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Sample records for blood spot samples

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    whole blood samples at low temperatures and rapid isolation of plasma and serum. Effects of different handling procedures for all markers studied are given. DBSS proved to be a robust and convenient way to handle samples for immunoassay analysis of inflammatory markers in whole blood Udgivelsesdato......The interests in monitoring inflammation by immunoassay determination of blood inflammatory markers call for information on the stability of these markers in relation to the handling of blood samples. The increasing use of stored biobank samples for such ventures that may have been collected and...... stored for other purposes, justifies the study hereof. Blood samples were stored for 0, 4, 24, and 48 h at 4 degrees C, room temperature (RT), and at 35 degrees C, respectively, before they were separated into serum or plasma and frozen. Dried blood spot samples (DBSS) were stored for 0, 1, 2, 3, 7, and...

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

    Science.gov (United States)

    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 (R2 > 0.80), and whole blood and DBS samples generally showed very good agreement. Distribution coefficients estimated using biologically-based models were near one and did not explain the observed distribution. Among the study population, median concentrations of several pesticides/CHCs and PBDEs exceeded levels reported in the 2007–2008 National Health and Nutrition Examination Survey, while levels of other OCPs and PBDEs were comparable or lower. Race and smoking status appeared to slightly affect plasma/blood concentration ratios for several POPs. The experimentally

  3. Genome-wide scans using archived neonatal dried blood spot samples

    Directory of Open Access Journals (Sweden)

    Wiuf Carsten

    2009-07-01

    Full Text Available Abstract Background Identification of disease susceptible genes requires access to DNA from numerous well-characterised subjects. Archived residual dried blood spot samples from national newborn screening programs may provide DNA from entire populations and medical registries the corresponding clinical information. The amount of DNA available in these samples is however rarely sufficient for reliable genome-wide scans, and whole-genome amplification may thus be necessary. This study assess the quality of DNA obtained from different amplification protocols by evaluating fidelity and robustness of the genotyping of 610,000 single nucleotide polymorphisms, using the Illumina Infinium HD Human610-Quad BeadChip. Whole-genome amplified DNA from 24 neonatal dried blood spot samples stored between 15 to 25 years was tested, and high-quality genomic DNA from 8 of the same individuals was used as reference. Results Using 3.2 mm disks from dried blood spot samples the optimal DNA-extraction and amplification protocol resulted in call-rates between 99.15% – 99.73% (mean 99.56%, N = 16, and conflicts with reference DNA in only three per 10,000 genotype calls. Conclusion Whole-genome amplified DNA from archived neonatal dried blood spot samples can be used for reliable genome-wide scans and is a cost-efficient alternative to collecting new samples.

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

    International Nuclear Information System (INIS)

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

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

    Science.gov (United States)

    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.

  6. Adiponectin levels measured in dried blood spot samples from neonates born small and appropriate for gestational age

    DEFF Research Database (Denmark)

    Klamer, A; Skogstrand, Kristin; Hougaard, D M;

    2007-01-01

    Adiponectin levels measured in neonatal dried blood spot samples (DBSS) might be affected by both prematurity and being born small for gestational age (SGA). The aim of the study was to measure adiponectin levels in routinely collected neonatal DBSS taken on day 5 (range 3-12) postnatal from...

  7. Role of therapeutic drug monitoring in pulmonary infections : use and potential for expanded use of dried blood spot samples

    NARCIS (Netherlands)

    Hofman, Susan; Bolhuis, Mathieu S.; Koster, Remco A.; Akkerman, Onno W.; van Assen, Sander; Stove, Christophe; Alffenaar, Jan-Willem C.

    2015-01-01

    Respiratory tract infections are among the most common infections in men. We reviewed literature to document their pharmacological treatments, and the extent to which therapeutic drug monitoring (TDM) is needed during treatment. We subsequently examined potential use of dried blood spots as sample p

  8. A sup 125 I-radioimmunoassay for measuring androstenedione in serum and in blood-spot samples from neonates

    Energy Technology Data Exchange (ETDEWEB)

    Thomson, S.; Wallace, A.M.; Cook, B. (Stobhill Hospital, Glasgow (England))

    1989-08-01

    We developed a radioimmunoassay with a gamma-emitting radioligand to measure androstenedione in human serum and in dried blood-spot samples from newborns. Antisera were raised in rabbits against androstenedione linked to bovine serum albumin at positions 3, 6, or 11 on the steroid nucleus. Radioligands were prepared by linking ({sup 125}I)iodohistamine at positions 3, 6, or 11. Linkages were through either carboxymethyloxime or hemisuccinate bridges. All label and antibody combinations were examined, and the most sensitive and specific combination (antiserum raised against androstenedione-3-carboxymethyloxime-bovine serum albumin with an androstenedione-carboxymethyloxime-({sup 125}I)iodohistamine label) was selected for full evaluation. We report the performance of these selected reagents in an immunoassay for androstenedione in both serum and dried blood-spot samples from neonates. We measured concentrations of androstenedione in serum under normal and pathological conditions such as congenital adrenal hyperplasia and polycystic ovarian disease. Diurnal variation in normal men was observed. Androstenedione was measured in blood spots from neonates born at term or prematurely, with respiratory distress syndrome, or with congenital adrenal hyperplasia.

  9. Evaluation of dried blood spots as sample matrix for gas chromatography/mass spectrometry based metabolomic profiling.

    Science.gov (United States)

    Kong, Sing Teang; Lin, Hai-Shu; Ching, Jianhong; Ho, Paul C

    2011-06-01

    We propose using dried blood spots (DBS) as sample matrix for gas chromatography/mass spectrometry (GC/MS) based metabolomic profiling for the benefits of higher sample stability, more convenient sample acquisition with DBS, higher analyte separation power, and more readily biomarker identification with GC/MS. To establish this proposition, the metabolomic profiles generated from DBS were compared with that obtained from the conventional whole blood and plasma matrixes and also with dried plasma spots (DPS) as another covariate control. Our findings indicated that whole blood produced the most number of detectable markers (866), whereas DPS yielded the least number (614). DBS and plasma matrix, on the other hand, produced the most similar numbers of detectable (695 vs 749) and identifiable markers (137 vs 147, matching with Fiehn library). From the analysis of the DBS and plasma metabolomic profiles, it was concluded that when l-lysine 2, iminodiacetic acid 2, dl-threo-beta-hydroxyaspartic acid, citric acid, or adenosine-5-monophosphate 2 are not involved as markers, DBS could be a suitable substitute for plasma for metabolomic profiling.

  10. Therapeutic drug monitoring in dried blood spots using liquid microjunction surface sampling and high resolution mass spectrometry.

    Science.gov (United States)

    Gaissmaier, Tanja; Siebenhaar, Markus; Todorova, Vanya; Hüllen, Volker; Hopf, Carsten

    2016-02-01

    Dried blood spots (DBS) are a versatile and stable tool for direct clinical blood analysis. Ambient high-resolution mass spectrometry is emerging as a method of choice for their quantitative analysis, for instance in therapeutic drug monitoring. Here, we coupled liquid microjunction surface sampling technology, a so-called Flowprobe, with an Orbitrap mass spectrometer and demonstrated the utility of this set-up for direct quantification of multiple drugs in DBS on filter paper. A three-layer set-up that we had introduced earlier enabled introduction of internal standards into DBS. We furthermore took an established point-of-care test system a step further and analyzed disposable test fields for blood glucose monitoring also for Flowprobe-based acetaminophen screening without additional sample preparation. Using as little as 2 μL blood, the method had an LOD of 1 μg mL(-1) (coefficient of variation of ≤15%) and acetaminophen recoveries of 82 to 119% for blinded samples, as assessed by LC-MS/MS. Half an hour after ingestions of a single 1000 mg acetaminophen dose, indistinguishable drug levels were measured in three healthy volunteers by LC-MS/MS and Flowprobe-Orbitrap MS analysis of DBS. Flowprobe analysis of DBS was 6- to 100-times more sensitive than corresponding desorption electrospray ionization MS analysis for four drugs. For instance, the LOD for salicylic acid analysis was 0.07 ng mL(-1) with Flowprobe measurement. Furthermore, we showed that multi-component analysis of five different substances, which may mimic polypharmacy in diabetes patients, in one blood sample for screening purposes was feasible. Taken together, our study suggests that microjunction surface sampling of DBS on filter paper and disposable point-of-care test fields may be developed into routine methods for near-patient multi-compound therapeutic drug monitoring that may advance blood screening analysis for patients with polypharmacy. PMID:26759833

  11. Archived neonatal dried blood spot samples can be used for accurate whole genome and exome-targeted next-generation sequencing

    DEFF Research Database (Denmark)

    Hollegaard, Mads Vilhelm; Grauholm, Jonas; Nielsen, Ronni;

    2013-01-01

    Dried blood spot samples (DBSS) have been collected and stored for decades as part of newborn screening programmes worldwide. Representing almost an entire population under a certain age and collected with virtually no bias, the Newborn Screening Biobanks are of immense value in medical studies...... can be used for accurate whole genome sequencing (WGS) and exome sequencing (WES). This study examined two individuals represented by three different types of samples each: whole-blood (reference samples), 3-year-old DBSS spotted with reference material (refDBSS), and 27- to 29-year-old archived...... neonatal DBSS (neoDBSS) stored at -20°C in the Danish Newborn Screening Biobank. The reference samples were genotyped using an Illumina Omni2.5M array, and all samples were sequenced on a HighSeq2000 Paired-End flow cell. First, we compared the array single nucleotide polymorphism (SNP) genotype data...

  12. Application of dried blood spots to determine vitamin D status in a large nutritional study with unsupervised sampling: the Food4Me project.

    Science.gov (United States)

    Hoeller, Ulrich; Baur, Manuela; Roos, Franz F; Brennan, Lorraine; Daniel, Hannelore; Fallaize, Rosalind; Forster, Hannah; Gibney, Eileen R; Gibney, Mike; Godlewska, Magdalena; Hartwig, Kai; Kolossa, Silvia; Lambrinou, Christina P; Livingstone, Katherine M; Lovegrove, Julie A; Macready, Anna L; Manios, Yannis; Marsaux, Cyril F M; Martinez, J Alfredo; Celis-Morales, Carlos; Moschonis, George; Navas-Carretero, Santiago; O'Donovan, Clare B; San-Cristobal, Rodrigo; Saris, Wim H M; Surwiłło, Agnieszka; Traczyk, Iwona; Tsirigoti, Lydia; Walsh, Marianne C; Woolhead, Clara; Mathers, John C; Weber, Peter

    2016-01-28

    An efficient and robust method to measure vitamin D (25-hydroxy vitamin D3 (25(OH)D3) and 25-hydroxy vitamin D2 in dried blood spots (DBS) has been developed and applied in the pan-European multi-centre, internet-based, personalised nutrition intervention study Food4Me. The method includes calibration with blood containing endogenous 25(OH)D3, spotted as DBS and corrected for haematocrit content. The methodology was validated following international standards. The performance characteristics did not reach those of the current gold standard liquid chromatography-MS/MS in plasma for all parameters, but were found to be very suitable for status-level determination under field conditions. DBS sample quality was very high, and 3778 measurements of 25(OH)D3 were obtained from 1465 participants. The study centre and the season within the study centre were very good predictors of 25(OH)D3 levels (Pnutritional study.

  13. Detecting 22q11.2 deletions by use of multiplex ligation-dependent probe amplification on DNA from neonatal dried blood spot samples

    DEFF Research Database (Denmark)

    Sørensen, Karina M; Agergaard, Peter; Olesen, Charlotte;

    2010-01-01

    of 22q11.2 deletions among certain manifestations, eg, congenital heart disease, on selected Danes, a multiplex ligation-dependant probe amplification (MLPA) analysis was designed. The analysis was planned to be performed on DNA extracted from dried blood spot samples (DBSS) obtained from Guthrie cards...... collected during neonatal screening programs. However, the DNA concentration necessary for a standard MLPA analysis (20 ng) could not be attained from DBSS, and a novel MLPA design was developed to permit for analysis on limited amounts of DNA (2 ng). A pilot study is reported here that validates the new...... MLPA design using nine patients diagnosed with the 22q11.2 deletion and 101 controls. All deletions were identified using DNA extracted from DBSS, and no copy number variations were detected in the controls, resulting in a specificity and sensitivity of 100%. It is thereby concluded that the novel MLPA...

  14. Detection of Streptococcus pneumoniae and Haemophilus influenzae type B by real-time PCR from dried blood spot samples among children with pneumonia: a useful approach for developing countries.

    Directory of Open Access Journals (Sweden)

    Laura Selva

    Full Text Available Dried blood spot (DBS is a reliable blood collection method for storing samples at room temperature and easily transporting them. We have previously validated a Real-Time PCR for detection of Streptococcus pneumoniae in DBS. The objective of this study was to apply this methodology for the diagnosis of S. pneumoniae and Haemophilus influenzae b (Hib in DBS samples of children with pneumonia admitted to two hospitals in Mozambique and Morocco.Ply and wzg genes of S. pneumoniae and bexA gene of Hib, were used as targets of Real-Time PCR. 329 DBS samples of children hospitalized with clinical diagnosis of pneumonia were tested.Real-Time PCR in DBS allowed for a significant increase in microbiological diagnosis of S. pneumoniae and Hib. When performing blood bacterial culture, only ten isolates of S. pneumoniae and none of Hib were detected (3·0% positivity rate, IC95% 1·4-5·5%. Real-Time PCR from DBS samples increased the detection yield by 4x fold, as 30 S. pneumoniae and 11 Hib cases were detected (12·4% positivity rate, IC95% 9·0-16·5%; P<0·001.Real-Time PCR applied in DBS may be a valuable tool for improving diagnosis and surveillance of pneumonia caused by S. pneumoniae or Hib in developing countries.

  15. Quantitation of 5-Methyltetrahydrofolic Acid in Dried Blood Spots and Dried Plasma Spots by Stable Isotope Dilution Assays.

    Directory of Open Access Journals (Sweden)

    Markus Kopp

    Full Text Available Because of minimal data available on folate analysis in dried matrix spots (DMSs, we combined the advantages of stable isotope dilution assays followed by LC-MS/MS analysis with DMS sampling to develop a reliable method for the quantitation of plasma 5-methyltetrahydrofolic acid in dried blood spots (DBSs and dried plasma spots (DPSs as well as for the quantitation of whole blood 5-methyltetrahydrofolic acid in DBSs. We focused on two diagnostically conclusive parameters exhibited by the plasma and whole blood 5-methyltetrahydrofolic acid levels that reflect both temporary and long-term folate status. The method is performed using the [2H4]-labeled isotopologue of the vitamin as the internal standard, and three steps are required for the extraction procedure. Elution of the punched out matrix spots was performed using stabilization buffer including Triton X-100 in a standardized ultrasonication treatment followed by enzymatic digestion (whole blood only and solid-phase extraction with SAX cartridges. This method is sensitive enough to quantify 27 nmol/L whole blood 5-methyltetrahydrofolic acid in DBSs and 6.3 and 4.4 nmol/L plasma 5-methyltetrahydrofolic acid in DBSs and DPSs, respectively. The unprecedented accurate quantification of plasma 5-methyltetrahydrofolic acid in DBSs was achieved by thermal treatment prior to ultrasonication, inhibiting plasma conjugase activity. Mass screenings are more feasible and easier to facilitate for this method in terms of sample collection and storage compared with conventional clinical sampling for the assessment of folate status.

  16. Quantification of hepatic carbohydrate metabolism in conscious mice using serial blood and urine spots

    NARCIS (Netherlands)

    van Dijk, TH; Boer, TS; Havinga, R; Stellaard, F; Kuipers, F; Reijngoud, DJ

    2003-01-01

    In vivo studies of hepatic carbohydrate metabolism in (genetically modified) conscious mice are hampered by limitations of blood and urine sample sizes. We developed and validated methods to quantify stable isotope dilution and incorporation in small blood and urine samples spotted onto filter paper

  17. Application of dried blood spot cards to determine olive oil phenols (hydroxytyrosol metabolites) in human blood.

    Science.gov (United States)

    de Las Hazas, María Carmen López; Motilva, Maria José; Piñol, Carme; Macià, Alba

    2016-10-01

    In this study, a fast and simple blood sampling and sample pre-treatment method based on the use of the dried blood spot (DBS) cards and ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for the quantification of olive oil phenolic metabolites in human blood was developed and validated. After validation, the method was applied to determine hydroxytyrosol metabolites in human blood samples after the acute intake of an olive oil phenolic extract. Using the FTA DMPK-A DBS card under optimum conditions, with 20µL as the blood solution volume, 100µL of methanol/Milli-Q water (50/50, v/v) as the extraction solvent and 7 disks punched out from the card, the main hydroxytyrosol metabolites (hydroxytyrosol-3-O-sulphate and hydroxytyrosol acetate sulphate) were identified and quantified. The developed methodology allowed detecting and quantifying the generated metabolites at low μM levels. The proposed method is a significant improvement over existing methods to determine phenolic metabolites circulating in blood and plasma samples, thus making blood sampling possible with the volunteer pricking their own finger, and the subsequent storage of the blood in the DBS cards prior to chromatographic analysis. PMID:27474297

  18. Manual versus automated blood sampling

    DEFF Research Database (Denmark)

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

    2014-01-01

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

  19. Dried saliva spot as a sampling technique for saliva samples.

    Science.gov (United States)

    Abdel-Rehim, Abbi; Abdel-Rehim, Mohamed

    2014-06-01

    For the first time, dried saliva spot (DSS) was used as a sampling technique for saliva samples. In the DSS technique 50 μL of saliva was collected on filter paper and the saliva was then extracted with an organic solvent. The local anesthetic lidocaine was used as a model compound, which was determined in the DSS using liquid chromatography and mass spectrometry. The results obtained for the determination of lidocaine in saliva using DSS were compared with those from a previous study using a microextraction by packed sorbent syringe as the sampling method for saliva. This study shows that DSS can be used for the analysis of saliva samples. The method is promising and very easy in terms of sampling and extraction procedures. The results from this study are in good agreement with those from our previous work on the determination of lidocaine in saliva. DSS can open a new dimension in the saliva handling process in terms of sampling, storing and transport.

  20. Dried blood spot analysis for therapeutic drug monitoring of pazopanib.

    Science.gov (United States)

    de Wit, Djoeke; den Hartigh, Jan; Gelderblom, Hans; Qian, Yanwen; den Hollander, Margret; Verheul, Henk; Guchelaar, Henk-Jan; van Erp, Nielka P

    2015-12-01

    Dried blood spot (DBS) sampling is potentially a more patient-friendly and flexible alternative to venous sampling of pazopanib. This study determined the agreement between pazopanib DBS and plasma concentrations to facilitate implementation of pazopanib DBS sampling into clinical practice. Paired DBS and plasma samples were collected in 12 patients. Pazopanib plasma concentrations were calculated from DBS concentrations using the formula: plasma concentration = DBSconcentration /(1 - hematocrit). Passing-Bablok and Bland-Altman analyses were used to determine the agreement between calculated and measured plasma concentrations. We predefined a clinical acceptance limit of 25% for the Bland-Altman analysis. Passing-Bablok analysis showed a small constant (intercept estimate, -8.53 [95%CI, -12.22 to -4.41]) and slightly proportional (slope estimate, 1.15 [95%CI, 1.04-1.24]) bias between calculated and measured concentrations. This bias was clinically nonrelevant, as shown by Bland-Altman analysis; the mean ratio of calculated to measured concentrations was 0.94 (95%CI, 0.65-1.23). The clinical acceptance limits were well within these 95% limits of agreement. More specifically, 92.6% of the data points were within the predefined acceptance limits. Pazopanib plasma concentrations can be accurately calculated from DBS concentrations. Although validation of DBS cards prepared by patients themselves is required, these results show that DBS sampling can be used to monitor pazopanib therapy in clinical practice. PMID:26032288

  1. The use of mass spectrometry to analyze dried blood spots.

    Science.gov (United States)

    Wagner, Michel; Tonoli, David; Varesio, Emmanuel; Hopfgartner, Gérard

    2016-01-01

    Dried blood spots (DBS) typically consist in the deposition of small volumes of capillary blood onto dedicated paper cards. Comparatively to whole blood or plasma samples, their benefits rely in the fact that sample collection is easier and that logistic aspects related to sample storage and shipment can be relatively limited, respectively, without the need of a refrigerator or dry ice. Originally, this approach has been developed in the sixties to support the analysis of phenylalanine for the detection of phenylketonuria in newborns using bacterial inhibition test. In the nineties tandem mass spectrometry was established as the detection technique for phenylalanine and tyrosine. DBS became rapidly recognized for their clinical value: they were widely implemented in pediatric settings with mass spectrometric detection, and were closely associated to the debut of newborn screening (NBS) programs, as a part of public health policies. Since then, sample collection on paper cards has been explored with various analytical techniques in other areas more or less successfully regarding large-scale applications. Moreover, in the last 5 years a regain of interest for DBS was observed and originated from the bioanalytical community to support drug development (e.g., PK studies) or therapeutic drug monitoring mainly. Those recent applications were essentially driven by improved sensitivity of triple quadrupole mass spectrometers. This review presents an overall view of all instrumental and methodological developments for DBS analysis with mass spectrometric detection, with and without separation techniques. A general introduction to DBS will describe their advantages and historical aspects of their emergence. A second section will focus on blood collection, with a strong emphasis on specific parameters that can impact quantitative analysis, including chromatographic effects, hematocrit effects, blood effects, and analyte stability. A third part of the review is dedicated to

  2. Dried blood spot analysis ; facing new challenges

    NARCIS (Netherlands)

    Koster, Remco A; Touw, Daan J; Alffenaar, Jan-willem C

    2015-01-01

    Over the last decade, DBS analysis has gained popularity for TDM because it’s a patient friendly sampling proce- dure [1-4]. Additional advantages are prolonged sample stability, lower risk of infections and transportation at ambient temperature [1-3,5]. These advantages may fa- cilitate implementat

  3. HEPATITIS B VIRUS DNA CAN BE AMPLIFIED DIRECTLY FROM DRIED BLOOD SPOT ON FILTER PAPER

    OpenAIRE

    Naglaa Fathy Alhusseini; Mamdouh Z. Abadeer; Sheref M. El-Taher

    2012-01-01

    Hepatitis B Virus (HBV) infections represent a major public health problem because of the ability of HBV to cause a chronic carrier state. Even though chronic carriers remain largely asymptomatic, a large number of these individuals subsequently develop cirrhosis and primary hepatocellular carcinomas. Dried Blood Spot (DBS) samples are a simple and inexpensive sampling method, especially useful for blood collection in resource poor settings with limited access to diagnostic facilities. The ma...

  4. A simple method to quantitate IP-10 in dried blood and plasma spots

    DEFF Research Database (Denmark)

    Aabye, Martine G; Eugen-Olsen, Jesper; Werlinrud, Anne Marie;

    2012-01-01

    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...

  5. Eliciting parental support for the use of newborn blood spots for pediatric research

    OpenAIRE

    Yeung, Edwina H.; Louis, Germaine Buck; Lawrence, David; Kannan, Kurunthachalam; McLain, Alexander C.; Caggana, Michele; Druschel, Charlotte; Bell, Erin

    2016-01-01

    Background Biomarkers of exposures such as infection or environmental chemicals can be measured in small volumes of blood extracted from newborn dried blood spots (DBS) underscoring their potential utility for population-based research. However, few studies have evaluated the feasibility and utility of this resource; particularly the factors associated with parental consent, and the ability to retrieve banked samples with sufficient remaining volume for epidemiologic research. Methods At 8 mo...

  6. Blood spots as an alternative to whole blood collection and the effect of a small monetary incentive to increase participation in genetic association studies

    Directory of Open Access Journals (Sweden)

    Ringer Danny

    2009-11-01

    Full Text Available Abstract Background Collection of buccal cells from saliva for DNA extraction offers a less invasive and convenient alternative to venipuncture blood collection that may increase participation in genetic epidemiologic studies. However, dried blood spot collection, which is also a convenient method, offers a means of collecting peripheral blood samples from which analytes in addition to DNA can be obtained. Methods To determine if offering blood spot collection would increase participation in genetic epidemiologic studies, we conducted a study of collecting dried blood spot cards by mail from a sample of female cancer cases (n = 134 and controls (n = 256 who were previously selected for a breast cancer genetics study and declined to provide a venipuncture blood sample. Participants were also randomized to receive either a $2.00 bill or no incentive with the blood spot collection kits. Results The average time between the venipuncture sample refusal and recruitment for the blood spot collection was 4.4 years. Thirty-seven percent of cases and 28% of controls provided a dried blood spot card. While the incentive was not associated with participation among controls (29% for $2.00 incentive vs. 26% for no incentive, p = 0.6, it was significantly associated with participation among the breast cancer cases (48% vs. 27%, respectively, p = 0.01. There did not appear to be any bias in response since no differences between cases and controls and incentive groups were observed when examining several demographic, work history and radiation exposure variables. Conclusion This study demonstrates that collection of dried blood spot cards in addition to venipuncture blood samples may be a feasible method to increase participation in genetic case-control studies.

  7. Analysis of the Stability of Urea in Dried Blood Spots Collected and Stored on Filter Paper

    Science.gov (United States)

    Lakshmy, Ramakrishnan; Mukhopadhyay, Ashok Kumar; Jailkhani, Bansi Lal

    2013-01-01

    The ability to use dry blood spots (DBSs) on filter paper for the analysis of urea levels could be an important diagnostic tool for areas that have limited access to laboratory facilities. We developed a method for the extraction and quantification of urea from DBSs that were stored on 3M Whatman filter paper and investigated the effect of long-term storage on the level of urea in DBSs. DBSs of 4.5 mm in diameter were used for our assay, and we determined the urea levels in blood using a commercially available enzymatic kit (UV GLDH-method; Randox laboratories Ltd., UK). The DBSs on filter discs were stored at 4℃ or at 37℃ for 120 days. The mean intra- and inter-assay coefficient of variance for our method of urea extraction from dried blood was 4.2% and 6.3%, respectively. We collected 75 fresh blood samples and compared the urea content of each fresh sample with the urea content of DBSs taken from corresponding fresh blood samples. Regression analysis reported a regression coefficient (r) value of 0.97 and a recovery of urea from dried spots was 102.2%. Urea concentrations in DBSs were stable for up to 120 and 90 days when stored at 4℃ and 37℃, respectively. Our results show that urea can be stored and quantitatively recovered from small volumes of blood that was collected on filter paper. PMID:23667845

  8. HEPATITIS B VIRUS DNA CAN BE AMPLIFIED DIRECTLY FROM DRIED BLOOD SPOT ON FILTER PAPER

    Directory of Open Access Journals (Sweden)

    Naglaa Fathy Alhusseini

    2012-01-01

    Full Text Available Hepatitis B Virus (HBV infections represent a major public health problem because of the ability of HBV to cause a chronic carrier state. Even though chronic carriers remain largely asymptomatic, a large number of these individuals subsequently develop cirrhosis and primary hepatocellular carcinomas. Dried Blood Spot (DBS samples are a simple and inexpensive sampling method, especially useful for blood collection in resource poor settings with limited access to diagnostic facilities. The main advantage of DBS samples over routine blood samples is that only a small quantity of blood, is required. They are easy to obtain, stable and can be transported to a reference laboratory at minimal cost. This study was to evaluate the feasibility of DBS samples for direct amplification of HBV DNA bypassing nucleic acid extraction. Results obtained from DBS samples were compared from those from plasma by routine molecular technique and also with those from whole blood. On the whole results for DBS, whole blood and plasma samples for HBV-DNA semi quantitative PCR monitoring, demonstrated very good agreement. This study also represents the first report in Egypt to evaluate the use of DBS to direct amplification of HBV-DNA and concluded that the use of DBS for direct amplification of HBV DNA without nucleic acid extraction was reliable, specific, sensitive, cheap and appropriate method to monitor the HBV infected patients."

  9. Whole-genome amplified DNA from stored dried blood spots is reliable in high resolution melting curve and sequencing analysis

    DEFF Research Database (Denmark)

    Winkel, Bo G; Hollegaard, Mads Vilhelm; Olesen, Morten S;

    2011-01-01

    The use of dried blood spots (DBS) samples in genomic workup has been limited by the relative low amounts of genomic DNA (gDNA) they contain. It remains to be proven that whole genome amplified DNA (wgaDNA) from stored DBS samples, constitutes a reliable alternative to gDNA.We wanted to compare m...

  10. Diagnosis of HIV-1 infection in infants using dried blood spots in Tamil Nadu, South India

    Directory of Open Access Journals (Sweden)

    D Anitha

    2011-01-01

    Full Text Available Introduction: Diagnosis of HIV infection in infants is difficult due to the presence of maternal antibodies; only nucleic acid assays are very helpful in early detection. Filter papers are especially useful for blood collection in resource-poor settings with limited access to diagnostic facilities. Materials & Methods: DBS samples were collected from the infants born to HIV seropositive mothers who had received single dose nevirapine at onset of labor. The samples were directly spotted onto the Whatman 903 cards from heel, big toe or finger prick depending on the age of the infants. A total of 766 infant samples were collected on dried blood spots (DBS and transported to the Department of Experimental Medicine (DEM, Chennai, for testing from different government hospitals of rural and urban parts of Tamil Nadu, South India. According to National AIDS Control Organization′s (NACO protocol DNA was extracted from all these DBS and PCR was performed using the Roche kit version 1.5. Results: Fifteen infants were found to be HIV positive and 751 were HIV negative; all these 15 positive infants and 49 negative infants who were in the age group between 10 and 18 months were repeated with another DBS and compared with whole blood. The DBS results were concordant with the whole blood method and the sensitivity and specificity were 100%.

  11. Age determination of blood spots in forensic medicine by force spectroscopy.

    Science.gov (United States)

    Strasser, Stefan; Zink, Albert; Kada, Gerald; Hinterdorfer, Peter; Peschel, Oliver; Heckl, Wolfgang M; Nerlich, Andreas G; Thalhammer, Stefan

    2007-07-20

    We present a new tool for the estimation of the age of bloodstains, which could probably be used during forensic casework. For this, we used atomic force microscopy (AFM) for high-resolution imaging of erythrocytes in a blood sample and the detection of elasticity changes on a nanometer scale. For the analytic procedure we applied a fresh blood spot on a glass slide and started the AFM detection after drying of the blood drop. In a first step, an overview image was generated showing the presence of several red blood cells, which could easily be detected due to their typical "doughnut-like" appearance. The consecutively morphological investigations in a timeframe of 4 weeks could not show any alterations. Secondly, AFM was used to test the elasticity by recording force-distance curves. The measurements were performed immediately after drying, 1.5 h, 30 h and 31 days. The conditions were kept constant at room temperature (20 degrees C) and a humidity of 30%. The obtained elasticity parameters were plotted against a timeline and repeated several times. The elasticity pattern showed a decrease over time, which are most probably influenced by the alteration of the blood spot during the drying and coagulation process. The preliminary data demonstrates the capacity of this method to use it for development of calibration curves, which can be used for estimation of bloodstain ages during forensic investigations.

  12. Use of Blood Smears and Dried Blood Spots for Polymerase Chain Reaction-Based Detection and Quantification of Bacterial Infection and Plasmodium falciparum in Severely Ill Febrile African Children.

    Science.gov (United States)

    Wihokhoen, Benchawan; Dondorp, Arjen M; Turner, Paul; Woodrow, Charles J; Imwong, Mallika

    2016-02-01

    Molecular approaches offer a means of testing archived samples stored as dried blood spots in settings where standard blood cultures are not possible. Peripheral blood films are one suggested source of material, although the sensitivity of this approach has not been well defined. Thin blood smears and dried blood spots from a severe pediatric malaria study were assessed using specific polymerase chain reaction (PCR) primers to detect non-typhoidal Salmonella (NTS; MisL gene), Streptococcus pneumoniae (lytA), and Plasmodium falciparum (18S rRNA). Of 16 cases of NTS and S. pneumoniae confirmed on blood culture, none were positive by PCR using DNA extracts from blood films or dried blood spots. In contrast, four of 36 dried blood spots and two of 178 plasma samples were PCR positive for S. pneumoniae, despite negative bacterial blood cultures, suggesting false positives. Quantitative assessment revealed that the effective concentration of P. falciparum DNA in blood films was three log orders of magnitude lower than for dried blood spots. The P. falciparum kelch13 gene could not be amplified from blood films. These findings question the value of blood PCR-based approaches for detection of NTS and S. pneumoniae, and show that stored blood films are an inefficient method of studying P. falciparum.

  13. Simultaneous measurement of 25 inflammatory markers and neurotrophins in neonatal dried blood spots by immunoassay with xMAP technology

    DEFF Research Database (Denmark)

    Skogstrand, Kristin; Thorsen, Poul; Nørgaard-Pedersen, Bent;

    2005-01-01

    BACKGROUND: Inflammatory reactions and other events in early life may be part of the etiology of late-onset diseases, including cerebral palsy, autism, and type 1 diabetes. Most neonatal screening programs for congenital disorders are based on analysis of dried blood spot samples (DBSS), and stored...

  14. Design of an online spot air sampling system

    International Nuclear Information System (INIS)

    In all the alpha handling facilities spot air sampling is essential while clearing a lab consequent to week end ventilation shut down or subsequent to completion of work/operations goes inside the facility. The sampling head is designed for 5 micron particles. Following pints are noticed over a period of such operational activities: Health Physicist goes inside a lab with a half face mask whose protection factor is 10, thereby useful for lab air activity up to ten DACs. Many times HP goes inside the lab when activity is more which is highly unsafe. On many occasions simultaneously a number of samples have to be taken by HP from a large number of laboratories inside the facility or for any special job where continuous radiological protection is required. Based on this it is proposed to design a sampling system which will overcome the above limitations. The design of the sampling head has been carried out which is for 5 micron particle size. The pump which is available in the existing facilities can be utilized. An innovative way of counting for large number of samples is fabricated in the RHC wing, RMD which can count ten samples at a time. Removal and counting of the sample may be carried out in a similar sampling carousel which is being used in RHC Unit, Radiometallurgy wing successfully with a little modification. In the proposed system, three samples can be operated in-line such that health physics intervention during the active operation would be minimum and during alarm situations (i.e. on the DAC level) proper protective equipment shall be advised by health physicist or he may suggest any other protective action. This type of online monitors will help in establishing the airborne activity inside the lab where special jobs are being carried out which will provide maximum protection to the lab personnel as well as to the health physicist who supervises the entire operation

  15. Evaluation of Amount of Blood in Dry Blood Spots: Ring-Disk Electrode Conductometry.

    Science.gov (United States)

    Kadjo, Akinde F; Stamos, Brian N; Shelor, C Phillip; Berg, Jordan M; Blount, Benjamin C; Dasgupta, Purnendu K

    2016-06-21

    A fixed area punch in dried blood spot (DBS) analysis is assumed to contain a fixed amount of blood, but the amount actually depends on a number of factors. The presently preferred approach is to normalize the measurement with respect to the sodium level, measured by atomic spectrometry. Instead of sodium levels, we propose electrical conductivity of the extract as an equivalent nondestructive measure. A dip-type small diameter ring-disk electrode (RDE) is ideal for very small volumes. However, the conductance (G) measured by an RDE depends on the depth (D) of the liquid below the probe. There is no established way of computing the specific conductance (σ) of the solution from G. Using a COMSOL Multiphysics model, we were able to obtain excellent agreement between the measured and the model predicted conductance as a function of D. Using simulations over a large range of dimensions, we provide a spreadsheet-based calculator where the RDE dimensions are the input parameters and the procedure determines the 99% of the infinite depth conductance (G99) and the depth D99 at which this is reached. For typical small diameter probes (outer electrode diameter ∼ <2 mm), D99 is small enough for dip-type measurements in extract volumes of ∼100 μL. We demonstrate the use of such probes with DBS extracts. In a small group of 12 volunteers (age 20-66), the specific conductance of 100 μL aqueous extracts of 2 μL of spotted blood showed a variance of 17.9%. For a given subject, methanol extracts of DBS spots nominally containing 8 and 4 μL of blood differed by a factor of 1.8-1.9 in the chromatographically determined values of sulfate and chloride (a minor and major constituent, respectively). The values normalized with respect to the conductance of the extracts differed by ∼1%. For serum associated analytes, normalization of the analyte value by the extract conductance can thus greatly reduce errors from variations in the spotted blood volume/unit area. PMID:27226021

  16. Pilot study for utilization of dried blood spots for screening of lead, mercury and cadmium in newborns.

    Science.gov (United States)

    Chaudhuri, Sanwat N; Butala, Steven J M; Ball, R Wayne; Braniff, Christopher T

    2009-03-01

    The exposure of pregnant women and young children to environmental pollutants is an ongoing concern of state and local public health departments. Of primary concern is the exposure to lead in lead-based paints, methyl mercury in contaminated fish and cadmium present at mining sites. The feasibility, utility and methodology of using blood spot cards collected for new born health screening purposes was studied for use in conducting routine state-wide surveillance of blood lead, mercury and cadmium levels in infants. Homogeneity of different lots of blank filter paper was examined. Mass measurements (weights) of filter paper punches were taken across three different lots of filter paper. Statistical analysis of the data was performed using one-way ANOVA, which indicated no significant difference in the means of all three lots, but high variances were noted. The three metals were examined in three different lots of filter papers purchased from the manufacturer. The lots had measurable amounts of cadmium and lead, but not mercury. Lead spike values were observed for roughly about 7% of the blank samples, indicating heterogeneous distribution of this metal. Statistical analysis of the data was also performed using a two-way ANOVA calculation with Tukey's pairwise comparisons. The results found that total mean metal loadings across the three lots were different. The concentration of the metals can be different from each other and the concentration of any one metal can differ across lots. Stability at different concentrations of the heavy metals in blood spotted onto filter paper with time and storage conditions was examined. Results indicate acceptable performance for at least 8.5 months for lead (near CDC's concern level) and for mercury (near NRC's concern level). The filter paper and blood spots were analyzed for metals using an acid extraction, followed by analysis using an inductively coupled plasma mass spectrometer (ICP-MS). Blood spot cards were studied from four

  17. HbA1c measurements from dried blood spots : validation and patient satisfaction

    NARCIS (Netherlands)

    Fokkema, Margaretha; Bakker, Andries J; de Boer, Fokje; Kooistra, Jeltsje; de Vries, Sifra; Wolthuis, Albert

    2009-01-01

    Background: This study evaluates HbA1c measurements from dried blood spots collected on filter paper and compares HbA1c from filter paper (capillary blood) with HbA1c measured in venous blood. Methods: Patient satisfaction was evaluated using a questionnaire. The performance with the filter paper me

  18. BloodSpot: a database of gene expression profiles and transcriptional programs for healthy and malignant haematopoiesis

    DEFF Research Database (Denmark)

    Bagger, Frederik Otzen; Sasivarevic, Damir; Hadi Sohi, Sina;

    2016-01-01

    largely inaccessible. Current databases provide information about gene-expression but fail to answer key questions regarding co-regulation, genetic programs or effect on patient survival. To address these shortcomings, we present BloodSpot (www.bloodspot.eu), which includes and greatly extends our...... the relationship between different cell types in the database. The database now includes 23 high-quality curated data sets relevant to normal and malignant blood formation and, in addition, we have assembled and built a unique integrated data set, BloodPool. Bloodpool contains more than 2000 samples assembled from...

