparation methods of plant hormones].

    PubMed

    Wu, Qian; Wang, Lus; Wu, Dapeng; Duan, Chunfeng; Guan, Yafeng

    2014-04-01

    Plant hormones are a group of naturally occurring trace substances which play a crucial role in controlling the plant development, growth and environment response. With the development of the chromatography and mass spectroscopy technique, chromatographic analytical method has become a widely used way for plant hormone analysis. Among the steps of chromatographic analysis, sample preparation is undoubtedly the most vital one. Thus, a highly selective and efficient sample preparation method is critical for accurate identification and quantification of phytohormones. For the three major kinds of plant hormones including acidic plant hormones & basic plant hormones, brassinosteroids and plant polypeptides, the sample preparation methods are reviewed in sequence especially the recently developed methods. The review includes novel methods, devices, extractive materials and derivative reagents for sample preparation of phytohormones analysis. Especially, some related works of our group are included. At last, the future developments in this field are also prospected.

  20. Interval sampling methods and measurement error: a computer simulation.

    PubMed

    Wirth, Oliver; Slaven, James; Taylor, Matthew A

    2014-01-01

    A simulation study was conducted to provide a more thorough account of measurement error associated with interval sampling methods. A computer program simulated the application of momentary time sampling, partial-interval recording, and whole-interval recording methods on target events randomly distributed across an observation period. The simulation yielded measures of error for multiple combinations of observation period, interval duration, event duration, and cumulative event duration. The simulations were conducted up to 100 times to yield measures of error variability. Although the present simulation confirmed some previously reported characteristics of interval sampling methods, it also revealed many new findings that pertain to each method's inherent strengths and weaknesses. The analysis and resulting error tables can help guide the selection of the most appropriate sampling method for observation-based behavioral assessments.

  1. Comparative analysis of RNA-Seq data from brain and blood samples of Parkinson's disease.

    PubMed

    Chatterjee, Paulami; Roy, Debjani

    2017-03-11

    Parkinson's disease (PD) is the second most common neurodegenerative disorders throughout the world. In order to search for PD biomarkers, we performed a system-level study of RNA-Seq data from PD brain and blood samples. Differentially expressed miRs of RNA-Seq data were subjected to generate the Co-expression networks. Three highly co-expressed clusters were identified based on their correlation coefficient values and fold change ratio. SM2miR drugs of the miRs contained in the three highly co-expressed clusters were identified, and drugs common among these clusters were selected. Co-expressed miRs not previously known to be associated with PD were identified from both the samples. Functional enrichment analyses of these miR targets were done, and the pathways common and unique to both the samples were identified. Thus, our study presents a comparative analysis of miRs, their associated pathways, and drugs from brain and blood samples of PD that may help in system level understanding of this disease. miRs identified from our study may serve as biomarkers for PD.

  2. Arterial line blood sampling: preventing hypoglycaemic brain injury 2014: the Association of Anaesthetists of Great Britain and Ireland.

    PubMed

    Woodcock, T E; Cook, T M; Gupta, K J; Hartle, A

    2014-04-01

    Drawing samples from an indwelling arterial line is the method of choice for frequent blood analysis in adult critical care areas. Sodium chloride 0.9% is the recommended flush solution for maintaining the patency of arterial catheters, but it is easy to confuse with glucose-containing bags on rapid visual examination. The unintentional use of a glucose-containing solution has resulted in artefactually high glucose concentrations in blood samples drawn from the arterial line, leading to insulin administration causing hypoglycaemia and fatal neuroglycopenic brain injury. Recent data show that it remains a common error for incorrect fluids to be administered as arterial line flush infusions. Adherence to the National Patient Safety Agency's 2008 Rapid Response Report on this topic may not be enough to prevent such errors. This guideline makes detailed recommendations on the prescription, checking and administration of arterial line infusions in adult practice. We also make recommendations about storage, arterial pressure monitoring and sampling systems and techniques. Finally, we make recommendations about glucose monitoring and insulin administration. It is intended that adherence to these guidelines will reduce the frequency of sample contamination errors in arterial line use and capture events, when they do occur, before they cause patient harm.

  3. [Weighted estimation methods for multistage sampling survey data].

    PubMed

    Hou, Xiao-Yan; Wei, Yong-Yue; Chen, Feng

    2009-06-01

    Multistage sampling techniques are widely applied in the cross-sectional study of epidemiology, while methods based on independent assumption are still used to analyze such complex survey data. This paper aims to introduce the application of weighted estimation methods for the complex survey data. A brief overview of basic theory is described, and then a practical analysis is illustrated to apply to the weighted estimation algorithm in a stratified two-stage clustered sampling data. For multistage sampling survey data, weighted estimation method can be used to obtain unbiased point estimation and more reasonable variance estimation, and so make proper statistical inference by correcting the clustering, stratification and unequal probability effects.

  4. A random spatial sampling method in a rural developing nation

    PubMed Central

    2014-01-01

    Background Nonrandom sampling of populations in developing nations has limitations and can inaccurately estimate health phenomena, especially among hard-to-reach populations such as rural residents. However, random sampling of rural populations in developing nations can be challenged by incomplete enumeration of the base population. Methods We describe a stratified random sampling method using geographical information system (GIS) software and global positioning system (GPS) technology for application in a health survey in a rural region of Guatemala, as well as a qualitative study of the enumeration process. Results This method offers an alternative sampling technique that could reduce opportunities for bias in household selection compared to cluster methods. However, its use is subject to issues surrounding survey preparation, technological limitations and in-the-field household selection. Application of this method in remote areas will raise challenges surrounding the boundary delineation process, use and translation of satellite imagery between GIS and GPS, and household selection at each survey point in varying field conditions. This method favors household selection in denser urban areas and in new residential developments. Conclusions Random spatial sampling methodology can be used to survey a random sample of population in a remote region of a developing nation. Although this method should be further validated and compared with more established methods to determine its utility in social survey applications, it shows promise for use in developing nations with resource-challenged environments where detailed geographic and human census data are less available. PMID:24716473

  5. Poppy seed consumption and toxicological analysis of blood and urine samples.

    PubMed

    Moeller, Manfred R; Hammer, Karin; Engel, Oliver

    2004-07-16

    Poppy seeds contain morphine in different amounts. Reported concentrations are up to 294 mg morphine/kg poppy seeds. Since penalties based on Street Traffic Law (parapgraph 24a StVG) in Germany (administrative offence) require definitive proof of morphine in blood samples, and the "Grenzwertkommission" in consultation with the Ministry of Transportation recommended a threshold of free morphine of 10 ng/mL, the question arose whether the consumption of poppy seeds can lead to a blood concentrations equal or higher than 10 ng/mL of free morphine. Therefore, five volunteers ate poppy seed products (50 mg morphine/kg poppy seeds). In urine, all on-site tests were enzyme immunologically positive for opiates and were positive to morphine by GC/MS. All the blood samples were negative to morphine by EIA and to free morphine by GC/MS. However, after hydrolysis, morphine was detected by GC/MS in all cases. Accordingly, in Germany, penalties based on parapgraph 24a StVG are not likely to cause road users any concerns should they have consumed poppy seeds. Driver Licensing Authorities, however, should be advised of this problem to avoid unjustified legal measures.

  6. An automatic algorithm for the detection of Trypanosoma cruzi parasites in blood sample images.

    PubMed

    Soberanis-Mukul, Roger; Uc-Cetina, Víctor; Brito-Loeza, Carlos; Ruiz-Piña, Hugo

    2013-12-01

    Chagas disease is a tropical parasitic disease caused by the flagellate protozoan Trypanosoma cruzi (T. cruzi) and currently affecting large portions of the Americas. One of the standard laboratory methods to determine the presence of the parasite is by direct visualization in blood smears stained with some colorant. This method is time-consuming, requires trained microscopists and is prone to human mistakes. In this article we propose a novel algorithm for the automatic detection of T. cruzi parasites, in microscope digital images obtained from peripheral blood smears treated with Wright's stain. Our algorithm achieved a sensitivity of 0.98 and specificity of 0.85 when evaluated against a dataset of 120 test images. Experimental results show the versatility of the method for parasitemia determination.

  7. A spectral boundary integral method for flowing blood cells

    NASA Astrophysics Data System (ADS)

    Zhao, Hong; Isfahani, Amir H. G.; Olson, Luke N.; Freund, Jonathan B.

    2010-05-01

    A spectral boundary integral method for simulating large numbers of blood cells flowing in complex geometries is developed and demonstrated. The blood cells are modeled as finite-deformation elastic membranes containing a higher viscosity fluid than the surrounding plasma, but the solver itself is independent of the particular constitutive model employed for the cell membranes. The surface integrals developed for solving the viscous flow, and thereby the motion of the massless membrane, are evaluated using an O(NlogN) particle-mesh Ewald (PME) approach. The cell shapes, which can become highly distorted under physiologic conditions, are discretized with spherical harmonics. The resolution of these global basis functions is, of course, excellent, but more importantly they facilitate an approximate de-aliasing procedure that stabilizes the simulations without adding any numerical dissipation or further restricting the permissible numerical time step. Complex geometry no-slip boundaries are included using a constraint method that is coupled into an implicit system that is solved as part of the time advancement routine. The implementation is verified against solutions for axisymmetric flows reported in the literature, and its accuracy is demonstrated by comparison against exact solutions for relaxing surface deformations. It is also used to simulate flow of blood cells at 30% volume fraction in tubes between 4.9 and 16.9 μm in diameter. For these, it is shown to reproduce the well-known non-monotonic dependence of the effective viscosity on the tube diameter.

  8. Near-infrared spectral methods for noninvasively measuring blood glucose

    NASA Astrophysics Data System (ADS)

    Fei, Sun; Kong, Deyi; Mei, Tao; Tao, Yongchun

    2004-05-01

    Determination of blood glucose concentrations in diabetic patients is a frequently occurring procedure and an important tool for diabetes management. Use of noninvasive detection techniques can relieve patients from the pain of frequent finger pokes and avoid the infection of disease via blood. This thesis discusses current research and analyzes the advantages and shortages of different measurement methods, including: optical methods (Transmission, Polarimetry and scattering), then, we give emphasis to analyze the technology of near-infrared (NIR) spectra. NIR spectral range 700 nm ~2300 nm was used because of its good transparency for biological tissue and presence of glucose absorption band. In this work, we present an outline of noninvasive blood glucose measurement. A near-infrared light beam is passed through the finger, and the spectral components of the emergent beam are measured using spectroscopic techniques. The device includes light sources having the wavelengths of 600 nm - 1800 nm to illuminate the tissue. Receptors associated with the light sources for receiving light and generating a transmission signal representing the light transmitted are also provided. Once a transmission signal is received by receptors, and the high and low values from each of the signals are stored in the device. The averaged values are then analyzed to determine the glucose concentration, which is displayed on the device.

  9. Blood typing

    MedlinePlus

    ... ABO blood typing; Blood group; Anemia - immune hemolytic blood type; ABO blood type; A blood type; AB blood type; O blood type ... The 2 steps above can accurately determine your blood type. Rh typing uses a method similar to ABO ...

  10. Integration of sample analysis method (SAM) for polychlorinated biphenyls

    SciTech Connect

    Monagle, M.; Johnson, R.C.

    1996-05-01

    A completely integrated Sample Analysis Method (SAM) has been tested as part of the Contaminant Analysis Automation program. The SAM system was tested for polychlorinated biphenyl samples using five Standard Laboratory Modules{trademark}: two Soxtec{trademark} modules, a high volume concentrator module, a generic materials handling module, and the gas chromatographic module. With over 300 samples completed within the first phase of the validation, recovery and precision data were comparable to manual methods. Based on experience derived from the first evaluation of the automated system, efforts are underway to improve sample recoveries and integrate a sample cleanup procedure. In addition, initial work in automating the extraction of semivolatile samples using this system will also be discussed.

  11. Improved age determination of blood and teeth samples using a selected set of DNA methylation markers

    PubMed Central

    Kamalandua, Aubeline

    2015-01-01

    Age estimation from DNA methylation markers has seen an exponential growth of interest, not in the least from forensic scientists. The current published assays, however, can still be improved by lowering the number of markers in the assay and by providing more accurate models to predict chronological age. From the published literature we selected 4 age-associated genes (ASPA, PDE4C, ELOVL2, and EDARADD) and determined CpG methylation levels from 206 blood samples of both deceased and living individuals (age range: 0–91 years). This data was subsequently used to compare prediction accuracy with both linear and non-linear regression models. A quadratic regression model in which the methylation levels of ELOVL2 were squared showed the highest accuracy with a Mean Absolute Deviation (MAD) between chronological age and predicted age of 3.75 years and an adjusted R2 of 0.95. No difference in accuracy was observed for samples obtained either from living and deceased individuals or between the 2 genders. In addition, 29 teeth from different individuals (age range: 19–70 years) were analyzed using the same set of markers resulting in a MAD of 4.86 years and an adjusted R2 of 0.74. Cross validation of the results obtained from blood samples demonstrated the robustness and reproducibility of the assay. In conclusion, the set of 4 CpG DNA methylation markers is capable of producing highly accurate age predictions for blood samples from deceased and living individuals PMID:26280308

  12. Method and sample spinning apparatus for measuring the NMR spectrum of an orientationally disordered sample

    DOEpatents

    Pines, Alexander; Samoson, Ago

    1990-01-01

    An improved NMR apparatus and method are described which substantially improve the resolution of NMR measurements made on powdered or amorphous or otherwise orientationally disordered samples. The apparatus spins the sample about an axis. The angle of the axis is mechanically varied such that the time average of two or more Legendre polynomials are zero.

  13. Measurement of carboxyhemoglobin in forensic blood samples using UV-visible spectrometry and improved principal component regression

    SciTech Connect

    Egan, William; Morgan, Stephen L. Brewer, William E.

    1999-02-01

    The forensic determination of carboxyhemoglobin (COHb) in blood was performed by using an improved principal component regression (PCR) technique applied to UV-visible spectra. Calibration data were decomposed into principal components, and the principal components useful for prediction were selected by their correlation with calibration spectra. Cross-validation of prediction results was done by leverage-corrected residuals. Confidence and prediction intervals derived from classical regression theory were found to be reasonable in size. The results compared favorably to a comparison study conducted by using a CO Oximeter method. In analysis of forensic case study samples, the improved PCR method allowed detection of abnormal samples and successfully predicted percentages of COHb and methemoglobin (MetHb), and provided error estimates for those predictions. {copyright} {ital 1999} {ital Society for Applied Spectroscopy}

  14. Straightforward and rapid determination of sulfadoxine and sulfamethoxazole in capillary blood on sampling paper with liquid chromatography and UV detection.

    PubMed

    Lindkvist, J; Malm, M; Bergqvist, Y

    2009-04-01

    A method for the determination of sulfadoxine and sulfamethoxazole in capillary blood on sampling paper has been developed and validated. The method is straightforward with minimal sample preparation, and is suitable for rural settings. Separation of sulfadoxine, sulfamethoxazole and internal standard was performed using a Purospher STAR RP-18 endcapped LC column (150x4.6mm) with a mobile phase consisting of acetonitrile:sodium acetate buffer pH 5.2, I=0.1 (1:2, v/v). For sulfadoxine, the within-day precision was 5.3% at 15micromol/l and 3.7% at 600micromol/l, while for sulfamethoxazole it was 5.7% at 15micromol/l and 3.8% at 600micromol/l. The lower limit of quantification was determined to 5micromol/l and precision was 5.5% and 5.0% for sulfadoxine and sulfamethoxazole, respectively.

  15. Molecular diagnosis of the tick-borne pathogen Anaplasma marginale in cattle blood samples from Nigeria using qPCR.

    PubMed

    Elelu, Nusirat; Ferrolho, Joana; Couto, Joana; Domingos, Ana; Eisler, Mark C

    2016-12-01

    Tick-borne diseases (TBDs) are some of the most important animal health and management problems in Africa, including Nigeria. This study aims to determine the prevalence of an important TBD, anaplasmosis, in a North-central region of Nigeria. Blood samples were collected from cattle and stored on Whatman FTA(®) cards. Information on village, age and sex associated with each cattle was also recorded. The packed red blood cell volume (PCV) for each blood sample was determined. After DNA extraction, pathogen presence was evaluated by TaqMan(®) based qPCR of which 75.9 % of the cattle tested positive for Anaplasma marginale. Statistical analysis revealed that the presence of A. marginale infection differed significantly between cattle age groups. However, there was no significant difference in the prevalence of this pathogen between the sexes or among cattle grouped by PCV level. Finally, using a highly sensitive molecular method our pioneer study contributes to the improvement of the current knowledge regarding tick-borne pathogens that seriously affect animal health in specific areas of Nigeria.

  16. Molecular RH blood group typing of serologically D-/CE+ donors: the use of a polymerase chain reaction-sequence-specific primer test kit with pooled samples.

    PubMed

    Londero, Donatella; Fiorino, Mauro; Miotti, Valeria; de Angelis, Vincenzo

    2011-01-01

    The known presence of RHD blood group alleles in apparently D– individuals who are positive for C or E antigens leads to an appropriate investigation for the RHD gene on the red blood cells (RBCs) of D– blood donors, thus preventing their RBCs from immunizing D– recipients. Ready-to-use polymerase chain reaction–sequence-specific primer (PCR-SSP) typing kits are available and allow single-sample results. The need to perform this testing on a large number of donors affiliated with the Transfusion Department of Udine (Northern Italy) led to the use of molecular genetic RH blood group typing with PCR-SSP test kits and DNA samples mixed in pools. From a population of 35,000 blood donors screened for D antigen by serologic typing, a total of 235 samples, distributed in pools of 5 DNA samples, were investigated. Positive results were reevaluated by opening the pools and retesting single samples. Validation of DNA-pool typing with commercial kits was done. Among 235 genotyped samples, 12 were found to be PCR positive (5.1%), exhibiting DEL genotype and RHD-CE-D hybrid alleles. Our data demonstrate that the use of a PCR-SSP commercial test kit with pooled samples is a helpful and valid method to correctly detect RHD alleles. As a consequence, we reclassified our donors as carriers of potentially immunogenic alleles.

  17. Methods for collection and analysis of water samples

    USGS Publications Warehouse

    Rainwater, Frank Hays; Thatcher, Leland Lincoln

    1960-01-01

    This manual contains methods used by the U.S. Geological Survey to collect, preserve, and analyze water samples. Throughout, the emphasis is on obtaining analytical results that accurately describe the chemical composition of the water in situ. Among the topics discussed are selection of sampling sites, frequency of sampling, field equipment, preservatives and fixatives, analytical techniques of water analysis, and instruments. Seventy-seven laboratory and field procedures are given for determining fifty-three water properties.

  18. Optimized method for dissolved hydrogen sampling in groundwater.

    PubMed

    Alter, Marcus D; Steiof, Martin

    2005-06-01

    Dissolved hydrogen concentrations are used to characterize redox conditions of contaminated aquifers. The currently accepted and recommended bubble strip method for hydrogen sampling (Wiedemeier et al., 1998) requires relatively long sampling times and immediate field analysis. In this study we present methods for optimized sampling and for sample storage. The bubble strip sampling method was examined for various flow rates, bubble sizes (headspace volume in the sampling bulb) and two different H2 concentrations. The results were compared to a theoretical equilibration model. Turbulent flow in the sampling bulb was optimized for gas transfer by reducing the inlet diameter. Extraction with a 5 mL headspace volume and flow rates higher than 100 mL/min resulted in 95-100% equilibrium within 10-15 min. In order to investigate the storage of samples from the gas sampling bulb gas samples were kept in headspace vials for varying periods. Hydrogen samples (4.5 ppmv, corresponding to 3.5 nM in liquid phase) could be stored up to 48 h and 72 h with a recovery rate of 100.1+/-2.6% and 94.6+/-3.2%, respectively. These results are promising and prove the possibility of storage for 2-3 days before laboratory analysis. The optimized method was tested at a field site contaminated with chlorinated solvents. Duplicate gas samples were stored in headspace vials and analyzed after 24 h. Concentrations were measured in the range of 2.5-8.0 nM corresponding to known concentrations in reduced aquifers.

  19. Capillary microextraction: A new method for sampling methamphetamine vapour.

    PubMed

    Nair, M V; Miskelly, G M

    2016-11-01

    Clandestine laboratories pose a serious health risk to first responders, investigators, decontamination companies, and the public who may be inadvertently exposed to methamphetamine and other chemicals used in its manufacture. Therefore there is an urgent need for reliable methods to detect and measure methamphetamine at such sites. The most common method for determining methamphetamine contamination at former clandestine laboratory sites is selected surface wipe sampling, followed by analysis with gas chromatography-mass spectrometry (GC-MS). We are investigating the use of sampling for methamphetamine vapour to complement such wipe sampling. In this study, we report the use of capillary microextraction (CME) devices for sampling airborne methamphetamine, and compare their sampling efficiency with a previously reported dynamic SPME method. The CME devices consisted of PDMS-coated glass filter strips inside a glass tube. The devices were used to dynamically sample methamphetamine vapour in the range of 0.42-4.2μgm(-3), generated by a custom-built vapour dosing system, for 1-15min, and methamphetamine was analysed using a GC-MS fitted with a ChromatoProbe thermal desorption unit. The devices showed good reproducibility (RSD<15%), and a curvilinear pre-equilibrium relationship between sampling times and peak area, which can be utilised for calibration. Under identical sampling conditions, the CME devices were approximately 30 times more sensitive than the dynamic SPME method. The CME devices could be stored for up to 3days after sampling prior to analysis. Consecutive sampling of methamphetamine and its isotopic substitute, d-9 methamphetamine showed no competitive displacement. This suggests that CME devices, pre-loaded with an internal standard, could be a feasible method for sampling airborne methamphetamine at former clandestine laboratories.

  20. First Molecular Identification of Dirofilaria repens in a Dog Blood Sample from Guanajuato, Mexico.

    PubMed

    Ramos-Lopez, Sandra; León-Galván, Ma Fabiola; Salas-Alatorre, Mariana; Lechuga-Arana, Alma A; Valencia-Posadas, Mauricio; Gutiérrez-Chávez, Abner J

    2016-11-01

    Dirofilaria repens is the causative zoonotic agent of subcutaneous dirofilariosis. Although the American continent is considered currently free of D. repens infections, in the past few years there have been some reports suggesting the presence of this agent in the Americas. In Guanajuato, Mexico, there is a report of the presence of D. repens. To determine whether this parasite is really present in Guanajuato, a total of 177 EDTA blood samples from dogs were tested using the modified Knott's technique. Using this technique, we found 15 positive samples, which were then analyzed using the PCR technique, with a panfilarial set of primers and a specific primer pair for D. repens. In one sample, the expected band was present and the corresponding amplicon, sequenced, displayed a 100% identity to D. repens, confirming the presence of this exotic and zoonotic filarial species in the municipality of Silao, Guanajuato, Mexico.

  1. Trace element levels in whole blood samples from residents of the city Badajoz, Spain.

    PubMed

    Moreno, M A; Marin, C; Vinagre, F; Ostapczuk, P

    1999-05-19

    Copper, lead, cadmium, and zinc were determined by anodic stripping voltammetry after sample digestion and potentiometric stripping analysis was used for Pb and Cd determination in original samples. Selenium was determined by cathodic stripping voltammetry or hydride generation AAS. Element levels found in the whole blood sample in a group of 82 people are for Cd: 0.98 +/- 0.94 ng/ml; for Pb: 46.7 +/- 28.6 ng/ml; for Cu: 1.07 +/- 0.12 micrograms/ml; for Zn: 6.95 +/- 1.08 micrograms/ml, and for Se: 116 +/- 25 ng/ml. Analytical data have been correlated to age, sex, smokers or non-smokers, drinking and food habits.

  2. Effects of repeated blood samplings on locomotor activity, evasion and wheel-running activity in mice.

    PubMed

    Pfeil, R

    1988-01-01

    The effects of serial blood sampling on nocturnal locomotor activity, evasion, wheel-running activity and body mass were studied in male NMRI mice aged 7-8 weeks. The erythrocyte count, haematocrit and haemoglobin concentration at the beginning and end of the study showed no difference in group 1 (two samples per week, 0.08 ml each) while there was a significant decrease in the group 2 values (three samples per week, 0.08 ml each). The total amount of nocturnal locomotor activity decreased in the animals bled repeatedly while the periods with locomotor activity increased. These alterations appeared particularly after bleeding. In the test-group animals evasion showed a decrease compared with the untreated control animals, but there was no evidence of a relation to the timing of the bleedings.

  3. Demonstration Report for Visual Sample Plan (VSP) Verification Sampling Methods at the Navy/DRI Site

    DTIC Science & Technology

    2011-08-01

    STATISTICAL VERIFICATION AND REMEDIATION SAMPLING METHODS (200837) August 2011 Pacific Northwest National Laboratory Brent Pulsipher...17. LIMIT ATIOH OF 1S. NUMSER 19~. NAME OS: RESPONSI’SLE PERSON ABSTRACT o• Brent Pulsipher ... ...., .. •. ’ · ’"" .... PAG .’ES uu 93 19b... Statistical Verification Sampling Methods in VSP ii August 2011 6.2.1  Transect Survey Design and Parameter Settings

  4. Blood group typing based on recording the elastic scattering of laser radiation using the method of digital imaging

    NASA Astrophysics Data System (ADS)

    Dolmashkin, A. A.; Dubrovskii, V. A.; Zabenkov, I. V.

    2012-05-01

    The possibility is demonstrated to determine the human blood group by recording the scattering of laser radiation with the help of the digital imaging method. It is experimentally shown that the action of a standing ultrasound wave leads to acceleration of the agglutination reaction of red blood cells, to formation of larger immune complexes of red blood cells, and, as a consequence, to acceleration of their sedimentation. In the absence of agglutination of red blood cells the ultrasound does not enhance the relevant processes. This difference in the results of ultrasound action on the mixture of blood and serum allows a method of blood typing to be offered. Theoretical modelling of the technique of the practical blood typing, carried out on the basis of the elastic light scattering theory, agrees well with the experimental results, which made it possible to plan further improvement of the proposed method. The studies of specific features of sedimentation of red blood cells and their immune complexes were aimed at the optimisation of the sample preparation, i.e., at the search for such experimental conditions that provide the maximal resolution of the method and the device for registering the reaction of red blood cells agglutination. The results of the study may be used in designing the instrumentation for blood group assessment in humans.

  5. Blood group typing based on recording the elastic scattering of laser radiation using the method of digital imaging

    SciTech Connect

    Dolmashkin, A A; Dubrovskii, V A; Zabenkov, I V

    2012-05-31

    The possibility is demonstrated to determine the human blood group by recording the scattering of laser radiation with the help of the digital imaging method. It is experimentally shown that the action of a standing ultrasound wave leads to acceleration of the agglutination reaction of red blood cells, to formation of larger immune complexes of red blood cells, and, as a consequence, to acceleration of their sedimentation. In the absence of agglutination of red blood cells the ultrasound does not enhance the relevant processes. This difference in the results of ultrasound action on the mixture of blood and serum allows a method of blood typing to be offered. Theoretical modelling of the technique of the practical blood typing, carried out on the basis of the elastic light scattering theory, agrees well with the experimental results, which made it possible to plan further improvement of the proposed method. The studies of specific features of sedimentation of red blood cells and their immune complexes were aimed at the optimisation of the sample preparation, i.e., at the search for such experimental conditions that provide the maximal resolution of the method and the device for registering the reaction of red blood cells agglutination. The results of the study may be used in designing the instrumentation for blood group assessment in humans.

  6. Nominal Weights Mean Equating: A Method for Very Small Samples

    ERIC Educational Resources Information Center

    Babcock, Ben; Albano, Anthony; Raymond, Mark

    2012-01-01

    The authors introduced nominal weights mean equating, a simplified version of Tucker equating, as an alternative for dealing with very small samples. The authors then conducted three simulation studies to compare nominal weights mean equating to six other equating methods under the nonequivalent groups anchor test design with sample sizes of 20,…

  7. Field Evaluation of Personal Sampling Methods for Multiple Bioaerosols

    PubMed Central

    Wang, Chi-Hsun; Chen, Bean T.; Han, Bor-Cheng; Liu, Andrew Chi-Yeu; Hung, Po-Chen; Chen, Chih-Yong; Chao, Hsing Jasmine

    2015-01-01

    Ambient bioaerosols are ubiquitous in the daily environment and can affect health in various ways. However, few studies have been conducted to comprehensively evaluate personal bioaerosol exposure in occupational and indoor environments because of the complex composition of bioaerosols and the lack of standardized sampling/analysis methods. We conducted a study to determine the most efficient collection/analysis method for the personal exposure assessment of multiple bioaerosols. The sampling efficiencies of three filters and four samplers were compared. According to our results, polycarbonate (PC) filters had the highest relative efficiency, particularly for bacteria. Side-by-side sampling was conducted to evaluate the three filter samplers (with PC filters) and the NIOSH Personal Bioaerosol Cyclone Sampler. According to the results, the Button Aerosol Sampler and the IOM Inhalable Dust Sampler had the highest relative efficiencies for fungi and bacteria, followed by the NIOSH sampler. Personal sampling was performed in a pig farm to assess occupational bioaerosol exposure and to evaluate the sampling/analysis methods. The Button and IOM samplers yielded a similar performance for personal bioaerosol sampling at the pig farm. However, the Button sampler is more likely to be clogged at high airborne dust concentrations because of its higher flow rate (4 L/min). Therefore, the IOM sampler is a more appropriate choice for performing personal sampling in environments with high dust levels. In summary, the Button and IOM samplers with PC filters are efficient sampling/analysis methods for the personal exposure assessment of multiple bioaerosols. PMID:25799419

  8. Methods for detection of West Nile virus antibodies in mosquito blood meals.

    PubMed

    Komar, Nicholas; Panella, Nicholas A; Young, Ginger R; Basile, Alison J

    2015-03-01

    We describe and compare 2 qualitative serologic techniques for detecting West Nile virus (WNV)-specific antibodies in mosquito blood meals. The techniques are the biotin microsphere immunoassay (b-MIA) and the inhibition platform of the VectorTest™ WNV antigen assay (VecTest-inhibition). To demonstrate the ability of these tests to detect WNV-neutralizing antibodies, we experimentally exposed feeding mosquitoes to blood containing 5 concentrations of 6B6C-1, a flavivirus-neutralizing monoclonal antibody. Antibody concentrations were quantified using the 90% plaque-reduction neutralization test (PRNT90). After 24 h of blood-meal digestion at 22.5°C, the threshold PRNT90 titer of detection was ≤18 for b-MIA and ≤50 for VecTest-inhibition. Both tests reliably detected antibodies in 3 of 3 blood meals that had been digested for up to 30 h, or were about 25% digested. The b-MIA was also applied to mosquitoes that had engorged on avian blood in Arizona following a WNV epidemic in 2010. There was no significant difference in the WNV antibody prevalence determined by b-MIA (52% of 71 avian blood meals) compared to the WNV-neutralizing antibody prevalence in birds determined by direct sampling (49% of 234 birds). VecTest-inhibition requires fewer resources and may be used in the field without a laboratory, but consumes the entire blood meal and relies on subjective interpretation of results. The b-MIA requires a laboratory and sophisticated equipment and reagents. Results for b-MIA are analyzed objectively and can be applied to mosquito blood meals with greater confidence than the VecTest-inhibition method and thus can contribute substantially to research and surveillance programs that would benefit from the detection of specific WNV antibodies in mosquito blood meals.