  19. Common criteria among States for storage and use of dried blood spot specimens after newborn screening

    Directory of Open Access Journals (Sweden)

    Carlo Petrini

    2012-06-01

    Full Text Available Biological samples collected in biobanks are a resource with significant research potential. The Italian Joint Group cNB - cNBBSV (National committee of Bioethics - National committee for Biosecurity, Biotechnologies and Life Sciences published a document reporting recommendations on storage and use of dried blood spot (DBS and on the development of a National Network of Regional Newborn Screening Repositories for collection of residual DBS. Several ethical questions (about consent, possible use of genetic information, unanticipated possible usages for research purposes rise from residual newborn screening specimens collections. Moreover, legal and ethical controversies are accentuated by the conflicts between the interests of sample donors, biobank holders, researchers and the public. To overcome these difficulties the identification of a few criteria for storage and research usage of DBS is crucial.

  20. Descriptive epidemiology of spot urine sodium-to-potassium ratio clarified close relationship with blood pressure level: the Nagahama study.

    OpenAIRE

    Tabara, Yasuharu; Takahashi, Yoshimitsu; Kumagai, Kyoko; Setoh, Kazuya; Kawaguchi, Takahisa; Takahashi, Meiko; Muraoka, Yuki; Tsujikawa, Akitaka; Gotoh, Norimoto; Terao, Chikashi; Yamada, Ryo; Kosugi, Shinji; Sekine, Akihiro; Yoshimura, Nagahisa; Nakayama, Takeo

    2015-01-01

    [Objectives]: We undertook descriptive epidemiology of spot urine sodium-to-potassium ratio (Na/K) in a population sample to clarify the close relationship between Na/K and blood pressure level independently of potential confounding factors. [Methods]: Study participants consisted of 9144 apparently healthy citizens (aged 54 ± 13 years). All clinical parameters were obtained at baseline. [Results]: Na/K was significantly higher in hypertensive individuals irrespective of antihypertensive medi...

  1. Blood parasites in Owls with conservation implications for the Spotted Owl (Strix occidentalis)

    Science.gov (United States)

    Ishak, H.D.; Dumbacher, J.P.; Anderson, N.L.; Keane, J.J.; Valkiunas, G.; Haig, S.M.; Tell, L.A.; Sehgal, R.N.M.

    2008-01-01

    The three subspecies of Spotted Owl (Northern, Strix occidentalis courina; California, S. o. occidentalis; and Mexican, S. o. lucida) are all threatened by habitat loss and range expansion of the Barred Owl (S. varia). An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp.) or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not been documented in the Spotted Owl. We screened 111 Spotted Owls, 44 Barred Owls, and 387 owls of nine other species for haemosporidian parasites (Leucocytozoon, Plasmodium, and Haemoproteus spp.). California Spotted Owls had the greatest number of simultaneous multi-species infections (44%). Additionally, sequencing results revealed that the Northern and California Spotted Owl subspecies together had the highest number of Leucocytozoon parasite lineages (n=17) and unique lineages (n=12). This high level of sequence diversity is significant because only one leucocytozoon species (L. danilewskyi) has been accepted as valid among all owls, suggesting that L. danilewskyi is a cryptic species. Furthermore, a Plasmodium parasite was documented in a Northern Spotted Owl for the first time. West Coast Barred Owls had a lower prevalence of infection (15%) when compared to sympatric Spotted Owls (S. o. caurina 52%, S. o. occidentalis 79%) and Barred Owls from the historic range (61%). Consequently, Barred Owls on the West Coast may have a competitive advantage over the potentially immune compromised Spotted Owls. ?? 2008 Ishak et al.

  2. Blood parasites in owls with conservation implications for the Spotted Owl (Strix occidentalis.

    Directory of Open Access Journals (Sweden)

    Heather D Ishak

    Full Text Available The three subspecies of Spotted Owl (Northern, Strix occidentalis caurina; California, S. o. occidentalis; and Mexican, S. o. lucida are all threatened by habitat loss and range expansion of the Barred Owl (S. varia. An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp. or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not been documented in the Spotted Owl. We screened 111 Spotted Owls, 44 Barred Owls, and 387 owls of nine other species for haemosporidian parasites (Leucocytozoon, Plasmodium, and Haemoproteus spp.. California Spotted Owls had the greatest number of simultaneous multi-species infections (44%. Additionally, sequencing results revealed that the Northern and California Spotted Owl subspecies together had the highest number of Leucocytozoon parasite lineages (n = 17 and unique lineages (n = 12. This high level of sequence diversity is significant because only one Leucocytozoon species (L. danilewskyi has been accepted as valid among all owls, suggesting that L. danilewskyi is a cryptic species. Furthermore, a Plasmodium parasite was documented in a Northern Spotted Owl for the first time. West Coast Barred Owls had a lower prevalence of infection (15% when compared to sympatric Spotted Owls (S. o. caurina 52%, S. o. occidentalis 79% and Barred Owls from the historic range (61%. Consequently, Barred Owls on the West Coast may have a competitive advantage over the potentially immune compromised Spotted Owls.

  3. Microwave-assisted on-spot derivatization for gas chromatography-mass spectrometry based determination of polar low molecular weight compounds in dried blood spots.

    Science.gov (United States)

    Sadones, Nele; Van Bever, Elien; Archer, John R H; Wood, David M; Dargan, Paul I; Van Bortel, Luc; Lambert, Willy E; Stove, Christophe P

    2016-09-23

    Dried blood spot (DBS) sampling and analysis is increasingly being applied in bioanalysis. Although the use of DBS has many advantages, it is also associated with some challenges. E.g. given the limited amount of available material, highly sensitive detection techniques are often required to attain sufficient sensitivity. In gas chromatography coupled to mass spectrometry (GC-MS), derivatization can be helpful to achieve adequate sensitivity. Because this additional sample preparation step is considered as time-consuming, we introduce a new derivatization procedure, i.e. "microwave-assisted on-spot derivatization", to minimize sample preparation of DBS. In this approach the derivatization reagents are directly applied onto the DBS and derivatization takes place in a microwave instead of via conventional heating. In this manuscript we evaluated the applicability of this new concept of derivatization for the determination of two polar low molecular weight molecules, gamma-hydroxybutyric acid (GHB) and gabapentin, in DBS using a standard GC-MS configuration. The method was successfully validated for both compounds, with imprecision and bias values within acceptance criteria (<20% at LLOQ, <15% at 3 other QC levels). Calibration lines were linear over the 10-100μg/mL and 1-30μg/mL range for GHB and gabapentin, respectively. Stability studies revealed no significant decrease of gabapentin and GHB in DBS upon storage at room temperature for at least 84 days. Furthermore, DBS-specific parameters, including hematocrit and volume spotted, were evaluated. As demonstrated by the analysis of GHB and gabapentin positive samples, "microwave-assisted on-spot derivatization" proved to be reliable, fast and applicable in routine toxicology. Moreover, other polar low molecular weight compounds of interest in clinical and/or forensic toxicology, including vigabatrin, beta-hydroxybutyric acid, propylene glycol, diethylene glycol, 1,4-butanediol and 1,2-butanediol, can also be

  4. Use of dried blood spots in doping control analysis of anabolic steroid esters.

    Science.gov (United States)

    Tretzel, Laura; Thomas, Andreas; Geyer, Hans; Gmeiner, Günter; Forsdahl, Guro; Pop, Valentin; Schänzer, Wilhelm; Thevis, Mario

    2014-08-01

    Dried blood spot (DBS) sampling, a technique for whole blood sampling on a piece of filter paper, has more than 50-years tradition, particularly in the diagnostic analysis of metabolic disorders in neonatal screening. Due to the minimal invasiveness, straightforwardness, robustness against manipulation and fastness DBS sampling recommends itself as an advantageous technique in doping control analysis. The present approach highlights the development of a screening assay for the analysis of eight anabolic steroid esters (nandrolone phenylpropionate, trenbolone enanthate, testosterone acetate, testosterone cypionate, testosterone isocaproate, testosterone phenylpropionate, testosterone decanoate and testosterone undecanoate) and nandrolone in DBS. The detection of the intact esters allows an unequivocal proof of the administration of conjugates of exogenous testosterone and its derivatives. Precise, specific and linear conditions were obtained by means of liquid chromatography high resolution/high accuracy mass spectrometry. Sensitivity in the low ppb range was accomplished by the preparation of the methyloxime derivatives of the target compounds. Labeled internal standards (d3-nandrolone, d3-nandrolone caproate and d3-nandrolone undecanoate) were applied to compensate for the broad range in chain length of the esters. The assay presented here outlines the application of DBS for the analysis of anabolic steroid esters in doping controls for the first time providing great potential to simplify the proof of exogenous administration of testosterone. PMID:24713476

  5. Blood Parasites in Owls with Conservation Implications for the Spotted Owl (Strix occidentalis)

    OpenAIRE

    Ishak, Heather D.; Dumbacher, John P.; Nancy L Anderson; Keane, John J.; Gediminas Valkiūnas; Haig, Susan M.; Tell, Lisa A.; Ravinder N M Sehgal

    2008-01-01

    The three subspecies of Spotted Owl (Northern, Strix occidentalis caurina; California, S. o. occidentalis; and Mexican, S. o. lucida) are all threatened by habitat loss and range expansion of the Barred Owl (S. varia). An unaddressed threat is whether Barred Owls could be a source of novel strains of disease such as avian malaria (Plasmodium spp.) or other blood parasites potentially harmful for Spotted Owls. Although Barred Owls commonly harbor Plasmodium infections, these parasites have not...

  6. A rapid liquid chromatography tandem mass spectrometry-based method for measuring propranolol on dried blood spots.

    Science.gov (United States)

    Della Bona, Maria Luisa; Malvagia, Sabrina; Villanelli, Fabio; Giocaliere, Elisa; Ombrone, Daniela; Funghini, Silvia; Filippi, Luca; Cavallaro, Giacomo; Bagnoli, Paola; Guerrini, Renzo; la Marca, Giancarlo

    2013-05-01

    Propranolol, a non-selective beta blocker drug, is used in young infants and newborns for treating several heart diseases; its pharmacokinetics has been extensively evaluated in adult patients using extrapolation to treat pediatric population. The purpose of the present study was to develop and validate a method to measure propranolol levels in dried blood spots. The analysis was performed by using liquid chromatography/tandem mass spectrometry operating in multiple reaction monitoring mode. The calibration curve in matrix was linear in the concentration range of 2.5-200 μg/L with correlation coefficient r=0.9996. Intra-day and inter-day precisions and biases were less than 8.0% (n=10) and 11.5% (n=10) respectively. The recoveries ranged from 94 to 100% and the matrix effect did not result in a severe signal suppression. Propranolol on dried blood spot showed a good stability at three different temperatures for one month. This paper describes a micromethod for measuring propranolol levels on dried blood spot, which determines a great advantage in neonates or young infants during pharmacokinetic studies because of less invasive sampling and small blood volume required.

  7. [Diagnosis of congenital cytomegalovirus infection in newborn dried blood spots on Guthrie cards. A promissory technique].

    Science.gov (United States)

    Distéfano, Angélica L; González, Cecilia A; Pardón, Fabián; Sarubi, María A; Canero Velazco, Cristina

    2008-04-01

    Laboratories play a crucial role in the diagnosis of congenital and perinatal cytomegalovirus infection, considering that other viral infections in newborn infants have similar clinical characteristics. The objectives of this work are to compare the results of the polymerase reaction in blood spots and urine as well as point out the relevance of the result in the Guthrie cards to differentiate congenital from perinatal infection. A total of 148 patients suspicious of CMVH infections were studied in the Congenital Perinatal Infections and Sexual Transmission Laboratory, at the National Institute "Carlos G. Malbrán". The dry blood samples (Guthrie cards) and urine of all patients were studied through the polymerase chain reaction. From the 148 patients, 3 presented other infections, 95 tested negative and 50 positive for cytomegalovirus: 35 had congenital infection and 15 perinatal. In the congenital cases, the polymerase reaction in dry blood was positive (sensitivity 100%, specificity 98.9%, VPP 98% and VPN 100%). Four of them with tardive symptoms were studied retrospectively. The urine specimens from the remaining 15 patients that were taken 15 days after birth were analyzed through the same methods, showing a sensitivity of 100%, the retrospective analysis of this dry blood group yielded negative results, so the infection was considered perinatal. Thus, the dry blood polymerase reaction of the newborn infants makes it a reliable assay for diagnosing congenital cytomegalovirus infection and could be used as an alternative method to urine polymerase reaction. In addition, this test is able to reveal whether the infection is congenital or perinatal in those cases of late symptom or other cases of controversial origin.

  8. Quantitation of brinzolamide in dried blood spots by a novel LC-QTOF-MS/MS method.

    Science.gov (United States)

    Foivas, Anargyros; Malenović, Anđelija; Kostić, Nađa; Božić, Marija; Knežević, Miroslav; Loukas, Yannis L; Dotsikas, Yannis

    2016-02-01

    In the current study, a rapid and sensitive LC-QTOF-MS/MS method for the determination of brinzolamide in dried blood spots (DBS) was developed and validated. This novel sample collection, storage and transfer technique was suitable for analyzing a drug with high distribution into red blood cells and negligible plasma levels. The method included an isocratic mobile phase consisting of methanol and 10mM ammonium formate (90:10, v/v) and detection in positive electrospray mode (ESI+). The flow rate was adjusted to 0.350mL/min yielding retention times of 1.7min for both brinzolamide and internal standard (IS) rabeprazole on a Cyano analytical column, respectively. The validation of the proposed method over the concentration range 0.500-20.0μg/mL was performed in compliance with EMEA and FDA guidelines, assessing all major performance characteristics. Inter- and intra- assay precisions were less than 14%, while inter- and intra- assay accuracies varied from 92.2 to 111%. No matrix effect was observed and the mean brinzolamide extraction recovery was 93.5%. The method was successfully applied to real DBS samples from patients in steady state condition, receiving brinzolamide ophthalmic suspension 1% (w/v) for several months. Initial concentrations were corrected due to hematocrit effect, using image processing algorithm written in Matlab. PMID:26669612

  9. A dried blood spot mass spectrometry metabolomic approach for rapid breast cancer detection

    Directory of Open Access Journals (Sweden)

    Wang Q

    2016-03-01

    Full Text Available Qingjun Wang,1,2,* Tao Sun,3,* Yunfeng Cao,1,2,4,5 Peng Gao,2,4,6 Jun Dong,2,4 Yanhua Fang,2 Zhongze Fang,2 Xiaoyu Sun,2 Zhitu Zhu1,2 1Oncology Department 2, The First Affiliated Hospital of Liaoning Medical University, 2Personalized Treatment and Diagnosis Research Center, The First Affiliated Hospital of Liaoning Medical University and Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Jinzhou, 3Department of Internal Medicine 1, Cancer Hospital of China Medical University, Liaoning Cancer Hospital & Insititute, Shenyang, 4CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 5Key Laboratory of Contraceptives and Devices Research (NPFPC, Shanghai Engineer and Technology Research Center of Reproductive Health Drug and Devices, Shanghai Institute of Planned Parenthood Research, Shanghai, 6Clinical Laboratory, Dalian Sixth People’s Hospital, Dalian, People’s Republic of China *These authors contributed equally to this work Objective: Breast cancer (BC is still a lethal threat to women worldwide. An accurate screening and diagnosis strategy performed in an easy-to-operate manner is highly warranted in clinical perspective. Besides the routinely focused protein markers, blood is full of small molecular metabolites with diverse structures and properties. This study aimed to screen metabolite markers with BC diagnosis potentials.Methods: A dried blood spot-based direct infusion mass spectrometry (MS metabolomic analysis was conducted for BC and non-BC differentiation. The targeted analytes included 23 amino acids and 26 acylcarnitines.Results: Multivariate analysis screened out 21 BC-related metabolites in the blood. Regression analysis generated a diagnosis model consisting of parameters Pip, Asn, Pro, C14:1/C16, Phe/Tyr, and Gly/Ala. Tested with another set of BC and non-BC samples, this model showed a sensitivity of 92.2% and a specificity

  10. Evaluation of dried blood spots with a multiplex assay for measuring recent HIV-1 infection.

    Directory of Open Access Journals (Sweden)

    Kelly A Curtis

    Full Text Available Laboratory-based HIV tests for recent infection (TRIs, which primarily measure a specific serological biomarker(s that distinguishes recent from long-term HIV infection, have facilitated the estimation of population-based incidence. Dried blood spots (DBS on filter paper are an attractive sample source for HIV surveillance, given the simplified and cost-effective methods of specimen collection, storage, and shipment. Here, we evaluated the use of DBS in conjunction with an in-house multiplex TRI, the HIV-1-specific Bio-Plex assay, which measures direct antibody binding and avidity to multiple HIV-1 analytes. The assay performance was comparable between matched plasma and DBS samples from HIV-1 infected individuals obtained from diverse sources. The coefficients of variation, comparing the median antibody reactivity for each analyte between plasma and DBS, ranged from 2.78% to 9.40% and the correlation coefficients between the two sample types ranged from 0.89 to 0.97, depending on the analyte. The correlation in antibody reactivity between laboratory and site-prepared DBS for each analyte ranged from 0.87 to 0.98 and from 0.90 to 0.97 between site-prepared DBS and plasma. The correlation in assay measures between plasma and DBS indicate that the sample types can be used interchangeably with the Bio-Plex format, without negatively impacting the misclassification rate of the assay.

  11. Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture

    Science.gov (United States)

    Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.

    2016-01-01

    Background Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4–90.1%) and 62.5% (95% CI 24.5–91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as

  12. Fetal scalp blood sampling during labor

    DEFF Research Database (Denmark)

    Chandraharan, Edwin; Wiberg, Nana

    2014-01-01

    Fetal cardiotocography is characterized by low specificity; therefore, in an attempt to ensure fetal well-being, fetal scalp blood sampling has been recommended by most obstetric societies in the case of a non-reassuring cardiotocography. The scientific agreement on the evidence for using fetal...... and Clinical Excellence guideline considers that fetal scalp blood sampling decreases instrumental delivery without differences in other outcome variables. The fetal scalp is supplied by vessels outside the skull below the level of the cranial vault, which is likely to be compressed during contractions...

  13. Microwave-assisted on-spot derivatization for gas chromatography-mass spectrometry based determination of polar low molecular weight compounds in dried blood spots.

    Science.gov (United States)

    Sadones, Nele; Van Bever, Elien; Archer, John R H; Wood, David M; Dargan, Paul I; Van Bortel, Luc; Lambert, Willy E; Stove, Christophe P

    2016-09-23

    Dried blood spot (DBS) sampling and analysis is increasingly being applied in bioanalysis. Although the use of DBS has many advantages, it is also associated with some challenges. E.g. given the limited amount of available material, highly sensitive detection techniques are often required to attain sufficient sensitivity. In gas chromatography coupled to mass spectrometry (GC-MS), derivatization can be helpful to achieve adequate sensitivity. Because this additional sample preparation step is considered as time-consuming, we introduce a new derivatization procedure, i.e. "microwave-assisted on-spot derivatization", to minimize sample preparation of DBS. In this approach the derivatization reagents are directly applied onto the DBS and derivatization takes place in a microwave instead of via conventional heating. In this manuscript we evaluated the applicability of this new concept of derivatization for the determination of two polar low molecular weight molecules, gamma-hydroxybutyric acid (GHB) and gabapentin, in DBS using a standard GC-MS configuration. The method was successfully validated for both compounds, with imprecision and bias values within acceptance criteria (weight compounds of interest in clinical and/or forensic toxicology, including vigabatrin, beta-hydroxybutyric acid, propylene glycol, diethylene glycol, 1,4-butanediol and 1,2-butanediol, can also be detected using this method. PMID:27578413

  14. Detection of IL28B SNP DNA from buccal epithelial cells, small amounts of serum, and dried blood spots.

    Directory of Open Access Journals (Sweden)

    Philippe Halfon

    Full Text Available BACKGROUND & AIMS: Point mutations in the coding region of the interleukin 28 gene (rs12979860 have recently been identified for predicting the outcome of treatment of hepatitis C virus infection. This polymorphism detection was based on whole blood DNA extraction. Alternatively, DNA for genetic diagnosis has been derived from buccal epithelial cells (BEC, dried blood spots (DBS, and genomic DNA from serum. The aim of the study was to investigate the reliability and accuracy of alternative routes of testing for single nucleotide polymorphism allele rs12979860CC. METHODS: Blood, plasma, and sera samples from 200 patients were extracted (400 µL. Buccal smears were tested using an FTA card. To simulate postal delay, we tested the influence of storage at ambient temperature on the different sources of DNA at five time points (baseline, 48 h, 6 days, 9 days, and 12 days. RESULTS: There was 100% concordance between blood, plasma, sera, and BEC, validating the use of DNA extracted from BEC collected on cytology brushes for genetic testing. Genetic variations in HPTR1 gene were detected using smear technique in blood smear (3620 copies as well as in buccal smears (5870 copies. These results are similar to those for whole blood diluted at 1/10. A minimum of 0.04 µL, 4 µL, and 40 µL was necessary to obtain exploitable results respectively for whole blood, sera, and plasma. No significant variation between each time point was observed for the different sources of DNA. IL28B SNPs analysis at these different time points showed the same results using the four sources of DNA. CONCLUSION: We demonstrated that genomic DNA extraction from buccal cells, small amounts of serum, and dried blood spots is an alternative to DNA extracted from peripheral blood cells and is helpful in retrospective and prospective studies for multiple genetic markers, specifically in hard-to-reach individuals.

  15. Laser cutting eliminates nucleic acid cross-contamination in dried-blood-spot processing.

    Science.gov (United States)

    Murphy, Sean C; Daza, Glenda; Chang, Ming; Coombs, Robert

    2012-12-01

    Dried blood spots (DBS) are useful for molecular assays but are prone to false positives from cross-contamination. In our malaria DBS assay, cross-contamination was encountered despite cleaning techniques suitable for HIV-1. We therefore developed a contact-free laser cutting system that effectively eliminated cross-contamination during DBS processing.

  16. Improving blood sample logistics using simulation

    DEFF Research Database (Denmark)

    Jørgensen, Pelle Morten Thomas; Jacobsen, Peter

    2012-01-01

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

  17. Superparamagnetic-bead Based Method: An Effective DNA Extraction from Dried Blood Spots (DBS) for Diagnostic PCR

    OpenAIRE

    Sirdah, Mahmoud Mohammed

    2014-01-01

    Introduction: Storing blood as dried spots on filter paper is a trustworthy approach used in genetic screening issues which justifies the necessity for a reliable DNA extraction method. The present work aims to investigate the effectiveness of superparamagnetic-bead based method in extracting DNA from dried blood spots (DBS).

  18. Blood and Dried Blood Spot Telomere Length Measurement by qPCR: Assay Considerations

    OpenAIRE

    Zanet, DeAnna L.; Sara Saberi; Laura Oliveira; Beheroze Sattha; Izabella Gadawski; Côté, Hélène C. F.

    2013-01-01

    Measurement of telomere length is crucial for the study of telomere maintenance and its role in molecular pathophysiology of diseases and in aging. Several methods are used to measure telomere length, the choice of which usually depends on the type and size of sample to be assayed, as well as cost and throughput considerations. The goal of this study was to investigate the factors that may influence the reliability of qPCR-based relative telomere length measurements in whole blood. Day to day...

  19. Fast LC-MS/MS analysis of tacrolimus, sirolimus, everolimus and cyclosporin A in dried blood spots and the influence of the hematocrit and immunosuppressant concentration on recovery

    NARCIS (Netherlands)

    Koster, Remco A.; Alffenaar, Jan-Willem C.; Greijdanus, Ben; Uges, Donald R. A.

    2013-01-01

    We developed a method for the analysis of four immunosuppressants in dried blood spot (DBS) samples to facilitate therapeutic drug monitoring for transplant patients outside the hospital. An 8 mm disc from the central part of the DBS was punched, extracted and followed by LC-MS/MS analysis. The meth

  20. The development and validation of dried blood spots for external quality assurance of syphilis serology

    Directory of Open Access Journals (Sweden)

    Smit Pieter W

    2013-02-01

    Full Text Available Abstract Background Syphilis causes up to 1,500,000 congenital syphilis cases annually. These could be prevented if all pregnant women were screened, and those with syphilis treated with a single dose of penicillin before 28 weeks gestation. In recent years, rapid point-of-care tests have allowed greater access to syphilis screening, especially in rural or remote areas, but the lack of quality assurance of rapid testing has been a concern. We determined the feasibility of using dried blood spots (DBS as specimens for quality assurance of syphilis serological assays. Methods We developed DBS extraction protocols for use with Treponema pallidum particle agglutination assay (TPPA, Treponema pallidum haemagglutination assay (TPHA and an enzyme immunoassay (EIA and compared the results with those using matching plasma samples from the same patient. Results Since DBS samples showed poor performance with TPHA and EIA (TPHA sensitivity was 50.5% (95% confidence interval: 39.9–61.2% and EIA specificity was 50.4% (95% CI: 43.7–57.1%, only the DBS TPPA was used in the final evaluation. DBS TPPA showed an sensitivity of 95.5% (95% CI: 91.3–98.0% and a specificity of 99.0% (95% CI: 98.1–99.5% compared to TPPA using plasma samples as a reference. Conclusion DBS samples can be recommended for use with TPPA, and may be of value for external quality assurance of point-of-care syphilis testing.

  1. Percutaneous ultrasound guided umbilical cord blood sampling

    International Nuclear Information System (INIS)

    This report describes a technique and the result of percutaneous ultrasound-guided umbilical cord blood sampling and its potential use in the management of diagnostic problems in the second and third trimester of pregnancy. This method has been employed in the prenatal assessment of 19 fetuses at risk for chromosomal disorders, fetal hypoxia and hematologic disorders. This simple and rapid procedure offers a safe access to the fetal circulation

  2. Free thyroxin measured in dried blood spots from normal, low-birth-weight, and hypothyroid neonates.

    Science.gov (United States)

    Lemonnier, F; Masson, J; Laroche, D; Travert, J; Travert, G

    1991-12-01

    We have adapted a new radioimmunoassay for free thyroxin (FT4) measurement in dried blood spots for use in neonatal screening for hypothyroidism. The method is easy, fast, and cheap. Within-assay and between-assay CVs are respectively 9.6% and 13.2%. In 997 neonates three days postpartum with normal thyrotropin concentrations, the mean FT4 concentration was 27.2 pmol/L (SD 7.3 pmol/L). There was no significant difference in mean FT4 concentration between boys and girls. FT4 concentrations increased linearly with birth weight or with gestational age, as expressed by multiple linear regression: FT4 (pmol/L) = 0.0016 birth weight (g) + 0.6931 gestational age (weeks) - 4.8772. Only gestational age significantly affected the FT4 value. For five hypothyroid infants tested on day three postpartum, FT4 values were all below the 1st percentile of values from healthy neonates. Thus, when the neonatal concentration of thyrotropin is above normal, FT4 measured in the same sample can provide a reliable earlier diagnosis of hypothyroidism. PMID:1764786

  3. Filter paper blood spot enzyme linked immunoassay for insulin and application in the evaluation of determinants of child insulin resistance.

    Directory of Open Access Journals (Sweden)

    Richard M Martin

    Full Text Available BACKGROUND: In large-scale epidemiology, bloodspot sampling by fingerstick onto filter paper has many advantages, including ease and low costs of collection, processing and transport. We describe the development of an enzyme-linked immunoassay (ELISA for quantifying insulin from dried blood spots and demonstrate its application in a large trial. METHODS: We adapted an existing commercial kit (Mercodia Human Insulin ELISA, 10-1113-01 to quantify insulin from two 3-mm diameter discs (≈6 µL of blood punched from whole blood standards and from trial samples. Paediatricians collected dried blood spots in a follow-up of 13,879 fasted children aged 11.5 years (interquartile range 11.3-11.8 years from 31 trial sites across Belarus. We quantified bloodspot insulin levels and examined their distribution by demography and anthropometry. RESULTS: Mean intra-assay (n = 157 coefficients of variation were 15% and 6% for 'low' (6.7 mU/L and 'high' (23.1 mU/L values, respectively; the respective inter-assay values (n = 33 were 23% and 11%. The intraclass correlation coefficient between 50 paired whole bloodspot versus serum samples, collected simultaneously, was 0.90 (95% confidence interval 0.85 to 0.95. Bloodspot insulin was stable for at least 31 months at -80°C, for one week at +30°C and following four freeze-thaw cycles. Paediatricians collected a median of 8 blood spots from 13,487 (97% children. The geometric mean insulin (log standard deviation concentrations amongst 12,812 children were 3.0 mU/L (1.1 in boys and 4.0 mU/L (1.0 in girls and were positively associated with pubertal stage, measures of central and peripheral adiposity, height and fasting glucose. CONCLUSIONS: Our simple and convenient bloodspot assay is suitable for the measurement of insulin in very small volumes of blood collected on filter paper cards and can be applied to large-scale epidemiology studies of the early-life determinants of circulating insulin.

  4. Stability of Blood Samples for Hemoglobin Electrophoresis

    Directory of Open Access Journals (Sweden)

    Yadira Valdés Fraser

    2013-07-01

    Full Text Available Background: the National Medical Genetics Center has conducted the prenatal screening for hemoglobinopathies in the province of Artemisa and the quality control of this program nationwide; reliability of the results is determined by the quality of the samples used. Objective: to describe the stability of whole blood samples using EDTAK2 and heparin as anticoagulants. Methods: a descriptive study of 100 samples of whole blood from pregnant women and their husbands was conducted at the National Medical Genetics Center. Hemoglobin electrophoresis with Hydrasis technology was performed using 10 % EDTAK2, 2.2 % and 5 % heparin, temperature at 4-8 0C and shelf-life of 7.15 and 30 days. Results: samples with EDTAK2 showed stability for a month with accuracy and repeatability in the electrophoresis runs. By using 5 % and 2.2 % heparin, problems were found in all periods analyzed. Conclusions: 10 % EDTAK2 anticoagulant is appropriate to ensure the reliability of the results in the screening for hemoglobinopathies. The results obtained in this study can be applied in all clinical, hematological and hemoglobin electrophoresis laboratories.

  5. Screening for late neonatal vitamin K deficiency by acarboxyprothrombin in dried blood spots.

    OpenAIRE

    Motohara, K.; Endo, F; Matsuda, I

    1987-01-01

    Acarboxyprothrombin (protein induced by vitamin K absence or antagonist-II (PIVKA-II] concentrations in dried blood spots were determined in 19,029 infants at about 1 month of age as an indicator of vitamin K deficiency. We observed 51 cases with raised blood concentrations of PIVKA-II (greater than 4 AU/ml), nine of whom showed very high concentrations (greater than 20 AU/ml). For infants who did not receive vitamin K prophylaxis at birth, the incidence of the PIVKA-II test yielding positive...

  6. Gene expression in archived newborn blood spots distinguishes infants who will later develop cerebral palsy from matched controls

    OpenAIRE

    Ho, Nhan Thi; Furge, Kyle; Fu, Wenjiang; Busik, Julia; Khoo, Sok Kean; Lu, Qing; Lenski, Madeleine; Wirth, Julia; Hurvitz, Edward; Dodge, Nancy; Resau, James; PANETH, Nigel

    2012-01-01

    Background Gene expression in archived newborn blood spots remaining from newborn screening may reflect pathophysiological disturbances useful in understanding the etiology of cerebral palsy (CP). Methods We quantified the expression of gene sets representing four physiological pathways hypothesized to contribute to CP in archived unfrozen residual newborn blood spot specimens from 53 children with CP and 53 age, gender, and gestational-age–matched controls. We selected four empirical and thr...

  7. Evaluation of market samples of ′Yashada bhasma′ using ′Namburi Phased Spot Test′

    Directory of Open Access Journals (Sweden)

    Santhosh Bhojashettar

    2011-01-01

    Full Text Available Yashada bhasma (Calx of Yashada i.e. Zinc which has its main indication in Prameha (Diabetes and Netra vikaras (Eye disorders was prepared according to the prescription in the Ayurvedic classics and subjected to various bhasma parikshas, including the Namburi Phased Spot Test (NPST, one of the qualitative tests described for various Ayurvedic preparations. NPST helps differentiate between, and thus identify, various bhasmas. It depends upon the pattern of the spot, which develops after a specific chemical reaction. Three market samples of Yashada bhasma, which were said to be Parada marita (incinerated using Mercury, were also subjected to the above tests and results compared. The various bhasmas exhibited marked differences in colour, and though NPST yielded desired results for all the samples, there were differences in their spot patterns and colour. The bhasma prepared in our department produced the most accurate results.

  8. Prediction of the hematocrit of dried blood spots via potassium measurement on a routine clinical chemistry analyzer.

    Science.gov (United States)

    Capiau, Sara; Stove, Veronique V; Lambert, Willy E; Stove, Christophe P

    2013-01-01

    The potential of dried blood spot (DBS) sampling as an alternative for classical venous sampling is increasingly recognized, with multiple applications in, e.g., therapeutic drug monitoring and toxicology. Although DBS sampling has many advantages, it is associated with several issues, the hematocrit (Hct) issue being the most widely discussed challenge, given its possible strong impact on DBS-based quantitation. Hitherto, no approaches allow Hct prediction from nonvolumetrically applied DBS. Following a simple and rapid extraction protocol, K(+) levels from 3 mm DBS punches were measured via indirect potentiometry, using the Roche Cobas 8000 routine chemistry analyzer. The extracts' K(+) concentrations were used to calculate the approximate Hct of the blood used to generate DBS. A linear calibration line was established, with a Hct range of 0.19 to 0.63 (lower limit of quantification, LLOQ, to upper limit of quantification, ULOQ). The procedure was fully validated; the bias and imprecision of quality controls (QCs) at three Hct levels and at the LLOQ and ULOQ was less than 5 and 12%, respectively. In addition, the influence of storage (pre- and postextraction), volume spotted, and punch homogeneity was evaluated. Application on DBS from patient samples (n = 111), followed by Bland and Altman, Passing and Bablok, and Deming regression analysis, demonstrated a good correlation between the "predicted Hct" and the "actual Hct". After correcting for the observed bias, limits of agreement of ±0.049 were established. Incurred sample reanalysis demonstrated assay reproducibility. In conclusion, potassium levels in extracts from 3 mm DBS punches can be used to get a good prediction of the Hct, one of the most important "unknowns" in DBS analysis.

  9. Rapid and sensitive LC-MS/MS method for the analysis of antibiotic linezolid on dried blood spot.

    Science.gov (United States)

    la Marca, Giancarlo; Villanelli, Fabio; Malvagia, Sabrina; Ombrone, Daniela; Funghini, Silvia; De Gaudio, Marina; Fallani, Stefania; Cassetta, Maria Iris; Novelli, Andrea; Chiappini, Elena; de Martino, Maurizio; Galli, Luisa

    2012-01-01

    Linezolid is a new drug from the oxazolidinone class of antibiotics used against mycobacteria and multi-drug resistant (MDR) Gram-positive bacterial infections, which may are also glycopeptide-resistant. The drug usage in pediatric age needs an accurate drug monitoring for effective patient management. The aim of this study was to evaluate the use of dried blood spot (DBS) specimens to determinate linezolid levels during treatment. Advantages of DBS include short collection time, low invasiveness, ease and low cost of sample collection, transport and storage. The analysis was performed in LC-MS/MS operating in positive ion mode and multiple reaction monitoring (MRM) mode. The calibration curve in matrix was linear in the concentration range of 1-100 mg/L with correlation coefficient value of 0.9987. Intraday and interday coefficients of variation were within 3.6% and 13.0%, respectively. We also tested the thermal and temporal drug stability in dried blood spots at four different temperatures to evaluate the risks of sample delivery in different conditions. The short term stability studies showed that linezolid concentration remained stable for at least one month under all the conditions tested. This new assay has favorable characteristics being highly precise and accurate and allows a fast linezolid analysis with a total run time 22 min long, in gradient analysis. Concentration data for plasma and DBS samples from patients after treatment were compared showing a good correlation. Correlation between DBS data and serum samples measured by HPLC-UV was satisfactory. The benefit for patients is the ability to monitor the treatment with a simple and convenient sample collection at home.

  10. Miniaturizing sample spots for matrix-assisted laser desorption/ionization mass spectrometry

    OpenAIRE

    Tu, Tingting; Gross, Michael L.

    2009-01-01

    The trend of miniaturization in bioanalytical chemistry is shifting from technical development to practical application. In matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), progress in miniaturizing sample spots has been driven by the needs to increase sensitivity and speed, to interface with other analytical microtechnologies, and to develop miniaturized instrumentation.

  11. Congener Production in Blood Samples During Preparation and Storage

    DEFF Research Database (Denmark)

    Felby, Søren; Nielsen, Erik

    1995-01-01

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

  12. DEVELOPMENT OF REAL-TIME MULTIPLEX PCR FOR THE QUANTITATIVE DETERMINATION OF TREC'S AND KREC'S IN WHOLE BLOOD AND IN DRIED BLOOD SPOTS

    Directory of Open Access Journals (Sweden)

    M. A. Gordukova

    2015-01-01

    Full Text Available Primary immunodeficiencies (PID such as severe combined immunodeficiency (SCID and X-linked agammaglobulinemia are characterized by the lack of functional Tand B-cells, respectively. Without early diagnosis and prompt treatment children with PID suffer from severe infectious diseases, leading to their death or disability. Our purpose was developing of simple, inexpensive, high throughput technique based on the quantitative determination of TREC and KREC molecules by real-time PCR, and its validation in a group of children with a verified diagnosis of SCID and X-linked agammaglobulinemia.In this study, we developed and validated multiplex real-time PCR for the TREC’s and KREC’s quantitative analysis. We have shown that linear range of Ct changes depending on the concentrations of targets with a correlation coefficient R2 not worse than 0.98 was observed at concentrations from 109 to 5 × 104 copies per ml. The lowest amount of targets reliably detected in a reaction volume was 10 TREC’s copies, 5 KREC ‘s copies and 5 copies of internal control (IL17RA. We determined the age-depended reference values of TRECs and KRECs in whole blood in 29 boys and 27 girls with normal immunological parameters. The normal cut-offs for TRECs and KRECs were defined in dry blood spots depending on the method of extraction.The proposed method showed 100% diagnostic sensitivity and specificity in the studied group. The method can be proposed as a screening tool for the diagnosis of SCID and X-linked agammaglobulinemia both in whole blood and in the dry blood spots. The further investigation is required with larger number of samples

  13. Rapid and simple LC-MS/MS screening of 64 novel psychoactive substances using dried blood spots.

    Science.gov (United States)

    Ambach, Lars; Hernández Redondo, Ana; König, Stefan; Weinmann, Wolfgang

    2014-04-01

    The range of novel psychoactive substances (NPS) including phenethylamines, cathinones, piperazines, tryptamines, etc. is continuously growing. Therefore, fast and reliable screening methods for these compounds are essential and needed. The use of dried blood spots (DBS) for a fast straightforward approach helps to simplify and shorten sample preparation significantly. DBS were produced from 10 µl of whole blood and extracted offline with 500 µl methanol followed by evaporation and reconstitution in mobile phase. Reversed-phase chromatographic separation and mass spectrometric detection (RP-LC-MS/MS) was achieved within a run time of 10 min. The screening method was validated by evaluating the following parameters: limit of detection (LOD), matrix effect, selectivity and specificity, extraction efficiency, and short-term and long-term stability. Furthermore, the method was applied to authentic samples and results were compared with those obtained with a validated whole blood method used for routine analysis of NPS. LOD was between 1 and 10 ng/ml. No interference from matrix compounds was observed. The method was proven to be specific and selective for the analytes, although with limitations for 3-FMC/flephedrone and MDDMA/MDEA. Mean extraction efficiency was 84.6 %. All substances were stable in DBS for at least a week when cooled. Cooling was essential for the stability of cathinones. Prepared samples were stable for at least 3 days. Comparison to the validated whole blood method yielded similar results. DBS were shown to be useful in developing a rapid screening method for NPS with simplified sample preparation.