  9. A cryopreservation method for Pasteurella multocida from wetland samples

    USGS Publications Warehouse

    Moore, Melody K.; Shadduck, D.J.; Goldberg, D.R.; Samuel, M.D.

    1998-01-01

    A cryopreservation method and improved isolation techniques for detection of Pasteurella multocida from wetland samples were developed. Wetland water samples were collected in the field, diluted in dimethyl sulfoxide (DMSO, final concentration 10%), and frozen at -180 C in a liquid nitrogen vapor shipper. Frozen samples were transported to the laboratory where they were subsequently thawed and processed in Pasteurella multocida selective broth (PMSB) to isolate P. multocida. This method allowed for consistent isolation of 2 to 18 organisms/ml from water seeded with known concentrations of P. multocida. The method compared favorably with the standard mouse inoculation method and allowed for preservation of the samples until they could be processed in the laboratory.

  10. Rapid Intrinsic Fluorescence Method for Direct Identification of Pathogens in Blood Cultures

    PubMed Central

    Walsh, John D.; Hyman, Jay M.; Borzhemskaya, Larisa; Bowen, Ann; McKellar, Caroline; Ullery, Michael; Mathias, Erin; Ronsick, Christopher; Link, John; Wilson, Mark; Clay, Bradford; Robinson, Ron; Thorpe, Thurman; van Belkum, Alex; Dunne, W. Michael

    2013-01-01

    ABSTRACT A positive blood culture is a critical result that requires prompt identification of the causative agent. This article describes a simple method to identify microorganisms from positive blood culture broth within the time taken to perform a Gram stain (<20 min). The method is based on intrinsic fluorescence spectroscopy (IFS) of whole cells and required development of a selective lysis buffer, aqueous density cushion, optical microcentrifuge tube, and reference database. A total of 1,121 monomicrobial-positive broth samples from 751 strains were analyzed to build a database representing 37 of the most commonly encountered species in bloodstream infections or present as contaminants. A multistage algorithm correctly classified 99.6% of unknown samples to the Gram level, 99.3% to the family level, and 96.5% to the species level. There were no incorrect results given at the Gram or family classification levels, while 0.8% of results were discordant at the species level. In 8/9 incorrect species results, the misidentified isolate was assigned to a species of the same genus. This unique combination of selective lysis, density centrifugation, and IFS can rapidly identify the most common microbial species present in positive blood cultures. Faster identification of the etiologic agent may benefit the clinical management of sepsis. Further evaluation is now warranted to determine the performance of the method using clinical blood culture specimens. PMID:24255123

  11. The effect of different blood sampling sites and analyses on the relationship between exercise intensity and 4.0 mmol.l-1 blood lactate concentration.

    PubMed

    Foxdal, P; Sjödin, A; Ostman, B; Sjödin, B

    1991-01-01

    The aim of the study was to examine whether the difference in lactate concentration in different blood fractions is of practical importance when using blood lactate as a test variable of aerobic endurance capacity. Ten male firefighters performed submaximally graded exercise on a cycle ergometer for 20-25 min. Venous and capillary blood samples were taken every 5 min for determination of haematocrit and lactate concentrations in plasma, venous and capillary blood. At the same time, expired air was collected in Douglas bags for determination of the oxygen consumption. A lactate concentration of 4.0 mmol.l-1 was used as the reference value to compare the oxygen consumption and exercise intensity when different types of blood specimen and sampling sites were used for lactate analysis. At this concentration the exercise intensity was 17% lower (P less than 0.01) when plasma lactate was compared to venous blood lactate, and 12% lower (P less than 0.05) when capillary blood lactate was used. Similar discrepancies were seen in oxygen consumption. The results illustrated the importance of standardizing sampling and handling of blood specimens for lactate determination to enable direct comparisons to be made among results obtained in different studies.

  12. Probability Sampling Method for a Hidden Population Using Respondent-Driven Sampling: Simulation for Cancer Survivors.

    PubMed

    Jung, Minsoo

    2015-01-01

    When there is no sampling frame within a certain group or the group is concerned that making its population public would bring social stigma, we say the population is hidden. It is difficult to approach this kind of population survey-methodologically because the response rate is low and its members are not quite honest with their responses when probability sampling is used. The only alternative known to address the problems caused by previous methods such as snowball sampling is respondent-driven sampling (RDS), which was developed by Heckathorn and his colleagues. RDS is based on a Markov chain, and uses the social network information of the respondent. This characteristic allows for probability sampling when we survey a hidden population. We verified through computer simulation whether RDS can be used on a hidden population of cancer survivors. According to the simulation results of this thesis, the chain-referral sampling of RDS tends to minimize as the sample gets bigger, and it becomes stabilized as the wave progresses. Therefore, it shows that the final sample information can be completely independent from the initial seeds if a certain level of sample size is secured even if the initial seeds were selected through convenient sampling. Thus, RDS can be considered as an alternative which can improve upon both key informant sampling and ethnographic surveys, and it needs to be utilized for various cases domestically as well.

  13. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    PubMed

    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.

  14. An enzyme-based DNA preparation method for application to forensic biological samples and degraded stains.

    PubMed

    Lounsbury, Jenny A; Coult, Natalie; Miranian, Daniel C; Cronk, Stephen M; Haverstick, Doris M; Kinnon, Paul; Saul, David J; Landers, James P

    2012-09-01

    Extraction of DNA from forensic samples typically uses either an organic extraction protocol or solid phase extraction (SPE) and these methods generally involve numerous sample transfer, wash and centrifugation steps. Although SPE has been successfully adapted to the microdevice, it can be problematic because of lengthy load times and uneven packing of the solid phase. A closed-tube enzyme-based DNA preparation method has recently been developed which uses a neutral proteinase to lyse cells and degrade proteins and nucleases [14]. Following a 20 min incubation of the buccal or whole blood sample with this proteinase, DNA is polymerase chain reaction (PCR)-ready. This paper describes the optimization and quantitation of DNA yield using this method, and application to forensic biological samples, including UV- and heat-degraded whole blood samples on cotton or blue denim substrates. Results demonstrate that DNA yield can be increased from 1.42 (±0.21)ng/μL to 7.78 (±1.40)ng/μL by increasing the quantity of enzyme per reaction by 3-fold. Additionally, there is a linear relationship between the amount of starting cellular material added and the concentration of DNA in the solution, thereby allowing DNA yield estimations to be made. In addition, short tandem repeat (STR) profile results obtained using DNA prepared with the enzyme method were comparable to those obtained with a conventional SPE method, resulting in full STR profiles (16 of 16 loci) from liquid samples (buccal swab eluate and whole blood), dried buccal swabs and bloodstains and partial profiles from UV or heat-degraded bloodstains on cotton or blue denim substrates. Finally, the DNA preparation method is shown to be adaptable to glass or poly(methyl methacrylate) (PMMA) microdevices with little impact on STR peak height but providing a 20-fold reduction in incubation time (as little as 60 s), leading to a ≥1 h reduction in DNA preparation time.

  15. A quantitative sampling method for Oncomelania quadrasi by filter paper.

    PubMed

    Tanaka, H; Santos, M J; Matsuda, H; Yasuraoka, K; Santos, A T

    1975-08-01

    Filter paper was found to attract Oncomelania quadrasi in waters the same way as fallen dried banana leaves, although less number of other species of snails was collected on the former than on the latter. Snails were collected in limited areas using a tube (85 cm2 area at cross-section) and a filter paper (20 X 20 CM) samplers. The sheet of filter paper was placed close to the spot where a tube sample was taken, and recovered after 24 hours. At each sampling, 30 samples were taken by each method in an area and sampling was made four times. The correlation of the number of snails collected by the tube and that by filter paper was studied. The ratio of the snail counts by the tube sampler to those by the filter paper was 1.18. A loose correlation was observed between snail counts of both methods as shown by the correlation coefficient r = 0.6502. The formulas for the regression line were Y = 0.77 X + 1.6 and X = 0.55 Y + 1.35 for 3 experiments where Y is the number of snails collected by tube sampling and X is the number of snails collected in the sheet of filter paper. The type of snail distribution was studied in the 30 samples taken by each method and this was observed to be nearly the same in both sampling methods. All sampling data were found to fit the negative binomial distribution with the values of the constant k varying very much from 0.5775 to 5.9186 in (q -- p)-k. In each experiment, the constant k was always larger in tube sampling than in filter paper sampling. This indicates that the uneven distribution of snails on the soil surface becomes more conspicuous by the filter paper sampling.

  16. The influence of sampling site and assay method on lactate concentration in response to rock climbing.

    PubMed

    Draper, Nick; Brent, Simon; Hale, Beverley; Coleman, Ian

    2006-11-01

    The sport of rock climbing has increased in popularity and as a focus for research. Previous studies have examined the physiological determinants for successful performance. Variation is evident between studies over lactate sampling sites and assay methods. The aim of this study was to examine the limits of agreement between the YSI 2300 analyser and the Lactate Pro for finger and ear capillary blood samples in a climbing context. Forty-five (31 males and 14 females) participants volunteered to complete the climbing trial. Blood samples were collected simultaneously from finger and ear pre, post and 5 min post climb. The repeatability results indicated a good agreement across samples. Modelling analysis indicated the use of a -0.175 mmol l(-1) adjustment to move from Lactate Pro to YSI finger concentrations. To move from finger to ear concentrations, using the Lactate Pro, modelling analysis suggested a regression equation of Y = 0.827x + 0.769 adjustment for pre climb samples and Y = 0.955x + 0.566 for post climb concentrations. To better understand the physiological demands of climbing further research on natural rock is required. Results from this study suggest the Lactate Pro and blood sampling from the ear lobe could be of benefit to future rock climbing field studies.

  17. Method for extending the useful shelf-life of refrigerated red blood cells by flushing with inert gas

    DOEpatents

    Bitensky, M.W.; Yoshida, Tatsuro

    1997-04-29

    A method is disclosed using oxygen removal for extending the useful shelf-life of refrigerated red blood cells. A cost-effective, 4 C storage procedure that preserves red cell quality and prolongs post-transfusion in vivo survival is described. Preservation of adenosine triphosphate levels and reduction in hemolysis and in membrane vesicle production of red blood cells stored at 4 C for prolonged periods of time is achieved by removing oxygen from the red blood cells at the time of storage; in particular, by flushing with an inert gas. Adenosine triphosphate levels of the stored red blood cells are boosted in some samples by addition of ammonium phosphate. 4 figs.

  18. Soil separator and sampler and method of sampling

    SciTech Connect

    O'Brien, Barry H; Ritter, Paul D

    2010-02-16

    A soil sampler includes a fluidized bed for receiving a soil sample. The fluidized bed may be in communication with a vacuum for drawing air through the fluidized bed and suspending particulate matter of the soil sample in the air. In a method of sampling, the air may be drawn across a filter, separating the particulate matter. Optionally, a baffle or a cyclone may be included within the fluidized bed for disentrainment, or dedusting, so only the finest particulate matter, including asbestos, will be trapped on the filter. The filter may be removable, and may be tested to determine the content of asbestos and other hazardous particulate matter in the soil sample.

  19. Method and apparatus for imaging a sample on a device

    DOEpatents

    Trulson, Mark; Stern, David; Fiekowsky, Peter; Rava, Richard; Walton, Ian; Fodor, Stephen P. A.

    1996-01-01

    The present invention provides methods and systems for detecting a labeled marker on a sample located on a support. The imaging system comprises a body for immobilizing the support, an excitation radiation source and excitation optics to generate and direct the excitation radiation at the sample. In response, labeled material on the sample emits radiation which has a wavelength that is different from the excitation wavelength, which radiation is collected by collection optics and imaged onto a detector which generates an image of the sample.

  20. System and method for measuring fluorescence of a sample

    DOEpatents

    Riot, Vincent J

    2015-03-24

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  1. Acetaminophen and meloxicam inhibit platelet aggregation and coagulation in blood samples from humans.

    PubMed

    Martini, Angela K; Rodriguez, Cassandra M; Cap, Andrew P; Martini, Wenjun Z; Dubick, Michael A

    2014-12-01

    Acetaminophen (Ace) and meloxicam (Mel) are the two types of analgesic and antipyretic medications. This study investigated the dose responses of acetaminophen and meloxicam on platelet aggregation and coagulation function in human blood samples. Blood samples were collected from six healthy humans and processed to make platelet-adjusted (100 × 10 cells/μl) blood samples. Acetaminophen (Tylenol, Q-PAP, 100 mg/ml) was added at the doses of 0 μg/ml (control), 214 μg/ml (the standard dose, 1 ×), 4 ×, 8 ×, 10 ×, 12 ×, 16 ×, and 20 ×. Similarly, meloxicam (Metacam, 5 mg/ml) was added at doses of 0 μg/ml (control), 2.85 μg/ml (the standard dose, 1 ×), 4 ×, 8 ×, 10 ×, 12 ×, 16 ×, and 20 ×. Fifteen minutes after the addition of acetaminophen and/or meloxicam, platelet aggregation was stimulated with collagen (2 μg/ml) or arachidonic acid (0.5 mmol/l) and assessed using a Chrono-Log 700 aggregometer. Coagulation function was assessed by prothrombin time (PT), activated partial thromboplastin time (aPTT), and using Rotem thrombelastogram. A robust inhibition by acetaminophen and/or meloxicam was observed in arachidonic acid-stimulated platelet aggregation starting at 1 × dose. Collagen-stimulated platelet aggregation was inhibited by ACE starting at 1 × (78 ± 10% of control), and by meloxicam starting at 4 × (72 ± 5% of control, both P < 0.05). The inhibitions by acetaminophen and meloxicam combined were similar to those by acetaminophen or meloxicam. aPTT was prolonged by meloxicam starting at 4 ×. No changes were observed in PT or any of Rotem measurements by acetaminophen and/or meloxicam. Acetaminophen and meloxicam compromised platelet aggregation and aPTT. Further effort is warranted to characterize the effects of acetaminophen and meloxicam on bleeding in vivo.

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

    USGS Publications Warehouse

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

    2003-01-01

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

  3. Convenient mounting method for electrical measurements of thin samples

    NASA Technical Reports Server (NTRS)

    Matus, L. G.; Summers, R. L.

    1986-01-01

    A method for mounting thin samples for electrical measurements is described. The technique is based on a vacuum chuck concept in which the vacuum chuck simultaneously holds the sample and established electrical contact. The mounting plate is composed of a glass-ceramic insulating material and the surfaces of the plate and vacuum chuck are polished. The operation of the vacuum chuck is examined. The contacts on the sample and mounting plate, which are sputter-deposited through metal masks, are analyzed. The mounting method was utilized for van der Pauw measurements.

  4. DOE methods for evaluating environmental and waste management samples

    SciTech Connect

    Goheen, S.C.; McCulloch, M.; Thomas, B.L.; Riley, R.G.; Sklarew, D.S.; Mong, G.M.; Fadeff, S.K.

    1994-10-01

    DOE Methods for Evaluating Environmental and Waste Management Samples (DOE Methods) is a resource intended to support sampling and analytical activities for the evaluation of environmental and waste management samples from U.S. Department of Energy (DOE) sites. DOE Methods is the result of extensive cooperation from all DOE analytical laboratories. All of these laboratories have contributed key information and provided technical reviews as well as significant moral support leading to the success of this document. DOE Methods is designed to encompass methods for collecting representative samples and for determining the radioisotope activity and organic and inorganic composition of a sample. These determinations will aid in defining the type and breadth of contamination and thus determine the extent of environmental restoration or waste management actions needed, as defined by the DOE, the U.S. Environmental Protection Agency, or others. The development of DOE Methods is supported by the Analytical Services Division of DOE. Unique methods or methods consolidated from similar procedures in the DOE Procedures Database are selected for potential inclusion in this document. Initial selection is based largely on DOE needs and procedure applicability and completeness. Methods appearing in this document are one of two types, {open_quotes}Draft{close_quotes} or {open_quotes}Verified{close_quotes}. {open_quotes}Draft{close_quotes} methods that have been reviewed internally and show potential for eventual verification are included in this document, but they have not been reviewed externally, and their precision and bias may not be known. {open_quotes}Verified{close_quotes} methods in DOE Methods have been reviewed by volunteers from various DOE sites and private corporations. These methods have delineated measures of precision and accuracy.

  5. Analytical and Biological Methods for Probing the Blood-Brain Barrier

    NASA Astrophysics Data System (ADS)

    Kuhnline, Sloan; Courtney, D.; Nandi, Pradyot; Linz, Thomas H.; Aldrich, Jane V.; Audus, Kenneth L.; Lunte, Susan M.

    2012-07-01

    The blood-brain barrier (BBB) is an important interface between the peripheral and central nervous systems. It protects the brain against the infiltration of harmful substances and regulates the permeation of beneficial endogenous substances from the blood into the extracellular fluid of the brain. It can also present a major obstacle in the development of drugs that are targeted for the central nervous system. Several methods have been developed to investigate the transport and metabolism of drugs, peptides, and endogenous compounds at the BBB. In vivo methods include intravenous injection, brain perfusion, positron emission tomography, and microdialysis sampling. Researchers have also developed in vitro cell-culture models that can be employed to investigate transport and metabolism at the BBB without the complication of systemic involvement. All these methods require sensitive and selective analytical methods to monitor the transport and metabolism of the compounds of interest at the BBB.

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

    PubMed

    Takei, Yoshio; Suzuki, Ippei; Wong, Marty Kwok-Shing; Milne, Ryan; Moss, Simon; Sato, Katsufumi; Hall, Ailsa

    2016-08-31

    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 >78% successful attempts to collect blood samples. Contamination of fluids in the tubing to the next blood sample was <1%, following the prior clearance of the tubing to a waste syringe. In captive harbor seals (Phoca vitulina) the automated blood sampling method was less stressful than direct blood withdrawal as plasma levels of stress hormones were lower in the former (p<0.05 for ACTH and p=0.078 for cortisol). HPLC analyses showed that both cortisol and cortisone were circulating in seal blood. Using the sampler, plasma levels of cardiovascular hormones, atrial natriuretic peptide (ANP), arginine vasopressin (AVP), and angiotensin II (AngII), 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 (AngII, 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 towards 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 semi-aquatic lifestyle.

  7. A new reference method for the determination of the oxygen content of blood.

    PubMed

    Dijkhuizen, P; Kwant, G; Zijlstra, W G

    1976-04-01

    To be able to determine the slight differences between the theoretical and the actual O2-binding capacity of human haemoglobin, a highly accurate method has been developed for measuring the O2 content of blood samples. It is an adaptation of an established O2 determination in organic microanalysis. The bound O2 of the blood is set free by conversion of HbO2 to Hi, and the O2 stripped from the blood is converted to CO by contact with granular carbon at 1120 degrees C. The CO is then converted to CO2 using CuO at 300 degrees C and the CO2 titrated in a solution of BaCl2, using NaOH of known strength. The measuring system was checked by analysis of 36 samples of air, yielding an O2 content of 20.93 +/- 0.06%. The coefficient of variation calculated from 62 duplicate determinations of samples of human blood was 0.65%.

  8. A Novel Ultrasound-Based Method to Evaluate Hemostatic Function of Whole Blood

    PubMed Central

    Viola, Francesco; Mauldin, F. William; Lin-Schmidt, Xiefan; Haverstick, Doris M.; Lawrence, Michael B.; Walker, William F.

    2009-01-01

    Background: Unregulated hemostasis represents a leading cause of mortality and morbidity in the developed world. Being able to recognize and quantify defects of the hemostatic process is critical to reduce mortality and implement appropriate treatment. Methods: We describe a novel ultrasound-based technology, named sonorheometry, which can assess hemostasis function from a small sample of blood. Sonorheometry uses the phenomenon of acoustic radiation force to measure the dynamic changes in blood viscoelasticity during clot formation and clot dissolution. We performed in vitro experiments using whole blood samples of 1ml to demonstrate that sonorheometry is indicative of hemostatic functions that depend on plasma coagulation factors, platelets, and plasma fibrinolytic factors. Results: Sonorheometry measurements show titration effects to compounds known to alter the coagulation factors (GPRP peptide, 0 to 8 mmol/l), platelets (abciximab, 0 to 12 ug/ml), and fibrinolytic factors (urokinase, 0 to 200U). Repeated measurements of blood samples from the same subjects yielded reproducibility errors on the order of 5%. Conclusions: These data indicate that sonorheometry accurately quantifies the functional role of the components of hemostasis in vitro. PMID:19861121

  9. Tests of a comparative method of dating plutonium samples

    NASA Astrophysics Data System (ADS)

    West, D.

    1987-04-01

    Tests of a comparative method of dating plutonium samples have been carried out using 241Pu in aqueous solution. The six samples were of known ages (between 0.25 and 15 yr) and with one exception the measured ages, using particular samples as standards, agreed with the stated ages. In one case the agreement was beter than 1% in age. Mixed-oxide fuel pins were also intercompared. In this case it was with some difficulty that a sample of known age was obtaine. Comparison using this sample and an older one gave the same value (within ±1%) for the separation date of the unknown sample on three occasions over a three year period.

  10. Evaluating Composite Sampling Methods of Bacillus Spores at Low Concentrations

    PubMed Central

    Hess, Becky M.; Amidan, Brett G.; Anderson, Kevin K.; Hutchison, Janine R.

    2016-01-01

    Restoring all facility operations after the 2001 Amerithrax attacks took years to complete, highlighting the need to reduce remediation time. Some of the most time intensive tasks were environmental sampling and sample analyses. Composite sampling allows disparate samples to be combined, with only a single analysis needed, making it a promising method to reduce response times. We developed a statistical experimental design to test three different composite sampling methods: 1) single medium single pass composite (SM-SPC): a single cellulose sponge samples multiple coupons with a single pass across each coupon; 2) single medium multi-pass composite: a single cellulose sponge samples multiple coupons with multiple passes across each coupon (SM-MPC); and 3) multi-medium post-sample composite (MM-MPC): a single cellulose sponge samples a single surface, and then multiple sponges are combined during sample extraction. Five spore concentrations of Bacillus atrophaeus Nakamura spores were tested; concentrations ranged from 5 to 100 CFU/coupon (0.00775 to 0.155 CFU/cm2). Study variables included four clean surface materials (stainless steel, vinyl tile, ceramic tile, and painted dry wallboard) and three grime coated/dirty materials (stainless steel, vinyl tile, and ceramic tile). Analysis of variance for the clean study showed two significant factors: composite method (p< 0.0001) and coupon material (p = 0.0006). Recovery efficiency (RE) was higher overall using the MM-MPC method compared to the SM-SPC and SM-MPC methods. RE with the MM-MPC method for concentrations tested (10 to 100 CFU/coupon) was similar for ceramic tile, dry wall, and stainless steel for clean materials. RE was lowest for vinyl tile with both composite methods. Statistical tests for the dirty study showed RE was significantly higher for vinyl and stainless steel materials, but lower for ceramic tile. These results suggest post-sample compositing can be used to reduce sample analysis time when

  11. Estimation of the Time Interval between the Administration of Heroin and the Sampling of Blood in Chronic Inhalers.

    PubMed

    Dubois, Nathalie; Hallet, Claude; Seidel, Laurence; Demaret, Isabelle; Luppens, David; Ansseau, Marc; Rozet, Eric; Albert, Adelin; Hubert, Philippe; Charlier, Corinne

    2015-05-01

    To develop a model for estimating the time delay between last heroin consumption and blood sampling in chronic drug users. Eleven patients, all heroin inhalers undergoing detoxification, were included in the study. Several plasma samples were collected during the detoxification procedure and analyzed for the heroin metabolites 6-acetylmorphine (6AM), morphine (MOR), morphine-6-glucuronide (M6G) and morphine-3-glucuronide (M3G), according to a UHPLC/MSMS method. The general linear mixed model was applied to time-related concentrations and a pragmatic four-step delay estimation approach was proposed based on the simultaneous presence of metabolites in plasma. Validation of the model was carried out using the jackknife technique on the 11 patients, and on a group of 7 test patients. Quadratic equations were derived for all metabolites except 6AM. The interval delay estimation was 2-4 days when only M3G present in plasma, 1-2 days when M6G and M3G were both present, 0-1 day when MOR, M6G and M3G were present and <2 h for all metabolites present. The 'jackknife' correlation between declared and actual estimated delays was 0.90. The overall precision of the delay estimates was 8-9 h. The delay between last heroin consumption and blood sampling in chronic drug users can be satisfactorily predicted from plasma heroin metabolites.

  12. Evaluation of potentially nonlethal sampling methods for monitoring mercury concentrations in smallmouth bass (Micropterus dolomieu)

    USGS Publications Warehouse

    Schmitt, C.J.; Brumbaugh, W.G.

    2007-01-01

    We evaluated three potentially nonlethal alternatives to fillet sampling for the determination of mercury (Hg) concentrations in smallmouth bass (Micropterus dolomieu). Fish (n = 62, 226-464 mm total length) from six sites in southern Missouri were captured by electrofishing. Blood samples (1 mL) from each fish were obtained by caudal veinipuncture with a heparinized needle and syringe. Biopsy needle (10 mm x 14 gauge; three cuts per fish; 10-20 mg total dry weight) and biopsy punch (7 mm x 5 mm in diameter, one plug per fish, 30-50 mg dry weight) samples were obtained from the area beneath the dorsal fin. Fillet samples were obtained from the opposite side of the fish. All samples were freeze-dried and analyzed for total Hg by combustion amalgamation atomic absorption spectrophotometry. Mean relative standard deviations (RSDs) of triplicate samples were similar for all four methods (2.2-2.4%), but the range of RSDs was greater for blood (0.4-5.5%) than for the muscle methods (1.8-4.0%). Total Hg concentrations in muscle were 0.0200-0.8809 ??g/g wet weight; concentrations in plug, needle, and fillet samples from each fish were nearly identical. Blood Hg concentrations were 0.0006-0.0812 ??g/mL and were highly correlated with muscle concentrations; linear regressions between log-transformed blood and fillet Hg concentrations were linear and statistically significant (p < 0.01), and explained 91-93% of the total variation. Correlations between fillet Hg concentrations and fish size and age were weak; together they explained ???37% of the total variation, and the relations differed among sites. Overall, any of the alternative methods could provide satisfactory estimates of fillet Hg in smallmouth bass; however, both blood and plug sampling with disposable instruments were easier to perform than needle sampling. The biopsy needle was the most difficult to use, especially on smaller fish, and its relative expense necessitates reuse and, consequently, thorough cleaning

  13. Blood

    MedlinePlus

    ... that die or are lost from the body. White Blood Cells White blood cells (WBCs, and also ... of severe pain. previous continue Diseases of the White Blood Cells Neutropenia (pronounced: new-truh-PEE-nee- ...

  14. Identification of food-derived bioactive peptides in blood and other biological samples.

    PubMed

    Sato, Kenji; Iwai, Koji; Aito-Inoue, Misako

    2008-01-01

    Recent studies have demonstrated the presence of food-derived peptides in human blood after ingestion of enzymatic hydrolysates of food proteins, while most peptides in food are degraded into amino acids during digestion and absorption. To capture and clarify the food-derived peptides in blood, solid-phase extraction (SPE) using a mini-spin column packed with a strong cation exchanger was developed. This technique allows the use of a nonvolatile acid such as trichloroacetic acid, a strong protein denaturant, for the deproteinizing procedure. To improve resolution of hydrophilic peptide and increase specificity and sensitivity in the detection of peptide by reversed-phase high-performance liquid chromatography (RP-HPLC) after subfractionation by size-exclusion chromatography (SEC), peptides are derivatized with phenyl isothiocyanate. The resultant phenyl thiocarbamyl (PTC)-peptides can be resolved with high resolution and sensitivity by RP-HPLC. By comparing chromatograms of PTC derivatives from blood before and after ingestion of a peptide sample, food-derived peptide can be detected. The isolated PTC-peptide can be applied to a peptide sequencer based on the Edman degradation reaction.