  14. The influence of the dried blood spot drying time on the recoveries of six immunosuppressants

    Directory of Open Access Journals (Sweden)

    Remco A. Koster

    2015-10-01

    Full Text Available Investigation of the drying time of dried blood spots (DBS is currently not included in DBS validations. The influence of the DBS drying time on the recovery of tacrolimus, ascomycin, sirolimus, everolimus, cyclosporin A and temsirolimus was evaluated by measuring DBS with a fixed blood volume at a hematocrit range between 0.1 and 0.6 L/L at 3, 24 and 48 hours of drying time. Results showed that the recovery of sirolimus, everolimus, temsirolimus and cyclosporin A was influenced by the DBS drying time, while the recovery of tacrolimus and ascomycin was not. A drying time of at least 24 hours is advised in order to stabilize hematocrit and concentration related recovery effects of sirolimus, everolimus, temsirolimus and cyclosporin A.

  15. A rapid fluorometric enzyme immunoassay for the determination of neonatal TSH from blood spots.

    Science.gov (United States)

    Tuuminen, T; Rakkolainen, A E; Welin, M G; Weber, T H; Nylander, P L; Käpyaho, K I

    1991-10-31

    We describe a novel method for the detection of thyrotropin from dried blood spots using a horseradish peroxidase-labelled sandwich enzyme immunoassay with fluorometric detection. The detection limit of the present assay is 1.25 mIU/l with within-run and between-run imprecision being in the range 5.2 to 11.4%. The results of the assay correlate well with two commercial methods: an enzyme immunoassay (r = 0.93) and a time-resolved fluorescence assay (r = 0.90). The blood spot values also show a good correlation (r = 0.93) with respective values obtained from plasma using a commercial immunoradiometric method. The assay may also be performed colorimetrically with sensitivity similar to the fluorometric assay. However, the latter provides a wider dynamic range with an upper limit of 400 mIU/l while the colorimetric method reaches a plateau at 25 mIU/l. Due to its simplicity and rapid performance (3 h), the fluorometric assay is suitable for the routine screening of congenital hypothyroidism. PMID:1814645

  16. Simultaneous measurement of 25 inflammatory markers and neurotrophins in neonatal dried blood spots by immunoassay with xMAP technology

    DEFF Research Database (Denmark)

    Skogstrand, Kristin; Thorsen, Poul; Nørgaard-Pedersen, Bent;

    2005-01-01

    BACKGROUND: Inflammatory reactions and other events in early life may be part of the etiology of late-onset diseases, including cerebral palsy, autism, and type 1 diabetes. Most neonatal screening programs for congenital disorders are based on analysis of dried blood spot samples (DBSS), and stored...... on flowmetric Luminex xMAP technology to measure inflammatory markers and neutrophins in DBSS. RESULTS: The high-capacity 25-plex multianalyte method measured 23 inflammatory and trophic cytokines, triggering receptor expressed on myeloid cells-1 (TREM-1), and C-reactive protein in two 3.2-mm punches from DBSS...... in DBSS models was 174%, mean intra- and interassay CVs were 6.2% and 16%, respectively, and the mean correlation between added and measured analytes was r2 = 0.91. In DBSS routinely collected on days 5-7 from 8 newborns with documented inflammatory reactions at birth, the method detected significantly...

  17. Development of an UPLC-MS/MS method for the determination of antibiotic ertapenem on dried blood spots.

    Science.gov (United States)

    la Marca, Giancarlo; Giocaliere, Elisa; Villanelli, Fabio; Malvagia, Sabrina; Funghini, Silvia; Ombrone, Daniela; Filippi, Luca; De Gaudio, Marina; De Martino, Maurizio; Galli, Luisa

    2012-03-01

    Ertapenem (Invanz) is a newly developed carbapenem β-lactam antimicrobial agent. The drug usage in pediatric age needs an accurate drug monitoring for effective patient management. The aim of this study was to evaluate the use of dried blood spot (DBS) specimens to measure ertapenem concentration during treatment. The analysis was performed by UPLC-MS/MS operating in multiple reaction monitoring (MRM) mode. The calibration curve in matrix was linear in the concentration range of 0.5-100 mg/L with correlation coefficient value higher than 0.997. Performance parameters of this method like lower limit of detection (LLOD, 0.2 mg/L), lower limit of quantification (LLOQ, 0.5 mg/L), matrix effect (20%), intra- and inter-day imprecision (CV within than 15%) and accuracy (between 94 and 155%) of drug concentrations have been evaluated. The drug stability at different temperatures was tested for one month, to evaluate the risks of sample delivery at different climatic conditions. The reported method allows now ertapenem analysis and offers many advantages for patients including the possibility of collecting samples at home. This new assay is both precise and accurate and is especially suitable for therapeutic drug monitoring and pharmacokinetic studies in neonates in whom obtaining larger blood samples is not convenient or possible.

  18. A Way to Improve Analytic Speed of Emergency Blood Sample

    Institute of Scientific and Technical Information of China (English)

    QI Zihui; LI Jun; LIU Zisheng

    2002-01-01

    The circulatory way of eccentric hatch of seperating emergency blood sample quickly and entirely, that is the blood sample is centrifugatened first; second, hatched at 37 ℃; then adopt the circulatory way of centrifugation which can seperate quickly emergency blood sample, and collect nonfibrin serum. The serum was placed paired, and was tested automatically by Corning 644 Electrolyte Analysor and Shimadzu CL- 7000 Biochemistry Analysor after placed naturely with sample 2 hours. There has not difference between two analytic result after T test of statistics. This way shortens the seperation time of serum of emergency blood sample, improves emergency analytic speed, and has a good value on sample seperation of common automatic analysis.

  19. The use of dried blood spots on filter paper for the diagnosis of HIV-1 in infants born to HIV seropositive women

    Directory of Open Access Journals (Sweden)

    Jacob S

    2008-01-01

    Full Text Available Polymerase chain reaction (PCR is the most sensitive test to diagnose HIV-1 infection among infants born to HIV seropositive mothers. The purpose of this study was to evaluate the use of dried blood spot (DBS specimens for PCR and to compare it with whole-blood stored in tubes for HIV-1 DNA PCR. Five hundred and seventy-seven whole-blood infant samples were tested using HIV-1 qualitative in-house nested DNA PCR. Three hundred and fifty-nine samples were from infants at 48 hours of birth and 218 samples at second month. All positive samples tested from whole-blood and every fifth negative sample were coated onto filter paper. DNA was extracted from the filter paper and was amplified using in-house nested PCR. Among the whole-blood samples tested using HIV-1 DNA PCR, 19 of 359 (5.29% samples were HIV-1 positive and 340 (94.7% were negative at 48 hours of birth. At second month, 19 (8.7% of the 218 samples were positive and 199 (91.2% were negative. Using dried filter paper, 18 samples (95% tested positive from 19 positive samples (using whole-blood and 1 tested negative at 48 hours of birth. The 68 negative samples tested using whole-blood were also negative in the DBS test (sensitivity 95% and specificity 100%. At second month, 19 were positive and 40 samples (every fifth sample of 199 were negative (sensitivity and specificity, 100%. PCR performed using DNA extracted from filter paper permits the diagnosis of HIV-1 infection among infants born to HIV-1 seropositive mothers. This assay is simple, rapid, sensitive and specific and can be used in resource limited settings.

  20. Validation of Cobas AmpliPrep/Cobas TaqMan HIV-1 Test on dried blood spots

    Directory of Open Access Journals (Sweden)

    N Ruiz

    2012-11-01

    Full Text Available The plasma specimen is the gold standard for viral load monitoring, the key method to assess the effect of antiviral chemotherapy and to monitor progression of the disease toward AIDS. Nevertheless, several works endorse the use of dried blood spots (DBS on filter paper for the reliable quantification of the levels needed to take therapeutic decisions, detect of treatment failure and monitor the occurrence of drug resistance. The purpose of this study was to validate the use of Cobas AmpliPrep/Cobas TaqMan HIV-1 test version 2.0, with DBS. To evaluate the performance of the above mentioned kit, three stages were involved: 1- Standardization of DBS working conditions, 2- Stability studies at three temperature conditions and 3- Performance evaluation of the kit using this alternative specimen. Additionally, the viral load was quantified in parallel (plasma and DBS to 43 genetically characterized samples, with different levels of viral load. The Pearson correlation coefficient was calculated and the prediction of the value of RNA in plasma starting from the obtained value in DBS was made. Linear regression analysis was performed and coefficients of variation in precision assays were calculated. The best conditions pickups to the work with DBS were: 100 µL of blood (2 spots/50 µl, dried time between 16 and 18 hours at room temperature and, elution of the blood, 2 hours, between 2 and 8°C; in TRIS-EDTA buffer. The samples on DBS proved to be stable during the study periods. A strong correlation was attained between the measurements of viral load in plasma and DBS samples (r=0.96. The detection rate was 90.7 and the coefficient of variation between the values obtained in plasma-DBS sample pairs averaged 3.42%. The CAP/CTM HIV-1 test provided a linear response in DBS, from 330 copies/mL to 420 000 copies/mL. Overall, coefficients of variation in precision tests were below 10%. Cobas AmpliPrep/Cobas TaqMan HIV-1 test version 2.0 had a good

  1. Identification and quantification of psychoactive drugs in whole blood using dried blood spot (DBS) by ultra-performance liquid chromatography tandem mass spectrometry.

    Science.gov (United States)

    Kyriakou, Chrystalla; Marchei, Emilia; Scaravelli, Giulia; García-Algar, Oscar; Supervía, August; Graziano, Silvia

    2016-09-01

    A procedure based on ultra-high-pressure liquid chromatography tandem mass spectrometry has been developed for the determination of twenty three psychoactive drugs and metabolites in whole blood using dried blood spot (DBS). Chromatographic separation was achieved at ambient temperature using a reverse-phase column and a linear gradient elution with two solvents: 0.1% formic acid in acetonitrile and 5mM ammonium formate at pH 3. The mass spectrometer was operated in positive ion mode, using multiple reaction monitoring via positive electro-spray ionization. The method was linear from the limit of quantification (5ng/ml for all the analytes apart from 15ng/ml for Δ-9-tetrahydrocannabinol and metabolites) to 500ng/ml, and showed good correlation coefficients (r(2)=0.990) for all substances. Analytical recovery of analytes under investigation was always higher than 75% and intra-assay and inter-assay precision and accuracy always better than 15%. Using the validated method, ten DBS samples, collected at the hospital emergency department in cases of acute drug intoxication, were found positive to one or more psychoactive drugs. Our data support the potential of DBS sampling for non invasive monitoring of exposure/intoxication to psychoactive drugs. PMID:27232151

  2. Show us your spots! Researchers need samples of bacterial leaf spots on celery, cilantro, parsley, and other crops.

    Science.gov (United States)

    Since 2002, a severe leaf spot disease on parsley has occurred throughout central coastal California and particularly in Monterey County. Three different bacterial pathogens (Pseudomonas syringae pv. apii, P. syringae pv. coriandricola and an organism very closely related to P. viridiflava) have bee...

  3. Segmentation and Analysis of Cancer Cells in Blood Samples

    Directory of Open Access Journals (Sweden)

    Arjun Nelikanti

    2015-10-01

    Full Text Available Blood cancer is an umbrella term for cancers that affect the blood, bone marrow and lymphatic system. Acute Lymphoblastic Leukemia (ALL is one of the kinds of blood cancer which can be affected at any age in the humans. The analysis of peripheral blood samples is an important test in the procedures for the diagnosis of leukemia. In this paper the blood sample images are used and implementing a clustering algorithm for detection of the cancer cells. This paper also implements morphological operations and feature extraction techniques using MATLAB for the analysis of cancer cells in the images.

  4. Bioanalysis of dried saliva spot (DSS) samples using detergent-assisted sample extraction with UHPLC-MS/MS detection.

    Science.gov (United States)

    Zheng, Naiyu; Zeng, Jianing; Ji, Qin C; Angeles, Aida; Aubry, Anne-Francoise; Basdeo, Shenita; Buzescu, Adela; Landry, Ishani Savant; Jariwala, Navin; Turley, Wesley; Burrell, Richard; Arnold, Mark E

    2016-08-31

    Dried saliva spot (DSS) sampling is a non-invasive sample collection technique for bioanalysis that can be potentially implemented at the patient's home. A UHPLC-MS/MS assay was developed using detergent-assisted sample extraction to quantify BMS-927711, a drug candidate in development for the treatment of migraines, in human DSS. By implementing DSS sampling at the patients' home, the bioanalytical sample collection for pharmacokinetic evaluation can be done at the time of the acute migraine attack without the need for clinical visits. DSS samples were prepared by spotting 15 μL of liquid saliva onto regular Whatman FTA™ DMPK-C cards and verified with a UV lamp (at λ 254 nm or 365 nm) during DSS punching. The 4-mm DSS punches in a 96-well plate were sonicated with 200 μL of [(13)C2, D4]-BMS-927711 internal standard (IS) solution in 20/80 MeOH/water for 10 min, followed by sonication with 50 μL of 100 mM NH4OAc with 1.0% Triton-X-100 (as detergent) prior to liquid-liquid extraction with 600 μL EtOAc/Hexane (90:10). UHPLC-MS/MS was performed with an Aquity(®) UPLC BEH C18 Column (2.1 × 50 mm, 1.7 μm) on a Triple Quad™ 5500 mass spectrometer. The assay was linear with a concentration range from 2.00 to 1000 ng mL(-1) for BMS-927711 in human saliva. The intra- and inter-assay precision was within 8.8% CV, and the accuracy was within ±6.7% Dev of the nominal concentration values. This UHPLC-MS/MS assay has been successfully applied to determine the drug's pharmacokinetics within a clinical study. For the first time, we observed BMS-927711 exposure in human DSS, confirming the suitability of this sampling technique for migraine patients to use at home. Detergent-assisted extraction with Triton-X-100 could be very useful in DSS or other dried matrix spot (DMS) assays to overcome low or inconsistent analyte recovery issues.

  5. Screening for late neonatal vitamin K deficiency by acarboxyprothrombin in dried blood spots.

    Science.gov (United States)

    Motohara, K; Endo, F; Matsuda, I

    1987-04-01

    Acarboxyprothrombin (protein induced by vitamin K absence or antagonist-II (PIVKA-II] concentrations in dried blood spots were determined in 19,029 infants at about 1 month of age as an indicator of vitamin K deficiency. We observed 51 cases with raised blood concentrations of PIVKA-II (greater than 4 AU/ml), nine of whom showed very high concentrations (greater than 20 AU/ml). For infants who did not receive vitamin K prophylaxis at birth, the incidence of the PIVKA-II test yielding positive results was significantly higher in those solely breast fed (0.51%) compared with those fed formula milk (0.18%). Among solely breast fed infants, the incidence of a very high result of the PIVKA-II test was 0.14% in those who had not received vitamin K prophylaxis at birth, 0.04% in those who received 2 mg orally, and 0.03% in those who received 2 mg orally plus a further dose of 2-4 mg orally at 7 days. Thus vitamin K prophylaxis at birth did not completely prevent vitamin K deficiency at 1 month. We administered vitamin K therapeutically to all infants whose PIVKA-II test yielded a positive result at 1 month. Only one infant with a positive result developed late neonatal intracranial haemorrhage.

  6. High-performance liquid chromatography determination of dapsone, monoacetyldapsone, and pyrimethamine in filter paper blood spots

    DEFF Research Database (Denmark)

    Rønn, A M; Lemnge, M M; Angelo, H R;

    1995-01-01

    A high-performance liquid chromatography method for the simultaneous analysis of dapsone (DDS), the major metabolite of DDS, monoacetyldapsone (MADDS), and pyrimethamine (PYR) was modified for capillary blood samples obtained by finger prick and dried on filter paper. Limit of quantitation using...

  7. Non-terminal blood sampling techniques in Guinea pigs

    DEFF Research Database (Denmark)

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

    2014-01-01

    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...

  8. Miniaturized Blood Sampling Techniques to Benefit Reduction in Mice and Refinement in Nonhuman Primates: Applications to Bioanalysis in Toxicity Studies with Antibody–Drug Conjugates

    OpenAIRE

    Caron, Alexis; Lelong, Christine; Pascual, Marie-Hélène; Benning, Véronique

    2015-01-01

    Minimizing the number of animals in regulatory toxicity studies while achieving study objectives to support the development of future medicines contributes to good scientific and ethical practices. Recent advances in technology have enabled the development of miniaturized blood sampling methods (including microsampling and dried blood spots) applicable to toxicokinetic determinations of small-molecule drugs. Implementation of miniaturized blood sampling methods in the context of biotherapeuti...

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

    Science.gov (United States)

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

    2014-01-01

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

  10. A study of correlations between crude oil spot and futures markets: A rolling sample test

    Science.gov (United States)

    Liu, Li; Wan, Jieqiu

    2011-10-01

    In this article, we investigate the asymmetries of exceedance correlations and cross-correlations between West Texas Intermediate (WTI) spot and futures markets. First, employing the test statistic proposed by Hong et al. [Asymmetries in stock returns: statistical tests and economic evaluation, Review of Financial Studies 20 (2007) 1547-1581], we find that the exceedance correlations were overall symmetric. However, the results from rolling windows show that some occasional events could induce the significant asymmetries of the exceedance correlations. Second, employing the test statistic proposed by Podobnik et al. [Quantifying cross-correlations using local and global detrending approaches, European Physics Journal B 71 (2009) 243-250], we find that the cross-correlations were significant even for large lagged orders. Using the detrended cross-correlation analysis proposed by Podobnik and Stanley [Detrended cross-correlation analysis: a new method for analyzing two nonstationary time series, Physics Review Letters 100 (2008) 084102], we find that the cross-correlations were weakly persistent and were stronger between spot and futures contract with larger maturity. Our results from rolling sample test also show the apparent effects of the exogenous events. Additionally, we have some relevant discussions on the obtained evidence.

  11. The pathology of facial vein blood sampling in mice

    DEFF Research Database (Denmark)

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

    2014-01-01

    vein blood sampling. Therefore, we investigated if this technique was associated with pathological changes of the jaw region. Methods: 43 NMRI mice were subjected to facial vein blood sampling by using the lancet method during 12 months, starting at the age of 8 weeks. The mice were restrained manually......, and the tissue of the jaw was evaluated. Results: In the 23 mice, from which blood samples had been taken 2 days previously, 5 mice had no signs of gross pathological changes, whereas 12 mice had signs of minimal local subcutaneous bleeding and 6 mice had moderate local subcutaneous bleeding. No additional gross...... pathological changes were observed. In the 23 mice, from which blood samples had been taken 4 weeks earlier, no hemorrhage or signs of scar tissue formation could be observed. Histological slides are currently being processed (HE staining) and will be evaluated and discussed....

  12. Measurements of purine derivatives and creatinine in spot urine samples of Chinese yellow cattle

    International Nuclear Information System (INIS)

    An experiment was conducted using 18 Chinese Yellow Cattle located in 5 farms to study how supplementation of fermentable energy to low quality straw-based rations would improve rumen microbial protein synthesis. Within each farm, the animals were fed on five diets. Diets 1-2 were typical rice straw + by-products used by farmers and were low in fermentable energy content; Diets 3- 5 were more balanced, containing a higher content of fermentable energy. Purine derivatives (PD) and creatinine in spot urine samples were measured. The results showed that the PD to creatinine ratio was significantly higher with Diets 3-5 than with Diets 1-2. Organic matter digestibility and thus organic matter intake was also higher with Diets 3-5 compared to Diets 1-2. The results indicted that the efficiency of microbial protein synthesis could be improved by balancing the diet. (author)

  13. Blood sampling and hemolysis affect concentration of plasma metabolites

    DEFF Research Database (Denmark)

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

    2012-01-01

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

  14. Temporal variability in urinary phthalate metabolite excretion based on spot, morning, and 24-h urine samples: Considerations for epidemiological studies

    DEFF Research Database (Denmark)

    Frederiksen, Hanne; Kranich, Selma K.; Jørgensen, Niels;

    2013-01-01

    healthy men each collected two spot, three first-morning, and three 24-h urine samples during a 3-month period. Samples were analyzed for the content of 12 urinary metabolites of 7 different phthalates. Variability was assessed as intraclass correlation coefficients (ICC). For the metabolites of diethyl...... of exposures for these two phthalates in population studies and hence an attenuation of the power to detect possible exposure-outcome associations. The only slightly higher ICCs for 24-h pools compared to first-morning and spot urine samples does not seem to justify the extra effort needed to collect 24-h...

  15. Parallel ultra high pressure liquid chromatography-mass spectrometry for the quantification of HIV protease inhibitors using dried spot sample collection format.

    Science.gov (United States)

    Watanabe, Kyoko; Varesio, Emmanuel; Hopfgartner, Gérard

    2014-08-15

    An assay was developed and validated for the quantification of eight protease inhibitors (indinavir (IDV), ritonavir (RTV), lopinavir (LPV), saquinavir (SQV), amprenavir (APV), nelfinavir (NFV), atazanavir (AZV) and darunavir (DRV)) in dried plasma spots using parallel ultra-high performance liquid chromatography and mass spectrometry detection in the multiple reaction monitoring mode. For each analyte an isotopically labeled internal standard was used and the assay based on liquid-solid extraction the area response ratio (analyte/IS) was found to be linear; from 0.025 μg/ml to 20 μg/ml for IDV, SQV, DRV, AZV, LPV, from 0.025 μg/ml to 10 μg/ml for NFV, APV and from 0.025 μg/ml to 5 μg/ml for RTV using 15 μl of plasma spotted on filter paper placed in a sample tube. The total analysis time was of 4 min and inter-assay accuracies and precisions were in the range of 87.7-109% and 2.5-11.8%, respectively. On dried plasma spots all analytes were found to be stable for at least 7 days. Practicability of the assay to blood was also demonstrated. The sample drying process could be reduced to 5 min using a commercial microwave system without any analyte degradation. Together with quantification, confirmatory analysis was performed on representative clinical samples. PMID:25049214

  16. Dried blood spots for viral load monitoring in Malawi: feasible and effective.

    Directory of Open Access Journals (Sweden)

    Sarah E Rutstein

    Full Text Available To evaluate the feasibility and effectiveness of dried blood spots (DBS use for viral load (VL monitoring, describing patient outcomes and programmatic challenges that are relevant for DBS implementation in sub-Saharan Africa.We recruited adult antiretroviral therapy (ART patients from five district hospitals in Malawi. Eligibility reflected anticipated Ministry of Health VL monitoring criteria. Testing was conducted at a central laboratory. Virological failure was defined as >5000 copies/ml. Primary outcomes were program feasibility (timely result availability and patient receipt and effectiveness (second-line therapy initiation.We enrolled 1,498 participants; 5.9% were failing at baseline. Median time from enrollment to receipt of results was 42 days; 79.6% of participants received results within 3 months. Among participants with confirmed elevated VL, 92.6% initiated second-line therapy; 90.7% were switched within 365 days of VL testing. Nearly one-third (30.8% of participants with elevated baseline VL had suppressed (4 years were more likely to be failing than participants on therapy 1-4 years (RR 1.7, 95% CI 1.0-2.8; older participants were less likely to be failing (RR 0.95, 95% CI 0.92-0.98. There was no difference in likelihood of failure based on clinical symptoms (RR 1.17, 95% CI 0.65-2.11.DBS for VL monitoring is feasible and effective in real-world clinical settings. Centralized DBS testing may increase access to VL monitoring in remote settings. Programmatic outcomes are encouraging, especially proportion of eligible participants switched to second-line therapy.

  17. Second-tier test for quantification of underivatized amino acids in dry blood spot for metabolic diseases in newborn screening.

    Science.gov (United States)

    Wang, Chunyan; Zhu, Hongbin; Zhang, Wenyan; Song, Fengrui; Liu, Zhiqiang; Liu, Shuying

    2013-02-01

    The quantitative analysis of amino acids (AAs) in single dry blood spot (DBS) samples is an important issue for metabolic diseases as a second-tier test in newborn screening. An analytical method for quantifying underivatized AAs in DBS was developed by using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The sample preparation in this method is simple and ion-pairing agent is not used in the mobile phase that could avoid ion suppression, which happens in mass spectrometry and avoids damage to the column. Through chromatographic separation, some isomeric compounds could be identified and quantified, which cannot be solved through only appropriate multiple reactions monitoring transitions by MS/MS. The concentrations of the different AAs were determined using non-deuterated internal standard. All calibration curves showed excellent linearity within test ranges. For most of the amino acids the accuracy of extraction recovery was between 85.3 and 115 %, and the precision of relative standard deviation was <7.0 %. The 35 AAs could be identified in DBS specimens by the developed LC-MS/MS method in 17-19 min, and eventually 24 AAs in DBS were quantified. The results of the present study prove that this method as a second-tier test in newborn screening for metabolic diseases could be performed by the quantification of free AAs in DBS using the LC-MS/MS method. The assay has advantages of high sensitive, specific, and inexpensive merits because non-deuterated internal standard and acetic acid instead of ion-pairing agent in mobile phase are used in this protocol.

  18. A novel dried blood spot-LCMS method for the quantification of methotrexate polyglutamates as a potential marker for methotrexate use in children.

    Directory of Open Access Journals (Sweden)

    Ahmed F Hawwa

    Full Text Available OBJECTIVE: Development and validation of a selective and sensitive LCMS method for the determination of methotrexate polyglutamates in dried blood spots (DBS. METHODS: DBS samples [spiked or patient samples] were prepared by applying blood to Guthrie cards which was then dried at room temperature. The method utilised 6-mm disks punched from the DBS samples (equivalent to approximately 12 µl of whole blood. The simple treatment procedure was based on protein precipitation using perchloric acid followed by solid phase extraction using MAX cartridges. The extracted sample was chromatographed using a reversed phase system involving an Atlantis T3-C18 column (3 µm, 2.1 × 150 mm preceded by Atlantis guard column of matching chemistry. Analytes were subjected to LCMS analysis using positive electrospray ionization. KEY RESULTS: The method was linear over the range 5-400 nmol/L. The limits of detection and quantification were 1.6 and 5 nmol/L for individual polyglutamates and 1.5 and 4.5 nmol/L for total polyglutamates, respectively. The method has been applied successfully to the determination of DBS finger-prick samples from 47 paediatric patients and results confirmed with concentrations measured in matched RBC samples using conventional HPLC-UV technique. CONCLUSIONS AND CLINICAL RELEVANCE: The methodology has a potential for application in a range of clinical studies (e.g. pharmacokinetic evaluations or medication adherence assessment since it is minimally invasive and easy to perform, potentially allowing parents to take blood samples at home. The feasibility of using DBS sampling can be of major value for future clinical trials or clinical care in paediatric rheumatology.

  19. Development of immune-affinity 96 spots monolith array for multiple mycotoxins detection in food samples.

    Science.gov (United States)

    Li, Li; Xia, Li-Ru; Zhao, Yong-Fu; Wang, He-Ye

    2016-09-01

    In this paper, a novel highly sensitive chemiluminescence immune-affinity 96 spots monolith array was developed to detect deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2), and fumonisin B1 (FB1) in corn samples. Firstly, the monolith array was prepared through on suit UV-initiated copolymerization using polyethylene glycol diacrylate (PEGDA) as cross-linker, glycidyl methacrylate (GMA) as functional monomer and polyethylene glycol 200 (PEG 200) as the porogen. Subsequently, the four mycotoxins immune-affinity monolith array was prepared by immobilization of DON, ZEN, T-2, and FB1 antibody. The mole ratio of PEGDA/GMA, UV exposure time, and the volume ratio of PEG 200/PEGDA were optimized to improve the performances of the immune-affinity monolith array. For the mycotoxins immune-affinity monolith array based on chemiluminescence detection, the limit of detection was 0.0036ng/mL (DON), 0.0048ng/mL (ZEN), 0.0039ng/mL (T-2), and 0.0017ng/mL (FB1), respectively. The linear response in the range of 0.01-0.1ng/mL (R(2)=0.98). The results showed that the proposed four mycotoxins immune-affinity monolith array was a stable, accurate, and highly sensitive method to determine levels of DON, ZEN, T-2, and FB1 in real samples. PMID:27423670

  20. Radioimmunoassay of ''free thyroxin'' in dried blood spots on filter paper - preliminary observations on the effective differentiation of subjects with congenital hypothyroidism from those with subnormal thyroxin-binding globulin and normal subjects

    International Nuclear Information System (INIS)

    In this sensitive, simple method for measuring ''free thyroxin'' (FT4) in eluates of dried blood spots on filter paper by use of a radioimmunoassay kit (Amerlex Free T4 RIA), the measurable range of FT4 is 1.8 to 57 ng/L (equivalent to the concentration in serum), or 7 to 237 fg/tube. The mean coefficients of variation for within assay-within spots, within assay-between spots, and between assays were 5.3%, 5.0%, and 6.2%, respectively. FT4 in blood spotted on filter paper is stable for at least a month when dried and kept at either -200C, 40C, room temperature (about 250C), or 370C. The results for FT4 in dried blood spots correlated closely with the free-T4 concentration in serum (r = 0.99). The method can be used to differentiate cases of primary and secondary hypothyroidism from normal subjects and those with subnormal thyroxin-binding globulin. This method may be useful in screening for congenital hypothyroidism, because sample-retesting is not necessary

  1. Exposure and risk factors to coxiella burnetii, spotted fever group and typhus group Rickettsiae, and Bartonella henselae among volunteer blood donors in Namibia.

    Directory of Open Access Journals (Sweden)

    Bruce H Noden

    Full Text Available The role of pathogen-mediated febrile illness in sub-Saharan Africa is receiving more attention, especially in Southern Africa where four countries (including Namibia are actively working to eliminate malaria. With a high concentration of livestock and high rates of companion animal ownership, the influence of zoonotic bacterial diseases as causes of febrile illness in Namibia remains unknown.The aim of the study was to evaluate exposure to Coxiella burnetii, spotted fever and typhus group rickettsiae, and Bartonella henselae using IFA and ELISA (IgG in serum collected from 319 volunteer blood donors identified by the Blood Transfusion Service of Namibia (NAMBTS. Serum samples were linked to a basic questionnaire to identify possible risk factors. The majority of the participants (64.8% had extensive exposure to rural areas or farms. Results indicated a C. burnetii prevalence of 26.1% (screening titre 1∶16, and prevalence rates of 11.9% and 14.9% (screening titre 1∶100 for spotted fever group and typhus group rickettsiae, respectively. There was a significant spatial association between C. burnetii exposure and place of residence in southern Namibia (P0.012, especially cattle (P>0.006, were also significantly associated with C. burnetii exposure. Males were significantly more likely than females to have been exposed to spotted fever (P<0.013 and typhus (P<0.011 group rickettsiae. Three (2.9% samples were positive for B. henselae possibly indicating low levels of exposure to a pathogen never reported in Namibia.These results indicate that Namibians are exposed to pathogenic fever-causing bacteria, most of which have flea or tick vectors/reservoirs. The epidemiology of febrile illnesses in Namibia needs further evaluation in order to develop comprehensive local diagnostic and treatment algorithms.

  2. Comparison of Different Blood Collection, Sample Matrix, and Immunoassay Methods in a Prenatal Screening Setting

    Directory of Open Access Journals (Sweden)

    Jeroen L. A. Pennings

    2014-01-01

    Full Text Available We compared how measurements of pregnancy-associated plasma protein A (PAPP-A and the free beta subunit of human chorionic gonadotropin (fβ-hCG in maternal blood are influenced by different methods for blood collection, sample matrix, and immunoassay platform. Serum and dried blood spots (DBS were obtained by venipuncture and by finger prick of 19 pregnant women. PAPP-A and fβ-hCG from serum and from DBS were measured by conventional indirect immunoassay on an AutoDELFIA platform and by antibody microarray. We compared methods based on the recoveries for both markers as well as marker levels correlations across samples. All method comparisons showed high correlations for both marker concentrations. Recovery levels of PAPP-A from DBS were 30% lower, while those of fβ-hCG from DBS were 50% higher compared to conventional venipuncture serum. The recoveries were not affected by blood collection or immunoassay method. The high correlation coefficients for both markers indicate that DBS from finger prick can be used reliably in a prenatal screening setting, as a less costly and minimally invasive alternative for venipuncture serum, with great logistical advantages. Additionally, the use of antibody arrays will allow for extending the number of first trimester screening markers on maternal and fetal health.

  3. Dried Saliva Spot (DSS) as a Convenient and Reliable Sampling for Bioanalysis: An Application for the Diagnosis of Diabetes Mellitus.

    Science.gov (United States)

    Numako, Masahiro; Takayama, Takahiro; Noge, Ichiro; Kitagawa, Yutaka; Todoroki, Kenichiro; Mizuno, Hajime; Min, Jun Zhe; Toyo'oka, Toshimasa

    2016-01-01

    This paper proposes the dried saliva spot (DSS) as a convenient sampling technique for bioanalysis. The analytical method with the DSS was used for the determination of D,L-lactic acid (D,L-LA) and the D/L ratio of diabetic patients and prediabetic persons for the simple screening of the disease. The D,L-LA in the DSS was labeled with a chiral reagent (DMT-3(S)-Apy) for carboxylic acids and determined by UPLC-ESI-MS/MS. The limits of detection (signal-to-noise ratio (S/N) = 3) for the DSS analysis were on the amol level (∼30 amol). Because good stability, recovery, accuracy, and precision of the D,L-LA for the DSS method was also obtained from the proposed procedure, the DSS method was applied to the determination of the D- and L-isomers of LA of diabetic patients, and prediabetic and healthy persons. The D/L-LA ratio by the present DSS method and the HbA1c value in blood were well-correlated to the serious diabetic patients, whereas the relation in the prediabetic persons was not very good. The reason seems to be due to the rough saliva sampling, and not to the DSS method, because strict regulation was not requested for the prediabetic and healthy persons. In order to have a successful DSS analysis, the stability of the target molecule, the detection sensitivity to the target molecule, and the validated determination method are important.

  4. Dried Saliva Spot (DSS) as a Convenient and Reliable Sampling for Bioanalysis: An Application for the Diagnosis of Diabetes Mellitus.

    Science.gov (United States)

    Numako, Masahiro; Takayama, Takahiro; Noge, Ichiro; Kitagawa, Yutaka; Todoroki, Kenichiro; Mizuno, Hajime; Min, Jun Zhe; Toyo'oka, Toshimasa

    2016-01-01

    This paper proposes the dried saliva spot (DSS) as a convenient sampling technique for bioanalysis. The analytical method with the DSS was used for the determination of D,L-lactic acid (D,L-LA) and the D/L ratio of diabetic patients and prediabetic persons for the simple screening of the disease. The D,L-LA in the DSS was labeled with a chiral reagent (DMT-3(S)-Apy) for carboxylic acids and determined by UPLC-ESI-MS/MS. The limits of detection (signal-to-noise ratio (S/N) = 3) for the DSS analysis were on the amol level (∼30 amol). Because good stability, recovery, accuracy, and precision of the D,L-LA for the DSS method was also obtained from the proposed procedure, the DSS method was applied to the determination of the D- and L-isomers of LA of diabetic patients, and prediabetic and healthy persons. The D/L-LA ratio by the present DSS method and the HbA1c value in blood were well-correlated to the serious diabetic patients, whereas the relation in the prediabetic persons was not very good. The reason seems to be due to the rough saliva sampling, and not to the DSS method, because strict regulation was not requested for the prediabetic and healthy persons. In order to have a successful DSS analysis, the stability of the target molecule, the detection sensitivity to the target molecule, and the validated determination method are important. PMID:26629726

  5. The validity of screening based on spot morning urine samples to detect subjects with microalbuminuria in the general population

    NARCIS (Netherlands)

    Gansevoort, RT; Verhave, JC; Hillege, HL; Burgerhof, JGM; Bakker, SJL; de Zeeuw, D; de Jong, PE

    2005-01-01

    Background. No study has yet investigated the validity of prescreening by albumin measurements in a spot morning urine sample to identify in the general population subjects with microalbuminuria. We therefore tested the diagnostic performance of urinary albumin concentration (UAC) and albumin-creati

  6. Temporal variability in urinary excretion of bisphenol A and seven other phenols in spot, morning, and 24-h urine samples

    DEFF Research Database (Denmark)

    Lassen, Tina Harmer; Frederiksen, Hanne; Jensen, Tina Kold;

    2013-01-01

    bisphenol A (BPA) and seven other phenols. All analytes were determined using TurboFlow-LC-MS/MS. Two spot, three first morning and three 24-h urine samples were collected from 33 young Danish men over a three months period. Temporal variability was estimated by means of intraclass correlation coefficients...

  7. From clinical sites to biorepositories: effectiveness in blood sample management.

    Science.gov (United States)

    Lefebvre, Céline; Tremblay, Nancy; Iverson, Bonnie; Wong, David; McWeeny, Kerri; Saghbini, Michael; Martinez, Heather; Hogan, Michael; Gaudet, Daniel; Arsenault, Steve

    2010-12-01

    Today's biobanks must work to take full advantage of collected samples, while maximizing sample quality and minimizing costs to sustain operations for a long period of time. This is a tall order that will require collaboration and compromise for both end-users and collection sites. This article discusses the efforts of the Génome Québec-Centre Hospitalier Affilié Universitaire Régional de Chicoutimi Biobank to fractionate blood samples for the simultaneous preservation of plasma and DNA-containing layers while minimizing resources required for shipping and transport. This article also describes methods for successful reproducible application of the plasma-depleted blood sample to GenPlates (GenVault, Carlsbad, CA).

  8. Single genome amplification of proviral HIV-1 DNA from dried blood spot specimens collected during early infant screening programs in Lusaka, Zambia.

    Science.gov (United States)

    Seu, Lillian; Mwape, Innocent; Guffey, M Bradford

    2014-07-01

    The ability to evaluate individual HIV-1 virions from the quasispecies of vertically infected infants was evaluated in a field setting at the Centre for Infectious Disease Research in Zambia. Infant heel-prick blood specimens were spotted onto dried blood spot (DBS) filter paper cards at government health clinics. Nucleic acid was extracted and used as a template for HIV-1 proviral DNA detection by a commercial Amplicor HIV-1 PCR test (Roche, version 1.5). On samples that tested positive by commercial diagnostic assay, amplification of DNA was performed using an in-house assay of the 5' and 3' region of the HIV-1 genome. Additionally, fragments covering 1200 nucleotides within pol (full length protease and partial reverse transcriptase) and 1400 nucleotides within env (variable 1-variable 5 region) were further analyzed by single genome amplification (SGA). In summary, we have demonstrated an in-house assay for amplifying the 5' and 3' proviral HIV-1 DNA as well as pol and env proviral DNA fragments from DBS cards collected and analyzed entirely in Zambia. In conclusion, this study shows the feasibility of utilizing DBS cards to amplify the whole proviral HIV-1 genome as well as perform SGA on key HIV-1 genes.