  15. Concentrations of persistent organic pollutants (POPs) in human blood samples from Mexico City, Mexico.

    PubMed

    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 (

  16. Blood Glucose Measurement in the Intensive Care Unit: What Is the Best Method?

    PubMed Central

    Le, Huong T.; Harris, Neil S.; Estilong, Abby J.; Olson, Arvid; Rice, Mark J.

    2013-01-01

    Abnormal glucose measurements are common among intensive care unit (ICU) patients for numerous reasons and hypoglycemia is especially dangerous because these patients are often sedated and unable to relate the associated symptoms. Additionally, wide swings in blood glucose have been closely tied to increased mortality. Therefore, accurate and timely glucose measurement in this population is critical. Clinicians have several choices available to assess blood glucose values in the ICU, including central laboratory devices, blood gas analyzers, and point-of-care meters. In this review, the method of glucose measurement will be reviewed for each device, and the important characteristics, including accuracy, cost, speed of result, and sample volume, will be reviewed, specifically as these are used in the ICU environment. Following evaluation of the individual measurement devices and after considering the many features of each, recommendations are made for optimal ICU glucose determination. PMID:23567008

  17. Method for breast cancer diagnosis by phase spectrophotometry of human blood plasma

    NASA Astrophysics Data System (ADS)

    Mintser, Ozar P.; Oliinychenko, B. P.

    2012-01-01

    The possibility of breast cancer diagnostics by means of phase structure measurements of laser radiation transformed by human blood plasma samples. The theoretical fundamentals of polarization filtration method for direct phase shifts measurements of microscopic images are provided. The optical model of polycrystalline networks of blood plasma proteins is suggested. The results of investigating the interrelation between the values of statistical (statistical moments of the 1st-4th order), correlation (correlation area, asymmetry coefficient and autocorrelation function excess) and fractal (dispersion of logarithmic dependencies of power spectra) parameters are presented. They characterize the coordinate distributions of phase shifts between the orthogonal components of the amplitude in the points of laser images of blood plasma smears and pathological changes in the mammary gland tissue. The diagnostic criteria of breast cancer nascency are determined.

  18. Method for breast cancer diagnosis by phase spectrophotometry of human blood plasma

    NASA Astrophysics Data System (ADS)

    Mintser, Ozar P.; Oliinychenko, B. P.

    2011-09-01

    The possibility of breast cancer diagnostics by means of phase structure measurements of laser radiation transformed by human blood plasma samples. The theoretical fundamentals of polarization filtration method for direct phase shifts measurements of microscopic images are provided. The optical model of polycrystalline networks of blood plasma proteins is suggested. The results of investigating the interrelation between the values of statistical (statistical moments of the 1st-4th order), correlation (correlation area, asymmetry coefficient and autocorrelation function excess) and fractal (dispersion of logarithmic dependencies of power spectra) parameters are presented. They characterize the coordinate distributions of phase shifts between the orthogonal components of the amplitude in the points of laser images of blood plasma smears and pathological changes in the mammary gland tissue. The diagnostic criteria of breast cancer nascency are determined.

  19. Evaluation of the Punch-it™ NA-Sample kit for detecting microbial DNA in blood culture bottles using PCR-reverse blot hybridization assay.

    PubMed

    Kim, Jungho; Wang, Hye-Young; Kim, Seoyong; Park, Soon Deok; Yu, Kwangmin; Kim, Hyo Youl; Uh, Young; Lee, Hyeyoung

    2016-09-01

    DNA extraction efficiency affects the success of PCR-based method applications. The Punch-it™ NA-Sample kit for extracting DNA by using paper chromatography is technically easy to use and requires just two reagents and only 10min to complete. The Punch-it™ NA-Sample kit could be offered as a rapid, accurate, and convenient method for extracting bacterial and fungal DNA from blood culture bottles. We compared the efficiencies of the commercial kit (Punch-it™ NA-Sample kit) and an in-house conventional boiling method with Chelex-100 resin for DNA extraction from blood culture bottles. The efficiency of the two DNA extraction methods was assessed by PCR-reverse blot hybridization assay (PCR-REBA, REBA Sepsis-ID) for detecting Gram positive (GP) bacteria, Gram negative (GN) bacteria, and Candida species with 196 positive and 200 negative blood culture bottles. The detection limits of the two DNA extraction methods were 10(3)CFU/mL for GP bacteria, 10(3)CFU/mL for GN bacteria, and 10(4)CFU/mL for Candida. The sensitivity and specificity of the Punch-it™ NA-Sample kit by REBA Sepsis-ID were 95.4% (187/196) and 100% (200/200), respectively. The overall agreement of the two DNA extraction methods was 98.9% (392/396). Three of four samples showing discrepant results between the two extraction methods were more accurately matched up with the Punch-it™ NA-Sample kit based on conventional culture methods. The results indicated that the Punch-it™ NA-Sample kit extracted bacterial and fungal DNA in blood culture bottles and allowed extracted DNA to be used in molecular assay.

  20. COMPARISON OF MACROINVERTEBRATE SAMPLING METHODS FOR NONWADEABLE STREAMS

    EPA Science Inventory

    The bioassessment of nonwadeable streams in the United States is increasing, but methods for these systems are not as well developed as for wadeable streams. In this study, we compared six benthic macroinvertebrate field sampling methods for nonwadeable streams based on those us...

  1. The Precision Efficacy Analysis for Regression Sample Size Method.

    ERIC Educational Resources Information Center

    Brooks, Gordon P.; Barcikowski, Robert S.

    The general purpose of this study was to examine the efficiency of the Precision Efficacy Analysis for Regression (PEAR) method for choosing appropriate sample sizes in regression studies used for precision. The PEAR method, which is based on the algebraic manipulation of an accepted cross-validity formula, essentially uses an effect size to…

  2. [Determination of Al, Be, Cd, Co, Cr, Mn, Ni, Pb, Se and Tl in whole blood by atomic absorption spectrometry without preliminary sample digestion].

    PubMed

    Ivanenko, N B; Ivanenko, A A; Solov'ev, N D; Navolotskiĭ, D V; Pavlova, O V; Ganeev, A A

    2014-01-01

    Methods of whole blood trace element determination by Graphite furnace atomic absorption spectrometry (in the variant of Zeeman's modulation polarization spectrometry) have been proposed. They do not require preliminary sample digestion. Furnace programs, modifiers and blood dilution factors were optimized. Seronorm™ human whole blood reference materials were used for validation. Dynamic ranges (for undiluted blood samples) were: Al 8 ¸ 210 мg/L; Be 0.3 ¸ 50 мg/L; Cd 0.2 ¸ 75 мg/L; Сo 5 ¸ 350 мg/L; Cr 10 ¸ 100 мg/L; Mn 6 ¸ 250 мg/L; Ni 10 ¸ 350 мg/L; Pb 3 ¸ 240 мg/L; Se 10 ¸ 500 мg/L; Tl 2 ¸ 600 мg/L. Precision (RSD) for the middle of dynamic range ranged from 5% for Mn to 11 for Se.

  3. Standard methods for sampling North American freshwater fishes

    USGS Publications Warehouse

    Bonar, Scott A.; Hubert, Wayne A.; Willis, David W.

    2009-01-01

    This important reference book provides standard sampling methods recommended by the American Fisheries Society for assessing and monitoring freshwater fish populations in North America. Methods apply to ponds, reservoirs, natural lakes, and streams and rivers containing cold and warmwater fishes. Range-wide and eco-regional averages for indices of abundance, population structure, and condition for individual species are supplied to facilitate comparisons of standard data among populations. Provides information on converting nonstandard to standard data, statistical and database procedures for analyzing and storing standard data, and methods to prevent transfer of invasive species while sampling.

  4. A multi-dimensional sampling method for locating small scatterers

    NASA Astrophysics Data System (ADS)

    Song, Rencheng; Zhong, Yu; Chen, Xudong

    2012-11-01

    A multiple signal classification (MUSIC)-like multi-dimensional sampling method (MDSM) is introduced to locate small three-dimensional scatterers using electromagnetic waves. The indicator is built with the most stable part of signal subspace of the multi-static response matrix on a set of combinatorial sampling nodes inside the domain of interest. It has two main advantages compared to the conventional MUSIC methods. First, the MDSM is more robust against noise. Second, it can work with a single incidence even for multi-scatterers. Numerical simulations are presented to show the good performance of the proposed method.

  5. A rapid wire-based sampling method for DNA profiling.

    PubMed

    Chen, Tong; Catcheside, David E A; Stephenson, Alice; Hefford, Chris; Kirkbride, K Paul; Burgoyne, Leigh A

    2012-03-01

    This paper reports the results of a commission to develop a field deployable rapid short tandem repeat (STR)-based DNA profiling system to enable discrimination between tissues derived from a small number of individuals. Speed was achieved by truncation of sample preparation and field deployability by use of an Agilent 2100 Bioanalyser(TM). Human blood and tissues were stabbed with heated stainless steel wire and the resulting sample dehydrated with isopropanol prior to direct addition to a PCR. Choice of a polymerase tolerant of tissue residues and cycles of amplification appropriate for the amount of template expected yielded useful profiles with a custom-designed quintuplex primer set suitable for use with the Bioanalyser(TM). Samples stored on wires remained amplifiable for months, allowing their transportation unrefrigerated from remote locations to a laboratory for analysis using AmpFlSTR(®) Profiler Plus(®) without further processing. The field system meets the requirements for discrimination of samples from small sets and retains access to full STR profiling when required.

  6. Development and validation of a dried blood spot LC-MS/MS assay to quantify ranitidine in paediatric samples.

    PubMed

    Yakkundi, Shirish; Millership, Jeff; Collier, Paul; Shields, Michael D; McElnay, James

    2011-12-15

    A novel approach has been developed to determine ranitidine in paediatric samples using dried blood spots (DBS) on Guthrie cards (Whatman 903). A selective and sensitive HPLC-MS/MS assay has been developed and validated using small volumes of blood (30 μl). A 6 mm disc was punched from each DBS and extracted with methanolic solution of the internal standard (IS) nizatidine. This was further subjected to solid phase extraction (SPE), followed by reversed phase HPLC separation, using a XBridge™ C18 column and mobile phase 10 mM ammonium acetate/methanol (98:2 v/v) with a flow rate of 0.3 mL/min. This was combined with multiple reaction monitoring (MRM) mass detection using electrospray ionisation (ESI). The calibration curve for ranitidine was found linear over the range 10-500 ng/mL (r=0.996). The limit of quantification (LOQ) of the method was validated at 10 ng/mL. Accuracy and precision values for within and between days were <20% at the LOQ and <15% at all other concentrations. The validated DBS method was successfully applied to a clinical study employing 81 samples from 36 paediatric patients.

  7. Finite element methods of studying mechanical factors in blood flow.

    PubMed

    Davids, N

    1981-01-01

    This paper reviews some biomechanical analyses of blood flow in large arteries based on a general computer modeling using the finite element method. We study the following question: What is the role played by the interrelated factors of mechanical stress, flow irregularities, and diffusion through the endothelium on the etiology of atherosclerosis or the aggravation of vascular injury. It presents the computational features of the method and stresses the physiological significance of the results, such as the effect of geometric complexities, material nonlinearities, and non-Newtonian rheology of the blood. The specific mechanical and fluid dynamic factors analyzed are wall shear stress, flow profiles, and pressure variations. After simulating tubes of circular cross section, we apply the analysis to a number of physiological situations of significance, including blood flow in the entrance region, at bifurcations, in the annular region between an inserted catheter of varying diameter and the vessel. A model study of pulsatile flow in a 60 degree bifurcated channel of velocity profiles provided corroborative measurements of these processes with special emphasis on reversed or distributed flow conditions. The corresponding analysis was extended to the situation in which flow separates and reverses in the neighborhood of stagnation points. This required developing the nonlinear expression for the convective velocity change in the medium. A computer algorithm was developed to handle simultaneous effects of pressure and viscous forces on velocity change across the element and applied to the canine prebranch arterial segment. For mean physiological flow conditions, low shear stresses (0-10 dynes/cm2) are predicted near the wall in the diverging plane, higher values (50 dynes/cm2) along the converging sides of the wall. Backflow is predicted along the outer wall, pressure recovery prior to and into the branches, and a peak shear at the divider lip.

  8. Efficiency of snake sampling methods in the Brazilian semiarid region.

    PubMed

    Mesquita, Paula C M D; Passos, Daniel C; Cechin, Sonia Z

    2013-09-01

    The choice of sampling methods is a crucial step in every field survey in herpetology. In countries where time and financial support are limited, the choice of the methods is critical. The methods used to sample snakes often lack objective criteria, and the traditional methods have apparently been more important when making the choice. Consequently researches using not-standardized methods are frequently found in the literature. We have compared four commonly used methods for sampling snake assemblages in a semiarid area in Brazil. We compared the efficacy of each method based on the cost-benefit regarding the number of individuals and species captured, time, and financial investment. We found that pitfall traps were the less effective method in all aspects that were evaluated and it was not complementary to the other methods in terms of abundance of species and assemblage structure. We conclude that methods can only be considered complementary if they are standardized to the objectives of the study. The use of pitfall traps in short-term surveys of the snake fauna in areas with shrubby vegetation and stony soil is not recommended.

  9. Beryllium Wipe Sampling (differing methods - differing exposure potentials)

    SciTech Connect

    Kerr, Kent

    2005-03-09

    This research compared three wipe sampling techniques currently used to test for beryllium contamination on room and equipment surfaces in Department of Energy facilities. Efficiencies of removal of beryllium contamination from typical painted surfaces were tested by wipe sampling without a wetting agent, with water-moistened wipe materials, and by methanol-moistened wipes. Analysis indicated that methanol-moistened wipe sampling removed about twice as much beryllium/oil-film surface contamination as water-moistened wipes, which removed about twice as much residue as dry wipes. Criteria at 10 CFR 850.30 and .31 were established on unspecified wipe sampling method(s). The results of this study reveal a need to identify criteria-setting method and equivalency factors. As facilities change wipe sampling methods among the three compared in this study, these results may be useful for approximate correlations. Accurate decontamination decision-making depends on the selection of appropriate wetting agents for the types of residues and surfaces. Evidence for beryllium sensitization via skin exposure argues in favor of wipe sampling with wetting agents that provide enhanced removal efficiency such as methanol when surface contamination includes oil mist residue.

  10. Pharmacokinetic Studies of Chinese Medicinal Herbs Using an Automated Blood Sampling System and Liquid Chromatography-mass Spectrometry

    PubMed Central

    Wu, Yu-Tse; Wu, Ming-Tsang; Lin, Chia-Chun; Chien, Chao-Feng; Tsai, Tung-Hu

    2012-01-01

    The safety of herbal products is one of the major concerns for the modernization of traditional Chinese medicine, and pharmacokinetic data of medicinal herbs guide us to design the rational use of the herbal formula. This article reviews the advantages of the automated blood sampling (ABS) systems for pharmacokinetic studies. In addition, three commonly used sample preparative methods, protein precipitation, liquid-liquid extraction and solid-phase extraction, are introduced. Furthermore, the definition, causes and evaluation of matrix effects in liquid chromatography-mass spectrometry (LC/MS) analysis are demonstrated. Finally, we present our previous works as practical examples of the application of ABS systems and LC/MS for the pharmacokinetic studies of Chinese medicinal herbs. PMID:24716112

  11. Method of high-precision microsampled blood and plasma mass densitometry

    NASA Technical Reports Server (NTRS)

    Hinghofer-Szalkay, H.