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

    Science.gov (United States)

    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%.

  10. The performance of five different dried blood spot cards for the analysis of six immunosuppressants

    NARCIS (Netherlands)

    Koster, Remco A.; Botma, Rixt; Greijdanus, Ben; Uges, Donald R. A.; Kosterink, Jos G. W.; Touw, Daan J.; Alffenaar, Jan-Willem C.

    2015-01-01

    Background: The relation between hematocrit, substance concentration, extraction recovery and spot formation of tacrolimus, sirolimus, everolimus, ascomycin, temsirolimus and cyclosporin A was investigated for Whatman 31 ET CHR, Whatman FTA DMPK-C, Whatman 903, Perkin Elmer 226 and Agilent Bond Elut

  11. The use of dried blood spot specimens for HIV-1 drug resistance genotyping in young children initiating antiretroviral therapy

    Science.gov (United States)

    Salimo, Anna T.; Ledwaba, Johanna; Coovadia, Ashraf; Abrams, Elaine J.; Technau, Karl-Günter; Kuhn, Louise; Morris, Lynn; Hunt, Gillian M.

    2015-01-01

    Paired plasma and dried blood spots (DBS) from 232 South African HIV-infected children initiating antiretroviral therapy (ART) were genotyped for drug resistance mutations, most of who had prior exposure to ART for prevention-of-mother-to-child-transmission. Non-nucleoside reverse transcriptase inhibitor mutations were most commonly detected in both specimen types, particularly Y181C/I and K103N/S. Resistance interpretation concordance was achieved in 97% of pairs with 7 children having mutations detected in DBS only. These results validate the preferential use of DBS specimens for HIVDR genotyping in this patient group. PMID:26192603

  12. Microbiological assessment along the fish production chain of the Norwegian pelagic fisheries sector - Results from a spot sampling programme

    OpenAIRE

    Svanevik, Cecilie Smith; Roiha, Irja Sunde; Levsen, Arne; Lunestad, Bjørn Tore

    2015-01-01

    Microbes play an important role in the degradation of fish products, thus better knowledge of the microbiological conditions throughout the fish production chain may help to optimise product quality and resource utilisation. This paper presents the results of a ten-year spot sampling programme (2005–2014) of the commercially most important pelagic fish species harvested in Norway. Fish-, surface-, and storage water samples were collected from fishing vessels and processing factories. Totally ...

  13. Comparison of Spot and Time Weighted Averaging (TWA Sampling with SPME-GC/MS Methods for Trihalomethane (THM Analysis

    Directory of Open Access Journals (Sweden)

    Don-Roger Parkinson

    2016-02-01

    Full Text Available Water samples were collected and analyzed for conductivity, pH, temperature and trihalomethanes (THMs during the fall of 2014 at two monitored municipal drinking water source ponds. Both spot (or grab and time weighted average (TWA sampling methods were assessed over the same two day sampling time period. For spot sampling, replicate samples were taken at each site and analyzed within 12 h of sampling by both Headspace (HS- and direct (DI- solid phase microextraction (SPME sampling/extraction methods followed by Gas Chromatography/Mass Spectrometry (GC/MS. For TWA, a two day passive on-site TWA sampling was carried out at the same sampling points in the ponds. All SPME sampling methods undertaken used a 65-µm PDMS/DVB SPME fiber, which was found optimal for THM sampling. Sampling conditions were optimized in the laboratory using calibration standards of chloroform, bromoform, bromodichloromethane, dibromochloromethane, 1,2-dibromoethane and 1,2-dichloroethane, prepared in aqueous solutions from analytical grade samples. Calibration curves for all methods with R2 values ranging from 0.985–0.998 (N = 5 over the quantitation linear range of 3–800 ppb were achieved. The different sampling methods were compared for quantification of the water samples, and results showed that DI- and TWA- sampling methods gave better data and analytical metrics. Addition of 10% wt./vol. of (NH42SO4 salt to the sampling vial was found to aid extraction of THMs by increasing GC peaks areas by about 10%, which resulted in lower detection limits for all techniques studied. However, for on-site TWA analysis of THMs in natural waters, the calibration standard(s ionic strength conditions, must be carefully matched to natural water conditions to properly quantitate THM concentrations. The data obtained from the TWA method may better reflect actual natural water conditions.

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

    Science.gov (United States)

    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.

  15. A simple method for the analysis by MS/MS of underivatized amino acids on dry blood spots from newborn screening.

    Science.gov (United States)

    Wang, Chunyan; Zhang, Wenyan; Song, Fengrui; Liu, Zhiqiang; Liu, Shuying

    2012-05-01

    The analysis by electrospray-ionization tandem mass spectrometry of amino acids with butyl esterification and isotopically labeled internal standard is routine in newborn screening laboratories worldwide. In the present study, we established a direct analysis method of higher accuracy that uses a non-deuterated internal standard. The automatic sampler and the pump of an LC apparatus were used to inject sample and mobile phase to MS, but no LC column was needed. The dry blood spot (DBS) material was prepared at levels of low, medium and high concentration; the running time was 1 min. In parallel to the new procedure, we applied the established method to analyze nine amino acids on DBS of healthy newborns and phenylketonuria newborns. The newly proposed method of product ion confirmation scan along with multiple reaction monitoring resulted in a very accurate identification of each amino acid. Our innovative protocol had high sensitivity and specificity in the analysis of cases of suspected metabolic diseases.

  16. Evaluation of Different Cytomegalovirus (CMV) DNA PCR Protocols for Analysis of Dried Blood Spots from Consecutive Cases of Neonates with Congenital CMV Infections▿

    Science.gov (United States)

    Soetens, Oriane; Vauloup-Fellous, Christelle; Foulon, Ina; Dubreuil, Pascal; De Saeger, Ben; Grangeot-Keros, Liliane; Naessens, Anne

    2008-01-01

    Two protocols for the extraction of cytomegalovirus (CMV) DNA and two methods for the amplification of CMV DNA in dried blood spots were evaluated for the retrospective diagnosis of congenital CMV infection. During the period from 1996 to 2006, a urine screening program detected 76 congenitally infected neonates. Stored Guthrie cards with blood from 55 cases and 12 controls were tested. Two spots of dried blood were cut from each card and evaluated in two centers. CMV DNA was extracted from a whole single spot. Center 1 used phenol-chloroform extraction and ethanol precipitation followed by a conventional PCR. Center 2 used the NucliSens easyMAG automated DNA/RNA extraction platform (bioMérieux) followed by a real-time PCR. For evaluation of the extraction method, DNA extracted from each blood spot was evaluated by the amplification method used by the collaborating center. The sensitivities were 66% for center 1 and 73% for center 2. None of the controls were positive. A sensitivity as high as 82% could be obtained by combining the most sensitive extraction method (the phenol-chloroform procedure) with the most sensitive PCR method (real-time PCR). The detection rate was not influenced by the duration of storage of the spots. The sensitivity was higher with blood from congenitally infected cases due to a primary maternal CMV infection, regardless of the protocol used. However, the difference reached significance only for the least-sensitive protocol (P = 0.036). PMID:18199787

  17. LC-MS/MS method for simultaneous determination on a dried blood spot of multiple analytes relevant for treatment monitoring in patients with tyrosinemia type I.

    Science.gov (United States)

    la Marca, Giancarlo; Malvagia, Sabrina; Materazzi, Serena; Della Bona, Maria Luisa; Boenzi, Sara; Martinelli, Diego; Dionisi-Vici, Carlo

    2012-01-17

    Tyrosinemia type 1 is caused by deficiency of fumarylacetoacetate hydrolase. The enzymatic defect impairs the conversion of fumarylacetoacetate to fumarate, causing accumulation of succinylacetone which induces severe liver and kidney dysfunction along with mutagenic changes and hepatocellular carcinoma. Treatment is based on nitisinone (NTBC), an enzymatic inhibitor which suppresses succinylacetone production. NTBC, which has dramatically changed the disease course improving liver and kidney functions and reducing risk of liver cancer, causes a side effect of the increase of tyrosine levels. Treatment is therefore based on the combination of NTBC with a protein-restricted diet to prevent the potential toxicity of excessive tyrosine accumulation. Long-term therapy requires a careful monitoring in blood of NTBC levels along with other disease biomarkers, which include succinylacetone, and a selected panel of circulating aminoacids. We have developed a straightforward and fast MS/MS method for the simultaneous determination of NTBC, succinylacetone, tyrosine, phenylalanine, and methionine on a dried blood spot requiring a 2 min run. A single assay suitable for quantitative evaluation of all biochemical markers is of great advance over conventional methods, especially in pediatric patients, since it reduces laboratory costs and blood sampling, is less invasive and particularly suitable for pediatric patients, and allows easier storage and shipping. PMID:22148291

  18. LC-MS/MS method for simultaneous determination on a dried blood spot of multiple analytes relevant for treatment monitoring in patients with tyrosinemia type I.

    Science.gov (United States)

    la Marca, Giancarlo; Malvagia, Sabrina; Materazzi, Serena; Della Bona, Maria Luisa; Boenzi, Sara; Martinelli, Diego; Dionisi-Vici, Carlo

    2012-01-17

    Tyrosinemia type 1 is caused by deficiency of fumarylacetoacetate hydrolase. The enzymatic defect impairs the conversion of fumarylacetoacetate to fumarate, causing accumulation of succinylacetone which induces severe liver and kidney dysfunction along with mutagenic changes and hepatocellular carcinoma. Treatment is based on nitisinone (NTBC), an enzymatic inhibitor which suppresses succinylacetone production. NTBC, which has dramatically changed the disease course improving liver and kidney functions and reducing risk of liver cancer, causes a side effect of the increase of tyrosine levels. Treatment is therefore based on the combination of NTBC with a protein-restricted diet to prevent the potential toxicity of excessive tyrosine accumulation. Long-term therapy requires a careful monitoring in blood of NTBC levels along with other disease biomarkers, which include succinylacetone, and a selected panel of circulating aminoacids. We have developed a straightforward and fast MS/MS method for the simultaneous determination of NTBC, succinylacetone, tyrosine, phenylalanine, and methionine on a dried blood spot requiring a 2 min run. A single assay suitable for quantitative evaluation of all biochemical markers is of great advance over conventional methods, especially in pediatric patients, since it reduces laboratory costs and blood sampling, is less invasive and particularly suitable for pediatric patients, and allows easier storage and shipping.

  19. Evaluation of Two Techniques for Viral Load Monitoring Using Dried Blood Spot in Routine Practice in Vietnam (French National Agency for AIDS and Hepatitis Research 12338)

    Science.gov (United States)

    Taieb, Fabien; Tram, Tran Hong; Ho, Hien Thi; Pham, Van Anh; Nguyen, Lan; Pham, Ban Hien; Tong, Linh An; Tuaillon, Edouard; Delaporte, Eric; Nguyen, Anh Tuan; Bui, Duc Duong; Do, NhanThi; Madec, Yoann

    2016-01-01

    Background. Although it is the best method to detect early therapeutic failure, viral load (VL) monitoring is still not widely available in many resource-limited settings because of difficulties in specimen transfer, personnel shortage, and insufficient laboratory infrastructures. Dried blood spot (DBS) use, which was introduced in the latest World Health Organization recommendations, can overcome these difficulties. This evaluation aimed at validating VL measurement in DBS, in a laboratory without previous DBS experience and in routine testing conditions. Methods. Human immunodeficiency virus (HIV)-infected adults were observed in a HIV care site in Hanoi, and each patient provided 2 DBS cards with whole blood spots and 2 plasma samples. Viral load was measured in DBS and in plasma using the COBAS Ampliprep/TaqMan and the Abbott RealTime assays. To correctly identify those with VL ≥ 1000 copies/mL, sensitivity and specificity were estimated. Results. A total of 198 patients were enrolled. With the Roche technique, 51 plasma VL were ≥1000 copies/mL; among these, 28 presented a VL in DBS that was also ≥1000 copies/mL (sensitivity, 54.9; 95% confidence interval [CI], 40.3–68.9). On the other hand, all plasma VL CI, 97.5–100). With the Abbott technique, 45 plasma VL were ≥1000 copies/mL; among these, 42 VL in DBS were also ≥1000 copies/mL (sensitivity, 93.3%; 95% CI, 81.7–98.6); specificity was 94.8 (95% CI, 90.0–97.7). Conclusions. The Abbott RealTime polymerase chain reaction assay provided adequate VL results in DBS, thus allowing DBS use for VL monitoring.

  20. Personalized monitoring of therapeutic salicylic acid in dried blood spots using a three-layer setup and desorption electrospray ionization mass spectrometry.

    Science.gov (United States)

    Siebenhaar, Markus; Küllmer, Kai; Fernandes, Nuno Miguel de Barros; Hüllen, Volker; Hopf, Carsten

    2015-09-01

    Desorption electrospray ionization (DESI) mass spectrometry is an emerging technology for direct therapeutic drug monitoring in dried blood spots (DBS). Current DBS methods require manual application of small molecules as internal standards for absolute drug quantification. With industrial standardization in mind, we superseded the manual addition of standard and built a three-layer setup for robust quantification of salicylic acid directly from DBS. We combined a dioctyl sodium sulfosuccinate weave facilitating sample spreading with a cellulose layer for addition of isotope-labeled salicylic acid as internal standard and a filter paper for analysis of the standard-containing sample by DESI-MS. Using this setup, we developed a quantification method for salicylic acid from whole blood with a validated linear curve range from 10 to 2000 mg/L, a relative standard deviation (RSD%) ≤14%, and determination coefficients of 0.997. The limit of detection (LOD) was 8 mg/L and the lower limit of quantification (LLOQ) was 10 mg/L. Recovery rates in method verification by LC-MS/MS were 97 to 101% for blinded samples. Most importantly, a study in healthy volunteers after administration of a single dose of Aspirin provides evidence to suggest that the three-layer setup may enable individual pharmacokinetic and endpoint testing following blood collection by finger pricking by patients at home. Taken together, our data suggests that DBS-based quantification of drugs by DESI-MS on pre-manufactured three-layer cartridges may be a promising approach for future near-patient therapeutic drug monitoring.

  1. Impact of blood sampling in very preterm infants

    DEFF Research Database (Denmark)

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

    2000-01-01

    In a prospective investigation, 99 very preterm infants (gestational age (GA) 24 32 weeks, birthweight 560-2,255 g) were studied during the first 4 weeks of life. The infants were divided into two groups: infants born extremely early (GA <28 weeks, n = 20) and infants of GA 28 - 32 weeks; the gro......In a prospective investigation, 99 very preterm infants (gestational age (GA) 24 32 weeks, birthweight 560-2,255 g) were studied during the first 4 weeks of life. The infants were divided into two groups: infants born extremely early (GA .../kg. For the extremely preterm infants a significant correlation between sampled and transfused blood volume was found (mean 37.1 and 33.3 ml/kg, respectively, r = + 0.71, p = 0.0003). The most frequently requested analyses were glucose, sodium and potassium. Few blood gas analyses were requested (1.9/ infant). No blood...... in extremely preterm, critically ill infants. Udgivelsesdato: 2000-Apr...

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

    OpenAIRE

    del Valle Mendoza, Juana; Silva Caso, Wilmer; 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...

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

    OpenAIRE

    Ana Flávia Belchior Andrade; Maria Emília Cambria Guimaro Siqueira; Luciano Chaves Arantes; Larissa Silva Queiroz; Rayane Luiza Viegas Silva; Eduardo Dias Ramalho

    2014-01-01

    Blood is the most common body fluid found at crime scenes. One-step presumptive tests have been designed as a rapid immunological test for the qualitative detection of human hemoglobin in stool samples (faecal occult blood) their usefulness for forensic purposes has been demonstrated before. In this study we compare Hexagon OBTI kit and FOB One-step Bioeasy kit sensitivity in the analysis of diluted blood samples. With Hexagon OBTI, positive test results are achieved in whole blood dilutions ...

  4. Evaluation of PCR-Based Assay for Diagnosis of Spotted Fever Group Rickettsiosis in Human Serum Samples

    OpenAIRE

    Choi, Yeon-Joo; Lee, Seung-Hyun; Park, Kyung-Hee; Koh, Young-Sang; Lee, Keun-Hwa; Baik, Hyung-Suk; Choi, Myung-Sik; Kim, Ik-Sang; Jang, Won-Jong

    2005-01-01

    A nested PCR assay was developed for the detection of spotted fever group (SFG) rickettsiae in serum samples. The assay was based on specific primers derived from the rickettsial outer membrane protein B gene (rompB) of Rickettsia conorii. An SFG rickettsia-specific signal is obtained from R. akari, R. japonica, R. sibirica, and R. conorii. Other bacterial species tested did not generate any signal, attesting to the specificity of the assay. As few as seven copies of the rompB gene of R. cono...

  5. Ag2S/CdS/TiO2 Nanotube Array Films with High Photocurrent Density by Spotting Sample Method

    OpenAIRE

    Sun, Hong; Zhao, Peini; Zhang, Fanjun; Liu, Yuliang; Hao, Jingcheng

    2015-01-01

    Ag2S/CdS/TiO2 hybrid nanotube array films (Ag2S/CdS/TNTs) were prepared by selectively depositing a narrow-gap semiconductor—Ag2S (0.9 eV) quantum dots (QDs)—in the local domain of the CdS/TiO2 nanotube array films by spotting sample method (SSM). The improvement of sunlight absorption ability and photocurrent density of titanium dioxide (TiO2) nanotube array films (TNTs) which were obtained by anodic oxidation method was realized because of modifying semiconductor QDs. The CdS/TNTs, Ag2S/TNT...

  6. Natural Antioxidants Improve Red Blood Cell “Survival” in Non-Leukoreduced Blood Samples

    Directory of Open Access Journals (Sweden)

    Yuliya V Kucherenko

    2015-03-01

    Full Text Available Background: Blood collected in an anticoagulant can be kept refrigerated in an unmodified state within 5 - 6 weeks. Oxidative damage is considered to be a one of the major factors contributing to the development of storage lesions. Lipid and membrane proteins oxidation results in changes in cation gradients that affect the cell survival. Aim: In the present study we used the natural antioxidants and ion channels blockers (L-carnosine, spermine, phloretin and their mixtures to prolong “survival” of red blood cells (RBCs, measured as the lack of PS exposure and cell hemolysis, in the Alsever's preservative solution upon hypothermic storage. Results: We show that the mixture of carnosine (20 mM, spermine (20 µM and phloretin (100 µM effectively blunted phosphatidylserine (PS exposure, Ca2+ accumulation and RBCs hemolysis in non-leukoreduced low (∼2% hematocrit samples after 36 days of storage as well as after 1 day of post-storage incubation of the stored cells in physiological saline solution. In addition, a slight but significant decrease in PS exposure was observed in non-leukoreduced high (∼20% hematocrit samples after 36 days of storage with the mixture of substances. Conclusion: We conclude that the use of the mixture of natural antioxidants (carnosine, spermine, and phloretin as an additive to blood preservative solution provides better RBCs storage and “survival”.

  7. Analysis of 145 cases of T SPOT.TB in peripheral blood of detecting tuberculosis infection%T-SPOT.TB外周全血检测结核感染145例分析

    Institute of Scientific and Technical Information of China (English)

    邱素娟; 叶峰山

    2013-01-01

    目的分析T—SPOT.TB检测结核感染的敏感度和特异性,其作为快速、准确的检测手段,给临床提供诊断依据,控制结核感染漫延新法。方法选择我院呼吸内科和消化内科,不能排除结核病289例,外周血检测T—SPOT.TB 145例,清晨痰培养加涂片144例,对照阳性率。T—SPOT.TB检测阳性率94.4%(137/145),痰培养加涂片41.6%(60/144),经统计学处理P<0.01,T—SPOT.TB检测优于痰培养加涂片。结果 T—SPOT.TB优于痰中找结核杆菌,T—SPOT.TB检测诊断,可在保持高灵性,高特异性的同时,进一步提高结核分枝检出率。结论对早期诊断可获得较好治疗效果的肺结核和肺外结核患者,该选择灵敏度高,特异性强,T—SPOT.TB快速检测有一定优势。%Objective analysis of T - SPOT.TB detection of tuberculosis infection of the sensitivity and specificity of purpose, as the fast, accurate detection means, to support clinical diagnosis, control of tuberculosis infection spread law.Methods in our hospital respiratory department of internal medicine and Department of internal medicine, 289 cases of tuberculosis could not be excluded, peripheral blood T and SPOT.TB in 145 cases, morning sputum culture and smear in 144 cases, the positive rate of control.T - SPOT.TB positive rate was 94.4% (137/145), sputum culture and smear of 41.6% (60/144), were treated by P<0.01, T - SPOT.TB test is better than that of sputum culture and smear.Result The results of Mycobacterium tuberculosis T - SPOT.TB is better than that of sputum, T - SPOT.TB detection and diagnosis, can be maintained in the high intelligence, high specificity at the same time, further improve the detection rate of Mycobacterium tuberculosis.Conclusion early diagnosis can obtain good therapeutic effect of pulmonary tuberculosis and pulmonary tuberculosis patients, the selection of high sensitivity, strong specificity, T - SPOT.TB rapid detection has

  8. Field evaluation of dried blood spots for routine HIV-1 viral load and drug resistance monitoring in patients receiving antiretroviral therapy in Africa and Asia.

    Science.gov (United States)

    Monleau, Marjorie; Aghokeng, Avelin F; Eymard-Duvernay, Sabrina; Dagnra, Anoumou; Kania, Dramane; Ngo-Giang-Huong, Nicole; Touré-Kane, Coumba; Truong, Lien X T; Chaix, Marie-Laure; Delaporte, Eric; Ayouba, Ahidjo; Peeters, Martine

    2014-02-01

    Dried blood spots (DBS) can be used in developing countries to alleviate the logistic constraints of using blood plasma specimens for viral load (VL) and HIV drug resistance (HIVDR) testing, but they should be assessed under field conditions. Between 2009 and 2011, we collected paired plasma-DBS samples from treatment-experienced HIV-1-infected adults in Burkina Faso, Cameroon, Senegal, Togo, Thailand, and Vietnam. The DBS were stored at an ambient temperature for 2 to 4 weeks and subsequently at -20°C before testing. VL testing was performed on the plasma samples and DBS using locally available methods: the Abbott m2000rt HIV-1 test, generic G2 real-time PCR, or the NucliSENS EasyQ version 1.2 test. In the case of virological failure (VF), i.e., a plasma VL of ≥1,000 copies/ml, HIVDR genotyping was performed on paired plasma-DBS samples. Overall, we compared 382 plasma-DBS sample pairs for DBS VL testing accuracy. The sensitivities of the different assays in different laboratories for detecting VF using DBS varied from 75% to 100% for the m2000rt test in labs B, C, and D, 91% to 93% for generic G2 real-time PCR in labs A and F, and 85% for the NucliSENS test in lab E. The specificities varied from 82% to 97% for the m2000rt and NucliSENS tests and reached only 60% for the generic G2 test. The NucliSENS test showed good agreement between plasma and DBS VL but underestimated the DBS VL. The lowest agreement was observed for the generic G2 test. Genotyping was successful for 96/124 (77%) DBS tested, and 75/96 (78%) plasma-DBS pairs had identical HIVDR mutations. Significant discrepancies in resistance interpretations were observed in 9 cases, 6 of which were from the same laboratory. DBS can be successfully used as an alternative to blood plasma samples for routine VL and HIVDR monitoring in African and Asian settings. However, the selection of an adequate VL measurement method and the definition of the VF threshold should be considered, and laboratory performance

  9. Does Vitamin D Supplementation Enhance Musculoskeletal Performance in Individuals Identified as Vitamin D Deficient through Blood Spot Testing?

    Science.gov (United States)

    Murphy, Kellie A.

    This thesis investigated possible changes in performance after one month of vitamin D supplementation in individuals found to be vitamin D deficient or insufficient through blood spot testing. Thirty-two males, ages 18-32, participated. Each subject visited the lab three times in one-month, completing four performance tests each session, including an isometric mid-thigh pull and a vertical jump on a force plate, a isometric 90-degree elbow flexion test using a load cell, and a psychomotor vigilance test on a palm pilot. The initial lab included blood spot tests to find vitamin D levels. In a single blind manner, 16 subjects were assigned vitamin D and 16 the placebo. Repeated measures ANOVA analysis did not reveal any main effects for time (F=2.626, p=0.364), treatment (vitamin D3 vs placebo; F=1.282, p=0.999), or interaction effects for treatment by time (F=0.304, p=0.999) for maximum force production during an isometric mid-thigh pull. Repeated measures ANOVA analysis did not reveal any main effects for time (F=1.323, p=0.999), treatment (vitamin D3 vs placebo; F=0.510, p=0.999), or interaction effects for treatment by time (F= 1.625, p=0.860) for rate of force production during a vertical jump. Repeated measures ANOVA analysis did not reveal any main effects for time (F=0.194, p=0.999), treatment (vitamin D3 vs placebo; F=2.452, p=0.513), or interaction effects for treatment by time (F= 1.179, p=0.999) for maximal force production during a 90-degree isometric elbow flexion. Repeated measures ANOVA analysis did not reveal any main effects for time (F=1.710, p=0.804), treatment (vitamin D3 vs placebo; F=1.471, p=0.94), or interaction effects for treatment by time (F= 0.293, p=0.999) for mean reaction time to random stimuli during the psychomotor vigilance test. Repeated measures ANOVA analysis did not reveal any main effects for time (F=0.530, p=0.999), treatment (vitamin D3 vs placebo; F=0.141, p=0.999), or interaction effects for treatment by time (F=0.784 p=0

  10. Butylated hydroxytoluene can protect polyunsaturated fatty acids in dried blood spots from degradation for up to 8 weeks at room temperature

    OpenAIRE

    Metherel, Adam H; Hogg, Ryan C; Buzikievich, Lindy M; Stark, Ken D.

    2013-01-01

    Background Dried blood spots (DBS) from fingertip prick blood can enable high throughput fatty acid profiling but may be prone to lipid peroxidation during storage. The use of butylated hydroxytoluene (BHT) on chromatography paper can prevent polyunsaturated fatty acid (PUFA) loss but examinations on the length of storage times possible are not comprehensive. Method In the first study, venous whole blood was saturated on paper strips pre-soaked with 0, 2.5 or 5.0 mg/mL BHT and exposed to air ...

  11. Microbiological assessment along the fish production chain of the Norwegian pelagic fisheries sector--Results from a spot sampling programme.

    Science.gov (United States)

    Svanevik, Cecilie Smith; Roiha, Irja Sunde; Levsen, Arne; Lunestad, Bjørn Tore

    2015-10-01

    Microbes play an important role in the degradation of fish products, thus better knowledge of the microbiological conditions throughout the fish production chain may help to optimise product quality and resource utilisation. This paper presents the results of a ten-year spot sampling programme (2005-2014) of the commercially most important pelagic fish species harvested in Norway. Fish-, surface-, and storage water samples were collected from fishing vessels and processing factories. Totally 1,181 samples were assessed with respect to microbiological quality, hygiene and food safety. We introduce a quality and safety assessment scheme for fresh pelagic fish recommending limits for heterotrophic plate counts (HPC), thermos tolerant coliforms, enterococci and Listeria monocytogenes. According to the scheme, in 25 of 41 samplings, sub-optimal conditions were found with respect to quality, whereas in 21 and 9 samplings, samples were not in compliance concerning hygiene and food safety, respectively. The present study has revealed that the quality of pelagic fish can be optimised by improving the hygiene conditions at some critical points at an early phase of the production chain. Thus, the proposed assessment scheme may provide a useful tool for the industry to optimise quality and maintain consumer safety of pelagic fishery products. PMID:26187839

  12. Multiplex Assay of Second-Line Anti-Tuberculosis Drugs in Dried Blood Spots Using Ultra-Performance Liquid Chromatography-Tandem Mass Spectrometry.

    Science.gov (United States)

    Lee, Kyunghoon; Jun, Sun Hee; Han, Minje; Song, Sang Hoon; Park, Jong Sun; Lee, Jae Ho; Park, Kyoung Un; Song, Junghan

    2016-09-01

    As dried blood spots (DBSs) have various advantages over conventional venous blood sampling, some assays for detection of one or two anti-tuberculosis (TB) drugs in DBSs have been developed. However, there are no assays currently available for the simultaneous measurement of three or more anti-TB drugs in DBSs. In this study, we developed and evaluated a multiplex method for detecting nine anti-TB drugs including streptomycin, kanamycin, clarithromycin, cycloserine, moxifloxacin, levofloxacin, para-aminosalicylic acid, prothionamide, and linezolid in DBSs by using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Seventy-nine patient samples of DBS were analyzed on the UPLC-MS/MS system. All drug concentrations were determined within 4 min, and assay performance was evaluated. All drugs were clearly separated without ion suppression. Within-run and between-run precisions were 1.7-13.0% and 5.7-17.0%, respectively, at concentrations representing low and high levels for the nine drugs. Lower limits of detection and quantification were 0.06-0.6 and 0.5-5.0 μg/mL, respectively. Linearity was acceptable at five level concentrations for each drug. Correlations between drug concentrations in plasma and DBSs by using Passing-Bablock regression and Pearson's rho (ρ 0.798-0.989) were acceptable. In conclusion, we developed a multiplex assay to measure nine second-line anti-TB drugs in DBSs successfully. This assay provided convenient and rapid drug quantification and could have applications in drug monitoring during treatment. PMID:27374716

  13. Improvement in CT image resolution due to the use of focal spot deflection and increased sampling.

    Science.gov (United States)

    Rubert, Nicholas; Szczykutowicz, Timothy; Ranallo, Frank

    2016-01-01

    When patient anatomy is positioned away from a CT scanner's isocenter, scans of limited diagnostic value may result. Yet in some cases, positioning of patient anatomy far from isocenter is unavoidable. This study examines the effect of posi-tion and reconstruction algorithm on image resolution achieved by a CT scanner operating in a high resolution (HR) scan mode which incorporates focal spot deflection and acquires an increased number of projections per rotation. Images of a metal bead contained in a phantom were acquired on a GE CT750 HD scanner with multiple reconstruction algorithms, in the normal and HR scan mode, and at two positions, scanner isocenter and 15 cm directly above isocenter. The images of the metal bead yielded two-dimensional point spread functions which were averaged along two perpendicular directions to yield line spread functions. Fourier transforms of the line spread functions yielded radial and azimuthal modulation transfer functions (MTFs). At isocenter, the radial and azimuthal MTFs were aver-aged. MTF improvement depended on image position and modulation direction. The results from a single algorithm, Edge, can be generalized to other algorithms. At isocenter, the 10% MTF cutoff was 14.4 cycles/cm in normal and HR mode. At 15 cm above isocenter, the 10% cutoff was 6.0 and 8.5 cycles/cm for the azimuthal and radial MTFs in normal mode. In HR mode, the azimuthal and radial MTF 10% cutoff was 8.3 and 10.3 cycles/cm. Our results indicate that the best image resolu-tion is achieved at scanner isocenter and that the azimuthal resolution degrades more significantly than the radial resolution. For the GE CT750 HD CT scanner, the resolution is significantly enhanced by the HR scan mode away from scanner isocenter, and the use of the HR scan mode has much more of an impact on image resolution away from isocenter than the choice of algorithm. PMID:27167276

  14. Development of a one-step probe based molecular assay for rapid immunodiagnosis of infection with M. tuberculosis using dried blood spots.

    Directory of Open Access Journals (Sweden)

    Thomas Blauenfeldt

    Full Text Available BACKGROUND: Antigen specific release of IP-10 is the most promising alternative marker to IFN-γ for infection with M. tuberculosis. Compared to Interferon-γ release assays (IGRA, IP-10 is released in high levels enabling novel approaches such as field friendly dried blood spots (DBS and molecular detection. AIM: To develop a robust IP-10 based molecular assay for the diagnosis of infection with M. tubercuolsis from whole blood and DBS. METHOD: We developed a one-step probe based multiplex RT-qPCR assay for detecting IP-10 and IFN-γ mRNA expression from whole blood and DBS samples. The assay was validated and applied for the diagnosis of M. tuberculosis infection in DBS samples from 43 patients with confirmed TB, 13 patients with latent TB and 96 presumed uninfected controls. In parallel, IP-10 and INF-γ levels were measured in Quantiferon (QFT-TB plasma supernatants. RESULTS: IP-10 mRNA upregulation was detectable at 4 hours after stimulation (6 fold upregulation peaking at 8 hours (108 fold upregulation. IFN-γ expression occurred in concert but levels were lower (peak 6.7 fold upregulation. IP-10 gene expression level was significantly higher in patients with tuberculosis (median 31.2, IQR 10.7-67.0 and persons with latent tuberculosis infection (LTBI (41.2, IQR 9.8-64.9 compared to healthy controls (1.6, IQR 1.1-2.4; p<0.0001. The IP-10 mRNA and protein based tests had comparable diagnostic accuracy to QFT-TB, sensitivity (85% and 88% vs 85% and specificity (96% and 96% vs 97%, p = ns.. CONCLUSION: We developed a rapid, robust and accurate molecular immunodiagnostic test for M. tuberculosis infection. By combining DBS based sample acquisition, mail or currier based sample transport with centralized molecular detection, this immunodiagnostic test concept can reduce the local technological requirements everywhere and make it possible to offer highly accurate immunodiagnostic tests in low resource settings.

  15. Development of a One-Step Probe Based Molecular Assay for Rapid Immunodiagnosis of Infection with M. tuberculosis Using Dried Blood Spots

    Science.gov (United States)

    Blauenfeldt, Thomas; Heyckendorf, Jan; Graff Jensen, Sidse; Lange, Christoph; Drabe, Camilla; Hermansen, Thomas S.; de Thurah, Lena; Lillebaek, Troels; Eugen-Olsen, Jesper; Seersholm, Niels; Hoff, Søren; Bonde, Jesper; Ruhwald, Morten

    2014-01-01

    Background Antigen specific release of IP-10 is the most promising alternative marker to IFN-γ for infection with M. tuberculosis. Compared to Interferon-γ release assays (IGRA), IP-10 is released in high levels enabling novel approaches such as field friendly dried blood spots (DBS) and molecular detection. Aim To develop a robust IP-10 based molecular assay for the diagnosis of infection with M. tubercuolsis from whole blood and DBS. Method We developed a one-step probe based multiplex RT-qPCR assay for detecting IP-10 and IFN-γ mRNA expression from whole blood and DBS samples. The assay was validated and applied for the diagnosis of M. tuberculosis infection in DBS samples from 43 patients with confirmed TB, 13 patients with latent TB and 96 presumed uninfected controls. In parallel, IP-10 and INF-γ levels were measured in Quantiferon (QFT-TB) plasma supernatants. Results IP-10 mRNA upregulation was detectable at 4 hours after stimulation (6 fold upregulation) peaking at 8 hours (108 fold upregulation). IFN-γ expression occurred in concert but levels were lower (peak 6.7 fold upregulation). IP-10 gene expression level was significantly higher in patients with tuberculosis (median 31.2, IQR 10.7–67.0) and persons with latent tuberculosis infection (LTBI) (41.2, IQR 9.8–64.9) compared to healthy controls (1.6, IQR 1.1–2.4; p<0.0001). The IP-10 mRNA and protein based tests had comparable diagnostic accuracy to QFT-TB, sensitivity (85% and 88% vs 85%) and specificity (96% and 96% vs 97%, p = ns.). Conclusion We developed a rapid, robust and accurate molecular immunodiagnostic test for M. tuberculosis infection. By combining DBS based sample acquisition, mail or currier based sample transport with centralized molecular detection, this immunodiagnostic test concept can reduce the local technological requirements everywhere and make it possible to offer highly accurate immunodiagnostic tests in low resource settings. PMID:25184553

  16. Development and Application of Zirconia Coated Paper Substrate for High Sensitivity Analysis of Therapeutic Drugs in Dried Blood Spots.

    Science.gov (United States)

    Zheng, Yajun; Wang, Qian; Wang, Xiaoting; Chen, Ying; Wang, Xuan; Zhang, Xiaoling; Bai, Zongquan; Han, Xiaoxiao; Zhang, Zhiping

    2016-07-19

    Paper spray mass spectrometry has been demonstrated to be promising for direct analysis of therapeutic drugs in dried blood spots (DBS); however, the strong hydrogen bond and van de Waals interactions between paper substrate and analytes containing polar functional groups (e.g., therapeutic drugs) affect greatly the elution behavior and analysis sensitivity of compounds of interest during paper spray. Herein, we developed a one-sided ZrO2 coated paper substrate through a facile vacuum filtration approach using commercial ZrO2 particles as coating material and soluble starch as adhesive agent. Owing to the unique surface properties, as-prepared ZrO2 paper substrate has been shown to have excellent performance for analysis of therapeutic drugs in DBS during paper spray mass spectrometry. In contrast to original cellulose paper substrates, improvements of 43-189-fold in lower limit of quantitation (LLOQ) were obtained for the tested drugs using ZrO2 coated paper for paper spray. In comparing with the previously reported grade SG81 paper and one-sided silica coated paper, the LLOQs of the tested drugs with as-prepared ZrO2 paper decreased 1.5-16.5-fold relative to those from the above two, revealing that ZrO2 coated paper is a good candidate for paper spray in high sensitivity analysis of therapeutic drugs in DBS. PMID:27314839

  17. Short communication: Tenofovir diphosphate in dried blood spots as an objective measure of adherence in HIV-infected women.

    Science.gov (United States)

    Castillo-Mancilla, Jose R; Searls, Kristina; Caraway, Patricia; Zheng, Jia-Hua; Gardner, Edward M; Predhomme, Julie; Bushman, Lane R; Anderson, Peter L; Meditz, Amie L

    2015-04-01

    Simple and reproducible tools to assess antiretroviral adherence are needed. A level of tenofovir diphosphate (TFV-DP) in dried blood spots (DBS) Pearson correlation coefficient. The average TFV-DP between the two visits (aTFV-DP) in DBS and PBMCs was 1,874 (706-3,776) fmol/punch and 125 (1-278) fmol/10(6) cells, respectively. AA women had lower levels of aTFV-DP in DBS compared to whites (1,660 vs. 1,970 fmol/punch; p=0.04), with a viremic patient having the lowest drug levels (706 fmol/punch). Days between pharmacy refills were 34 (30-54) vs. 30 (26-40) in women with TFV-DP in DBS <1,250 vs. ≥1,250 fmol/punch (p=0.006). TFV-DP in DBS was negatively correlated with an increasing number of days between refills (r=-0.56, p=0.002). TFV-DP DBS was a reliable and objective measure of adherence in HIV-infected women based on a strong inverse relationship with pharmacy refill adherence.

  18. Short communication: Tenofovir diphosphate in dried blood spots as an objective measure of adherence in HIV-infected women.

    Science.gov (United States)

    Castillo-Mancilla, Jose R; Searls, Kristina; Caraway, Patricia; Zheng, Jia-Hua; Gardner, Edward M; Predhomme, Julie; Bushman, Lane R; Anderson, Peter L; Meditz, Amie L

    2015-04-01

    Simple and reproducible tools to assess antiretroviral adherence are needed. A level of tenofovir diphosphate (TFV-DP) in dried blood spots (DBS) Pearson correlation coefficient. The average TFV-DP between the two visits (aTFV-DP) in DBS and PBMCs was 1,874 (706-3,776) fmol/punch and 125 (1-278) fmol/10(6) cells, respectively. AA women had lower levels of aTFV-DP in DBS compared to whites (1,660 vs. 1,970 fmol/punch; p=0.04), with a viremic patient having the lowest drug levels (706 fmol/punch). Days between pharmacy refills were 34 (30-54) vs. 30 (26-40) in women with TFV-DP in DBS <1,250 vs. ≥1,250 fmol/punch (p=0.006). TFV-DP in DBS was negatively correlated with an increasing number of days between refills (r=-0.56, p=0.002). TFV-DP DBS was a reliable and objective measure of adherence in HIV-infected women based on a strong inverse relationship with pharmacy refill adherence. PMID:25328112

  19. Evaluation of spot and passive sampling for monitoring, flux estimation and risk assessment of pesticides within the constraints of a typical regulatory monitoring scheme.

    Science.gov (United States)

    Zhang, Zulin; Troldborg, Mads; Yates, Kyari; Osprey, Mark; Kerr, Christine; Hallett, Paul D; Baggaley, Nikki; Rhind, Stewart M; Dawson, Julian J C; Hough, Rupert L

    2016-11-01

    In many agricultural catchments of Europe and North America, pesticides occur at generally low concentrations with significant temporal variation. This poses several challenges for both monitoring and understanding ecological risks/impacts of these chemicals. This study aimed to compare the performance of passive and spot sampling strategies given the constraints of typical regulatory monitoring. Nine pesticides were investigated in a river currently undergoing regulatory monitoring (River Ugie, Scotland). Within this regulatory framework, spot and passive sampling were undertaken to understand spatiotemporal occurrence, mass loads and ecological risks. All the target pesticides were detected in water by both sampling strategies. Chlorotoluron was observed to be the dominant pesticide by both spot (maximum: 111.8ng/l, mean: 9.35ng/l) and passive sampling (maximum: 39.24ng/l, mean: 4.76ng/l). The annual pesticide loads were estimated to be 2735g and 1837g based on the spot and passive sampling data, respectively. The spatiotemporal trend suggested that agricultural activities were the primary source of the compounds with variability in loads explained in large by timing of pesticide applications and rainfall. The risk assessment showed chlorotoluron and chlorpyrifos posed the highest ecological risks with 23% of the chlorotoluron spot samples and 36% of the chlorpyrifos passive samples resulting in a Risk Quotient greater than 0.1. This suggests that mitigation measures might need to be taken to reduce the input of pesticides into the river. The overall comparison of the two sampling strategies supported the hypothesis that passive sampling tends to integrate the contaminants over a period of exposure and allows quantification of contamination at low concentration. The results suggested that within a regulatory monitoring context passive sampling was more suitable for flux estimation and risk assessment of trace contaminants which cannot be diagnosed by spot

  20. Comparison of Measurements of Autoantibodies to Glutamic Acid Decarboxylase and Islet Antigen-2 in Whole Blood Eluates from Dried Blood Spots Using the RSR-Enzyme Linked Immunosorbent Assay Kits and In-House Radioimmunoassays

    Directory of Open Access Journals (Sweden)

    Anders Persson

    2010-01-01

    Full Text Available To evaluate the performance of dried blood spots (DBSs with subsequent analyses of glutamic acid decarboxylase (GADA and islet antigen-2 (IA-2A with the RSR-ELISAs, we selected 80 children newly diagnosed with type 1 diabetes and 120 healthy women. DBSs from patients and controls were used for RSR-ELISAs while patients samples were analysed also with in-house RIAs. The RSR-ELISA-GADA performed well with a specificity of 100%, albeit sensitivity (46% was lower compared to in RIA (56%; P=.008. No prozone effect was observed after dilution of discrepant samples. RSR-ELISA-IA-2A achieved specificity of 69% and sensitivity was lower (59% compared with RIA (66%; P<.001. Negative or low positive patients and control samples in the RSR-ELISA-IA-2A increased after dilution. Eluates from DBS can readily be used to analyse GADA with the RSR-ELISA, even if low levels of autoantibodies were not detected. Some factor could disturb RSR-ELISA-IA-2A analyses.

  1. Two novel nonradioactive polymerase chain reaction-based assays of dried blood spots, genomic DNA, or whole cells for fast, reliable detection of Z and S mutations in the alpha 1-antitrypsin gene

    DEFF Research Database (Denmark)

    Andresen, B S; Knudsen, I; Jensen, P K;

    1992-01-01

    Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples...... from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele...

  2. Do "Instant Polls" Hit the Spot? Phone-In vs. Random Sampling of Public Opinion.

    Science.gov (United States)

    Bates, Benjamin; Harmon, Mark

    1993-01-01

    Compares television phone-in polls to random sample polling. Finds significant differences between the two types of opinion indicators. Shows that persons with strongly held opinions and a pro-change, activist stance are more likely to respond in phone-in polls. (SR)

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

    Directory of Open Access Journals (Sweden)

    Juana del Valle Mendoza

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

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

    DEFF Research Database (Denmark)

    Seemann, T. L.; Nybo, M.

    2015-01-01

    . As suggested by the EFLM working group on the preanalytical phase we introduced continuous quality control of the blood sampling procedure using a structured observation scheme to monitor the quality of blood sampling performed on an everyday basis. Materials and methods: Based on our own routines the EFLM...... auditing questionnaire was altered giving an observation scheme containing 19 observation issues. Using this scheme three blood samplings from two phlebotomists was observed twice a week (at the blood sampling unit and at a hospital ward, respectively), giving a total of 12 blood drawings observed per week....... Conclusion: It is possible to establish a continuous quality control on blood sampling. It has been well accepted by the staff and we have already been able to identify critical areas in the sampling process. We find that continuous auditing increase focus on the quality of blood collection which ensures...

  5. Validation and Application of a Dried Blood Spot Assay for Biofilm-Active Antibiotics Commonly Used for Treatment of Prosthetic Implant Infections.

    Science.gov (United States)

    Knippenberg, Ben; Page-Sharp, Madhu; Salman, Sam; Clark, Ben; Dyer, John; Batty, Kevin T; Davis, Timothy M E; Manning, Laurens

    2016-08-01

    Dried blood spot (DBS) antibiotic assays can facilitate pharmacokinetic (PK)/pharmacodynamic (PD) studies in situations where venous blood sampling is logistically difficult. We sought to develop, validate, and apply a DBS assay for rifampin (RIF), fusidic acid (FUS), and ciprofloxacin (CIP). These antibiotics are considered active against organisms in biofilms and are therefore commonly used for the treatment of infections associated with prosthetic implants. A liquid chromatography-mass spectroscopy DBS assay was developed and validated, including red cell partitioning and thermal stability for each drug and the rifampin metabolite desacetyl rifampin (Des-RIF). Plasma and DBS concentrations in 10 healthy adults were compared, and the concentration-time profiles were incorporated into population PK models. The limits of quantification for RIF, Des-RIF, CIP, and FUS in DBS were 15 μg/liter, 14 μg/liter, 25 μg/liter, and 153 μg/liter, respectively. Adjusting for hematocrit, red cell partitioning, and relative recovery, DBS-predicted plasma concentrations were comparable to measured plasma concentrations for each antibiotic (r > 0.95; P < 0.0001), and Bland-Altman plots showed no significant bias. The final population PK estimates of clearance, volume of distribution, and time above threshold MICs for measured and DBS-predicted plasma concentrations were comparable. These drugs were stable in DBSs for at least 10 days at room temperature and 1 month at 4°C. The present DBS antibiotic assays are robust and can be used as surrogates for plasma concentrations to provide valid PK and PK/PD data in a variety of clinical situations, including therapeutic drug monitoring or studies of implant infections.

  6. Dried Blood Spot Test for HIV Exposed Infants and Children and Their Anti-Retro Viral Treatment Status in Selected Hospitals in Ethiopia

    OpenAIRE

    Wondafrash, Beyene; Hiko, Desta

    2016-01-01

    Background Infants and children living with HIV receive antiretroviral treatment often late, are exposed to opportunistic infection and quickly develop AIDS. Few hospitals are providing ART service after Dried Blood Spot (DBS)test.The objective of this study is to assess the status of infants and children linked to ART. Methods Descriptive cross-sectional study was conducted in hospitals. Data of 138 infants and children exposed to HIV were collected from registration books and data bases fro...

  7. What is the right blood hematocrit preparation procedure for standards and quality control samples for dried blood spot analysis?

    NARCIS (Netherlands)

    Koster, Remco A.; Alffenaar, Jan-Willem C.; Botma, Rixt; Greijdanus, Ben; Touw, Daan J.; Uges, Donald R. A.; Kosterink, Jos G. W.

    2015-01-01

    Remco Koster is a research analyst and PhD candidate at the University Medical Center Groningen and University of Groningen. He has been working in the field of bioanalysis for over 13 years, where he has developed numerous analytical methods using LC-MS/MS. His main research focus is the influence

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

    Energy Technology Data Exchange (ETDEWEB)

    Anon.

    2012-07-01

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

  9. Typing of Plasmodium falciparum DNA from 2 years old Giemsa-stained dried blood spots using nested polymerase chain reaction assay.

    Science.gov (United States)

    Kumar, D; Dhiman, S; Rabha, B; Goswami, D; Yadav, K; Deka, M; Veer, V; Baruah, I

    2016-01-01

    A panel of 129 Giemsa-stained thick blood spots (TBS) confirmed for Plasmodium falciparum infection having different levels of parasite density were collected from a malaria endemic area. DNA was extracted and nested polymerase chain reaction (PCR) assay was performed to amplify P. falciparum DNA. Nested PCR assay successfully amplified P. falciparum DNA at a very low parasitaemia of ~10 parasites/μl of blood. Current PCR assay is very simple and can be used retrospectively to monitor the invasion and prevalence of different Plasmodium species in endemic areas. PMID:27080775

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

    Directory of Open Access Journals (Sweden)

    Ana Flávia Belchior Andrade

    2014-08-01

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

  11. Popliteal Vein Blood Sampling and the Postmortem Redistribution of Diazepam, Methadone, and Morphine.

    Science.gov (United States)

    Lemaire, Eric; Schmidt, Carl; Denooz, Raphael; Charlier, Corinne; Boxho, Philippe

    2016-07-01

    Postmortem redistribution (PMR) refers to the site- and time-related blood drug concentration variations after death. We compared central blood (cardiac and subclavian) with peripheral blood (femoral and popliteal) concentrations of diazepam, methadone, and morphine. To our knowledge, popliteal blood has never been compared with other sites. Intracardiac blood (ICB), subclavian blood (SB), femoral blood (FB), and popliteal blood (PB) were sampled in 30 cases. To assess PMR, mean concentrations and ratios were compared. Influence of postmortem interval on mean ratios was also assessed. Results show that popliteal mean concentrations were lower than those for other sites for all three drugs, even lower than femoral blood; mean ratios suggested that the popliteal site was less subject to PMR, and estimated postmortem interval did not influence ratios except for diazepam and methadone FB/PB. In conclusion, our study is the first to explore the popliteal site and suggests that popliteal blood is less prone to postmortem redistribution. PMID:27364283

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    months period and analyzed for 37 variables in 27 professional soccer players from the best Danish league. Additionally, players were tested for body composition, VO2max and physical performance by the Yo-Yo intermittent endurance sub-max test (IE2).Multiple variations in blood parameters occurred during...

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

    Science.gov (United States)

    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. PMID:22273184

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

    OpenAIRE

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

    2012-01-01

    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.

  15. Quantitative photoacoustic blood oxygenation measurement of whole porcine blood samples using a multi-wavelength semiconductor laser system

    Science.gov (United States)

    Friedrich, Claus-Stefan; Mienkina, Martin P.; Brenner, Carsten; Gerhardt, Nils C.; Jörger, Manfred; Strauß, Andreas; Beckmann, Martin F.; Schmitz, Georg; Hofmann, Martin R.

    2011-07-01

    We present a photoacoustic measurement system based on semiconductor lasers for blood oxygenation measurements. It permits to use four different optical wavelengths (650nm, 808nm, 850nm, 905nm) to generate photoacoustic signals. As the optical extinction coefficient of oxygenated hemoglobin and deoxygenated hemoglobin is different at specific wavelengths, a blood oxygenation measurement by a multi-wavelength photoacoustic laser system is feasible. Especially at 650nm, the clear difference between the extinction coefficients of the two hemoglobin derivates permits to determine the blood oxygenation in combination with other near infrared wavelengths. A linear model based on tabulated values of extinction coefficients for fully oxygenated and fully deoxygenated hemoglobin is presented. We used heparin stabilized whole porcine blood samples to model the optical behavior of human blood, as the optical absorption behavior of porcine hemoglobin does not differ significantly from human hemoglobin. To determine the real oxygen saturation values of the blood samples, we measured the partial oxygen pressure with an IRMA Trupoint Blood Analysis System. The oxygen saturation values were calculated from a dissociation curve for porcine blood. The results of the photoacoustic measurement are in qualitatively good agreement with the predicted linear model. Further, we analyze the abilities and the limitations of quantitative oxygenation measurements.

  16. Using dried blood spots collected under field condition to determine HIV-1 diversity and drug resistance mutations in resource limited Tanzania

    Directory of Open Access Journals (Sweden)

    James Kimaro

    2014-11-01

    Full Text Available Introduction: A dried blood spot (DBS on filter paper has been used for different tests globally and has gained popularities in resource limited settings especially during HIV/AIDS epidemic. We assessed the efficiency of molecular characterization of HIV-1 subtypes using DBS collected under field conditions in northern Tanzania. Materials and Methods: In 2011 and 2012, 60 DBS samples were collected under field conditions from exposed and newly diagnosed HIV-1 infected children from Kilimanjaro (n=20, Arusha (n=20, Tanga (n=10 and Manyara (n=10. Results and discussion: Of 60 DBS analyzed at both Protease (PR and Reverse Transcriptase (RT regions, 45 (75% were analyzed, including 17 (85% from Kilimanjaro, 15 (75% from Arusha, 8 (80% from Tanga, and 5 (50% from Manyara region. All 45 DBS characterized had viral load above 1000 copies/mL with mean log10 viral loads of 3.87 copies/mL (SD 0.995. The phylogenetic results indicated presence of subtype and circulating recombinant form (CRF. In which, 24 were subtype A1 (53.33%, 16 were subtype C (35.55%, 3 were subtype D (6.67% and 2 were CRF10_CD (4.35%. All major mutations were detected in the RT region, none from protease (PR region. The mutations detected were Y181C (n=8, K103 (n=4 and G190A (n=1, conferring resistance to non-nucleoside reverse transcriptase inhibitors (NNRTIs, and M184V (n=1, conferring resistance to lamivudine and emtricitabine. Conclusions: Our results indicate that DBS collected from field conditions in resource scarcity areas can be used to determine the phylogeny of the virus and drug resistance mutations in areas with diverse HIV-1 group M subtypes.

  17. Liver spots

    Science.gov (United States)

    Sun-induced skin changes - liver spots; Senile or solar lentigines; Skin spots - aging; Age spots ... your skin by using skin bleaching lotions or creams. Most bleaching lotions use hydroquinone. This medicine is ...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1984-01-01

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

  19. Minimally invasive blood sampling method for genetic studies on Gopherus tortoises

    OpenAIRE

    García–Feria, L. M.; Ureña–Aranda, C. A.; Espinosa de los Monteros, A.

    2015-01-01

    Obtaining good quality tissue samples is the first hurdle in any molecular study. This is especially true for studies involving management and conservation of wild fauna. In the case of tortoises, the most common sources of DNA are blood samples. However, only a minimal amount of blood is required for PCR assays. Samples are obtained mainly from the brachial and jugular vein after restraining the animal chemically, or from conscious individuals by severe handling methods and clamping. Herein,...

  20. The efficacy of field techniques for obtaining and storing blood samples from fishes.

    Science.gov (United States)

    Clark, T D; Donaldson, M R; Drenner, S M; Hinch, S G; Patterson, D A; Hills, J; Ives, V; Carter, J J; Cooke, S J; Farrell, A P

    2011-11-01

    Prompted by the dramatic increase in the use of blood analyses in fisheries research and monitoring, this study investigated the efficacy of common field techniques for sampling and storing blood from fishes. Three questions were addressed: (1) Do blood samples taken via rapid caudal puncture (the 'grab-and-stab' technique) yield similar results for live v. sacrificed groups of fishes? (2) Do rapidly obtained caudal blood samples accurately represent blood properties of fishes prior to capture? (3) Does storage of whole blood in an ice slurry for a working day (8·5 h) modify the properties of the plasma? It was shown that haematocrit, plasma ions, metabolites, stress hormones and sex hormones of caudal blood samples were statistically similar when taken from live v. recently sacrificed groups of adult coho salmon Oncorhynchus kisutch. Moreover, this study confirmed by using paired blood samples from cannulated O. kisutch that blood acquired through the caudal puncture technique (mean ±s.e. 142 ± 26 s after capture) was representative of fish prior to capture. Long-term (8·5 h) cold storage of sockeye salmon Oncorhynchus nerka whole blood caused significant decreases in plasma potassium and chloride, and a significant increase in plasma glucose. Previous research has suggested that these changes largely result from net movements of ions and molecules between the plasma and erythrocytes, movements that can occur within minutes of storage. Thus, blood samples from fishes should be centrifuged as quickly as practicable in the field for separation of plasma and erythrocytes to prevent potentially misleading data. PMID:22026608

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

    Science.gov (United States)

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

    2016-08-01

    Since driving under the influence of drugs (DUID) is as dangerous as drink-driving, many countries regulate DUID by law. However, laws against the use of drugs while driving are not yet established in Korea. In order to investigate the type and frequency of drugs used by drivers in Korea, we analyzed controlled and non-controlled drugs in alcohol-positive blood samples. Total 275 blood samples were taken from Korean drivers, which were positive in roadside alcohol testing. The following analyses were performed: blood alcohol concentrations by GC; screening for controlled drugs by immunoassay and confirmation for positive samples by GC-MS. For the detection of DUID related drugs in blood samples, a total of 49 drugs were selected and were examined by GC-MS. For a rapid detection of these drugs, an automated identification software called "DrugMan" was used. Concentrations of alcohol in 275 blood samples ranged from 0.011 to 0.249% (average 0.119%). Six specimens showed positive results by immunoassay: one methamphetamine and five benzodiazepines I. By GC-MS confirmation, only benzodiazepines in four cases were identified, while methamphetamine and benzodiazepine in two cases were not detected from the presumptive positive blood samples. Using DrugMan, four drugs were detected; chlorpheniramine (5)*, diazepam (4), dextromethorphan (1) and doxylamine (1). In addition, ibuprofen (1), lidocaine (1) and topiramate (1) were also detected as general drugs in blood samples ('*' indicates frequency). The frequency of drug abuse by Korean drivers was relatively low and a total 14 cases were positive in 275 blood samples with a ratio of 5%. However it is necessary to analyze more samples including alcohol negative blood, and to expand the range of drug lists to get the detailed information. PMID:27015372

  2. Minimally invasive blood sampling method for genetic studies on Gopherus tortoises

    Directory of Open Access Journals (Sweden)

    García–Feria, L. M.

    2015-04-01

    Full Text Available Obtaining good quality tissue samples is the first hurdle in any molecular study. This is especially true for studies involving management and conservation of wild fauna. In the case of tortoises, the most common sources of DNA are blood samples. However, only a minimal amount of blood is required for PCR assays. Samples are obtained mainly from the brachial and jugular vein after restraining the animal chemically, or from conscious individuals by severe handling methods and clamping. Herein, we present a minimally invasive technique that has proven effective for extracting small quantities of blood, suitable for genetic analyses. Furthermore, the samples obtained yielded better DNA amplification than other cell sources, such as cloacal epithelium cells. After two years of use on wild tortoises, this technique has shown to be harmless. We suggest that sampling a small amount of blood could also be useful for other types of analyses, such as physiologic and medical monitoring.

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

    Science.gov (United States)

    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.

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

    NARCIS (Netherlands)

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

    2011-01-01

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

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

    DEFF Research Database (Denmark)

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

    2001-01-01

    in humans. The overall recovery of radioiron from blood is more than 90%, and the coefficient of variation, as judged by the variation in the ratio Fe-55/Fe-59 is in the order of 4%. Combined with whole-body counting of 59Fe and direct gamma -counting of Fe-59 on blood samples, this method represents...

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

    Directory of Open Access Journals (Sweden)

    Thaís Heloisa Vaz Farias

    2016-05-01

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

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

    Institute of Scientific and Technical Information of China (English)

    Reza Mirnejad; Mozafar mohamadi; Vahbeh Piranfar; Seied Mojtaba Mortazavi; Reza Kachuei

    2013-01-01

    Objective: To design a duplex PCR for rapid and simultaneous detection of Brucella species. in human blood samples. Methods: Fifty-two peripheral bloods samples were collected from suspicious patients with brucellosis. Following DNA extraction, PCR assay were performed, using three primers that could simultaneously identify and differentiate three major species of pathogenic Brucella in humans and animals. Results: Of the 52 peripheral bloods samples tested, 25 sample (48%) showed positive reactions in PCR. Twelve samples were positive for Brucella abortus (B. abortus) (23%), 13 for Brucella melitensis (B. melitensis) (25%) and 0 for Brucella ovis (B. ovis) (0%). Conclusions: This work de=monstrates that in case where specific primers were utilized, duplex PCR has proved to be a simple, fast, and relatively inexpensive method for simultaneous detection of important species of Brucella in clinical samples.

  8. Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study

    OpenAIRE

    Williams Adam R; Mondala Tony S; Robison Elizabeth H; Head Steven R; Salomon Daniel R; Kurian Sunil M

    2009-01-01

    Abstract Background Whole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a veno...

  9. Laser beam induced nanoscale spot through nonlinear “thick” samples: A multi-layer thin lens self-focusing model

    International Nuclear Information System (INIS)

    Self-focusing is a well-researched phenomenon. Nanoscale spots can be achieved through self-focusing, which is an alternative method for achieving high-density data storage, high-resolution light imaging, and maskless nanolithography. Several research groups have observed that self-focusing spots can be reduced to nanoscale levels via incident laser power manipulation. Self-focusing spots can be analyzed by solving the nonlinear Schrödinger equation and the finite difference time domain method. However, both procedures are complex and time-consuming. In the present work, a multi-layer thin-lens self-focusing model that considers diffraction effects and changes of refractive index along the radial and film thickness directions is proposed to analyze the self-focusing behavior and traveling process of light beams intuitively. The self-focusing behaviors of As2S3 are simulated, and results show that a nanoscale self-focusing spot with a radius of about 0.12 μm can be formed at the bottom of nonlinear sample when the incident laser power exceeds 4.25 mW. Our findings are basically consistent with experimental reports and provide a good method for analyzing and understanding the self-focusing process. An appropriate application schematic design is also provided

  10. Effects of storage conditions on forensic examinations of blood samples and bloodstains stored for 20 years.

    Science.gov (United States)

    Hara, M; Nakanishi, H; Yoneyama, K; Saito, K; Takada, A

    2016-01-01

    The effects of various storage conditions on blood identification tests, DNA degradation, and short tandem repeat (STR) typing were evaluated. Bloodstains stored at room temperature, 4 °C, -20 °C, and -80 °C for 20 years; blood samples stored at -20 °C and -80 °C for 20 years; and fresh blood samples were analyzed. Leuco-malachite-green testing, anti-human hemoglobin (Hb) testing (using immunochromatography), and tests for hemoglobin-beta (HBB) mRNA were performed as blood identification tests. DNA degradation was evaluated by quantifying the ratios of 305 and 129 base pair (bp) fragments to 41 bp fragments. STR typing was performed using an AmpFlSTR® Identifiler™ Plus PCR Amplification Kit. All samples were positive in leuco-malachite-green staining and anti-human Hb assays. HBB was not detected in blood samples stored at -20 °C or -80 °C, although this marker was detected in all bloodstains. As indicated by the ratio of 129:41 bp and 305:41 bp DNA fragments, DNA from bloodstains stored at room temperature or 4 °C were significantly degraded compared to DNA from all other samples. STR typing analyses revealed that a portion of the loci was undetected in bloodstains stored at room temperature. Therefore, to prevent DNA degradation during long-term storage, it is recommended that bloodstains and blood be stored at below -20 °C. In addition, because bloodstains are more suitable for detection of blood-specific mRNAs than blood sample, it is desirable that blood is stored as bloodstain for this method.

  11. Dengue-3 outbreak in Paraguay: investigations using capillary blood samples on filter paper.

    Science.gov (United States)

    Matheus, Severine; Meynard, Jean-Baptiste; Lavergne, Anne; Girod, Romain; Moua, David; Labeau, Bhety; Dussart, Philippe; Lacoste, Vincent; Deparis, Xavier

    2008-11-01

    During a dengue-3 outbreak in Paraguay at the beginning of 2007, capillary blood samples absorbed onto filter papers were collected from 44 suspected cases. These samples were subjected to three molecular and serologic tests, and 31 of the 44 samples gave a positive result by at least one of the techniques used. Molecular analyses detected the dengue-3 serotype in 22 patients and additionally the dengue-2 serotype in two patients. Therefore two different serotypes were co-circulating during this outbreak. Overall, this study validates the use of dried-blood samples for field screening investigations. Indeed, all types of laboratory studies of dengue were possible with samples consisting of a few drops of dried blood from finger pricks.

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

    DEFF Research Database (Denmark)

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

    2006-01-01

    and that it might take an extra hour to recover from it. CO2 anaesthesia seemed unable to prevent the increase in blood pressure and the fluctuations in body temperature induced by blood sampling, and up to 10 h after sampling, the rats were still affected by CO2 anaesthesia. Rats anaesthetized with isoflurane...... to be the method from which rats most rapidly recover when compared with periorbital puncture and tail vein puncture, and that for anaesthesia, isoflurane is recommended in preference to CO2....... for rats sampled from the tail vein, which showed fluctuations in body temperature in excess of 30 h after sampling. Increases in heart rate and blood pressure within the first hours after sampling indicated that periorbital puncture was the method that had the largest acute impact on the rats...

  13. Time impact on non-activated and kaolin-activated blood samples in thromboelastography

    OpenAIRE

    Durila, Miroslav; Lukáš, Pavel; Bronský, Jiří; Cvachovec, Karel

    2015-01-01

    Background The correct methodology of thrombelastography might be influenced by elapsing time. In our study we investigated kaolin activated citrated samples together with non-activated citrated samples in relation to the elapsed times of 0, 15 and 30 minutes to compare both methods and to find out if there is an impact of time on results of thrombelastography. Methods Blood samples obtained from 10 healthy volunteers were analyzed after 0, 15 and 30 minutes from sampling with kaolin activati...

  14. Measurement of cerebral blood flow the blood sampling method using {sup 99m}Tc-ECD. Simultaneous scintigram scanning of arterial blood samples and the brain with a gamma camera

    Energy Technology Data Exchange (ETDEWEB)

    Hachiya, Takenori; Inugami, Atsushi [Rehabilitation Center for Physically Disabled Persons and Medical Center for Mental Health-Akita, Kyowa (Japan); Iida, Hidehiro; Mizuta, Yoshihiko; Kawakami, Takeshi; Inoue, Minoru

    1999-01-01

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

  15. Comparison of drug concentrations in blood and oral fluid collected with the Intercept sampling device.

    Science.gov (United States)

    Gjerde, Hallvard; Mordal, Jon; Christophersen, Asbjørg S; Bramness, Jørgen G; Mørland, Jørg

    2010-05-01

    The aim of the study was to determine drug concentration ratios between oral fluid collected with the Intercept device and whole blood. Samples of blood and oral fluid were obtained from patients admitted to acute psychiatric treatment and drivers suspected of drugged driving. Samples were analyzed for illegal drugs, benzodiazepines, opioids, carisoprodol, and meprobamate. Drugs were detected in samples of both blood and oral fluid from 59 subjects; altogether, 17 different drugs were found. Concentration ratios between oral fluid and blood were determined for all cases. The distributions of drug concentration ratios were wide for most drugs and do not allow reliable estimations of drug concentrations in blood using concentrations in oral fluid. The median oral fluid/blood drug concentration ratios for the most prevalent drugs were 0.036 diazepam, 0.027 nordiazepam, 7.1 amphetamine, 2.9 methamphetamine, 5.4 codeine, 1.9 morphine, and 4.7 tetrahydrocannabinol. The correlation coefficients between drug concentrations in oral fluid and blood ranged from 0.15 to 0.96 for the six most prevalent drugs. PMID:20465866

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

    Directory of Open Access Journals (Sweden)

    Bölenius Karin

    2012-01-01

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

  17. Heel blood sampling in European neonatal intensive care units: compliance with pain management guidelines

    DEFF Research Database (Denmark)

    Losacco, Valentina; Cuttini, Marina; Greisen, Gorm;

    2011-01-01

    Objective To describe the use of heel blood sampling and non-pharmacological analgesia in a large representative sample of neonatal intensive care units (NICUs) in eight European countries, and compare their self-reported practices with evidence-based recommendations. Methods Information on use...... with published guidelines is needed for clinical and ethical reasons....

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

    Science.gov (United States)

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

    2015-11-01

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

  19. Alcohol levels in cerebrospinal fluid and blood samples from patients under pathological conditions.

    Science.gov (United States)

    Agapejev, S; Vassilieff, I; Curi, P R

    1992-11-01

    We measured alcohol levels by the Cordebard method in 148 CSF samples from individuals who had abstained from alcohol for at least 7 days prior to the beginning of the study. Each blood sample was accompanied by a CSF sample from the same patient. CSF samples found to be normal after analysis were used as controls. Mean alcohol concentration in blood did not differ significantly between the control group and the groups with altered CSF. The group with altered CSF had statistically higher alcohol levels in CSF than in blood. CSF lactate, glucose and protein levels were not correlated with alcohol level. The results suggest the presence of endogenous alcohol in the CSF, with levels increasing in the presence of pathological processes involving the nervous system.

  20. Determination of optimal sampling times for a two blood sample clearance method using (51)Cr-EDTA in cats.

    Science.gov (United States)

    Vandermeulen, Eva; De Sadeleer, Carlos; Piepsz, Amy; Ham, Hamphrey R; Dobbeleir, André A; Vermeire, Simon T; Van Hoek, Ingrid M; Daminet, Sylvie; Slegers, Guido; Peremans, Kathelijne Y

    2010-08-01

    Estimation of the glomerular filtration rate (GFR) is a useful tool in the evaluation of kidney function in feline medicine. GFR can be determined by measuring the rate of tracer disappearance from the blood, and although these measurements are generally performed by multi-sampling techniques, simplified methods are more convenient in clinical practice. The optimal times for a simplified sampling strategy with two blood samples (2BS) for GFR measurement in cats using plasma (51)chromium ethylene diamine tetra-acetic acid ((51)Cr-EDTA) clearance were investigated. After intravenous administration of (51)Cr-EDTA, seven blood samples were obtained in 46 cats (19 euthyroid and 27 hyperthyroid cats, none with previously diagnosed chronic kidney disease (CKD)). The plasma clearance was then calculated from the seven point blood kinetics (7BS) and used for comparison to define the optimal sampling strategy by correlating different pairs of time points to the reference method. Mean GFR estimation for the reference method was 3.7+/-2.5 ml/min/kg (mean+/-standard deviation (SD)). Several pairs of sampling times were highly correlated with this reference method (r(2) > or = 0.980), with the best results when the first sample was taken 30 min after tracer injection and the second sample between 198 and 222 min after injection; or with the first sample at 36 min and the second at 234 or 240 min (r(2) for both combinations=0.984). Because of the similarity of GFR values obtained with the 2BS method in comparison to the values obtained with the 7BS reference method, the simplified method may offer an alternative for GFR estimation. Although a wide range of GFR values was found in the included group of cats, the applicability should be confirmed in cats suspected of renal disease and with confirmed CKD. Furthermore, although no indications of age-related effect were found in this study, a possible influence of age should be included in future studies. PMID:20452793

  1. Use of Dried Blood Spots for Estimating Children?s Exposures to Heavy Metals in Epidemiological Research

    Science.gov (United States)

    Background: Children’s exposures to arsenic (As), lead (Pb), mercury (Hg), and cadmium (Cd) are of particular concern in early-life. Exposures to heavy metals are traditionally measured in whole venous blood, which is costly and invasive. As an alternative we describe a met...

  2. Effect of blood sampling on apomorphine-induced penile tumescence in erectile impotence: a case report.

    OpenAIRE

    Kiely, M E; Thavundayil, J X; Lal, S

    1995-01-01

    Apomorphine HCl (Apo) (0.5 mg sc), but not placebo, induced an erectile response (monitored with a mercury strain gauge) lasting 40 min in an impotent hyperprolactinemic patient. Serial blood sampling modified the 40 min erectile response. Prompt detumescence followed by complete or partial restoration of tumescence occurred each time blood was drawn. This observation points to the sensitivity of the Apo-erectile response to experimental procedures subjectively perceived as anxiogenic.

  3. Tracer input for kinetic modelling of liver physiology determined without sampling portal venous blood in pigs

    International Nuclear Information System (INIS)

    Quantification of hepatic tracer kinetics by PET requires measurement of tracer input from the hepatic artery (HA) and portal vein (PV). We wished to develop a method for estimating dual tracer input without the necessity to sample PV blood. Pigs weighing 40 kg were given bolus doses of C15O (CO), 2-[18F]fluoro-2-deoxy-D-glucose (FDG), [11C]-methylglucose (MG), 2-[18F]fluoro-2-deoxy-D-galactose (FDGal) or H215O (H2O). Tracer concentration 3-min time courses were measured in the femoral artery and PV by blood sampling. Blood flow was measured in the HA and PV using flow-meters. A model for transfer of tracer through the splanchnic circulation was used to estimate values of a tracer-specific model parameter β. Tracer-specific mean values of β were used to estimate tracer concentration time courses in the PV from the measured arterial concentration. A model-derived dual-input was calculated using the mean HA flow fraction (0.25) and validated by comparison of the use of the measured dual-input and a kinetic model with a fixed ''true'' K1true, i.e. clearance of tracer from blood to liver cells. The rank order of the means of β was CO 2O, reflecting their different splanchnic mean transit times. Estimated K1est was not significantly different from ''true'' K1true. The hepatic dual tracer input, which is of great importance for the assessment of processes such as transfer across the plasma-hepatocyte membrane or hepatic blood perfusion, can be well approximated in pigs without the necessity to sample PV blood and measure hepatic blood flow; only arterial blood sampling is needed. (orig.)

  4. Genetic Characterization of Atypical Mansonella (Mansonella) ozzardi Microfilariae in Human Blood Samples from Northeastern Peru

    OpenAIRE

    Marcos, Luis A.; Arrospide, Nancy; Recuenco, Sergio; Cabezas, Cesar; Gary J Weil; Fischer, Peter U.

    2012-01-01

    DNA sequence comparisons are useful for characterizing proposed new parasite species or strains. Microfilariae with an atypical arrangement of nuclei behind the cephalic space have been recently described in human blood samples from the Amazon region of Peru. Three blood specimens containing atypical microfilariae were genetically characterized using three DNA markers (5S ribosomal DNA, 12S ribosomal DNA, and cytochrome oxidase I). All atypical microfilariae were clustered into the Mansonella...

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

    Science.gov (United States)

    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.

  6. Biomarkers for monitoring pre-analytical quality variation of mRNA in blood samples.

    Directory of Open Access Journals (Sweden)

    Hui Zhang

    Full Text Available 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.

  7. Detection of micrometastasis in peripheral blood by multi-sampling in patients with colorectal cancer

    Institute of Scientific and Technical Information of China (English)

    Xi-Wei Zhang; Hong-Yu Yang; Ping Fan; Li Yang; Guo-Yu Chen

    2005-01-01

    AIM: To evaluate the reverse transcriptase-PCR assay and multiple sampling for detection of cytokeratin-positive cells in peripheral blood of colorectal carcinoma patients and to investigate the clinical significance of micrometastasis in peripheral blood.METHODS: The expression of CK20 mRNA by RT-PCR was investigated in bone marrow, portal vein and peripheral blood in 58 colorectal cancer patients and 12 controls without known cancer. The peripheral blood was sampled twice at intervals of 3 d before operation. All the patients were followed up for one year.RESULTS: There was no positive expression of CK20mRNA in 12 volunteers. The positive expression of CK20mRNA was 77.6% (45/58) in bone marrow, and that in portal vein was 74.1% (43/58) of colorectal carcinoma patients.The positive expression of CK20mRNA cells in peripheral blood rose from 44.8% (26/58) to 69.0% (40/58) (P<0.01).The total positivity of CK20mRNA expression in peripheral blood was similar to the positivity of CK20mRNA in bone marrow and portal vein. The positive rates became higher in later clinical stages than in early stages. The CK20mRNA positive patients had a higher relapse rate within one year than the CK20mRNA negative patients.CONCLUSION: Multiple blood sampling can increase the detection of tumor cells in peripheral blood by RT-PCR for CK20mRNA in colorectal carcinoma patients and it is as sensitive and specific as that of bone marrow and portal vein. This technique may be reliable and convenient to diagnose micrometastasis of colorectal carcinoma and has an important significance in determining the prognosis of cancer patients.

  8. The future of doping control in athletes. Issues related to blood sampling.

    Science.gov (United States)

    Birkeland, K I; Hemmersbach, P

    1999-07-01

    When current antidoping programmes were developed, the most frequently used doping agents were xenobiotics, such as stimulants and anabolic steroids, that are readily detectable in urine with the use of gas chromatography and mass spectrometry. As control of traditional doping agents became effective, some athletes turned to other means to improve performance, including blood doping and the application of recombinant peptide hormones such as erythropoietin and growth hormone. Doping with these agents is not easily detected in urine samples, and therefore new strategies must be developed as a supplement to those already in use. Such strategies will probably include analysing blood samples, as several of the most promising methods that are able to detect modern doping agents use blood as the analytical matrix. Non-autologous blood doping results in an admixture of self and foreign red blood cells that can be detected in a blood sample with the methods available. Methods to indicate doping with erythropoietin include the indirect finding of an elevated level of soluble transferrin receptor in serum, or a direct demonstration of a shift from the normal to an abnormal spectrum of erythropoietin isoforms. To indicate doping with growth hormone, a set of serum parameters including insulin growth factors and their binding proteins are under investigation as indirect evidence. A direct method using isotopic differences between endogenous and recombinant growth hormones is being investigated. A similar method has been established to detect the administration of testosterone esters. Several legal and ethical questions must be solved before blood sampling can become a part of routine doping control, but the major ethical question is whether sport can continue as today without proper methods to detect many modern doping agents.