    1986-01-01

    The reliability of the mechanical oscillator technique for blood and plasma density measurements on samples of volumes less than 0.1 ml is examined, and a precision of 0.001 g/l is found if plasma-isodensic heparin solution and siliconized densitometers are employed. Sources of measurement errors in the density determinations include storage of plasma samples, inhomogeneity of blood samples, and density reading before adequate temperature equilibration. In tests of plasma sample storage, the best reproducibility was obtained with samples kept at 4 C. Linear correlations were found between plasma density and plasma protein concentration, blood density and blood hemoglobin concentration, and erythrocyte density and MCHC.

  12. Was the Driver Drunk? An Instrumental Methods Experiment for the Determination of Blood Alcohol Content

    NASA Astrophysics Data System (ADS)

    Zabzdyr, Jennifer L.; Lillard, Sheri J.

    2001-09-01

    Introducing forensic scenarios into the instrumental laboratory is a simple yet effective strategy to give students the opportunity to perform realistic experiments and to learn proper analytical techniques. In this laboratory experiment, which is designed for upper-division students in an instrumental methods course, unknown ethanol concentrations are quantitated in simulated serum samples using headspace gas chromatography (GC) with flame ionization detection. For the quantitative determination of blood alcohol content (BAC) in cases where drunk driving is suspected, GC is considered the most reliable method. Furthermore, headspace sampling helps to eliminate interferences from biological matrices such as blood or serum. An unknown sample of ethanol in serum, which brackets the 0.080 g/mL legal limit (e.g., BAC 0.075-0.087 g/mL), is given to the student for measurement. Both external and internal standardization methods are used for calibration. The students calculate serum alcohol concentration based on the measurement of their unknown sample using each calibration curve, and the results from each calibration method are compared and discussed in terms of accuracy. They then use a conversion factor to calculate BAC from serum alcohol concentration and use their results to determine if the "suspected driver" was driving under the influence.

    See Letter re: this article.

  13. Environmental contaminants in Texas, USA, wetland reptiles: Evaluation using blood samples

    USGS Publications Warehouse

    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.

  14. Capillary blood sampling from the ear of dogs and cats and use of portable meters to measure glucose concentration.

    PubMed

    Wess, G; Reusch, C

    2000-02-01

    Two new methods for collection of capillary blood from the ear of dogs and cats for the measurement of blood glucose concentration using portable blood glucose meters (PBGMs) are described. The first method uses a lancing device after pre-warming the ear, while the second employs a vacuum lancing device. Both methods generated blood drops of adequate size, although the latter method was faster and easier to perform. Accuracy of the two PBGMs was evaluated clinically and statistically. Although assessment of statistical accuracy revealed differences between the PBGMs and the reference method, all of the PBGM readings were within clinically acceptable ranges. Measurement of capillary blood glucose concentration is easy to perform, inexpensive and fast. It may be used by owners to determine blood glucose concentrations at home, and could serve as a new tool for monitoring diabetic dogs and cats.

  15. Simple and rapid analytical method for detection of amino acids in blood using blood spot on filter paper, fast-GC/MS and isotope dilution technique.

    PubMed

    Kawana, Shuichi; Nakagawa, Katsuhiro; Hasegawa, Yuki; Yamaguchi, Seiji

    2010-11-15

    A simple and rapid method for quantitative analysis of amino acids, including valine (Val), leucine (Leu), isoleucine (Ile), methionine (Met) and phenylalanine (Phe), in whole blood has been developed using GC/MS. In this method, whole blood was collected using a filter paper technique, and a 1/8 in. blood spot punch was used for sample preparation. Amino acids were extracted from the sample, and the extracts were purified using cation-exchange resins. The isotope dilution method using ²H₈-Val, ²H₃-Leu, ²H₃-Met and ²H₅-Phe as internal standards was applied. Following propyl chloroformate derivatization, the derivatives were analyzed using fast-GC/MS. The extraction recoveries using these techniques ranged from 69.8% to 87.9%, and analysis time for each sample was approximately 26 min. Calibration curves at concentrations from 0.0 to 1666.7 μmol/l for Val, Leu, Ile and Phe and from 0.0 to 333.3 μmol/l for Met showed good linearity with regression coefficients=1. The method detection limits for Val, Leu, Ile, Met and Phe were 24.2, 16.7, 8.7, 1.5 and 12.9 μmol/l, respectively. This method was applied to blood spot samples obtained from patients with phenylketonuria (PKU), maple syrup urine disease (MSUD), hypermethionine and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), and the analysis results showed that the concentrations of amino acids that characterize these diseases were increased. These results indicate that this method provides a simple and rapid procedure for precise determination of amino acids in whole blood.

  16. A Simple Method to Quantitate IP-10 in Dried Blood and Plasma Spots

    PubMed Central

    Aabye, Martine G.; Eugen-Olsen, Jesper; Werlinrud, Anne Marie; Holm, Line Lindebo; Tuuminen, Tamara; Ravn, Pernille; Ruhwald, Morten

    2012-01-01

    Background Antigen specific release of IP-10 is an established marker for infection with M.tuberculosis. Compared to IFN-γ, IP-10 is released in 100-fold higher concentrations enabling the development of novel assays for detection. Dried blood spots are a convenient sample for high throughput newborn screening. Aim To develop a robust and sensitive ELISA-based assay for IP-10 detection in plasma, dried blood spots (DBS) and dried plasma spots (DPS); to validate the ELISA in clinically relevant samples; and to assess the performance of the assay for detection of Cytomegalovirus (CMV) and M.tuberculosis specific immune responses. Method We raised mice and rat monoclonal antibodies against human IP-10 and developed an ELISA. The assay was validated and applied to the detection of CMV and M.tuberculosis specific responses in 18 patients with immune reactivity towards M.tuberculosis and 32 healthy controls of which 22 had immune reactivity towards CMV and none towards M.tuberculosis. We compared the performance of this new assay to IFN-γ. Results The ELISA was reliable for IP-10 detection in both plasma and filter paper samples. The linear range of the ELISA was 2.5–600 pg/ml. IFN-γ was not readily detectable in DPS samples. IP-10 was stabile in filter paper samples for at least 4 weeks at 37°C. The correlation between IP-10 detected in plasma, DPS and DBS samples was excellent (r2>0.97). Conclusions This newly developed assay is reliable for IP-10 quantification in plasma, DBS and DPS samples from antigen stimulated and non-stimulated whole blood. The filter paper assays enable easy sample acquisition and transport at ambient temperature e.g. via the postal system. The system can potentially simplify diagnostic assays for M.tuberculosis and CMV infection. PMID:22761744

  17. Comparison of Two Methods for the Determination of the Effects of Ionizing Radiation on Blood Cell Counts in Mice

    PubMed Central

    Romero-Weaver, Ana L.; Kennedy, Ann R.

    2012-01-01

    A reliable technique is needed to determine the effect of ionizing radiation on white blood cell (WBC) counts. Facilities that utilize automated methods can provide this service. However, utilizing external facilities can introduce additional variables, such as differences between time of sample collection and time of sample processing, which may affect the results. The purpose of the present study was to determine whether an automated method at an external facility can accurately determine radiation-induced changes in total WBC, lymphocyte and granulocyte counts when samples are analyzed at periods of time up to 24 hours after collection and stored either at room temperature or at 4°C. To accomplish this, we compared automated blood cell counts determined at an external facility with our manual blood cell counts processed immediately after sample collection or 24 h after sample collection and stored either at room temperature or 4°C from mice exposed to 2 Gy proton or 2 Gy gamma radiation. Our results show a close correlation and good agreement between the two methods, indicating that neither a delay of 24 hours in sample processing nor storage temperature affected white blood cell counts. Analysis of the effects of radiation on blood cell counts by either manual or automated cell counts revealed a statistically significant decrease in lymphocyte and granulocyte counts at different days post-irradiation, with no statistically significant difference between the methods employed; therefore both manual and automated blood cell counts are reliable methods to determine the effects of ionizing radiation in blood cells. PMID:23450807

  18. Self-contained cryogenic gas sampling apparatus and method

    DOEpatents

    McManus, G.J.; Motes, B.G.; Bird, S.K.; Kotter, D.K.

    1996-03-26

    Apparatus for obtaining a whole gas sample, is composed of: a sample vessel having an inlet for receiving a gas sample; a controllable valve mounted for controllably opening and closing the inlet; a valve control coupled to the valve for opening and closing the valve at selected times; a portable power source connected for supplying operating power to the valve control; and a cryogenic coolant in thermal communication with the vessel for cooling the interior of the vessel to cryogenic temperatures. A method is described for obtaining an air sample using the apparatus described above, by: placing the apparatus at a location at which the sample is to be obtained; operating the valve control to open the valve at a selected time and close the valve at a selected subsequent time; and between the selected times maintaining the vessel at a cryogenic temperature by heat exchange with the coolant. 3 figs.

  19. Self-contained cryogenic gas sampling apparatus and method

    DOEpatents

    McManus, Gary J.; Motes, Billy G.; Bird, Susan K.; Kotter, Dale K.

    1996-01-01

    Apparatus for obtaining a whole gas sample, composed of: a sample vessel having an inlet for receiving a gas sample; a controllable valve mounted for controllably opening and closing the inlet; a valve control coupled to the valve for opening and closing the valve at selected times; a portable power source connected for supplying operating power to the valve control; and a cryogenic coolant in thermal communication with the vessel for cooling the interior of the vessel to cryogenic temperatures. A method of obtaining an air sample using the apparatus described above, by: placing the apparatus at a location at which the sample is to be obtained; operating the valve control to open the valve at a selected time and close the valve at a selected subsequent time; and between the selected times maintaining the vessel at a cryogenic temperature by heat exchange with the coolant.

  20. Examination of Hydrate Formation Methods: Trying to Create Representative Samples

    SciTech Connect

    Kneafsey, T.J.; Rees, E.V.L.; Nakagawa, S.; Kwon, T.-H.

    2011-04-01

    Forming representative gas hydrate-bearing laboratory samples is important so that the properties of these materials may be measured, while controlling the composition and other variables. Natural samples are rare, and have often experienced pressure and temperature changes that may affect the property to be measured [Waite et al., 2008]. Forming methane hydrate samples in the laboratory has been done a number of ways, each having advantages and disadvantages. The ice-to-hydrate method [Stern et al., 1996], contacts melting ice with methane at the appropriate pressure to form hydrate. The hydrate can then be crushed and mixed with mineral grains under controlled conditions, and then compacted to create laboratory samples of methane hydrate in a mineral medium. The hydrate in these samples will be part of the load-bearing frame of the medium. In the excess gas method [Handa and Stupin, 1992], water is distributed throughout a mineral medium (e.g. packed moist sand, drained sand, moistened silica gel, other porous media) and the mixture is brought to hydrate-stable conditions (chilled and pressurized with gas), allowing hydrate to form. This method typically produces grain-cementing hydrate from pendular water in sand [Waite et al., 2004]. In the dissolved gas method [Tohidi et al., 2002], water with sufficient dissolved guest molecules is brought to hydrate-stable conditions where hydrate forms. In the laboratory, this is can be done by pre-dissolving the gas of interest in water and then introducing it to the sample under the appropriate conditions. With this method, it is easier to form hydrate from more soluble gases such as carbon dioxide. It is thought that this method more closely simulates the way most natural gas hydrate has formed. Laboratory implementation, however, is difficult, and sample formation is prohibitively time consuming [Minagawa et al., 2005; Spangenberg and Kulenkampff, 2005]. In another version of this technique, a specified quantity of gas

  1. Characterizing lentic freshwater fish assemblages using multiple sampling methods

    USGS Publications Warehouse

    Fischer, Jesse R.; Quist, Michael

    2014-01-01

    Characterizing fish assemblages in lentic ecosystems is difficult, and multiple sampling methods are almost always necessary to gain reliable estimates of indices such as species richness. However, most research focused on lentic fish sampling methodology has targeted recreationally important species, and little to no information is available regarding the influence of multiple methods and timing (i.e., temporal variation) on characterizing entire fish assemblages. Therefore, six lakes and impoundments (48–1,557 ha surface area) were sampled seasonally with seven gear types to evaluate the combined influence of sampling methods and timing on the number of species and individuals sampled. Probabilities of detection for species indicated strong selectivities and seasonal trends that provide guidance on optimal seasons to use gears when targeting multiple species. The evaluation of species richness and number of individuals sampled using multiple gear combinations demonstrated that appreciable benefits over relatively few gears (e.g., to four) used in optimal seasons were not present. Specifically, over 90 % of the species encountered with all gear types and season combinations (N = 19) from six lakes and reservoirs were sampled with nighttime boat electrofishing in the fall and benthic trawling, modified-fyke, and mini-fyke netting during the summer. Our results indicated that the characterization of lentic fish assemblages was highly influenced by the selection of sampling gears and seasons, but did not appear to be influenced by waterbody type (i.e., natural lake, impoundment). The standardization of data collected with multiple methods and seasons to account for bias is imperative to monitoring of lentic ecosystems and will provide researchers with increased reliability in their interpretations and decisions made using information on lentic fish assemblages.

  2. A Novel, Nondestructive, Dried Blood Spot-Based Hematocrit Prediction Method Using Noncontact Diffuse Reflectance Spectroscopy.

    PubMed

    Capiau, Sara; Wilk, Leah S; Aalders, Maurice C G; Stove, Christophe P

    2016-06-21

    Dried blood spot (DBS) sampling is recognized as a valuable alternative sampling strategy both in research and in clinical routine. Although many advantages are associated with DBS sampling, its more widespread use is hampered by several issues, of which the hematocrit effect on DBS-based quantitation remains undoubtedly the most widely discussed one. Previously, we developed a method to derive the approximate hematocrit from a nonvolumetrically applied DBS based on its potassium content. Although this method yielded good results and was straightforward to perform, it was also destructive and required sample preparation. Therefore, we now developed a nondestructive method which allows to predict the hematocrit of a DBS based on its hemoglobin content, measured via noncontact diffuse reflectance spectroscopy. The developed method was thoroughly validated. A linear calibration curve was established after log/log transformation. The bias, intraday and interday imprecision of quality controls at three hematocrit levels and at the lower and upper limit of quantitation (0.20 and 0.67, respectively) were less than 11%. In addition, the influence of storage and the volume spotted was evaluated, as well as DBS homogeneity. Application of the method to venous DBSs prepared from whole blood patient samples (n = 233) revealed a good correlation between the actual and the predicted hematocrit. Limits of agreement obtained after Bland and Altman analysis were -0.076 and +0.018. Incurred sample reanalysis demonstrated good method reproducibility. In conclusion, mere scanning of a DBS suffices to derive its approximate hematocrit, one of the most important variables in DBS analysis.

  3. Capillary compared to venous blood sampling in clozapine treatment: patients׳ and healthcare practitioners׳ experiences with a point-of-care device.

    PubMed

    Bogers, Jan P A M; Bui, Hong; Herruer, Martien; Cohen, Dan

    2015-03-01

    Underuse of the antipsychotic clozapine for schizophrenia is an impediment to improving outcomes for patients. Because of its possible severe side effects, including granulocytopenia or even agranulocytosis, clozapine treatment entails regular WBC monitoring, which can be a major drawback for patients and practitioners. The HemoCue WBC DIFF system is a point-of-care device using capillary blood sampling which provides WBC counts with differentials, including granulocytes. We investigated if capillary sampling instead of conventional venous sampling might diminish the burden for patients and practitioners and motivate them to continue clozapine treatment. A randomized cross-over trial design was used to compare the two sampling methods. Patients׳ subjective experiences of various aspects of blood sampling were rated on a 10-cm visual analogue scale (VAS). Patients and practitioners were also asked if they had any preference for venous or capillary sampling and patients were asked if the sampling method influenced their motivation to continue clozapine treatment. Seventy-three patients were included in this study. Three dropped out before completion. The VAS ratings on all five aspects and the total burden experienced showed a consistent pattern favouring capillary blood sampling (p<0.001). This pattern was more pronounced for inpatients than for outpatients. Patients strongly preferred capillary testing (p<0.001). The method used moderately influenced their motivation for clozapine therapy. In this study patients tolerated capillary blood testing with a point-of-care device better than traditional venous sampling at a laboratory and practitioners also preferred it. Using this method might therefore boost clozapine prescription rates.

  4. Automatic method for quantitation of mercury in blood, plasma and urine.

    PubMed

    Vesterberg, O

    1991-01-01

    Here we report our experience of quantification of mercury in blood, plasma and urine by using modifications of a procedure for cold vapour atomic absorption. We have tried: (1) modifications of the instrumentation including the tower, the cell and apparatus for measurement; (2) to increase the volume of sample, avoiding problems caused by foaming and background to arrive at a reliable method with low detection limit. Blood and plasma samples were digested overnight in a mixture of nitric acid and perchloric acid (1:5). Recovery of known additions of mercury was close to 100%. Coefficients of variation (CV) within runs and between runs was for B-Hg 4.7 and 9.5, respectively at 20 nmol/l, and for U-Hg 1.8 and 5.2, respectively at 57 nmol/l. The same detection limit of 5 nmol/l was obtained with blood, plasma and urine. This is in the lower range of non-occupationally exposed normal subjects. The results, including those obtained in sample exchange with other laboratories and with reference materials, indicate that the accuracy of this method for quantification of mercury is good.

  5. Compressive sampling in computed tomography: Method and application

    NASA Astrophysics Data System (ADS)

    Hu, Zhanli; Liang, Dong; Xia, Dan; Zheng, Hairong

    2014-06-01

    Since Donoho and Candes et al. published their groundbreaking work on compressive sampling or compressive sensing (CS), CS theory has attracted a lot of attention and become a hot topic, especially in biomedical imaging. Specifically, some CS based methods have been developed to enable accurate reconstruction from sparse data in computed tomography (CT) imaging. In this paper, we will review the progress in CS based CT from aspects of three fundamental requirements of CS: sparse representation, incoherent sampling and reconstruction algorithm. In addition, some potential applications of compressive sampling in CT are introduced.

  6. [Progress in sample preparation and analytical methods for trace polar small molecules in complex samples].

    PubMed

    Zhang, Qianchun; Luo, Xialin; Li, Gongke; Xiao, Xiaohua

    2015-09-01

    Small polar molecules such as nucleosides, amines, amino acids are important analytes in biological, food, environmental, and other fields. It is necessary to develop efficient sample preparation and sensitive analytical methods for rapid analysis of these polar small molecules in complex matrices. Some typical materials in sample preparation, including silica, polymer, carbon, boric acid and so on, are introduced in this paper. Meanwhile, the applications and developments of analytical methods of polar small molecules, such as reversed-phase liquid chromatography, hydrophilic interaction chromatography, etc., are also reviewed.

  7. Comparison of pigment content of paint samples using spectrometric methods.

    PubMed

    Trzcińska, Beata; Kowalski, Rafał; Zięba-Palus, Janina

    2014-09-15

    The aim of the paper was to evaluate the influence of pigment concentration and its distribution in polymer binder on the possibility of colour identification and paint sample comparison. Two sets of paint samples: one containing red and another one green pigment were prepared. Each set consisted of 13 samples differing gradually in the concentration of pigment. To obtain the sets of various colour shades white paint was mixed with the appropriate pigment in the form of a concentrated suspension. After solvents evaporation the samples were examined using spectrometric methods. The resin and main filler were identified by IR method. Colour and white pigments were identified on the base of Raman spectra. Colour of samples were compared based on Vis spectrometry according to colour theory. It was found that samples are homogenous (parameter measuring colour similarity ΔE<3). The values of ΔE between the neighbouring samples in the set revealed decreasing linear function and between the first and following one--a logarithmic function.

  8. A Review of Methods for Detecting Melamine in Food Samples.

    PubMed

    Lu, Yang; Xia, Yinqiang; Liu, Guozhen; Pan, Mingfei; Li, Mengjuan; Lee, Nanju Alice; Wang, Shuo

    2017-01-02

    Melamine is a synthetic chemical used in the manufacture of resins, pigments, and superplasticizers. Human beings can be exposed to melamine through various sources such as migration from related products into foods, pesticide contamination, and illegal addition to foods. Toxicity studies suggest that prolonged consumption of melamine could lead to the formation of kidney stones or even death. Therefore, reliable and accurate detection methods are essential to prevent human exposure to melamine. Sample preparation is of critical importance, since it could directly affect the performance of analytical methods. Some methods for the detection of melamine include instrumental analysis, immunoassays, and sensor methods. In this paper, we have summarized the state-of-the-art methods used for food sample preparation as well as the various detection techniques available for melamine. Combinations of multiple techniques and new materials used in the detection of melamine have also been reviewed. Finally, future perspectives on the applications of microfluidic devices have also been provided.

  9. RAPID METHOD FOR DETERMINATION OF RADIOSTRONTIUM IN EMERGENCY MILK SAMPLES

    SciTech Connect

    Maxwell, S.; Culligan, B.

    2008-07-17

    A new rapid separation method for radiostrontium in emergency milk samples was developed at the Savannah River Site (SRS) Environmental Bioassay Laboratory (Aiken, SC, USA) that will allow rapid separation and measurement of Sr-90 within 8 hours. The new method uses calcium phosphate precipitation, nitric acid dissolution of the precipitate to coagulate residual fat/proteins and a rapid strontium separation using Sr Resin (Eichrom Technologies, Darien, IL, USA) with vacuum-assisted flow rates. The method is much faster than previous method that use calcination or cation exchange pretreatment, has excellent chemical recovery, and effectively removes beta interferences. When a 100 ml sample aliquot is used, the method has a detection limit of 0.5 Bq/L, well below generic emergency action levels.

  10. Fungal DNA detected in blood samples of patients who received contaminated methylprednisolone injections reveals increased complexity of causative agents.

    PubMed

    Zhao, Yanan; Armeanu, Emilian; DiVerniero, Richard; Lewis, Terri A; Dobson, Richard C; Kontoyiannis, Dimitrios P; Roilides, Emmanuel; Walsh, Thomas J; Perlin, David S

    2014-06-01

    Using Exserohilum rostratum-specific and panfungal real-time PCR, we studied 24 blood samples and 2 synovial fluid specimens from 20 patients with persistent or worsening pain following injections of contaminated methylprednisolone. Seven blood specimens from 6 patients were significantly positive for fungal DNA by panfungal PCR, with multiple fungal species identified.

  11. Evaluation of toxic metals in biological samples (scalp hair, blood and urine) of steel mill workers by electrothermal atomic absorption spectrometry.

    PubMed

    Afridi, Hassan I; Kazi, Tasneem G; Jamali, Mohammad K; Kazi, Gul H; Arain, Mohammad B; Jalbani, Nusrat; Shar, Ghulam Q; Sarfaraz, Raja A

    2006-10-01

    The determination of toxic metals in the biological samples of human beings is an important clinical screening procedure. This study aimed to assess the possible influence of environmental exposure on production workers (PW) and quality control workers (QCW) of a steel mill, all male subjects aged 25-55 years. In this investigation, the concentrations of Pb, Cd, Ni and Cr were determined in biological samples (blood, urine and scalp hair samples) from these steel mill workers in relation to controlled unexposed healthy subjects of the same age group. After pre-treatment with nitric acid-hydrogen peroxide, the samples were digested via a microwave oven, and for comparison purposes, the same samples were digested by the conventional wet acid digestion method. The samples digested were subjected to graphite furnace atomic absorption spectrometry (GFAAS). To assess the reliability of these methods, critical factors, such as detection limit(s), calibration range(s), accuracy and precision, were studied. Quality control for these procedures was established with certified sample of human hair, urine and whole blood. The results indicate that the level of lead, cadmium and nickel in scalp hair, blood and urine samples were significantly higher in both groups of exposed workers (QW and PW) than those of the controls. The possible connection of these elements with the etiology of disease is discussed. The results also show the need for immediate improvements in workplace ventilation and industrial hygiene practices.

  12. A comparison of sampling methods for examining the laryngeal microbiome

    PubMed Central

    Hanshew, Alissa S.; Jetté, Marie E.; Tadayon, Stephanie; Thibeault, Susan L.

    2017-01-01

    Shifts in healthy human microbial communities have now been linked to disease in numerous body sites. Noninvasive swabbing remains the sampling technique of choice in most locations; however, it is not well known if this method samples the entire community, or only those members that are easily removed from the surface. We sought to compare the communities found via swabbing and biopsied tissue in true vocal folds, a location that is difficult to sample without causing potential damage and impairment to tissue function. A secondary aim of this study was to determine if swab sampling of the false vocal folds could be used as proxy for true vocal folds. True and false vocal fold mucosal samples (swabbed and biopsied) were collected from six pigs and used for 454 pyrosequencing of the V3–V5 region of the 16S rRNA gene. Most of the alpha and beta measures of diversity were found to be significantly similar between swabbed and biopsied tissue samples. Similarly, the communities found in true and false vocal folds did not differ considerably. These results suggest that samples taken via swabs are sufficient to assess the community, and that samples taken from the false vocal folds may be used as proxies for the true vocal folds. Assessment of these techniques opens an avenue to less traumatic means to explore the role microbes play in the development of diseases of the vocal folds, and perhaps the rest of the respiratory tract. PMID:28362810

  13. NEW COLUMN SEPARATION METHOD FOR EMERGENCY URINE SAMPLES

    SciTech Connect

    Maxwell, S; Brian Culligan, B

    2007-08-28

    The Savannah River Site Environmental Bioassay Lab participated in the 2007 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2007. A new rapid column separation method was applied directly to the NRIP 2007 emergency urine samples, with only minimal sample preparation to reduce preparation time. Calcium phosphate precipitation, previously used to pre-concentrate actinides and Sr-90 in NRIP 2006 urine and water samples, was not used for the NRIP 2007 urine samples. Instead, the raw urine was acidified and passed directly through the stacked resin columns (TEVA+TRU+SR Resins) to separate the actinides and strontium from the NRIP urine samples more quickly. This improvement reduced sample preparation time for the NRIP 2007 emergency urine analyses significantly. This approach works well for small volume urine samples expected during an emergency response event. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and strontium-90 analyses for NRIP 2007 urine samples.

  14. Assessment of DDT and DDE levels in soil, dust, and blood samples from Chihuahua, Mexico.

    PubMed

    Martínez, Fernando Díaz-Barriga; Trejo-Acevedo, Antonio; Betanzos, Angel F; Espinosa-Reyes, Guillermo; Alegría-Torres, Jorge Alejandro; Maldonado, Iván Nelinho Pérez

    2012-02-01

    The aim of this study was to assess levels of DDT and DDE in two environmental matrices (soil and dust) and to investigate the blood levels of these insecticides in exposed children living in a north Mexican state (Chihuahua) where DDT was sprayed several years ago during (1) health campaigns for the control of malaria and (2) agricultural activities. DDT and DDE were analyzed by gas chromatography/mass spectrometry. In general, lower levels were found in household outdoor samples. The levels in outdoor samples ranged from 0.001 to 0.788 mg/kg for DDT and from 0.001 to 0.642 mg/kg for DDE. The levels in indoor samples ranged from 0.001 to 15.47 mg/kg for DDT and from 0.001 to 1.063 mg/kg for DDE. Similar results to those found in indoor soil were found in dust, in which the levels ranged from 0.001 to 95.87 mg/kg for DDT and from 0.001 to 0.797 mg/kg for DDE. Moreover, blood levels showed that all of the communities studied had been exposed to DDT and/or DDE, indicating a general past or present exposure to DDT. It is important to note that the quotient DDT/DDE in all matrices was always >1. Whether the people living in our study area are at risk is an issue that deserves further analysis. However, applying precautionary principles, it is important to initiate a risk-reduction program to decrease exposure to DDT and its metabolites in people living in this area.

  15. Quantitation of fluoride ion released sarin in red blood cell samples by gas chromatography-chemical ionization mass spectrometry using isotope dilution and large-volume injection.

    PubMed

    Jakubowski, E M; McGuire, J M; Evans, R A; Edwards, J L; Hulet, S W; Benton, B J; Forster, J S; Burnett, D C; Muse, W T; Matson, K; Crouse, C L; Mioduszewski, R J; Thomson, S A

    2004-01-01

    A new method for measuring fluoride ion released isopropyl methylphosphonofluoridate (sarin, GB) in the red blood cell fraction was developed that utilizes an autoinjector, a large-volume injector port (LVI), positive ion ammonia chemical ionization detection in the SIM mode, and a deuterated stable isotope internal standard. This method was applied to red blood cell (RBC) and plasma ethyl acetate extracts from spiked human and animal whole blood samples and from whole blood of minipigs, guinea pigs, and rats exposed by whole-body sarin inhalation. Evidence of nerve agent exposure was detected in plasma and red blood cells at low levels of exposure. The linear method range of quantitation was 10-1000 pg on-column with a detection limit of approximately 2-pg on-column. In the course of method development, several conditions were optimized for the LVI, including type of injector insert, injection volume, initial temperature, pressure, and flow rate. RBC fractions had advantages over the plasma with respect to assessing nerve agent exposure using the fluoride ion method especially in samples with low serum butyrylcholinesterase activity.

  16. On the Exploitation of Sensitivity Derivatives for Improving Sampling Methods

    NASA Technical Reports Server (NTRS)

    Cao, Yanzhao; Hussaini, M. Yousuff; Zang, Thomas A.