  9. CONFIRMATION OF HEREDITARY TYROSINEMIA TYPE 1 IN NEONATAL STAGE USING SPECTROPHOTOMETRIC MICROASSAY BASED ON THE DETERMINATION OF SUCCINYLACETONE LEVEL IN DRIED-BLOOD SPOTS

    Directory of Open Access Journals (Sweden)

    Gautam Kumar

    2014-05-01

    Full Text Available Hereditary Tyrosinemia Type 1 (HT1 is a metabolic disorder below to the class of autosomal recessive inheritance caused by the dearth of enzyme fumaryl acetoacetase the last enzyme in the tyrosine catabolic pathway . Affected individuals show increased tyrosine and succinyl acetone concentration (SA in blood. Patients also excrete increased concentration of SA in urine. The disorder is characterized by progressive liver disease and renal tubular defects with accompanying hypophosphatemic rickets. Symptoms of HT1 usually appear in the first few months of life and include failure to gain weight and grow at the expected rate, diarrhea, vomiting, yellowing of the skin and whites of the eyes (jaundice. It may also lead to liver and kidney failure and an increased risk of liver cancer. Liver transplantation is the only effective treatment for hereditary tyrosinemia type 1. In the present study, Succinyl Acetone is measured by using its inhibitory property on d-aminolaevulinate dehydratase enzyme for the diagnosis of HT1 in dried blood spots.

  10. Monitoring and evaluation of lymphatic filariasis interventions: an improved PCR-based pool screening method for high throughput Wuchereria bancrofti detection using dried blood spots

    OpenAIRE

    Plichart, Catherine; Lemoine, Aurore

    2013-01-01

    Background Effective diagnostic tools are necessary to monitor and evaluate interruption of Lymphatic Filariasis (LF) transmission. Accurate detection of Wuchereria bancrofti (Wb) microfilaria (mf) is essential to measure the impact of community treatment programmes. PCR-based assays are specific, highly sensitive tools allowing the detection of Wuchereria bancrofti DNA in human blood samples. However, current protocols describing the pool screening approach, use samples of less than 60 μl of...

  11. Sample pretreatment microfluidic chip for DNA extraction from rat peripheral blood

    Institute of Scientific and Technical Information of China (English)

    CHEN Xing; CUI Dafu; LIU Changchun; LI Hui; ZHAO Weixing

    2007-01-01

    A sample pretreatment microfluidic chip was described based on the principle of solid phase extraction and micro electro mechanical system technology.Oxidized porous silicon with the large surface area as the solid phase matrix for absorption of DNA from a biological sample can greatly improve the DNA yield.The factors that could affect the DNA yield were analyzed and the preparation technology and the experiment procedure were improved.The DNA purification process from the rat peripheral blood can be achieved and the DNA yield is 24 ng/(μL whole blood),which can reach the level of the commercial DNA purification kits.Furthermore,the DNA extracted from the whole blood can be amplified by polymerase chain reaction,which can achieve a high efficiency of the amplification.

  12. Evaluation of different sized blood sampling tubes for thromboelastometry, platelet function, and platelet count

    DEFF Research Database (Denmark)

    Andreasen, Jo Bønding; Pistor-Riebold, Thea Unger; Knudsen, Ingrid Hell;

    2014-01-01

    and compared three blood sampling tubes of different size: 1.8, 2.7, and 3.6 mL. All tubes were made of plastic and contained 3.2% sodium-citrate as anticoagulant. Platelet aggregation was investigated in 12 healthy individuals employing the Multiplate® Analyser comparing tubes of 3.6 mL and 1.8 mL. Platelet...... be preferred for RoTEM® analyses in order to minimise the volume of blood drawn. With regard to platelet aggregation analysed by impedance aggregometry tubes of different size cannot be used interchangeably. If platelet count is determined later than 10 min after blood sampling using tubes containing citrate...

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

    Science.gov (United States)

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

    1990-01-01

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

  14. Theorical and practical bases for blood sample collection from the heel of newborns for neonatal screening

    Directory of Open Access Journals (Sweden)

    Marcela Vela-Amieva

    2014-07-01

    collected in a special filter paper (Guthrie’s card. Despite its apparent simplicity, NBS laboratories commonly receive a large number of samples collected incorrectly and technically unsuitable for perfor4ming biochemical determinations. The aim of the present paper is to offer recommendations based on scientific evidence, for the properly blood collection on filter paper for NBS programs.

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

    Science.gov (United States)

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

    2009-12-21

    This paper describes a microsystem for automated blood sampling from laboratory mice used in pharmacokinetic studies. Intended to be mounted as a "backpack" on a mouse, it uses a microneedle, reservoir, and an actuator to instantaneously prick the animal for a time-point sample, eliminating the need for a tethered catheter with large dead volume. The blood is collected by capillary effect through a 31-33 gauge microneedle (250-210 microm OD) into a approximately 1 microL micromachined steel reservoir. The voice coil actuator provides a peak force of approximately 300 mN, which amply exceeds the measured piercing force of mouse skin (i.e., 60-85 mN for a 31-gauge needle with 12 degrees bevel). The sampling system was tested in vitro using a mock vessel with adjustable pressure; the reservoir was filled in electropolishing the inner surface to make it more hydrophilic or using a polymer wire insert to increase the surface area. The steel surface of the reservoir is also coated with silicon oxynitride by plasma-enhanced chemical vapor deposition to improve its hydrophilicity. Blood from fresh bovine tissue was collected into the reservoir to simulate interstitial fluid sampling. In vivo tests on live, anesthetized mice resulted in successful collection of blood into the reservoir. The possible integration of the device in microanalytical systems and the device scalability for multisampling are discussed. PMID:20024028

  16. Barrier screens: a method to sample blood-fed and host-seeking exophilic mosquitoes

    Directory of Open Access Journals (Sweden)

    Burkot Thomas R

    2013-02-01

    Full Text Available Abstract Background Determining the proportion of blood meals on humans by outdoor-feeding and resting mosquitoes is challenging. This is largely due to the difficulty of finding an adequate and unbiased sample of resting, engorged mosquitoes to enable the identification of host blood meal sources. This is particularly difficult in the south-west Pacific countries of Indonesia, the Solomon Islands and Papua New Guinea where thick vegetation constitutes the primary resting sites for the exophilic mosquitoes that are the primary malaria and filariasis vectors. Methods Barrier screens of shade-cloth netting attached to bamboo poles were constructed between villages and likely areas where mosquitoes might seek blood meals or rest. Flying mosquitoes, obstructed by the barrier screens, would temporarily stop and could then be captured by aspiration at hourly intervals throughout the night. Results In the three countries where this method was evaluated, blood-fed females of Anopheles farauti, Anopheles bancroftii, Anopheles longirostris, Anopheles sundaicus, Anopheles vagus, Anopheles kochi, Anopheles annularis, Anopheles tessellatus, Culex vishnui, Culex quinquefasciatus and Mansonia spp were collected while resting on the barrier screens. In addition, female Anopheles punctulatus and Armigeres spp as well as male An. farauti, Cx. vishnui, Cx. quinquefasciatus and Aedes species were similarly captured. Conclusions Building barrier screens as temporary resting sites in areas where mosquitoes were likely to fly was an extremely time-effective method for collecting an unbiased representative sample of engorged mosquitoes for determining the human blood index.

  17. Whole genome transcript profiling from fingerstick blood samples: a comparison and feasibility study

    Directory of Open Access Journals (Sweden)

    Williams Adam R

    2009-12-01

    Full Text Available Abstract Background Whole genome gene expression profiling has revolutionized research in the past decade especially with the advent of microarrays. Recently, there have been significant improvements in whole blood RNA isolation techniques which, through stabilization of RNA at the time of sample collection, avoid bias and artifacts introduced during sample handling. Despite these improvements, current human whole blood RNA stabilization/isolation kits are limited by the requirement of a venous blood sample of at least 2.5 mL. While fingerstick blood collection has been used for many different assays, there has yet to be a kit developed to isolate high quality RNA for use in gene expression studies from such small human samples. The clinical and field testing advantages of obtaining reliable and reproducible gene expression data from a fingerstick are many; it is less invasive, time saving, more mobile, and eliminates the need of a trained phlebotomist. Furthermore, this method could also be employed in small animal studies, i.e. mice, where larger sample collections often require sacrificing the animal. In this study, we offer a rapid and simple method to extract sufficient amounts of high quality total RNA from approximately 70 μl of whole blood collected via a fingerstick using a modified protocol of the commercially available Qiagen PAXgene RNA Blood Kit. Results From two sets of fingerstick collections, about 70 uL whole blood collected via finger lancet and capillary tube, we recovered an average of 252.6 ng total RNA with an average RIN of 9.3. The post-amplification yields for 50 ng of total RNA averaged at 7.0 ug cDNA. The cDNA hybridized to Affymetrix HG-U133 Plus 2.0 GeneChips had an average % Present call of 52.5%. Both fingerstick collections were highly correlated with r2 values ranging from 0.94 to 0.97. Similarly both fingerstick collections were highly correlated to the venous collection with r2 values ranging from 0.88 to 0

  18. Fabrication and Characterization of a Microfluidic Device to Ultrapurify Blood Samples

    KAUST Repository

    Tallerico, Marco

    2015-05-04

    The improvement of blood cell sorting techniques in recent years have attracted the attention of many researchers due to the possible benefits that these methods can lead in biology, regenerative medicine, materials science and therapeutic area. In this work a cell sorting technique based on filtration is described. The separation occurs by means of a microfluidic device, suitably designed, manufactured and tested, that is connected to an external experimental set-up. The fabrication process can be divided in two parts: at first it is described the manufacturing process of a filtering membrane, with holes of specific size that allow the passage of only certain cell types. Following the microfluidic device is fabricated through the mechanical micromilling. The membrane and the microdevice are suitably bonded and tested by means of an external connection with syringe pumps that inject blood samples at specific flow rates. The device is designed to separate blood cells and tumor cells only by using differences in size and shape. In particular during the first experiments red blood cells and platelets are sorted from white blood cells; in the other experiments red blood cells and platelets are separated from white blood cells and tumor cells. The microdevice has proven to be very efficient, in fact a capture efficiency of 99% is achieved. For this reason it could be used in identification and isolation of circulating tumor cells, a very rare cancer cell type whose presence in the bloodstream could be symptom of future solid tumor formation. The various experiments have also demonstrated that tumor cells survive even after the separation treatment, and then the suffered stress during the sorting process does not harm the biological sample.

  19. Midazolam sedates Passeriformes for field sampling but affects multiple venous blood analytes

    Directory of Open Access Journals (Sweden)

    Heatley JJ

    2015-01-01

    Full Text Available J Jill Heatley,1 Jennifer Cary,2,3 Lyndsey Kingsley,1 Hughes Beaufrere,4 Karen E Russell,5 Gary Voelker2,3 1Department of Small Animal Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, 2Department of Wildlife and Fisheries Sciences, 3Texas A&M Biodiversity Research and Teaching Collections, Texas A&M University, College Station, TX, USA; 4Health Sciences Centre, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada; 5Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, College Station, TX, USA Abstract: Feasibility and effect of midazolam administration on blood analytes and for sedation of Passeriformes being collected in a larger study of genetic biodiversity was assessed. Midazolam (5.6±2.7 mg/kg was administered intranasally prior to sampling, euthanasia, and specimen preparation of 104 passerine birds. Each bird was assessed for sedation score and then multiple analytes were determined from jugular blood samples using the i-STAT® point of care analyzer at “bird side”. Most birds were acceptably sedated, sedation became more pronounced as midazolam dose increased, and only a single bird died. Electrolyte concentrations and venous blood gas analytes were affected by midazolam administration while blood pH, packed cell volume, hemoglobin, and calculated hematocrit were not. Intranasal midazolam gives adequate sedation and is safe for short-term use in free-living Passeriformes. Based on venous blood analyte data, sedation of Passeriformes prior to handling appears to reduce stress but also produces venous blood gas differences consistent with hypoventilation relative to birds which were not given midazolam. Further study is recommended to investigate midazolam's continued use in free-living avian species. Studies should include safety, reversal and recovery, effect upon additional endogenous analytes, and compatibility with studies of ecology and toxicology

  20. Evaluation of a nested-pcr for Mycobacterium tuberculosis detection in blood and urine samples

    Directory of Open Access Journals (Sweden)

    Heidi Lacerda Alves da Cruz

    2011-03-01

    Full Text Available The polymerase chain reaction (PCR and its variations, such as the nested-PCR, have been described as promising techniques for rapid diagnosis of tuberculosis (TB. With the aim of evaluating the usefulness of a nested-PCR method on samples of blood and urine of patients suspected of tuberculosis we analyzed 192 clinical samples, using as a molecular target the insertion element IS6110 specific of M. tuberculosis genome. Nested-PCR method showed higher sensitivity in patients with extrapulmonary tuberculosis (47.8% and 52% in blood and urine when compared to patients with the pulmonary form of the disease (sensitivity of 29% and 26.9% in blood and urine, regardless of the type of biological sample used. The nested-PCR is a rapid technique that, even if not showing a good sensitivity, should be considered as a helpful tool especially in the extrapulmonary cases or in cases where confirmatory diagnosis is quite difficult to be achieved by routine methods. The performance of PCR-based techniques should be considered and tested in future works on other types of biological specimens besides sputum, like blood and urine, readily obtainable in most cases. The improving of M. tuberculosis nested-PCR detection in TB affected patients will give the possibility of an earlier detection of bacilli thus interrupting the transmission chain of the disease.

  1. Performance of an Early Infant Diagnostic Test, AmpliSens DNA-HIV-FRT, Using Dried Blood Spots Collected from Children Born to Human Immunodeficiency Virus-Infected Mothers in Ukraine

    Science.gov (United States)

    Shanmugam, Vedapuri; Azarskova, Marianna; Nguyen, Shon; Hurlston, Mackenzie; Sabatier, Jennifer; Zhang, Guoqing; Osmanov, Saladin; Ellenberger, Dennis; Yang, Chunfu; Vitek, Charles; Liulchuk, Maria; Nizova, Natalya

    2015-01-01

    An accurate accessible test for early infant diagnosis (EID) is crucial for identifying HIV-infected infants and linking them to treatment. To improve EID services in Ukraine, dried blood spot (DBS) samples obtained from 237 HIV-exposed children (≤18 months of age) in six regions in Ukraine in 2012 to 2013 were tested with the AmpliSens DNA-HIV-FRT assay, the Roche COBAS AmpliPrep/COBAS TaqMan (CAP/CTM) HIV-1 Qual test, and the Abbott RealTime HIV-1 Qualitative assay. In comparison with the paired whole-blood results generated from AmpliSens testing at the oblast HIV reference laboratories in Ukraine, the sensitivity was 0.99 (95% confidence interval [CI], 0.95 to 1.00) for the AmpliSens and Roche CAP/CTM Qual assays and 0.96 (95% CI, 0.90 to 0.98) for the Abbott Qualitative assay. The specificity was 1.00 (95% CI, 0.97 to 1.00) for the AmpliSens and Abbott Qualitative assays and 0.99 (95% CI, 0.96 to 1.00) for the Roche CAP/CTM Qual assay. McNemar analysis indicated that the proportions of positive results for the tests were not significantly different (P > 0.05). Cohen's kappa (0.97 to 0.99) indicated almost perfect agreement among the three tests. These results indicated that the AmpliSens DBS and whole-blood tests performed equally well and were comparable to the two commercially available EID tests. More importantly, the performance characteristics of the AmpliSens DBS test meets the World Health Organization EID test requirements; implementing AmpliSens DBS testing might improve EID services in resource-limited settings. PMID:26447114

  2. Performance of an Early Infant Diagnostic Test, AmpliSens DNA-HIV-FRT, Using Dried Blood Spots Collected from Children Born to Human Immunodeficiency Virus-Infected Mothers in Ukraine.

    Science.gov (United States)

    Chang, Joy; Tarasova, Tetyana; Shanmugam, Vedapuri; Azarskova, Marianna; Nguyen, Shon; Hurlston, Mackenzie; Sabatier, Jennifer; Zhang, Guoqing; Osmanov, Saladin; Ellenberger, Dennis; Yang, Chunfu; Vitek, Charles; Liulchuk, Maria; Nizova, Natalya

    2015-12-01

    An accurate accessible test for early infant diagnosis (EID) is crucial for identifying HIV-infected infants and linking them to treatment. To improve EID services in Ukraine, dried blood spot (DBS) samples obtained from 237 HIV-exposed children (≤18 months of age) in six regions in Ukraine in 2012 to 2013 were tested with the AmpliSens DNA-HIV-FRT assay, the Roche COBAS AmpliPrep/COBAS TaqMan (CAP/CTM) HIV-1 Qual test, and the Abbott RealTime HIV-1 Qualitative assay. In comparison with the paired whole-blood results generated from AmpliSens testing at the oblast HIV reference laboratories in Ukraine, the sensitivity was 0.99 (95% confidence interval [CI], 0.95 to 1.00) for the AmpliSens and Roche CAP/CTM Qual assays and 0.96 (95% CI, 0.90 to 0.98) for the Abbott Qualitative assay. The specificity was 1.00 (95% CI, 0.97 to 1.00) for the AmpliSens and Abbott Qualitative assays and 0.99 (95% CI, 0.96 to 1.00) for the Roche CAP/CTM Qual assay. McNemar analysis indicated that the proportions of positive results for the tests were not significantly different (P > 0.05). Cohen's kappa (0.97 to 0.99) indicated almost perfect agreement among the three tests. These results indicated that the AmpliSens DBS and whole-blood tests performed equally well and were comparable to the two commercially available EID tests. More importantly, the performance characteristics of the AmpliSens DBS test meets the World Health Organization EID test requirements; implementing AmpliSens DBS testing might improve EID services in resource-limited settings. PMID:26447114

  3. Development of a Melting Curve-Based Allele-Specific PCR of Apolipoprotein E (APOE) Genotyping Method for Genomic DNA, Guthrie Blood Spot, and Whole Blood.

    Science.gov (United States)

    Chen, Chia-Hsiang

    2016-01-01

    Genetic polymorphisms of apolipoprotein E (APOE) are associated with various health conditions and diseases, such as Alzheimer's disease, cardiovascular diseases, type 2 diabetes, etc. Hence, genotyping of APOE has broad applications in biomedical research and clinical settings, particularly in the era of precision medicine. The study aimed to develop a convenient and accurate method with flexible throughput to genotype the APOE polymorphisms. A melting curve-based allele-specific PCR method was developed to genotype two single nucleotide polymorphisms (SNPs) of APOE, i.e. rs429358 at codon 112 and rs7412 at codon 158. These two SNPs determine the genotype of APOE2, E3, and E4. PCR-based Sanger sequencing was used as the reference method for APOE genotyping. A 100% concordance rate was obtained in 300 subjects between the melting curve-based allele-specific PCR method and the Sanger sequencing method. This method was applied to a genetic association analysis of APOE and schizophrenia consisting of 711 patients with schizophrenia and 665 control subjects from Taiwan. However, no significant differences in the allele and genotype frequencies were detected between these two groups. Further experiments showed that DNA dissolved from blood collected on Guthrie filter paper and total blood cell lysate without DNA extraction can be used in the melting curve-based allele-specific PCR method. Thus, we suggest that this is a fast, accurate and robust APOE genotyping method with a flexible throughput and suitable for DNA template from different preparations. This convenient method shall meet the different needs of various research and clinical laboratories. PMID:27078154

  4. Tracer input for kinetic modelling of liver physiology determined without sampling portal venous blood in pigs

    Energy Technology Data Exchange (ETDEWEB)

    Winterdahl, Michael; Alstrup, Aage Kristian Olsen; Munk, Ole Lajord [Aarhus University Hospital, PET Centre, Aarhus C (Denmark); Keiding, Susanne; Soerensen, Michael [Aarhus University Hospital, PET Centre, Aarhus C (Denmark); Aarhus University Hospital, Department of Hepato-Gastroenterology V, Aarhus C (Denmark); Mortensen, Frank Viborg [Aarhus University Hospital, Department of Surgery L, Aarhus C (Denmark)

    2011-02-15

    Quantification of hepatic tracer kinetics by PET requires measurement of tracer input from the hepatic artery (HA) and portal vein (PV). We wished to develop a method for estimating dual tracer input without the necessity to sample PV blood. Pigs weighing 40 kg were given bolus doses of C{sup 15}O (CO), 2-[{sup 18}F]fluoro-2-deoxy-D-glucose (FDG), [{sup 11}C]-methylglucose (MG), 2-[{sup 18}F]fluoro-2-deoxy-D-galactose (FDGal) or H{sub 2} {sup 15}O (H{sub 2}O). Tracer concentration 3-min time courses were measured in the femoral artery and PV by blood sampling. Blood flow was measured in the HA and PV using flow-meters. A model for transfer of tracer through the splanchnic circulation was used to estimate values of a tracer-specific model parameter {beta}. Tracer-specific mean values of {beta} were used to estimate tracer concentration time courses in the PV from the measured arterial concentration. A model-derived dual-input was calculated using the mean HA flow fraction (0.25) and validated by comparison of the use of the measured dual-input and a kinetic model with a fixed ''true'' K{sub 1} {sup true}, i.e. clearance of tracer from blood to liver cells. The rank order of the means of {beta} was CO < FDG {approx} MG < FDGal < H{sub 2}O, reflecting their different splanchnic mean transit times. Estimated K{sub 1} {sup est} was not significantly different from ''true'' K{sub 1} {sup true}. The hepatic dual tracer input, which is of great importance for the assessment of processes such as transfer across the plasma-hepatocyte membrane or hepatic blood perfusion, can be well approximated in pigs without the necessity to sample PV blood and measure hepatic blood flow; only arterial blood sampling is needed. (orig.)

  5. Genetic characterization of atypical Mansonella (Mansonella) ozzardi microfilariae in human blood samples from northeastern Peru.

    Science.gov (United States)

    Marcos, Luis A; Arrospide, Nancy; Recuenco, Sergio; Cabezas, Cesar; Weil, Gary J; Fischer, Peter U

    2012-09-01

    DNA sequence comparisons are useful for characterizing proposed new parasite species or strains. Microfilariae with an atypical arrangement of nuclei behind the cephalic space have been recently described in human blood samples from the Amazon region of Peru. Three blood specimens containing atypical microfilariae were genetically characterized using three DNA markers (5S ribosomal DNA, 12S ribosomal DNA, and cytochrome oxidase I). All atypical microfilariae were clustered into the Mansonella group and indistinguishable from M. ozzardi based on these DNA markers. PMID:22826497

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

    DEFF Research Database (Denmark)

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

    2008-01-01

    impact on the quality of results, we wanted to study which combination of transport conditions could fulfil our pre-defined goals for maximum allowable error. METHODS: Samples from 406 patients from nine general practitioners (GPs) in two Danish counties were sent to two hospitals for analyses, during......, centrifuged and separated at the doctor's office within 45-60 min. This sample was considered as the best estimate of a comparison value. RESULTS: The pre-set quality goals were fulfilled for all the investigated components for samples transported to hospital by courier either as whole blood or as "on gel...

  7. Hot-spot detection and calibration of a scanning thermal probe with a noise thermometry gold wire sample

    NARCIS (Netherlands)

    Gaitas, A.; Wolgast, S.; Covington, E.; Kurdak, C.

    2013-01-01

    Measuring the temperature profile of a nanoscale sample using scanning thermal microscopy is challenging due to a scanning probe's non-uniform heating. In order to address this challenge, we have developed a calibration sample consisting of a 1-μm wide gold wire, which can be heated electrically by

  8. An investigation of dust lead sampling locations and children's blood lead levels.

    Science.gov (United States)

    Wilson, Jonathan; Dixon, Sherry; Galke, Warren; McLaine, Patricia

    2007-01-01

    The objective of this study is to provide guidance on where to collect dust lead wipe samples in homes to best characterize the risk of a resident child having a blood lead level at or above the CDC level of concern (10 microg/dl). In 1998, the Milwaukee Health Department enrolled 72 children living in pre-1950 buildings: 34 had elevated (i.e., > or = 10 microg/dl) blood lead levels (EBL); and 38 had non-elevated blood lead levels (non-EBL). This study explored dust lead sampling locations by examining loading differences between homes where children with EBL and non-EBL lived. Floor, windowsill, and window trough samples were collected in the living room, kitchen, bathroom, and child's bedroom and play area. Floor samples were collected at four locations: room entry; center of the room; under a window; and against the wall opposite the window (perimeter). Geometric mean floor dust lead levels were generally two to three times higher in homes of EBL children than homes of non-EBL children. Sampling the floor at the room entry or center is preferable to sampling under the window or from the perimeter of the room. When the central floor average was used, the room combinations that had the greatest differences between homes of EBL children and non-EBL children all included a sample from the child's bedroom and excluded the bathroom. When the entry floor average was used, the greatest differences also excluded bathrooms, but otherwise included a mix of all of the other rooms. Window samples did not distinguish where children with EBLs versus non-EBLs resided. This paper is based on Milwaukee alone, so generalizing results to other locations should be done with caution. PMID:16823397

  9. Highly Effective DNA Extraction Method from Fresh, Frozen, Dried and Clotted Blood Samples

    Directory of Open Access Journals (Sweden)

    Jaleh Barar

    2011-09-01

    Full Text Available Introduction: Today, with the tremendous potential of genomics and other recent advances in science, the role of science to improve reliable DNA extraction methods is more relevant than ever before. The ideal process for genomic DNA extraction demands high quantities of pure, integral and intact genomic DNA (gDNA from the sample with minimal co-extraction of inhibitors of downstream processes. Here, we report the development of a very rapid, less-hazardous, and high throughput protocol for extracting of high quality DNA from blood samples. Methods: Dried, clotted and ethylene diamine tetra-acetic acid (EDTA treated fresh and frozen blood samples were extracted using this method in which the quality and integrity of the extracted DNA were corroborated by agarose gel electrophoresis, PCR reaction and DNA digestion using restricted enzyme. The UV spectrophotometric and gel electrophoresis analysis resulted in high A260/A280 ratio (>1.8 with high intactness of DNA. Results: PCR and DNA digestion experiments indicated that the final solutions of extracted DNA contained no inhibitory substances, which confirms that the isolated DNA is of good quality. Conclusion: The high quality and quantity of current method, no enzymatic processing and accordingly its low cost, make it appropriate for DNA extraction not only from human but also from animal blood samples in any molecular biology labs.

  10. Is intrapartum fetal blood sampling a gold standard diagnostic tool for fetal distress?

    Science.gov (United States)

    Mahendru, Amita A; Lees, Christoph C

    2011-06-01

    Developed in 1960s, cardiotocography is a screening test and fetal blood sampling (FBS) is an adjunctive, diagnostic technique to detect fetal hypoxia. A fetal blood sample pH value of less than 7.20 has a higher specificity than a pathological CTG to predict low Apgar score at 1 min. Though with a pathological CTG and despite a normal FBS pH value the risk of delivering a hypoxic infant is 30-50%, FBS has assumed considerable importance in purportedly reducing unnecessary obstetric intervention. The evidence for this is weak: the use of FBS with CTG has been shown to reduce operative vaginal deliveries though not Caesarean sections due to fetal distress. There is no difference in the umbilical artery pH at delivery with the use of intermittent FBS with CTG compared to CTG alone. FBS is an invasive procedure: obtaining an adequate blood sample is often difficult and the pH results are affected by handling of the sample, aerobic contamination and processing. Validation of intrapartum FBS requires that the pH and other values obtained are compared to a 'gold standard' technique. Although FBS has been compared to other tests such as scalp lactate, pulse oximetry, fetal ECG waveform analysis, and central haemodynamics in labouring rhesus monkeys, none of these can be considered as 'gold standard'. In the light of the existing evidence, the role of intrapartum FBS as a gold standard diagnostic technique is unproven. PMID:21300427

  11. Transcutaneous monitoring of blood gases: is it comparable with arterialized earlobe sampling?

    Science.gov (United States)

    Dawson, S; Cave, C; Pavord, I; Potter, J F

    1998-03-01

    Researchers are increasingly looking for reliable non-invasive methods of assessing blood gas concentrations, and several new techniques have recently become available. Values derived using arterialized earlobe samples have been found to be comparable with conventional arterial samples, and recent studies have compared transcutaneous blood gas analysis with the traditional arterial samples and found a reasonable level of agreement in particular for the partial pressure of carbon dioxide. There are no data comparing oxygen and carbon dioxide partial pressures (pO2, pCO2) derived from arterialized samples with one of the newer transcutaneous techniques. We therefore simultaneously studied arterialized earlobe blood gas samples and values for pO2 and pCO2 obtained by a transcutaneous monitor (TINA, Radiometer, Copenhagen) in 26 subjects with varying blood gas values. There was a close agreement between the two methods for assessment of pCO2 [mean difference (95% C.I.) between transcutaneous and earlobe values 0.25 kPa (-0.004, 0.5 kPa)], but not for pO2 [1.71 kPa (0.35, 3.07 kPa)]. Similarly, the limits of agreement were narrow for pCO2 compared to those for pO2 (-0.98, 1.47 kPa and -6.44, 3.02 kPa respectively). We conclude that transcutaneous measurement of pCO2 using the TINA is acceptable in the research setting, whereas assessment of pO2 cannot reliably be made using this technique. PMID:9692127

  12. DETERMINATION OF LEPTIN EXPRESSION IN BEEF CATTLE BLOOD SAMPLES USED BY RTQ PCR

    Directory of Open Access Journals (Sweden)

    Miroslava Kačániová

    2011-08-01

    Full Text Available The aim of our study was to detect the presence and concentration of leptin in different breeds of cattle by PCR and Real time PCR method. Blood of different breeds of bulls was used as biological material in our experiments: Slovak pied cattle (10 samples, Blondaquitane × Pinzgau breed (10 samples and Holstein breed (10 samples. The presence of leptin was detected in all samples based on the results of molecular-genetic detection of leptin gene. The average concentration of leptinin 30 samples of beef cattle was 22.1477 μg.μl-1. Differences in leptin concentrations were statistically significant between Holstein breed and Slovak pied cattle and between Slovak pied cattle and Blondaquitane × Pinzgau breed.

  13. Dried blood spots versus plasma for the quantitation of HIV-1 RNA using a real-Time PCR, m2000rt assay.

    Science.gov (United States)

    Vidya, Madhavan; Saravanan, Shanmugam; Rifkin, Samara; Solomon, Sunil S; Waldrop, Greer; Mayer, Kenneth H; Solomon, Suniti; Balakrishnan, Pachamuthu

    2012-05-01

    High costs and stringent requirements for storage and transport of plasma, often prohibit the availability of HIV viral load quantification in resource-limited settings. Dried blood spots (DBS) represent a better method of specimen collection that removes many of these logistical and technical limitations. The present study aimed to assess the performance of the Abbott m2000rt assay for quantitation of HIV-1 RNA in DBS specimens using plasma as a "gold standard" for comparison. One hundred paired DBS and plasma specimens were collected from patients infected with HIV, who were 18 years and older during routine visits to a private tertiary-care clinic in Chennai, India. HIV-1 RNA was extracted manually and then detected using the m2000rt assay. The mean plasma and DBS viral loads were 4.27 (95% CI: 2.65, 5.88) and 4.14 (95% CI: 1.96, 6.32) log copies/mL, respectively. The overall sensitivity of DBS reached 95%; with sensitivities of 62%, 88% and 100% when stratified by viral load ranges of ≤1000, 1000-3000 and >3000 copies/mL, respectively. An over quantitation of the viral load with DBS was observed in pairs with plasma viral loadfailure in resource-limited settings. PMID:22401801

  14. Lead and cadmium determinations by atomic absorption technique in biological samples: blood, placenta and umbilical cord

    International Nuclear Information System (INIS)

    In order to determine the possibility contamination of lead and cadmium in pregnant women living in the mining-smelting city of La Oroya in Peru, lead and cadmium concentrations were assessed in maternal blood (pre-birth), umbilical cord blood and placental tissue. Forty deliveries with normal evolution were evaluated between October 2002 and January 2003. Samples were analyzed by atomic absorption on a graphite furnace at the Peruvian Institute of Nuclear Energy (IPEN) laboratories. Results are summarized as follows: a) Mean lead concentrations in maternal blood (MB), umbilical cord blood (UCB) and placental tissue (PT) were 27.23 μg/dL, 18.48 μg/dL and 363.97 μg/100g, respectively; b) Mean cadmium concentrations in MB, UCB and PT were 8.82 μg/dL, 12,0 μg/dL and 104,44 μg/100g, respectively; c) The correlation coefficient between lead concentration in maternal blood and umbilical cord was 0.122; d). The correlation coefficient of cadmium concentration between MB and UCB was 0.223; e). The correlation coefficient of lead concentration between MB and PT was 0.189; f). The correlation coefficient of cadmium concentration between MB and PT was 0.633. Trans-placental transport of lead was 67.84% (27,23 μg/dL in MB vs. 18.48 μg/dL in UCB); whereas in the case of cadmium, the concentration in UC (12,00 μg/dL) was greater than in MB (8.82 μg/dL.). These results could indicate that the placenta acts as a barrier trapping lead and cadmium. This barrier is efficient for lead since the concentration in cord blood is inferior to maternal blood but it is less efficient for cadmium. (author)

  15. Performance evaluation of continuous blood sampling system for PET study. Comparison of three detector-systems

    Energy Technology Data Exchange (ETDEWEB)

    Matsumoto, Keiichi; Shinoda, Masaki; Sakamoto, Setsu; Senda, Michio [Inst. of Biomedical Research and Innovation, Kobe (Japan); Yamamoto, Seiichi [Kobe City Coll. of Technology (Japan); Tarutani, Kazumasa; Minato, Kotaro [Nara Inst. of Science and Technology, Ikoma (Japan). Graduate School of Information Science

    2002-11-01

    To measure cerebral blood flow with {sup 15}O PET, it is necessary to measure the time course of arterial blood radioactivity. We examined the performance of three different types of continuous blood sampling system. Three kinds of continuous blood sampling system were used: a plastic scintillator-based beta detector (conventional beta detector (BETA)), a bismuth germinate (BGO)-based coincidence gamma detector (Pico-count flow-through detector (COINC)) and a Phoswich detector (PD) composed by a combination of plastic scintillator and BGO scintillator. Performance of these systems was evaluated for absolute sensitivity, count rate characteristic, sensitivity to background gamnra photons, and reproducibility for nylon tube geometry. The absolute sensitivity of the PD was 0.21 cps/Bq for {sup 68}Ga positrons at the center of the detector. This was approximately three times higher than BETA, two times higher than COINC. The value measured with BETA was stable, even when background radioactivity was increased. The count rate characteristic of the PD and COINC was linear up to 8 kcps. The reproducibility of sensitivity for nylon tube geometry of COINC was the smallest (coefficient of variation (C.V.)=1.00%) among the three. PD was the weights the least (3.5 kg) among the three, which is convenient for clinical use. Each detector has unique characteristics derived from its own structure. Although the performance of all three detectors meets clinical requirement, PD had the highest physical performance. (author)

  16. Performance evaluation of continuous blood sampling system for PET study. Comparison of three detector-systems

    CERN Document Server

    Matsumoto, K; Sakamoto, S; Senda, M; Yamamoto, S; Tarutani, K; Minato, K

    2002-01-01

    To measure cerebral blood flow with sup 1 sup 5 O PET, it is necessary to measure the time course of arterial blood radioactivity. We examined the performance of three different types of continuous blood sampling system. Three kinds of continuous blood sampling system were used: a plastic scintillator-based beta detector (conventional beta detector (BETA)), a bismuth germinate (BGO)-based coincidence gamma detector (Pico-count flow-through detector (COINC)) and a Phoswich detector (PD) composed by a combination of plastic scintillator and BGO scintillator. Performance of these systems was evaluated for absolute sensitivity, count rate characteristic, sensitivity to background gamnra photons, and reproducibility for nylon tube geometry. The absolute sensitivity of the PD was 0.21 cps/Bq for sup 6 sup 8 Ga positrons at the center of the detector. This was approximately three times higher than BETA, two times higher than COINC. The value measured with BETA was stable, even when background radioactivity was incre...

  17. Bier spots

    Directory of Open Access Journals (Sweden)

    Ahu Yorulmaz,

    2015-10-01

    Full Text Available Also called as physiologic anemic macules, Bier spots are small, hypopigmented irregularly shaped macules against a background of diffuse erythema, which creates an appearance of speckled vascular mottling of the skin. Bier spots most commonly appear on distal portions of the limbs though there are case reports describing diffuse involvement, which also affect trunk and mucous membranes of the patient. Although the exact pathophysiological mechanisms underlying Bier spots still need to be elucidated, Bier spots have been suggested to be a vascular anomaly caused by vasoconstriction of small vessels. In addition, several diseases have been proposed to be associated with Bier spots, including scleroderma renal crisis, cryoglobulinemia, Peutz-Jeghers syndrome, alopecia areata and hypoplasia of the aorta, although it has not been shown whether these associations are casual or coincidental. The clinical presentation of Bier spots is quite typical. These tiny whitish macules easily become prominent when the affected limb is placed in a dependent position and fade away when the limb is raised. Here we report a case of Bier spots in a 32-year-old male patient with characteristical clinical manifestations.

  18. Methods for extracting genomic DNA from whole blood samples: current perspectives

    OpenAIRE

    Griffiths, Lyn

    2014-01-01

    Diego Chacon-Cortes, Lyn R Griffiths Genomics Research Centre, Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, QLD, Australia Abstract: Deoxyribonucleic acid (DNA) extraction has considerably evolved since it was initially performed back in 1869. It is the first step required for many of the available downstream applications used in the field of molecular biology. Whole blood samples are one of the main sources used to obtain DNA, and there a...

  19. Identification of Nocardiopsis dassonvillei in a Blood Sample from a Child

    Directory of Open Access Journals (Sweden)

    Flavio Lejbkowicz

    2005-01-01

    Full Text Available Nocardiopsis dassonvillei is an environmental aerobic actinomycete producing a funguslike mycelium and aerial hyphae. Here we report the first Nocardiopsis dassonvillei isolated from a blood sample from a 3-year-old child hospitalized with fever, respiratory difficulty and cough. To the best of our knowledge this is the first time this organism has been detected in the BacT/Alert system. This Nocardiopsis was designated Nocardiopsis dassonvillei based on morphological and physiological tests.

  20. ANALYSIS OF CHROMIUM BIOACCUMULATION IN BLOOD, URINE AND NAIL SAMPLES OF CHROMATE FACTORY WORKERS

    Directory of Open Access Journals (Sweden)

    M. JOB GOPINATH

    2006-01-01

    Full Text Available Industrial play a major role in country's economy. Air pollution is an occupational health problem in industries. Generaly, there are more man made pollution in the air of heavy industries and major cities around the industries, Exposure as inhalation represents one of the major routes by which the body can be exposed by accident or design to foreign materials. One having entered to the respiratory tract, inhaled materails may be readly absorbed or may react directly with the alveolar epithelium and entered in to the blood steam. These air pollution induce many changes in physiological and biochemical process. The present study was contucted on chromate industrial workers as these workers were repeatedly exposed to pollutants in chromium factors (Tamil Nadu Chromate and Chemical Ltd., Ranipet industrial Town, Tamil Nadu. The bioaccumulation of chromium was estimated from one hundred samples of blood and urine as well as twenty five samples of nails collected from TCC industrial workers and control subject. The workers and control subject were randomly selected. Estimation was carried out by Atomic Absorption spectrometry. From the results it was obeserved tha the accumulation of heavy metal chromium in the blood. urine and nail samples of chromate factory worker are above the normal ranges. Results are statistically significant.

  1. Temporal variability in urinary excretion of bisphenol A and seven other phenols in spot, morning, and 24-h urine samples

    DEFF Research Database (Denmark)

    Lassen, Tina Harmer; Frederiksen, Hanne; Jensen, Tina Kold;

    2013-01-01

    (ICCs). More than 70% of the urine samples had detectable levels of BPA, triclosan (TCS), benzophenone-3 (BP-3) and sum of 2,4-dichlorophenol and 2,5-dichlorophenol (σDCP). We found low to moderate ICCs for BPA (0.10-0.42) and σDCP (0.39-0.72), whereas the ICCs for BP-3 (0.69-0.80) and TCS (0...

  2. Application of Atomic Dielectric Resonance Spectroscopy for the screening of blood samples from patients with clinical variant and sporadic CJD

    Directory of Open Access Journals (Sweden)

    Ironside James W

    2007-08-01

    Full Text Available Abstract Background Sub-clinical variant Creutzfeldt-Jakob disease (vCJD infection and reports of vCJD transmission through blood transfusion emphasise the need for blood screening assays to ensure the safety of blood and transplanted tissues. Most assays aim to detect abnormal prion protein (PrPSc, although achieving required sensitivity is a challenge. Methods We have used innovative Atomic Dielectric Resonance Spectroscopy (ADRS, which determines dielectric properties of materials which are established by reflectivity and penetration of radio/micro waves, to analyse blood samples from patients and controls to identify characteristic ADR signatures unique to blood from vCJD and to sCJD patients. Initial sets of blood samples from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors were screened as training samples to determine group-specific ADR characteristics, and provided a basis for classification of blinded sets of samples. Results Blood sample groups from vCJD, sCJD, non-CJD neurological diseases and normal healthy adults (blood donors screened by ADRS were classified with 100% specificity and sensitivity, discriminating these by a co-variance expert analysis system. Conclusion ADRS appears capable of recognising and discriminating serum samples from vCJD, sCJD, non-CJD neurological diseases, and normal healthy adults, and might be developed to provide a system for primary screening or confirmatory assay complementary to other screening systems.

  3. Sensitivity of PCR assays for murine gammaretroviruses and mouse contamination in human blood samples.

    Directory of Open Access Journals (Sweden)

    Li Ling Lee

    Full Text Available Gammaretroviruses related to murine leukemia virus (MLV have variously been reported to be present or absent in blood from chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME patients and healthy controls. Using subjects from New York State, we have investigated by PCR methods whether MLV-related sequences can be identified in nucleic acids isolated from whole blood or from peripheral blood mononuclear cells (PBMCs or following PBMC culture. We have also passaged the prostate cancer cell line LNCaP following incubation with plasma from patients and controls and assayed nucleic acids for viral sequences. We have used 15 sets of primers that can effectively amplify conserved regions of murine endogenous and exogenous retrovirus sequences. We demonstrate that our PCR assays for MLV-related gag sequences and for mouse DNA contamination are extremely sensitive. While we have identified MLV-like gag sequences following PCR on human DNA preparations, we are unable to conclude that these sequences originated in the blood samples.

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

    Directory of Open Access Journals (Sweden)

    Dekker Aldo

    2011-02-01

    Full Text Available Abstract Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 individual dogs, representing 73 different breeds and a group of 40 mixed breed dogs, categorized into healthy dogs and dogs with internal and hematological diseases, and dogs that underwent a surgical procedure. GeNorm analysis revealed that a combination of 5 to 6 of the most stably expressed genes constituted a stable normalizing factor. Evaluation of the expression revealed different ranking of reference genes in Normfinder and GeNorm. The disease category and the white blood cell count significantly affected reference gene expression. Conclusions The discrepancy between the ranking of reference genes in this study by Normfinder and Genorm can be explained by differences between the experimental groups such as "disease category" and "WBC count". This stresses the importance of assessing the expression stability of potential reference genes for gene experiments in canine whole blood anew for each specific experimental condition.

  5. A Simple, Inexpensive and Safe Method for DNA Extraction of Frigid and Clotted Blood Samples

    Directory of Open Access Journals (Sweden)

    Nasrin Mohammadi

    2015-07-01

    Full Text Available Background: Extraction of blood genomicDNAis one of the main approaches for clinical and molecular biology studies. Although several methods have been developed for extraction of blood genomic DNA, most of these methods consume long time and use expensive chemicals such as proteinase K and toxic organic solvent such as phenol and chloroform. The objective of this study was to developed easy and safe method forDNAextraction from clotted and frozen whole blood. This method has many advantages: time reducing, using inexpensive materials, without phenol and chloroform, achieving of high molecular weight and good quality genomicDNA.Materials and Methods: DNA extraction was performed by two methods (new and phenol-chloroform method. Then quantity and quality parameters were evaluated by 1% agarose gel electrophoresis, Nano drop analysis and efficiency of Polymerase Chain Reaction (PCR.Results: Extracted DNA from 500μL of blood samples were 457.7ng/μl and 212ng/μL and their purity (OD260/OD280 were 1.8 and 1.81 for new recommended and phenol–chloroform methods respectively. The PCR results indicated that D16S539 and CSF1PO loci were amplified.Conclusion: These results shown that this method is simple, fast, safe and most economical.

  6. An improved, PCR-based strategy for the detection of Trypanosoma cruzi in human blood samples.

    Science.gov (United States)

    Ribeiro-dos-Santos, G; Nishiya, A S; Sabino, E C; Chamone, D F; Saez-Alquézar, A

    1999-10-01

    Attempts were made to improve the PCR-based detection of Trypanosoma cruzi in blood samples, primarily for screening blood donors. Samples were obtained from candidate donors who were reactive in one or two of three serological tests for Chagas disease (and therefore considered 'indeterminate') or in all three tests (3+). Each sample was then examined using three different, PCR-based techniques: 'PCR-I' (in which the target DNA is a nuclear repetitive sequence); 'PCR-II' [amplifying a conserved region of the T. cruzi kinetoplast DNA (kDNA)]; and 'PCR-III' (a new strategy in which the target kDNA is amplified by 'nested' PCR). Among the samples from 3+ individuals, PCR-I, PCR-II and PCR-III amplified two (3.8%) out of 52, four (4.5%) out of 88, and 27 (25.7%) out of 105 samples tested, respectively. Seven, 69 and 70 samples from 'indeterminate' subjects were tested by PCR-I, PCR-II and PCR-III, respectively; there was not a single positive result by PCR-I or PCR-II, but three (4.3%) of the samples tested by PCR-III were positive. In a reconstruction experiment, in conditions in which PCR-I and PCR-II could not detect 10,000 parasites/ml, PCR-III was able to detect one parasite/ml. Although all three PCR-based strategies examined had rather poor sensitivities, PCR-III was far more sensitive than PCR-I or PCR-II. PMID:10715696

  7. Behavior of optical properties of coagulated blood sample at 633 nm wavelength

    Science.gov (United States)

    Morales Cruzado, Beatriz; Vázquez y Montiel, Sergio; Delgado Atencio, José Alberto

    2011-03-01

    Determination of tissue optical parameters is fundamental for application of light in either diagnostics or therapeutical procedures. However, in samples of biological tissue in vitro, the optical properties are modified by cellular death or cellular agglomeration that can not be avoided. This phenomena change the propagation of light within the biological sample. Optical properties of human blood tissue were investigated in vitro at 633 nm using an optical setup that includes a double integrating sphere system. We measure the diffuse transmittance and diffuse reflectance of the blood sample and compare these physical properties with those obtained by Monte Carlo Multi-Layered (MCML). The extraction of the optical parameters: absorption coefficient μa, scattering coefficient μs and anisotropic factor g from the measurements were carried out using a Genetic Algorithm, in which the search procedure is based in the evolution of a population due to selection of the best individual, evaluated by a function that compares the diffuse transmittance and diffuse reflectance of those individuals with the experimental ones. The algorithm converges rapidly to the best individual, extracting the optical parameters of the sample. We compare our results with those obtained by using other retrieve procedures. We found that the scattering coefficient and the anisotropic factor change dramatically due to the formation of clusters.

  8. Development of a One-Step Probe Based Molecular Assay for Rapid Immunodiagnosis of Infection with M. tuberculosis Using Dried Blood Spots

    DEFF Research Database (Denmark)

    Blauenfeldt, Thomas; Heyckendorf, Jan; Graff Jensen, Sidse;

    2014-01-01

    detection. AIM: To develop a robust IP-10 based molecular assay for the diagnosis of infection with M. tubercuolsis from whole blood and DBS. METHOD: We developed a one-step probe based multiplex RT-qPCR assay for detecting IP-10 and IFN-γ mRNA expression from whole blood and DBS samples. The assay......RNA upregulation was detectable at 4 hours after stimulation (6 fold upregulation) peaking at 8 hours (108 fold upregulation). IFN-γ expression occurred in concert but levels were lower (peak 6.7 fold upregulation). IP-10 gene expression level was significantly higher in patients with tuberculosis (median 31.......2, IQR 10.7-67.0) and persons with latent tuberculosis infection (LTBI) (41.2, IQR 9.8-64.9) compared to healthy controls (1.6, IQR 1.1-2.4; p

  9. Analysis of hemoglobin adducts from acrylamide, glycidamide, and ethylene oxide in paired mother/cord blood samples from Denmark

    DEFF Research Database (Denmark)

    von Stedingk, Hans; Vikström, Anna C; Rydberg, Per;

    2011-01-01

    for analysis of Hb adducts by liquid chromatography-mass spectrometry, the adduct FIRE procedure, was applied to measurements of adducts from these compounds in maternal blood samples (n = 87) and umbilical cord blood samples (n = 219). The adduct levels from the three compounds, acrylamide, glycidamide.......20-0.73) for glycidamide, and 0.43 (range 0.17-1.34) for ethylene oxide. In vitro studies with acrylamide and glycidamide showed a lower (0.38-0.48) rate of adduct formation with Hb in cord blood than with Hb in maternal blood, which is compatible with the structural differences in fetal and adult Hb. Together...... of the measured adducts in cord blood with gestational age was observed, which may reflect the gestational age-related change of the cord blood Hb composition toward a higher content of adult Hb. The results suggest that the Hb adduct levels measured in cord blood reflect the exposure to the fetus during...

  10. Physiological and Pathological Impact of Blood Sampling by Retro-Bulbar Sinus Puncture and Facial Vein Phlebotomy in Laboratory Mice

    DEFF Research Database (Denmark)

    Teilmann, Anne Charlotte; Nygaard Madsen, Andreas; Holst, Birgitte;

    2014-01-01

    time points, and the samples were analyzed for plasma corticosterone. Body weights were measured at the day of blood sampling and the day after blood sampling, and the food consumption was recorded automatically during the 24 hours post-procedure. At the end of study, cheeks and orbital regions were...... weight following blood sampling, but the body weight loss was higher in mice subjected to facial vein phlebotomy. The food consumption was not significantly different between the two groups. At gross necropsy, subcutaneous hematomas were found in both groups and the histopathological analyses revealed...

  11. Evaluation of Chromosomal Disorders in Tissue and Blood Samples in Patients with Oral Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    A. Parvaneroo

    2004-12-01

    Full Text Available Statement of Problem: Many studies have indicated that genetic disturbances are common findings in patients with Oral Squamous Cell Carcinoma (OSCC. Identification of these changes can be helpful in diagnostic procedures of these tumors.Purpose: The aim of this study was to appraise the chromosomal disorders in blood and tissue patients with OSCC.Methods and Materials: In this descriptive study, the study group consisted of all OSCC patients who were referred to the Faculty of Dentistry, Tehran University of Medical Sciences, Maxillofacial Surgery Clinic of Shariati Hospital, and Amir Aalam Hospital fromSeptember 2000 to November 2002. In order to study chromosomal disorders in the peripheral blood lymphocytes, 5 mL of blood was obtained from each patient In patients with the large lesion, a piece of involved tissue were obtained and cultured for 24 hours.This led to 29 blood samples and 16 tissue specimens and any relation between OSCC and age, sex, smoking and alcohol use were evaluated.Results: In this study, OSCC was more common in males than in females (3 to 5. 31% of our patients were smokers, and one had a history of alcoholic consumption. There was an increase in incidence of OSCC with age. In this study, all patients had numerical(aneuploidy, polyploidy and structural chromosomal disorders (double minute, fragment,breakage and dicentric. There was significant difference between blood and tissue chromosomal disorders (aneuploidy, polyploidy,breakage in OSCC patients.Conclusion: It can be concluded that chromosomes in patients with OSCC might show some genetic aberration and evaluation of involved tissue might be better way for determining this disorders.

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

    DEFF Research Database (Denmark)

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

    2012-01-01

    to handle 1,073 of 1,092 (98.3%) samples of whole blood from forensic material, including postmortem samples, without any need for repeating sample preparation. Only three samples required special treatment such as dilution. The addition of internal and calibration standards were validated by pipetting...

  13. Comparison of Ahlstrom grade 226, Munktell TFN, and Whatman 903 filter papers for dried blood spot specimen collection and subsequent HIV-1 load and drug resistance genotyping analysis.

    Science.gov (United States)

    Rottinghaus, Erin; Bile, Ebi; Modukanele, Mosetsanagape; Maruping, Maruping; Mine, Madisa; Nkengasong, John; Yang, Chunfu

    2013-01-01

    Dried blood spots (DBS) collected onto filter paper have eased the difficulty of blood collection in resource-limited settings. Currently, Whatman 903 (W-903) filter paper is the only filter paper that has been used for HIV load and HIV drug resistance (HIVDR) testing. We therefore evaluated two additional commercially available filter papers, Ahlstrom grade 226 (A-226) and Munktell TFN (M-TFN), for viral load (VL) testing and HIVDR genotyping using W-903 filter paper as a comparison group. DBS specimens were generated from 344 adult patients on antiretroviral therapy (ART) in Botswana. The VL was measured with NucliSENS EasyQ HIV-1 v2.0, and genotyping was performed for those specimens with a detectable VL (≥ 2.90 log(10) copies/ml) using an in-house method. Bland-Altman analysis revealed a strong concordance in quantitative VL analysis between W-903 and A-226 (bias = -0.034 ± 0.246 log(10) copies/ml [mean difference ± standard deviation]) and W-903 and M-TFN (bias = -0.028 ± 0.186 log(10) copies/ml) filter papers, while qualitative VL analysis for virological failure determination, defined as a VL of ≥ 3.00 log(10) copies/ml, showed low sensitivities for A-266 (71.54%) and M-TFN (65.71%) filter papers compared to W-903 filter paper. DBS collected on M-TFN filter paper had the highest genotyping efficiency (100%) compared to W-903 and A-226 filter papers (91.7%) and appeared more sensitive in detecting major HIVDR mutations. DBS collected on A-226 and M-TFN filter papers performed similarly to DBS collected on W-903 filter paper for quantitative VL analysis and HIVDR detection. Together, the encouraging genotyping results and the variability observed in determining virological failure from this small pilot study warrant further investigation of A-226 and M-TFN filter papers as specimen collection devices for HIVDR monitoring surveys.

  14. Capillary blood sampling: national recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

    Science.gov (United States)

    Krleza, Jasna Lenicek; Dorotic, Adrijana; Grzunov, Ana; Maradin, Miljenka

    2015-01-01

    Capillary blood sampling is a medical procedure aimed at assisting in patient diagnosis, management and treatment, and is increasingly used worldwide, in part because of the increasing availability of point-of-care testing. It is also frequently used to obtain small blood volumes for laboratory testing because it minimizes pain. The capillary blood sampling procedure can influence the quality of the sample as well as the accuracy of test results, highlighting the need for immediate, widespread standardization. A recent nationwide survey of policies and practices related to capillary blood sampling in medical laboratories in Croatia has shown that capillary sampling procedures are not standardized and that only a small proportion of Croatian laboratories comply with guidelines from the Clinical Laboratory Standards Institute (CLSI) or the World Health Organization (WHO). The aim of this document is to provide recommendations for capillary blood sampling. This document has been produced by the Working Group for Capillary Blood Sampling within the Croatian Society of Medical Biochemistry and Laboratory Medicine. Our recommendations are based on existing available standards and recommendations (WHO Best Practices in Phlebotomy, CLSI GP42-A6 and CLSI C46-A2), which have been modified based on local logistical, cultural, legal and regulatory requirements. We hope that these recommendations will be a useful contribution to the standardization of capillary blood sampling in Croatia.

  15. Identification of a suitable internal control for fluorescence analysis on canine peripheral blood samples.

    Science.gov (United States)

    Riondato, F; Martini, V; Poggi, A; Rota, A; Comazzi, S; Sulce, M; Bruno, B; Borrelli, A; Miniscalco, B

    2016-04-01

    Reliable detection of fluorescence intensity (FI) by flow cytometry (FC) is fundamental. FI depends on instrument settings and sample processing procedures: thus, measurements should be done using internal controls with known FI. Commercially available beads-based standards are expensive, thus reducing their usability in the veterinary practice. Cell subsets with stable mean FI (MFI) within the population have been proposed as acceptable surrogates in human medicine. In veterinary medicine, no data exist about stability of antigen expression among different subjects or upon sample storage. The aim of the present study was to evaluate MFI variability of main lymphocytes antigens among the lymphoid cells within each subject, among different subjects, and upon 24-h storage, in order to identify the antigen most suitable as stable internal control in MFI analyses. Peripheral blood samples from 18 healthy dogs were analysed by FC within 3h from sampling to assess the expression of CD3, CD5, CD4, CD8, CD21 and cyCD79b using conjugated monoclonal antibodies. Analyses were restricted to the lymphoid population. Fluorescent microbeads were added to each tube, and antigen MFI was calculated as Relative Fluorescence Intensity RFI (CD/beads). Fluorescence histogram CV (fhCV) for each CD was regarded as an index of the variability of expression among lymphocytes within each subject (cell-to-cell variability); whereas the CV of RFI was regarded as an index of inter-subjects variability (dog-to-dog variability). In 11 cases, FC analyses were repeated after 24h storage at 4°C and RFI and CVs of fresh and stored samples were compared to assess variability linked to storage. CD4 was identified as the best antigen to be used as an internal control for MFI analyses in canine peripheral blood samples because of low cell-to-cell and dog-to-dog variability, and optimal stability upon 24-h storage. Blood samples from a second group of 21 healthy dogs were labelled only with CD4, in order

  16. Concentrations of cadmium, lead, and zinc in fish from mining-influenced waters of northeastern Oklahoma: sampling of blood, carcass, and liver for aquatic biomonitoring.

    Science.gov (United States)

    Brumbaugh, William G; Schmitt, Christopher J; May, Thomas W

    2005-07-01

    The Tri-States Mining District (TSMD) of Missouri (MO), Kansas (KS), and Oklahoma (OK), USA, was mined for lead (Pb) and zinc (Zn) for more than a century. Mining ceased more than 30 years ago, but wastes remain widely distributed in the region, and there is evidence of surface- and groundwater contamination in the Spring River-Neosho River (SR-NR) system of northeastern OK. In October 2001, we collected a total of 74 fish from six locations in the SR-NR system that included common carp (Cyprinus carpio), channel- and flathead catfish (Ictalurus punctatus and Pylodictis olivaris), largemouth- and spotted bass (Micropterus salmoides and Micropterus punctulatus), and white crappie (Pomoxis annularis). We obtained additional fish from locations in MO that included three reference sites and one site that served as a "positive control" (heavily contaminated by Pb). Blood, carcass (headed, eviscerated, and scaled) and liver (carp only) samples were analyzed for cadmium (Cd), Pb, and Zn. Our objectives were to assess the degree to which fish from the OK portion of the SR-NR system are contaminated by these elements and to evaluate fish blood sampling for biomonitoring. Concentrations of Cd and Pb in carp and catfish from OK sites were elevated and Pb concentrations of some approached those of the highly contaminated site in MO, but concentrations in bass and crappie were relatively low. For Zn, correlations were weak among concentrations in the three tissues and none of the samples appeared to reflect site contamination. Variability was high for Cd in all three tissues of carp; differences between sites were statistically significant (p < 0.05) only for blood even though mean liver concentrations were at least 100-fold greater than those in blood. Blood concentrations of Cd and Pb were positively correlated (r2 = 0.49 to 0.84) with the concentration of the same element in carp and catfish carcasses or in carp livers, and the corresponding multiple regression models were

  17. Trace elements in blood samples of workers in Atbara railways foundry

    International Nuclear Information System (INIS)

    This study was conducted to determine trace elements and toxic substances in biological samples (blood samples) of humans. The aim of the current study was to determine the concentration of iron (Fe), copper (Cu), (Pb), lead, and zinc (Zn) in biological samples of workers employed in the industrial workshops in the River Nile state to assess the potential impact of exposure to the work environmental factors. For the purpose of comparison biological samples were collected from the same group of workers exposed to the elements of the work environment and workers not exposed to the elements of the work environment. The analysis of all elements in biological samples was done by x-ray fluorescence technique (X RF). There were no statistically significant differences between the analytical results for the exposed group and non-exposed group, using the same technique. The results showed that the concentrations of the four elements copper, lead, iron, and zinc in all biological samples from workers exposed were not much higher than those not exposed, it could be argued that there was a possible link between these elements with different causes of physiological disorder. The results also showed that need for an attention for improvements in hygiene practice in the workplace and industrial ventilation.(Author)

  18. Automated processing of whole blood samples for the determination of immunosuppressants by liquid chromatography tandem-mass spectrometry

    OpenAIRE

    Vogeser, Michael; Spöhrer, Ute

    2006-01-01

    Background: Liquid chromatography tandem-mass spectrometry (LC-MS/MS) is an efficient technology for routine determination of immunosuppressants in whole blood; however, time-consuming manual sample preparation remains a significant limitation of this technique. Methods: Using a commercially available robotic pipetting system (Tecan Freedom EVO), we developed an automated sample-preparation protocol for quantification of tacrolimus in whole blood by LC-MS/MS. Barcode reading, sample resuspens...

  19. Is liquid heparin comparable to dry balanced heparin for blood gas sampling in intensive care unit?

    Directory of Open Access Journals (Sweden)

    Viswas Chhapola

    2014-01-01

    Full Text Available Introduction: Blood gas (BG analysis is required for management of critically ill patients in emergency and intensive care units. BG parameters can be affected by the type of heparin formulations used-liquid heparin (LH or dry balanced heparin (DBH. This study was conducted to determine whether blood gas, electrolyte, and metabolite estimations performed by using DBH and LH are comparable. Materials and Methods: A prospective study was conducted at pediatric intensive care unit (PICU of a tertiary care hospital. Paired venous samples were collected from 35 consecutive children in commercially prepared DBH syringes and custom-prepared LH syringes. Samples were immediately analyzed by blood gas analyzer and compared for pH, pCO 2 , pO 2 , HCO 3 - , Na + , K + , Cl - , and lactate. Paired comparisons were done and agreement was assessed by Bland-Altman difference plots. The 95% limits of absolute agreement (LOA were compared with the specifications for total allowable error (TEa. Results: The P values were significant for all measured parameters, with the exception of pCO 2 and K +. Bland-Altman difference plots showed wide LOA for pCO 2 , pO 2 , HCO3 - , Na + , K + , and Cl - when compared against TEa. For pCO 2 , HCO3 - , Na + , K + , and Cl - , 40%, 23%, 77%, 34%, and 54% of samples were outside the TEa limits, respectively, with LH. Conclusion: Our study showed that there is poor agreement between LH and DBH for the BG parameters pCO2, pO2, HCO3 - , K + , Na + , and Cl - and, thus, are not comparable. But for pH and lactate, LH and DBH can be used interchangeably.

  20. Effect of concentrate supplementation on feed consumption, nutrient utilization and blood metabolite profile in captive spotted deer (Axis axis) fed oat (Avena sativa) and berseem (Trifolium alexandrinum) fodders based diet.

    Science.gov (United States)

    Suresh, C; Das, A; Katole, Shrikant; Saini, Mohini; Swarup, D

    2013-03-01

    This experiment was conducted to determine the optimum level of a maize-soybean meal-wheat bran concentrate supplement fed to captive spotted deer fed an oat and berseem fodder-based diet. Twelve adult spotted deer [64-76 kg body weight (BW)] were distributed into three groups of four each and were housed individually. A diet consisting of 5 kg of oat fodder and 5.5 kg of berseem fodder was offered to each one of the experimental animals. The animal in group I received no supplementary concentrate, whereas, those in groups II and III received 0.5 and 1 kg of supplementary concentrate, respectively. A 60 days digestibility trial was conducted with a 5 days collection period on Days 55-59 of the trial. Blood samples were collected from all animals on Day 60 of the experiment. Average daily dry matter intake (DMI) was 1,224, 1,613, and 1,574 g/day in groups I, II, and III, respectively. Dry matter (DM) and organic matter (OM) intake was lowest (P < 0.01) in group I. Intake of P, Cu, and Zn was highest (P < 0.01) in group III, followed by groups II and I. Digestibility of neutral detergent fiber was highest (P < 0.05) in group II. Digestibility of OM and CP was lowest (P < 0.05) in group I. Digestibility of gross energy was highest (P < 0.01) in group III (74.9%), followed by groups II (69.3%) and I (66.2%). Digestible energy (DE) intake (kcal/kg BW(0.75) ) was highest (P < 0.01) in group III (195.4), followed by groups II (180.9) and I (129.8). Initial BW was 72.7, 72.5, and 71.0 kg, whereas, final BW was 71.0, 72.7, and 73.5 kg, in groups I, II and III, respectively. Average daily change in body mass was significantly (P < 0.01) different among the groups. The body mass was lost (-29.2 g/day), maintained (4.1 g/day) and gained (41.6 g/day) in groups I, II, and III, respectively. Blood glucose and cholesterol concentration was highest (P < 0.05) in group III, followed by groups II and I. Serum concentration of Cu and

  1. Optimization of single plate-serial dilution spotting (SP-SDS with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples

    Directory of Open Access Journals (Sweden)

    Pious Thomas

    2015-12-01

    Full Text Available We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single plate-serial dilution spotting (SP-SDS with the prime recommendation of sample anchoring (100 stocks. For pure cultures, serial dilutions were prepared from 0.1 OD (100 stock and 20 μl aliquots of six dilutions (101–106 were applied as 10–15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 100–105 dilutions, and for colloidal suspensions and solid samples (10% w/v, 101–106 dilutions were used. Following incubation, at least one dilution level yielded 6–60 cfu per sector comparable to the standard method involving 100 μl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae, SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.

  2. Screening for genetic haemochromatosis in blood samples with raised alanine aminotransferase

    OpenAIRE

    Bhavnani, M; Lloyd, D; Bhattacharyya, A.; Marples, J; Elton, P; Worwood, M.

    2000-01-01

    BACKGROUND—In the UK approximately 1 in 140 people are homozygous for the C282Y mutation of the HFE gene and are at risk from iron overload caused by genetic haemochromatosis (GH). Early detection can prevent organ damage secondary to iron deposition and increase life expectancy.
AIM—To screen for GH in all blood samples sent to the laboratory for routine liver function tests in which raised serum alanine aminotransferase (ALT) activity was detected.
METHODS—ALT was measured in sera sent to t...

  3. Eosinophilia in routine blood samples as a biomarker for solid tumor development

    DEFF Research Database (Denmark)

    Andersen, Christen Bertel L; Siersma, V.D.; Hasselbalch, H.C.;

    2014-01-01

    eosinophilia in routine blood samples as a potential biomarker of solid tumor development in a prospective design. MATERIAL AND METHODS: From the Copenhagen Primary Care Differential Count (CopDiff) Database, we identified 356 196 individuals with at least one differential cell count (DIFF) encompassing the...... tumors within the first three years following the DIFF. Using multivariable logistic regression, odds ratios (OR) were calculated and adjusted for previous eosinophilia, sex, age, year, month, C-reactive protein, previous cancer and Charlson's Comorbidity Index. RESULTS: The risk of bladder cancer was...

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

    Science.gov (United States)

    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. Quantitative Analysis of Trace Chromium in Blood Samples. Combination of the Advanced Oxidation Process with Catalytic Adsorptive Stripping Voltammetry

    OpenAIRE

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

    2006-01-01

    A new method for pretreating blood samples for trace Cr analysis is described. The Advanced Oxidation Process (AOP with H2O2 and 5.5-W 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 (CAdSV) at a bismuth film electrode (BiFE) gives Cr concentration of 6.0 ± 0.3 ppb in the blood samples. This c...

  6. Performance testing of a semi-automatic card punch system, using direct STR profiling of DNA from blood samples on FTA™ cards.

    Science.gov (United States)

    Ogden, Samantha J; Horton, Jeffrey K; Stubbs, Simon L; Tatnell, Peter J

    2015-01-01

    The 1.2 mm Electric Coring Tool (e-Core™) was developed to increase the throughput of FTA(™) sample collection cards used during forensic workflows and is similar to a 1.2 mm Harris manual micro-punch for sampling dried blood spots. Direct short tandem repeat (STR) DNA profiling was used to compare samples taken by the e-Core tool with those taken by the manual micro-punch. The performance of the e-Core device was evaluated using a commercially available PowerPlex™ 18D STR System. In addition, an analysis was performed that investigated the potential carryover of DNA via the e-Core punch from one FTA disc to another. This contamination study was carried out using Applied Biosystems AmpflSTR™ Identifiler™ Direct PCR Amplification kits. The e-Core instrument does not contaminate FTA discs when a cleaning punch is used following excision of discs containing samples and generates STR profiles that are comparable to those generated by the manual micro-punch. PMID:25407399

  7. Multiplexed flow cytometric sensing of blood electrolytes in physiological samples using fluorescent bulk optode microspheres.

    Science.gov (United States)

    Xu, Chao; Wygladacz, Katarzyna; Retter, Robert; Bell, Michael; Bakker, Eric

    2007-12-15

    Polymeric bulk optode microsphere ion sensors in combination with suspension array technologies such as analytical flow cytometry may become a power tool for measuring electrolytes in physiological samples. In this work, the methodology for the direct measurement of common blood electrolytes in physiological samples using bulk optode microsphere sensors was explored. The simultaneous determination of Na(+), K(+), and Ca(2+) in diluted sheep blood plasma was demonstrated for the first time, using a random suspension array containing three types of mixed microsphere bulk optodes of similar size, fabricated from the same chromoionophore without additional labeling. Sodium ionophore X, potassium ionophore III, and grafted AU-1 in poly(butyl acrylate) were the ionophores used in the bulk optode microsphere ion sensors for Na(+), K(+), and Ca(2+), respectively, in combination with the cation-exchanger NaTFPB (sodium tetrakis-[3,5-bis(trifluoromethyl)phenyl]borate) and the same concentration of the chromoionophore ETH 5294 (9-(di-ethylamino)-5-octadecanoylimino-5H-benzo[a]phen-oxazine) in plasticized poly(vinyl chloride). Excellent reproducibility was achieved for the sensing of potassium ions. The effect of sample pH was relatively small at near-physiological pH and followed theoretical predictions, yet the sample temperature was found to influence the sensor response to a larger extent. Multiplexed ion sensing was achieved by taking advantage of the chemical tunability of the sensor response, adjusting the sensor compositions so that the three types of ion sensors responded with distinct levels of protonation of the chromoionophore. Consequently, three well-resolved peaks were simultaneously observed in the single-channel histogram during the multiplexed calibration as well as in the subsequent measurement of the three cations in 10-fold-diluted sheep plasma. The assigned peak positions corresponded very well to the physiological range of the measured ions. PMID

  8. A STUDY OF METALLO-BETA-LACTAMASE PRODUCING PSEUDOMONAS AERUGINOSA IN BLOOD SAMPLES OF BURNED PATIENTS

    Directory of Open Access Journals (Sweden)

    Piyali

    2014-11-01

    Full Text Available : BACKGROUND: Septicaemia is a life threatening complication of severely burned patients. Among many organisms invading blood stream Pseudomonas aeruginosa is a well-known for its powerful antibiotic resistance mechanisms which increasingly limit the choices for treatment. Among many such resistance mechanisms it is the metallo-beta-lactamase (MBL which confers resistance to Carbapenem group of antibiotics, one of the final resorts to fight them. The present study was undertaken to detect MBL producing P. aeruginosa using phenotypic method from blood samples of burned patients as well as to know their drug sensitivity pattern. MATERIALS AND METHODS: For this purpose 67 Pseudomonas aeruginosa isolates from blood samples of admitted burned patients were subjected to susceptibility testing to antipseudomonal drugs by disc diffusion test and those found to be Carbapenem resistant were subjected to Imipenem - EDTA combined disk synergy test for MBL detection. RESULT: Out of 67 isolates of P.aeruginosa, 19 (28.4% were found to be Carbapenem resistant and 11 (16.4% were MBL producers. A particularly important feature was that the MBL producers were highly resistant to the antibiotics tested than the non-producers. However all of them were susceptible to Colistin and Polymixin B. CONCLUSION: This study has made us to think that a constant vigil and careful selection of antibiotics are necessary to keep prevalence of MBL producing P.aeruginosa in check. The accurate identification and reporting of MBL producing P. aeruginosa will aid infection control practitioners in preventing the spread of these multidrug-resistant isolates

  9. Environmental contaminants in Texas, USA, wetland reptiles: Evaluation using blood samples

    Science.gov (United States)

    Clark, D.R., Jr.; 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.

  10. Age Spots

    Science.gov (United States)

    ... for treating age spots include: Improved appearance. Enhanced self-esteem. Promotion of better skin health. What you need ... 480px View Render 320px View Connect with ASDS: Facebook LinkedIn YouTube Twitter Quick Links About ASDS Advocacy ...

  11. Concentrations of environmental contaminants in blood samples collected from Sharp-shinned hawks (Accipiter striatus) from the Eastern Flyway

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — Table 1 provides the results of organochlorine and mercury analysis on plasma and whole blood samples (respectively) collected from 20 sharp-shinned hawks at HMS...

  12. Potentiating day-old blood samples for detection of interferon-gamma responses following infection with Mycobacterium avium subsp. paratuberculosis

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Nielsen, Søren Saxmose; Jungersen, Gregers

    The interferon gamma (IFN-γ) test measuring specific cell-mediated immune responses in whole blood can be used for diagnosis at an early stage of Mycobacterium avium subsp. paratuberculosis (MAP) infection. A major obstacle for the practical use of IFN-γ testing is the recommended maximum 8 hour...... result in production of IFN-γ in samples previously exposed to MAP antigens. Whole blood samples were collected from heifers in a Danish dairy herd known to be infected with MAP. The samples were collected on three sample dates, and on each date the blood samples were stimulated with PPDj and recombinant....... This aspect needs to be further investigated. Optimisation of the IFN-γ test on day-old samples for diagnosing MAP infected herds would facilitate the large scale use of this diagnostic test....

  13. DNA damage focus analysis in blood samples of minipigs reveals acute partial body irradiation.

    Directory of Open Access Journals (Sweden)

    Andreas Lamkowski

    Full Text Available Radiation accidents frequently involve acute high dose partial body irradiation leading to victims with radiation sickness and cutaneous radiation syndrome that implements radiation-induced cell death. Cells that are not lethally hit seek to repair ionizing radiation (IR induced damage, albeit at the expense of an increased risk of mutation and tumor formation due to misrepair of IR-induced DNA double strand breaks (DSBs. The response to DNA damage includes phosphorylation of histone H2AX in the vicinity of DSBs, creating foci in the nucleus whose enumeration can serve as a radiation biodosimeter. Here, we investigated γH2AX and DNA repair foci in peripheral blood lymphocytes of Göttingen minipigs that experienced acute partial body irradiation (PBI with 49 Gy (± 6% Co-60 γ-rays of the upper lumbar region. Blood samples taken 4, 24 and 168 hours post PBI were subjected to γ-H2AX, 53BP1 and MRE11 focus enumeration. Peripheral blood lymphocytes (PBL of 49 Gy partial body irradiated minipigs were found to display 1-8 DNA damage foci/cell. These PBL values significantly deceed the high foci numbers observed in keratinocyte nuclei of the directly γ-irradiated minipig skin regions, indicating a limited resident time of PBL in the exposed tissue volume. Nonetheless, PBL samples obtained 4 h post IR in average contained 2.2% of cells displaying a pan-γH2AX signal, suggesting that these received a higher IR dose. Moreover, dispersion analysis indicated partial body irradiation for all 13 minipigs at 4 h post IR. While dose reconstruction using γH2AX DNA repair foci in lymphocytes after in vivo PBI represents a challenge, the DNA damage focus assay may serve as a rapid, first line indicator of radiation exposure. The occurrence of PBLs with pan-γH2AX staining and of cells with relatively high foci numbers that skew a Poisson distribution may be taken as indicator of acute high dose partial body irradiation, particularly when samples are available

  14. Effect of Different Blood Sampling Methods on Blood Lead Detection%不同采血方法对血铅检测结果的影响

    Institute of Scientific and Technical Information of China (English)

    周宇华; 海日姑·加帕

    2015-01-01

    目的:探究不同采血方法对血铅检测结果的影响。方法选取2013年3月-2015年3月前来该中心进行职业性健康检查的工人及其子女120名作为研究对象,并于同时期采集工人的静脉血与手指末梢血,对其中的血铅含量进行检测。结果所有受检工人中,手指血与静脉血的血铅含量平均为77μg/L和58μg/L,A、B、C 3组受检工人中手指末端血与静脉血中的血铅含量比较差异有统计学意义(P0.05)。结论在采血过程中,对血液中铅含量的影响因素很多,对静脉血进行采血可以有效提高检测结果的准确性,应引起采血检测人员的高度重视。%Objective To explore the effect of different blood sampling methods on blood lead detection results. Methods In 2013 March 2015 March came to my center for occupational health examination of workers and their children 120 as the research object and at the same time acquisition workers venous blood and peripheral finger blood, on the blood lead levels were detected. Results All the workers under the test, blood lead content in finger blood and venous blood in average 77μg/L and 58μg/L, a, B, C three groups by workers in finger tip blood and venous blood in the blood lead content difference was significant (P0.05). Conclusion In blood of the influencing factors of blood lead levels many, on venous blood can effectively improve the accuracy of detection results, which should be attached great importance to the blood testing personnel.

  15. MalHaploFreq: A computer programme for estimating malaria haplotype frequencies from blood samples

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    Smith Thomas A

    2008-07-01

    Full Text Available Abstract Background Molecular markers, particularly those associated with drug resistance, are important surveillance tools that can inform policy choice. People infected with falciparum malaria often contain several genetically-distinct clones of the parasite; genotyping the patients' blood reveals whether or not the marker is present (i.e. its prevalence, but does not reveal its frequency. For example a person with four malaria clones may contain both mutant and wildtype forms of a marker but it is not possible to distinguish the relative frequencies of the mutant and wildtypes i.e. 1:3, 2:2 or 3:1. Methods An appropriate method for obtaining frequencies from prevalence data is by Maximum Likelihood analysis. A computer programme has been developed that allows the frequency of markers, and haplotypes defined by up to three codons, to be estimated from blood phenotype data. Results The programme has been fully documented [see Additional File 1] and provided with a user-friendly interface suitable for large scale analyses. It returns accurate frequencies and 95% confidence intervals from simulated dataset sets and has been extensively tested on field data sets. Additional File 1 User manual for MalHaploFreq. Click here for file Conclusion The programme is included [see Additional File 2] and/or may be freely downloaded from 1. It can then be used to extract molecular marker and haplotype frequencies from their prevalence in human blood samples. This should enhance the use of frequency data to inform antimalarial drug policy choice. Additional File 2 executable programme compiled for use on DOS or windows Click here for file

  16. Combining rapid diagnostic tests and dried blood spot assays for point-of-care testing of human immunodeficiency virus, hepatitis B and hepatitis C infections in Burkina Faso, West Africa.