    2003-01-01

    Many application codes, such as finite-element structural analyses and computational fluid dynamics codes, are capable of producing many sensitivity derivatives at a small fraction of the cost of the underlying analysis. This paper describes a simple variance reduction method that exploits such inexpensive sensitivity derivatives to increase the accuracy of sampling methods. Three examples, including a finite-element structural analysis of an aircraft wing, are provided that illustrate an order of magnitude improvement in accuracy for both Monte Carlo and stratified sampling schemes.

  17. Two-dimensional signal reconstruction: The correlation sampling method

    SciTech Connect

    Roman, H. E.

    2007-12-15

    An accurate approach for reconstructing a time-dependent two-dimensional signal from non-synchronized time series recorded at points located on a grid is discussed. The method, denoted as correlation sampling, improves the standard conditional sampling approach commonly employed in the study of turbulence in magnetoplasma devices. Its implementation is illustrated in the case of an artificial time-dependent signal constructed using a fractal algorithm that simulates a fluctuating surface. A statistical method is also discussed for distinguishing coherent (i.e., collective) from purely random (noisy) behavior for such two-dimensional fluctuating phenomena.

  18. Evaluation of Stress Loaded Steel Samples Using Selected Electromagnetic Methods

    SciTech Connect

    Chady, T.

    2004-02-26

    In this paper the magnetic leakage flux and eddy current method were used to evaluate changes of materials' properties caused by stress. Seven samples made of ferromagnetic material with different level of applied stress were prepared. First, the leakage magnetic fields were measured by scanning the surface of the specimens with GMR gradiometer. Next, the same samples were evaluated using an eddy current sensor. A comparison between results obtained from both methods was carried out. Finally, selected parameters of the measured signal were calculated and utilized to evaluate level of the applied stress. A strong coincidence between amount of the applied stress and the maximum amplitude of the derivative was confirmed.

  19. Improvement and Evaluation of Loop-Mediated Isothermal Amplification for Rapid Detection of Toxoplasma gondii Infection in Human Blood Samples

    PubMed Central

    Sun, Xi-meng; Ji, Yong-sheng; Liu, Xian-yong; Xiang, Mei; He, Guang; Xie, Li; Suo, Jing-xia; Suo, Xun

    2017-01-01

    Loop-mediated isothermal amplification (LAMP), an attractive DNA amplification method, was developed as a valuable tool for the rapid detection of Toxoplasma gondii. In this study, species-specific LAMP primers were designed by targeting the AF146527 sequence, which was a conserved sequence of 200- to 300-fold repetitive 529 bp fragment of T.gondii. LAMP reaction system was optimized so that it could detect the minimal DNA sample such as a single tachyzoite or 10 copies of recombinant plasmid. No cross-reactivity was found when using DNA from other parasites as templates. Subsequently, a total of 200 human blood samples were directly investigated by two diagnostic methods, LAMP and conventional PCR. Fourteen of 200 (7%) samples were positive for Toxoplasma by LAMP (the primers developed in this study), whereas only 5 of 200 (2.5%) were proved positive by conventional PCR. The procedure of the LAMP assay was very simple, as the reaction would be carried out in a single tube under isothermal conditions at 64°C and the result would be read out with 1 h (as early as 35 min with loop primers). Thus, this method has the advantages of rapid amplification, simple operation, and easy detection and would be useful for rapid and reliable clinical diagnosis of acute toxoplasmosis, especially in developing countries. PMID:28056092

  20. Comparison of sampling methods for radiocarbon dating of carbonyls in air samples via accelerator mass spectrometry

    NASA Astrophysics Data System (ADS)

    Schindler, Matthias; Kretschmer, Wolfgang; Scharf, Andreas; Tschekalinskij, Alexander

    2016-05-01

    Three new methods to sample and prepare various carbonyl compounds for radiocarbon measurements were developed and tested. Two of these procedures utilized the Strecker synthetic method to form amino acids from carbonyl compounds with either sodium cyanide or trimethylsilyl cyanide. The third procedure used semicarbazide to form crystalline carbazones with the carbonyl compounds. The resulting amino acids and semicarbazones were then separated and purified using thin layer chromatography. The separated compounds were then combusted to CO2 and reduced to graphite to determine 14C content by accelerator mass spectrometry (AMS). All of these methods were also compared with the standard carbonyl compound sampling method wherein a compound is derivatized with 2,4-dinitrophenylhydrazine and then separated by high-performance liquid chromatography (HPLC).

  1. A molecular method to assess Phytophthora diversity in environmental samples.

    PubMed

    Scibetta, Silvia; Schena, Leonardo; Chimento, Antonio; Cacciola, Santa O; Cooke, David E L

    2012-03-01

    Current molecular detection methods for the genus Phytophthora are specific to a few key species rather than the whole genus and this is a recognized weakness of protocols for ecological studies and international plant health legislation. In the present study a molecular approach was developed to detect Phytophthora species in soil and water samples using novel sets of genus-specific primers designed against the internal transcribed spacer (ITS) regions. Two different rDNA primer sets were tested: one assay amplified a long product including the ITS1, 5.8S and ITS2 regions (LP) and the other a shorter product including the ITS1 only (SP). Both assays specifically amplified products from Phytophthora species without cross-reaction with the related Pythium s. lato, however the SP assay proved the more sensitive and reliable. The method was validated using woodland soil and stream water from Invergowrie, Scotland. On-site use of a knapsack sprayer and in-line water filters proved more rapid and effective than centrifugation at sampling Phytophthora propagules. A total of 15 different Phytophthora phylotypes were identified which clustered within the reported ITS-clades 1, 2, 3, 6, 7 and 8. The range and type of the sequences detected varied from sample to sample and up to three and five different Phytophthora phylotypes were detected within a single sample of soil or water, respectively. The most frequently detected sequences were related to members of ITS-clade 6 (i.e. P. gonapodyides-like). The new method proved very effective at discriminating multiple species in a given sample and can also detect as yet unknown species. The reported primers and methods will prove valuable for ecological studies, biosecurity and commercial plant, soil or water (e.g. irrigation water) testing as well as the wider metagenomic sampling of this fascinating component of microbial pathogen diversity.

  2. RAPID SEPARATION METHOD FOR EMERGENCY WATER AND URINE SAMPLES

    SciTech Connect

    Maxwell, S.; Culligan, B.

    2008-08-27

    The Savannah River Site Environmental Bioassay Lab participated in the 2008 NRIP Emergency Response program administered by the National Institute for Standards and Technology (NIST) in May, 2008. A new rapid column separation method was used for analysis of actinides and {sup 90}Sr the NRIP 2008 emergency water and urine samples. Significant method improvements were applied to reduce analytical times. As a result, much faster analysis times were achieved, less than 3 hours for determination of {sup 90}Sr and 3-4 hours for actinides. This represents a 25%-33% improvement in analysis times from NRIP 2007 and a {approx}100% improvement compared to NRIP 2006 report times. Column flow rates were increased by a factor of two, with no significant adverse impact on the method performance. Larger sample aliquots, shorter count times, faster cerium fluoride microprecipitation and streamlined calcium phosphate precipitation were also employed. Based on initial feedback from NIST, the SRS Environmental Bioassay Lab had the most rapid analysis times for actinides and {sup 90}Sr analyses for NRIP 2008 emergency urine samples. High levels of potential matrix interferences may be present in emergency samples and rugged methods are essential. Extremely high levels of {sup 210}Po were found to have an adverse effect on the uranium results for the NRIP-08 urine samples, while uranium results for NRIP-08 water samples were not affected. This problem, which was not observed for NRIP-06 or NRIP-07 urine samples, was resolved by using an enhanced {sup 210}Po removal step, which will be described.

  3. Foetal scalp blood sampling during labour for pH and lactate measurements.

    PubMed

    Carbonne, Bruno; Pons, Kelly; Maisonneuve, Emeline

    2016-01-01

    Second-line methods of foetal monitoring have been developed in an attempt to reduce unnecessary interventions due to continuous cardiotocography (CTG), and to better identify foetuses that are at risk of intrapartum asphyxia. Very few studies directly compared CTG with foetal scalp blood (FBS) and CTG only. Only one randomised controlled trial (RCT) was published in the 1970s and had limited power to assess neonatal outcome. Direct and indirect comparisons conclude that FBS could reduce the number of caesarean deliveries associated with the use of continuous CTG. The main drawbacks of FBS are its invasive and discontinuous nature and the need for a sufficient volume of foetal blood for analysis, especially for pH measurement, resulting in failure rates reaching 10%. FBS for lactate measurement became popular with the design of test-strip devices, requiring <0.5 mL of foetal blood. RCTs showed similar outcomes with the use of FBS for lactates compared with pH in terms of obstetrical interventions and neonatal outcomes. In conclusion, there is some evidence that FBS reduces the need for operative deliveries. However, the evidence is limited with regard to actual standards, and large RCTs, directly comparing CTG only with CTG with FBS, are still needed.

  4. Extra-corporeal blood access, sensing, and radiation methods and apparatuses

    NASA Technical Reports Server (NTRS)

    Castle, Kent D. (Inventor)

    1993-01-01

    The described invention is related to extra-corporeal blood access and radiation methods and apparatuses and, in particular, to subjecting flowing blood to energy in variety of forms, including radiation, electromagnetic force fields or atomic particles. It is directed to methods and apparatuses for accessing flowing blood and for subjecting the blood to electrical conductive, electrostatic or electromagnetic fields or for radiating the blood with some type of radiation, e.g., radio waves, ultrasonic or audio waves, microwaves, IR rays, visible light, UV radiation, x-rays, alpha, beta or gamma rays. An apparatus is employed which includes one or more access ports or windows for radiating blood and/or for sensing/analyzing blood. This invention is useful for killing viruses and bacteria in blood, monitoring blood for medical purposes, genetic modification of blood, and analyzing and/or treating blood components.

  5. Application of monolithic supports to online extraction and LC-MS analysis of benzodiazepines in whole blood samples.

    PubMed

    Bugey, Aurélie; Staub, Christian

    2007-11-01

    A column-switching (CS) LC-MS method allowing high-speed determination of benzodiazepines (BZDs) in whole blood is presented. After protein precipitation with ACN followed by evaporation and reconstitution with the loading mobile phase, the online sample clean-up was carried out using a CS device. Two extractive precolumns were evaluated: a conventional restricted access material (RAM) sorbent and a monolithic silica support. Separation was achieved using a Chromolith Performance RP-18e (100 mmx4.6 mm id) monolithic silica column, and detection was performed by atmospheric pressure chemical ionization (APCI) MS. The method with both supports has been fully validated according to an accuracy profile approach. Finally, the monolithic silica column, demonstrating better validation data and a higher robustness than the RAM sorbent, was used for the analysis of several real forensic cases.

  6. A new method to measure air-borne pyrogens based on human whole blood cytokine response.

    PubMed

    Kindinger, Ilona; Daneshian, Mardas; Baur, Hans; Gabrio, Thomas; Hofmann, Andreas; Fennrich, Stefan; von Aulock, Sonja; Hartung, Thomas

    2005-03-01

    Air-borne microorganisms, as well as their fragments and components, are increasingly recognized to be associated with pulmonary diseases, e.g. organic dust toxic syndrome, humidifier lung, building-related illness, "Monday sickness." We have previously described and validated a new method for the detection of pyrogenic (fever-inducing) microbial contaminations in injectable drugs, based on the inflammatory reaction of human blood to pyrogens. We have now adapted this test to evaluate the total inflammatory capacity of air samples. Air was drawn onto PTFE membrane filters, which were incubated with human whole blood from healthy volunteers inside the collection device. Cytokine release was measured by ELISA. The test detects endotoxins and non-endotoxins, such as fungal spores, Gram-positive bacteria and their lipoteichoic acid moiety and pyrogenic dust particles with high sensitivity, thus reflecting the total inflammatory capacity of a sample. When air from different surroundings such as working environments and animal housing was assayed, the method yielded reproducible data which correlated with other parameters of microbial burden tested. We further developed a standard material for quantification and showed that this assay can be performed with cryopreserved as well as fresh blood. The method offers a test to measure the integral inflammatory capacity of air-borne microbial contaminations relevant to humans. It could thus be employed to assess air quality in different living and work environments.

  7. Methods to Detect Nitric Oxide and its Metabolites in Biological Samples

    PubMed Central

    Bryan, Nathan S.; Grisham, Matthew B.

    2007-01-01

    Nitric oxide (NO) methodology is a complex and often confusing science and the focus of many debates and discussion concerning NO biochemistry. NO is involved in many physiological processes including regulation of blood pressure, immune response and neural communication. Therefore its accurate detection and quantification is critical to understanding health and disease. Due to the extremely short physiological half life of this gaseous free radical, alternative strategies for the detection of reaction products of NO biochemistry have been developed. The quantification of NO metabolites in biological samples provides valuable information with regards to in vivo NO production, bioavailability and metabolism. Simply sampling a single compartment such as blood or plasma may not always provide an accurate assessment of whole body NO status, particularly in tissues. Therefore, extrapolation of plasma or blood NO status to specific tissues of interest is no longer a valid approach. As a result, methods continue to be developed and validated which allow the detection and quantification of NO and NO-related products/metabolites in multiple compartments of experimental animals in vivo. The methods described in this review is not an exhaustive or comprehensive discussion of all methods available for the detection of NO but rather a description of the most commonly used and practical methods which allow accurate and sensitive quantification of NO products/metabolites in multiple biological matrices under normal physiological conditions. PMID:17664129

  8. Blood cell counting and classification by nonflowing laser light scattering method

    NASA Astrophysics Data System (ADS)

    Yang, Ye; Zhang, Zhenxi; Yang, Xinhui; Jiang, Dazong; Yeo, Joon Hock

    1999-11-01

    A new non-flowing laser light scattering method for counting and classifying blood cells is presented. A linear charge- coupled device with 1024 elements is used to detect the scattered light intensity distribution of the blood cells. A pinhole plate is combined with the CCD to compete the focusing of the measurement system. An isotropic sphere is used to simulate the blood cell. Mie theory is used to describe the scattering of blood cells. In order to inverse the size distribution of blood cells from their scattered light intensity distribution, Powell method combined with precision punishment method is used as a dependent model method for measurement red blood cells and blood plates. Non-negative constraint least square method combined with Powell method and precision punishment method is used as an independent model for measuring white blood cells. The size distributions of white blood cells and red blood cells, and the mean diameter of red blood cells are measured by this method. White blood cells can be divided into three classes: lymphocytes, middle-sized cells and neutrocytes according to their sizes. And the number of blood cells in unit volume can also be measured by the linear dependence of blood cells concentration on scattered light intensity.

  9. Proteome Analysis of Human Perilymph using an Intraoperative Sampling Method.

    PubMed

    Schmitt, Heike Andrea; Pich, Andreas; Schröder, Anke; Scheper, Verena; Lilli, Giorgio; Reuter, Günter; Lenarz, Thomas

    2017-03-10

    The knowledge about the etiology and pathophysiology of sensorineural hearing loss (SNHL) is still very limited. The project aims at the improvement of understanding different types of SNHL by proteome analysis of human perilymph. Sampling of perilymph has been established during inner ear surgeries (cochlear implant and vestibular sch