    Science.gov (United States)

    Kania, D; Bekalé, A M; Nagot, N; Mondain, A-M; Ottomani, L; Meda, N; Traoré, M; Ouédraogo, J B; Ducos, J; Van de Perre, P; Tuaillon, E

    2013-12-01

    People screened for human immunodeficiency virus (HIV) using rapid diagnostic tests (RDTs) in Africa remain generally unaware of their status for hepatitis B (HBV) and hepatitis C (HCV) infections. We evaluated a two-step screening strategy in Burkina Faso, using both HIV RDTs and Dried Blood Spot (DBS) assays to confirm an HIV-positive test, and to test for HBV and HCV infections. HIV counselling and point-of-care testing were performed at a voluntary counselling and testing centre with HBV, HCV status and HIV confirmation using DBS specimens, being assessed at a central laboratory. Serological testing on plasma was used as the reference standard assay to control for the performance of DBS assays. Nineteen out of 218 participants included in the study were positive for HIV using RDTs. A fourth-generation HIV ELISA and immunoblot assays on DBS confirmed HIV status. Twenty-four out of 25 participants infected with HBV were found positive for hepatitis B surface antigen (HBsAg) using DBS. One sample with a low HBsAg concentration on plasma was not detected on DBS. Five participants tested positive for HCV antibodies were confirmed positive with an immunoblot assay using DBS specimens. Laboratory results were communicated within 7 days to participants with no loss to follow up of participants between the first and second post-test counselling sessions. In conclusion, DBS collection during HIV point-of-care testing enables screening and confirmation of HBV, HCV and HIV infections. Diagnosis using DBS may assist with implementation of national programmes for HBV, HCV and HIV screening and clinical care in middle- to low-income countries. PMID:23902574

  17. Putative Epimutagens in Maternal Peripheral and Cord Blood Samples Identified Using Human Induced Pluripotent Stem Cells

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    Yoshikazu Arai

    2015-01-01

    Full Text Available The regulation of transcription and genome stability by epigenetic systems are crucial for the proper development of mammalian embryos. Chemicals that disturb epigenetic systems are termed epimutagens. We previously performed chemical screening that focused on heterochromatin formation and DNA methylation status in mouse embryonic stem cells and identified five epimutagens: diethyl phosphate (DEP, mercury (Hg, cotinine, selenium (Se, and octachlorodipropyl ether (S-421. Here, we used human induced pluripotent stem cells (hiPSCs to confirm the effects of 20 chemicals, including the five epimutagens, detected at low concentrations in maternal peripheral and cord blood samples. Of note, these individual chemicals did not exhibit epimutagenic activity in hiPSCs. However, because the fetal environment contains various chemicals, we evaluated the effects of combined exposure to chemicals (DEP, Hg, cotinine, Se, and S-421 on hiPSCs. The combined exposure caused a decrease in the number of heterochromatin signals and aberrant DNA methylation status at multiple gene loci in hiPSCs. The combined exposure also affected embryoid body formation and neural differentiation from hiPSCs. Therefore, DEP, Hg, cotinine, Se, and S-421 were defined as an “epimutagen combination” that is effective at low concentrations as detected in maternal peripheral and cord blood.

  18. Identification of malaria infected red blood samples by digital holographic quantitative phase microscope

    Science.gov (United States)

    Patel, Nimit R.; Chhaniwal, Vani K.; Javidi, Bahram; Anand, Arun

    2015-07-01

    Development of devices for automatic identification of diseases is desired especially in developing countries. In the case of malaria, even today the gold standard is the inspection of chemically treated blood smears through a microscope. This requires a trained technician/microscopist to identify the cells in the field of view, with which the labeling chemicals gets attached. Bright field microscopes provide only low contrast 2D images of red blood cells and cell thickness distribution cannot be obtained. Quantitative phase contrast microscopes can provide both intensity and phase profiles of the cells under study. The phase information can be used to determine thickness profile of the cell. Since cell morphology is available, many parameters pertaining to the 3D shape of the cell can be computed. These parameters in turn could be used to decide about the state of health of the cell leading to disease diagnosis. Here the investigations done on digital holographic microscope, which provides quantitative phase images, for comparison of parameters obtained from the 3D shape profile of objects leading to identification of diseased samples is described.

  19. Preliminary Blood Pressure Screening in a Representative Sample of Extremely Obese Kuwaiti Adolescents

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    Rima Abdul Razzak

    2013-01-01

    Full Text Available A relationship between blood pressure (BP and obesity has been found in young adults, but no data are available for adolescents in Kuwait. 257 adolescent (11–19 years participants were categorized into two groups according to their BMI; 48 nonobese (21 males: 43.7% and 27 females: 56.3% with mean age of years and 209 obese (128 males: 61.25% and 81 females: 38.75% with mean age of years. The mean BMI was  kg/m2 for the nonobese group and  kg/m3 for the obese group. Most BP measures based on a single screening were significantly higher in the obese group. The prevalence of elevated BP was significantly higher in the obese subjects (nonobese: 13%; obese: 63%; . In the obese group, there was a significant positive correlation between total sample BMI and all BP measures except the pulse pressure. There was a similar rate of elevated blood pressure between males and females (64% versus 60%; . For both isolated systolic elevated BP and isolated diastolic elevated BP, the prevalences were comparable between the males (systolic: 42%; diastolic: 5% and females (systolic: 34%; diastolic: 14%. Only systolic BP was positively correlated with BMI in obese adolescent males (Spearman ; , with a significant correlation between BMI with diastolic (Spearman ; and mean BP (Spearman ; in females.

  20. Antidepressants detection and quantification in whole blood samples by GC-MS/MS, for forensic purposes.

    Science.gov (United States)

    Truta, Liliana; Castro, André L; Tarelho, Sónia; Costa, Pedro; Sales, M Goreti F; Teixeira, Helena M

    2016-09-01

    Depression is among the most prevalent psychiatric disorders of our society, leading to an increase in antidepressant drug consumption that needs to be accurately determined in whole blood samples in Forensic Toxicology Laboratories. For this purpose, this work presents a new gas chromatography tandem mass spectrometry (GC-MS/MS) method targeting the simultaneous and rapid determination of 14 common Antidepressants in whole blood: 13 Antidepressants (amitriptyline, citalopram, clomipramine, dothiepin, fluoxetine, imipramine, mianserin, mirtazapine, nortryptiline, paroxetine, sertraline, trimipramine and venlafaxine) and 1 Metabolite (N-desmethylclomipramine). Solid-phase extraction was used prior to chromatographic separation. Chromatographic and MS/MS parameters were selected to improve sensitivity, peak resolution and unequivocal identification of the eluted analyte. The detection was performed on a triple quadrupole tandem MS in selected ion monitoring (SIM) mode in tandem, using electronic impact ionization. Clomipramine-D3 and trimipramine-D3 were used as deutered internal standards. The validation parameters included linearity, limits of detection, lower limit of quantification, selectivity/specificity, extraction efficiency, carry-over, precision and robustness, and followed internationally accepted guidelines. Limits of quantification and detection were lower than therapeutic and sub-therapeutic concentration ranges. Overall, the method offered good selectivity, robustness and quick response (<16min) for typical concentration ranges, both for therapeutic and lethal levels. PMID:27376459

  1. Production of dendritic cells and cytokine-induced killer cells from banked umbilical cord blood samples

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    Phuc Van Pham

    2015-11-01

    Full Text Available Umbilical cord blood (UCB is considered to be a source of hematopoietic stem cells (HSCs. All UCB banks have recently become interested in the isolation and storage of HSCs for the treatment of hematological diseases. However, UCB was also recently confirmed as a source of immune cells for immunotherapy such as dendritic cells (DCs and cytokine-induced killer cells (CIKs. This study aimed to exploit this source of immune cells in banked UCB samples. After collection of UCB samples, mononuclear cells (MNCs containing stem cells, progenitor cells, and mature cells were isolated by Ficoll-Hypaque-based centrifugation. The MNCs were subjected to freezing and thawing according to a previously published protocol. The banked MNCs were used to produce DCs and CIKs. To produce DCs, MNCs were induced in RPMI 1640 medium supplemented with GM-CSF (50 ng/ml and IL-4 (40 ng/ml for 14 days. To produce CIKs, MNCs were induced in RPMI 1640 medium supplemented an anti-CD3 monoclonal antibody, IL-3, and GMC-SF for 21 and ndash;28 days. Both DCs and CIKs were evaluated for their phenotypes and functions according to previously published protocols. The results showed that banked UCB samples can be successfully used to produce functional DCs and CIKs. These samples are valuable sources of immune cells for immunotherapy. The present results suggest that banked UCB samples are useful not only for stem cell isolation, but also for immune cell production. [Biomed Res Ther 2015; 2(11.000: 402-408

  2. PERT: a method for expression deconvolution of human blood samples from varied microenvironmental and developmental conditions.

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    Wenlian Qiao

    Full Text Available The cellular composition of heterogeneous samples can be predicted using an expression deconvolution algorithm to decompose their gene expression profiles based on pre-defined, reference gene expression profiles of the constituent populations in these samples. However, the expression profiles of the actual constituent populations are often perturbed from those of the reference profiles due to gene expression changes in cells associated with microenvironmental or developmental effects. Existing deconvolution algorithms do not account for these changes and give incorrect results when benchmarked against those measured by well-established flow cytometry, even after batch correction was applied. We introduce PERT, a new probabilistic expression deconvolution method that detects and accounts for a shared, multiplicative perturbation in the reference profiles when performing expression deconvolution. We applied PERT and three other state-of-the-art expression deconvolution methods to predict cell frequencies within heterogeneous human blood samples that were collected under several conditions (uncultured mono-nucleated and lineage-depleted cells, and culture-derived lineage-depleted cells. Only PERT's predicted proportions of the constituent populations matched those assigned by flow cytometry. Genes associated with cell cycle processes were highly enriched among those with the largest predicted expression changes between the cultured and uncultured conditions. We anticipate that PERT will be widely applicable to expression deconvolution strategies that use profiles from reference populations that vary from the corresponding constituent populations in cellular state but not cellular phenotypic identity.

  3. PERT: a method for expression deconvolution of human blood samples from varied microenvironmental and developmental conditions.

    Science.gov (United States)

    Qiao, Wenlian; Quon, Gerald; Csaszar, Elizabeth; Yu, Mei; Morris, Quaid; Zandstra, Peter W

    2012-01-01

    The cellular composition of heterogeneous samples can be predicted using an expression deconvolution algorithm to decompose their gene expression profiles based on pre-defined, reference gene expression profiles of the constituent populations in these samples. However, the expression profiles of the actual constituent populations are often perturbed from those of the reference profiles due to gene expression changes in cells associated with microenvironmental or developmental effects. Existing deconvolution algorithms do not account for these changes and give incorrect results when benchmarked against those measured by well-established flow cytometry, even after batch correction was applied. We introduce PERT, a new probabilistic expression deconvolution method that detects and accounts for a shared, multiplicative perturbation in the reference profiles when performing expression deconvolution. We applied PERT and three other state-of-the-art expression deconvolution methods to predict cell frequencies within heterogeneous human blood samples that were collected under several conditions (uncultured mono-nucleated and lineage-depleted cells, and culture-derived lineage-depleted cells). Only PERT's predicted proportions of the constituent populations matched those assigned by flow cytometry. Genes associated with cell cycle processes were highly enriched among those with the largest predicted expression changes between the cultured and uncultured conditions. We anticipate that PERT will be widely applicable to expression deconvolution strategies that use profiles from reference populations that vary from the corresponding constituent populations in cellular state but not cellular phenotypic identity.

  4. Effects of two kinds of femoral vein blood sampling methods on blood samples of newborn%两种股静脉采血方法对新生儿血标本的影响

    Institute of Scientific and Technical Information of China (English)

    宋力艳; 海冬; 王彩芳

    2016-01-01

    Objective Effects of 2 kinds of methods to explore the application of disposable blood collecting needle essence of neonatal femoral vein blood sample collection and traditional syringe oblique femoral vein blood sample collection of blood samples in blood coagulation, hemolysis, blood volume in 3 aspects. Methods Our department from January to May 2014, 60 cases of the neonates were divided into observation group and control group with 30 cases in each group, the 2 group objects are the newborn, blood sampling sites were the femoral vein, the observation group used a disposable blood taking needle oblique femoral venous retention method to take blood samples, the control group used the traditional syringe inclined thorn femoral vein leaving method blood samples, will affect the 2 groups of blood sampling methods on blood samples of newborn compared. Results The observation group used a disposable blood taking needle oblique femoral venous blood specimens in anti coagulation, anti hemolysis, blood volume reached 3 aspects of statistical difference is significant. Conclusion The observation group used a disposable blood taking needle oblique femoral venous blood specimens in anti coagulation, anti hemolysis, blood volume reached 3 aspects of statistical difference is significant.%目的:探讨新生儿应用一次性采血针斜刺股静脉采集血标本与传统注射器斜刺股静脉采集血标本2种方法对血标本在凝血、溶血、采血量3个方面的影响。方法本科室将2014年1月至5月60例新生儿分为观察组和对照组各30例,2组采血对象均为新生儿,采血部位均为股静脉,观察组采用一次性采血针斜刺股静脉留取血标本的方法,对照组采用传统注射器斜刺股静脉留取血标本的方法,将2组采血方法对新生儿血标本的影响进行比较。结果观察组采用一次性采血针斜刺股静脉留取血标本在防凝血、防溶血、达到采血量3个方面统计学的

  5. Systematic assessment of reduced representation bisulfite sequencing to human blood samples

    DEFF Research Database (Denmark)

    Wang, Li; Sun, Jihua; Wu, Honglong;

    2012-01-01

    Complementary to the time- and cost-intensive direct bisulfite sequencing, we applied reduced representation bisulfite sequencing (RRBS) to the human peripheral blood mononuclear cells (PBMC) from YH, the Asian individual whose genome and epigenome has been deciphered in the YH project...... and systematically assessed the genomic coverage, coverage depth and reproducibility of this technology as well as the concordance of DNA methylation levels measured by RRBS and direct bisulfite sequencing for the detected CpG sites. Our result suggests that RRBS can cover more than half of CpG islands and promoter...... between the two methods is high. It can be concluded that RRBS is a time and cost-effective sequencing method for unbiased DNA methylation profiling of CpG islands and promoter regions in a genome-wide scale and it is the method of choice to assay certain genomic regions for multiple samples in a rapid...

  6. An atypical microfilaria in blood samples from inhabitants of Brazilian Amazon.

    Science.gov (United States)

    Adami, Y L; Moraes, M A P; Lanfredi, R M; Maia-Herzog, M

    2008-12-01

    An unidentified microfilaria sharing characteristics with Mansonella ozzardi and Onchocerca volvulus was detected in blood samples from seven human volunteers, inhabitants of a community in the border of Amazonas and Acre State. They were detected during epidemiological studies carried out in some communities along Antimary, Acre, and Purus Rivers in the Brazilian Amazon. The most striking difference was presented in the shape of the cephalic space from this microfilaria which was different from those of M. ozzardi and with similarities to O. volvulus in this region, but no remarkable differences were observed at the caudal region. More accurate studies are being carried out in order to provide additional data and supporting evidences before establishment of a new species can be done. PMID:18779979

  7. Determination of appropriate sampling frequency and time of multiple blood sampling dual exponential method with {sup 99m}Tc-DTPA for calculating GFR

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Chung Ho; O, Joo Hyun; Chung, Yong An; Yoo, Le Ryung; Sohn, Hyung Sun; Kim, Sung Hoon; Chung, Soo Kyo; Lee, Hyoung Koo [Catholic University of Korea, Seoul (Korea, Republic of)

    2006-02-15

    To determine appropriate sampling frequency and time of multiple blood sampling dual exponential method with {sup 99m}Tc-DTPA for calculating glomerular filtration rate (GFR). Thirty four patients were included in this study. Three mCi of {sup 99m}Tc-DTPA was intravenously injected and blood sampling at 9 different times, 5 ml each, were done. Using the radioactivity of serum, measured by gamma counter, the GFR was calculated using dual exponential method and corrected with the body surface area. Using spontaneously chosen 2 data points of serum radioactivity, 15 collections of 2-sample GFR were calculated. And 10 collections of 3-sample GFR and 12 collections of 4-sample GFR were also calculated. Using the 9-sample GFR as a reference value, degree of agreement was analyzed with Kendall's {tau} correlation coefficients, mean difference and standard deviation. Although some of the 2-sample GFR showed high correlation coefficient, over or underestimation had evolved as the renal function change. The 10-120-240 min 3-sample GFR showed a high correlation coefficient {tau} =0.93), minimal difference (Mean{+-}SD= -1.784{+-}3.972), and no over or underestimation as the renal function changed. Th 4-sample GFR showed no better accuracy than the 3-sample GFR. Int the wide spectrum or renal function, the 10-120-240 min 3-sample GFR could be the best choice for estimating the patients' renal function.

  8. Decreased mitochondrial DNA content in blood samples of patients with stage I breast cancer

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    Fokas Emmanouil

    2009-12-01

    Full Text Available Abstract Background Alterations of mitochondrial DNA (mtDNA have been implicated in carcinogenesis. We developed an accurate multiplex quantitative real-time PCR for synchronized determination of mtDNA and nuclear DNA (nDNA. We sought to investigate whether mtDNA content in the peripheral blood of breast cancer patients is associated with clinical and pathological parameters. Methods Peripheral blood samples were collected from 60 patients with breast cancer and 51 age-matched healthy individuals as control. DNA was extracted from peripheral blood for the quantification of mtDNA and nDNA, using a one-step multiplex real-time PCR. A FAM labeled MGB probe and primers were used to amplify the mtDNA sequence of the ATP 8 gene, and a VIC labeled MGB probe and primers were employed to amplify the glyceraldehyde-3-phosphate-dehydrogenase gene. mtDNA content was correlated with tumor stage, menstruation status, and age of patients as well as lymph node status and the expression of estrogen receptor (ER, progesterone receptor (PR and Her-2/neu protein. Results The content of mtDNA in stage I breast cancer patients was significantly lower than in other stages (overall P = 0.023. Reduced mtDNA was found often in post menopausal cancer group (P = 0.024. No difference in mtDNA content, in regards to age (p = 0.564, lymph node involvement (p = 0.673, ER (p = 0.877, PR (p = 0.763, and Her-2/neu expression (p = 0.335, was observed. Conclusion Early detection of breast cancer has proved difficult and current detection methods are inadequate. In the present study, decreased mtDNA content in the peripheral blood of patients with breast cancer was strongly associated with stage I. The use of mtDNA may have diagnostic value and further studies are required to validate it as a potential biomarker for early detection of breast cancer.

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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. PMID:23806567

  11. Quantification de la Charge Virale et tests de résistance du VIH-1 aux ARV à partir d’échantillons DBS (Dried Blood Spots chez des patients Guinéens sous traitement antirétroviral

    Directory of Open Access Journals (Sweden)

    Nestor Bangoura

    2015-05-01

    antiretroviral treatment.Problem: As in several countries of the South, the virological monitoring of patients undergoing antiretroviral treatment (ARVT in Guinea is low or non-existent in some locations. The aim ofthis study was to assess the technical and logistical feasibility of the use of (dried blood spots DBSs in viral load (VL and genotyping tests.Method: From September 2010 to October 2010, DBS were prepared from blood samples of adult patients under ARVT. The samples had to be sent to the reference laboratory within 30 days after the sample had been done at ambient temperature. The VL was quantified and the samples of patients with virological failure (CV ≥ 3 log10 copies/mL were genotyped according to the ANRS protocol. The Stanford algorithm, version 6.0.8, was used to analyse and interpret the resistance mutations.Results: Amongst the 136 included patients, 129 and 7 were under first and second line treatment respectively, and monitored for an average of 35 months [IQR: 6-108]. Virological failure was noticed among 33 patients. Among them, 84.8% (n = 28/33 benefited from genotyping. The global resistance rate was 14% (n = 19/136. CRF02_AG was the most prevalent viral subtype (82%; n = 23.Conclusion: In addition to demonstrating the technical and logistic feasibility of VL and genotyping tests from DBSs, these results show the relevance of their use in the virological monitoring of patients under ARVT. Also, this study made it possible to provide informationon virological failure, ARV resistance and the HIV-1 genetic diversity in Guinea.

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

    Science.gov (United States)

    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.

  13. Rat dried blood spot analysis of (R,S)-(-)- and (S,R)-(+)- enantiomers of emtricitabin on immobilized tris-(3,5-dimethylphenyl carbamate) amylose silica as a chiral stationary phase.

    Science.gov (United States)

    Rao, Ramisetti Nageswara; Santhakumar, Kondapalli; Naidu, Challa Gangu

    2015-10-01

    An enantioselective high performance liquid chromatography method has been developed and validated by evaluating the suitability of newly introduced immobilized polysaccharide chiral stationary phases, the effect of different organic modifiers and temperature including the entropy and enthalpy on resolution of the (R,S)-(-) & (S,R)-(+) emtricitabine enantiomers on rat dried blood spots. Both the enantiomers were extracted from dried blood spots using ethanol: methanol (80:20 v/v) mixture and separated on an immobilized amylose tris-(3,5-dimethyl phenyl carbamate) chiral stationary phase using n-hexane:ethanol (65:35 v/v) as a mobile phase at a flow rate of 0.8mL/min. The detection was carried out at 280nm using photo diode array detector connected to a polarimeter in series to determine their order of eluton. The method was validated with respect to limits of detection and quantification, linearity, accuracy and precision. The calibration curves were linear over the concentration range of 0.5-500μg/mL for both enantiomers and the correlation coefficient (r(2)) was >0.998. The overall recovery of (R,S)- & (S,R)-enantiomers of emtricitabin from DBS were 90.4 and 90.6%, respectively. The limits of detection and quantification of enantiomers were 0.26, 0.30 and 0.85, 0.92μg/mL for (R,S)- and (S,R)-emtricitabin enantiomers, respectively. The assay was specific and precise (RSD <10%). The stability of emtricitabin was also performed and the results were found to be well within the limits. The effect of hematocrit on extraction of emtricitabin enantiomers from dried blood spots was evaluated and no interference from endogenous substances was observed.

  14. Blood culture

    Science.gov (United States)

    Culture - blood ... A blood sample is needed . The site where blood will be drawn is first cleaned with an antiseptic such ... organism from the skin getting into (contaminating) the blood sample and causing a false-positive result (see ...

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

    International Nuclear Information System (INIS)

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

  16. Antigen-induced cytokine and chemokine release test for tuberculosis infection using adsorption of stimulated whole blood on filter paper and multiplex analysis

    DEFF Research Database (Denmark)

    Skogstrand, Kristin; Thysen, Anna H; Jørgensen, Charlotte S;

    2012-01-01

    (ELISA), as described in the kit insert. In parallel, activated blood was spotted on filter paper (Schleicher & Schuell) and dried. The dried blood spot samples were analysed for 21 inflammatory markers with an in-house assay based on Luminex technology. Results: Our multiplex measurements...

  17. Effect of Familiar Olfactory Stimulus on Responses to Blood Sampling Pain in Neonates

    Directory of Open Access Journals (Sweden)

    A. Sadathosseini

    2011-04-01

    Full Text Available Introduction & Objective: Pain in neonates can lead to various risks. So, it seems essential to find a simple, safe, and acceptable method for relieving pain. The objective of this study was to assess the effectiveness of olfactory stimuli (familiar and unfamiliar on physiological and behavioral responses to the pain of arterial blood draws in term neonates. Materials & Methods: In this quasi-experimental clinical trial, according to the conditions of the study 135 term neonates were chosen by convenience sampling and were assigned to three groups. During the procedure, familiar odor group was presented with the vanilla smell with which they had been familiarized prior to the procedure for 9 hours. Unfamiliar odor group was presented with the vanilla smell to which they had not been previously exposed, and the control group was presented with no odor. The heart rate and O2 saturation levels were measured before, after inserting and after removing the needle. Also, their cry duration was measured from onset until a crying free interval of more than five seconds. Results: The infants exposed to the familiar odor cried significantly less during the procedure compared to the unfamiliar odor and no odor group (P<0.001. Moreover, there was no statistically significant difference in the heart rate among the groups after inserting and removing the needle and in the O2 saturation rate after inserting the needle. The O2 saturation rate was significantly higher in the familiar odor group compared with the other groups (p<0.05 after the needle removal. Conclusion: A familiar odor is effective in reducing crying during arterial blood draws in neonates, but does not affect on physiological parameters. (Sci J Hamadan Univ Med Sci 2011;18(1:10-19

  18. Rapid detection of Candida albicans by polymerase spiral reaction assay in clinical blood samples

    Directory of Open Access Journals (Sweden)

    Xiaoqun eJiang

    2016-06-01

    Full Text Available 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 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.

  19. Mongolian spots

    Directory of Open Access Journals (Sweden)

    Divya Gupta

    2013-01-01

    Full Text Available Mongolian spots (MS are birthmarks that are present at birth and their most common location is sacrococcygeal or lumbar area. Lesions may be single or multiple and usually involve < 5% total body surface area. They are macular and round, oval or irregular in shape. The color varies from blue to greenish, gray, black or a combination of any of the above. The size varies from few to more than 20 centimetres. Pigmentation is most intense at the age of one year and gradually fades thereafter. It is rarely seen after the age of 6 years. Aberrant MS over occiput, temple, mandibular area, shoulders and limbs may be confused with other dermal melanocytoses and bruises secondary to child abuse, thus necessitating documentation at birth. Although regarded as benign, recent data suggest that MS may be associated with inborn errors of metabolism and neurocristopathies. Mongolian spots usually resolve by early childhood and hence no treatment is generally needed if they are located in the sacral area. However, sometimes it may be required for extrasacral lesions for cosmesis.

  20. Systemic Metabolomic Changes in Blood Samples of Lung Cancer Patients Identified by Gas Chromatography Time-of-Flight Mass Spectrometry

    OpenAIRE

    Suzanne Miyamoto; Taylor, Sandra L.; Barupal, Dinesh K; Ayumu Taguchi; Gert Wohlgemuth; Wikoff, William R.; Yoneda, Ken Y.; Gandara, David R.; Samir M. Hanash; Kyoungmi Kim; Oliver Fiehn

    2015-01-01

    Lung cancer is a leading cause of cancer deaths worldwide. Metabolic alterations in tumor cells coupled with systemic indicators of the host response to tumor development have the potential to yield blood profiles with clinical utility for diagnosis and monitoring of treatment. We report results from two separate studies using gas chromatography time-of-flight mass spectrometry (GC-TOF MS) to profile metabolites in human blood samples that significantly differ from non-small cell lung cancer ...

  1. Single blood-Hg samples can result in exposure misclassification: temporal monitoring within the Japanese community (United States

    Directory of Open Access Journals (Sweden)

    Tsuchiya Ami

    2012-06-01

    Full Text Available Abstract Background The most prominent non-occupational source of exposure to methylmercury is the consumption of fish. In this study we examine a fish consuming population to determine the extent of temporal exposure and investigate the extent to which single time estimates of methylmercury exposure based on blood-Hg concentration can provide reliable estimates of longer-term average exposure. Methods Blood-mercury levels were obtained from a portion of the Arsenic Mercury Intake Biometric Study (AMIBS cohort. Specifically, 56 Japanese women residing in the Puget Sound area of Washington State, US were sampled on three occasions across a one-year period. Results An average of 135 days separated samples, with mean blood-mercury levels for the visits being 5.1, 6.6 and 5.0 μg/l and geometric means being 2.7, 4.5 and 3.1 μg/l. The blood-mercury levels in this group exceed national averages with geometric means for two of the visits being between the 90th and 95th percentiles of nationally observed levels and the lowest geometric mean being between the 75th and 90th percentile. Group means were not significantly different across sampling periods suggesting that exposure of combined subjects remained relatively constant. Comparing intra-individual results over time did not reveal a strong correlation among visits (r = 0.19, 0.50, 0.63 between 1st and 2nd, 2nd and 3rd, and 1st and 3rd sample results, respectively. In comparing blood-mercury levels across two sampling interval combinations (1st and 2nd, 2nd and 3rd, and 1st and 3rd visits, respectively, 58% (n = 34, 53% (n = 31 and 29% (n = 17 of the individuals had at least a 100% difference in blood-Hg levels. Conclusions Point estimates of blood-mercury, when compared with three sample averages, may not reflect temporal variability and individual exposures estimated on the basis of single blood samples should be treated with caution as indicators of long-term exposure

  2. Comparative determination of methyl mercury in whole blood samples using GC-ICP-MS and GC-MS techniques.

    Science.gov (United States)

    Hippler, J; Hoppe, H W; Mosel, F; Rettenmeier, A W; Hirner, A V

    2009-08-15

    Two methods for the determination of methyl mercury (MeHg) in whole blood samples based on different mass spectrometric detection techniques are compared. The methods were employed in two studies in which the internal exposure of a group of mercury-exposed workers to total mercury and MeHg was investigated. Blood samples of these workers were analysed for MeHg independently from each other in two laboratories using similar extraction procedures but different detection techniques, viz. coupled GC-EI-MS/ICP-MS and GC-MS using D(3)-MeHg as internal standard. MeHg was detected in all blood samples in concentrations ranging from 0.3 to 9.0 microg/L. Though different detection techniques were employed, the results obtained by the two laboratories were in relatively good agreement.

  3. Trace samples of human blood in mosquitoes as a forensic investigation tool.

    Science.gov (United States)

    Rabêlo, K C N; Albuquerque, C M R; Tavares, V B; Santos, S M; Souza, C A; Oliveira, T C; Oliveira, N C L; Crovella, S

    2015-01-01

    Investigations of any type of crime invariably starts at the crime scene by collecting evidence. Thus, the purpose of this research was to collect and analyze an entomological trace from an environment that is similar to those of indoor crime scenes. Hematophagous mosquitoes were collected from two residential units; saliva of volunteers that were residents in the units was also collected for genetic analysis as reference samples. We examined the allele frequencies of 15 short tandem repeat loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) and amelogenin. A total of 26 female hematophagous mosquitoes were identified as Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus; we were able to obtain 11 forensically valid genetic profiles, with a minimum of 0.028203 ng/μL of human DNA. Thus, the results of this study showed that it was possible to correlate human genetic information from mosquitoes with the volunteer reference samples, which validates the use of this information as forensic evidence. Furthermore, we observed mixed genetic profiles from one mosquito. Therefore, it is clearly important to collect these insects indoors where crimes were committed, because it may be possible to find intact genetic profiles of suspects in the blood found in the digestive tract of hematophagous mosquitoes for later comparison to identify an offender and/or exclude suspects. PMID:26600546

  4. Trace samples of human blood in mosquitoes as a forensic investigation tool.

    Science.gov (United States)

    Rabêlo, K C N; Albuquerque, C M R; Tavares, V B; Santos, S M; Souza, C A; Oliveira, T C; Oliveira, N C L; Crovella, S

    2015-11-23

    Investigations of any type of crime invariably starts at the crime scene by collecting evidence. Thus, the purpose of this research was to collect and analyze an entomological trace from an environment that is similar to those of indoor crime scenes. Hematophagous mosquitoes were collected from two residential units; saliva of volunteers that were residents in the units was also collected for genetic analysis as reference samples. We examined the allele frequencies of 15 short tandem repeat loci (D8S1179, D21S11, D7S820, CSF1PO, D3S1358, TH01, D13S317, D16S539, D2S1338, D19S433, vWA, TPOX, D18S51, D5S818, and FGA) and amelogenin. A total of 26 female hematophagous mosquitoes were identified as Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus; we were able to obtain 11 forensically valid genetic profiles, with a minimum of 0.028203 ng/μL of human DNA. Thus, the results of this study showed that it was possible to correlate human genetic information from mosquitoes with the volunteer reference samples, which validates the use of this information as forensic evidence. Furthermore, we observed mixed genetic profiles from one mosquito. Therefore, it is clearly important to collect these insects indoors where crimes were committed, because it may be possible to find intact genetic profiles of suspects in the blood found in the digestive tract of hematophagous mosquitoes for later comparison to identify an offender and/or exclude suspects.

  5. Flow cytometric comparison of platelets from a whole blood and finger-prick sample: impact of 24 hours storage.

    Science.gov (United States)

    Swanepoel, Albe C; Stander, Andre; Pretorius, Etheresia

    2013-03-01

    In this study, we investigate the validity and laboratory utility of flow cytometry when analyzing platelet activation by studying CD41, CD42b, CD62P and CD63. We compare flow cytometry results from citrated whole-blood and finger-prick samples directly after collection and also after storing both a finger-prick and whole-blood sample for 24 hours. Citrated whole-blood and finger-prick samples were taken from three healthy individuals on two occasions, and a total of 60,000 cells were analyzed for each of the four phycoerythrin-labeled monoclonal antibodies. Half of each sample was analyzed immediately after sampling while the other half was kept in the fridge at 6 °C for 24 hours before analysis. No significant difference was found between the sampling methods or the period of time before analysis. Results therefore suggest that an appropriately prepared finger-prick sample can be used for platelet function analysis, and samples can be stored for 24 hours in the fridge at 6 °C before analysis. PMID:23320994

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

    Science.gov (United States)

    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 (pGHB in blood and urine samples. Regarding the new formation of GHB in vitro both 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, although there was no significant increases of GHB levels throughout the period of investigation, the lowest increases were found both in blood and urine at -20°C, therefore we recommend the latter as optimal storage temperature.

  7. The identification of menstrual blood in forensic samples by logistic regression modeling of miRNA expression.

    Science.gov (United States)

    Hanson, Erin K; Mirza, Mohid; Rekab, Kamel; Ballantyne, Jack

    2014-11-01

    We report the identification of sensitive and specific miRNA biomarkers for menstrual blood, a tissue that might provide probative information in certain specialized instances. We incorporated these biomarkers into qPCR assays and developed a quantitative statistical model using logistic regression that permits the prediction of menstrual blood in a forensic sample with a high, and measurable, degree of accuracy. Using the developed model, we achieved 100% accuracy in determining the body fluid of interest for a set of test samples (i.e. samples not used in model development). The development, and details, of the logistic regression model are described. Testing and evaluation of the finalized logistic regression modeled assay using a small number of samples was carried out to preliminarily estimate the limit of detection (LOD), specificity in admixed samples and expression of the menstrual blood miRNA biomarkers throughout the menstrual cycle (25-28 days). The LOD was blood was identified only during the menses phase of the female reproductive cycle in two donors.

  8. Sampling blood from big brown bats (Eptesicus fuscus) in the field with and without anesthesia: Impacts on survival

    Science.gov (United States)

    Ellison, L.E.; O'Shea, T.J.; Wimsatt, J.; Pearce, R.D.; Neubaum, D.J.; Neubaum, M.A.; Bowen, R.A.

    2006-01-01

    Blood was collected from wild big brown bats (Eptesicus fuscus) with and without anesthesia in Fort Collins, Colorado in 2004 to assess the impacts of these procedures on short-term survival and 1-yr return rates. Short-term survival and 1-yr return rates after release were passively monitored using PIT tag detection hoops placed at selected buildings. Comparison of 14-day maximum likelihood survival estimates from bats not bled (142 adult females, 62 volant juveniles), and bats sampled for blood with anesthesia (96 adult females, 23 volant juveniles) and without anesthesia (112 adult females, 22 volant juveniles) indicated no adverse effects of either treatment (juveniles: X2=53.38, df=41, P=0.09; adults: X2=39.09, df=44, P=0.68). Return rates of bats one year after sampling were similar among adult female controls (75.4%, n=142, 95% CI=67.4-82.2%), females sampled for blood with anesthesia (83.0%, n=112, 95% CI=74.8-89.5%), and females sampled without anesthesia (87.5%, n=96, 95% CI=79.2-93.4%). Lack of an effect was also noted in 1-yr return rates of juvenile females. These data suggest that the use of anesthesia during sampling of blood has no advantages in terms of enhancement of survival in big brown bats. ?? Wildlife Disease Association 2006.

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

    Directory of Open Access Journals (Sweden)

    Paul F. Rühle

    2016-08-01

    Full Text Available 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.

  10. A Comparative Study of Blood Culture Sampling from Umbilical Catheter Line versus Peripheral Site

    Directory of Open Access Journals (Sweden)

    Abdolkarim Hamedi

    2010-08-01

    Full Text Available Neonatal sepsis is an important cause of death and morbidity in newborns and is diagnosed by isolation of organism in blood culture. In several reports,reliablity of blood cultures were done from umbi lical catheters,have been demonstrated. The objective of the present study was to determine,wether an inde welling umbilical catheter, could be an alternative site for blood culture. In a prospective study over 6 months during 2006,141 paired blood cultures from 134 infant,were done simultaneously from peripheral site and umbilical catheter (mostly U. V. C,during the first four days of life. Majority of these infants were preterm and admitted to NICU for special care. these infants had indwelling umbilical line and had indication of sepsis workup. A total of 141 pairs of blood cultures were obtained from 134 infants. In 16 infants blood culture pairs were positive for one organism in both peripheral vein and umbilical site. 71. 6% of total cultures (n=11pairs were negative in boths site. A total of 22 pairs were positive in one site only,with 5 positive from peripheral vein only and the other 17 from umblical site. Two pairs were positve in boths site with two different organism. In over all 16 infant (11%of blood were considered to be contaminated. Contamination rate were 2. 4% and 9. 2% for peripheral and umbilical catheter site. Contamination rate increased after 48 hours of age in umbilical catheter. The result showed that after 2 days contamination rate for blood culture taken from catheter line increased and specifity decreased. We recommended that blood culture via umblical catheter in first 2 days in sick neonates with indwelling catheter can be a alternate site of blood culture sampelling.

  11. Using CF11 cellulose columns to inexpensively and effectively remove human DNA from Plasmodium falciparum-infected whole blood samples

    Directory of Open Access Journals (Sweden)

    Venkatesan Meera

    2012-02-01

    Full Text Available Abstract Background Genome and transcriptome studies of Plasmodium nucleic acids obtained from parasitized whole blood are greatly improved by depletion of human DNA or enrichment of parasite DNA prior to next-generation sequencing and microarray hybridization. The most effective method currently used is a two-step procedure to deplete leukocytes: centrifugation using density gradient media followed by filtration through expensive, commercially available columns. This method is not easily implemented in field studies that collect hundreds of samples and simultaneously process samples for multiple laboratory analyses. Inexpensive syringes, hand-packed with CF11 cellulose powder, were recently shown to improve ex vivo cultivation of Plasmodium vivax obtained from parasitized whole blood. This study was undertaken to determine whether CF11 columns could be adapted to isolate Plasmodium falciparum DNA from parasitized whole blood and achieve current quantity and purity requirements for Illumina sequencing. Methods The CF11 procedure was compared with the current two-step standard of leukocyte depletion using parasitized red blood cells cultured in vitro and parasitized blood obtained ex vivo from Cambodian patients with malaria. Procedural variations in centrifugation and column size were tested, along with a range of blood volumes and parasite densities. Results CF11 filtration reliably produces 500 nanograms of DNA with less than 50% human DNA contamination, which is comparable to that obtained by the two-step method and falls within the current quality control requirements for Illumina sequencing. In addition, a centrifuge-free version of the CF11 filtration method to isolate P. falciparum DNA at remote and minimally equipped field sites in malaria-endemic areas was validated. Conclusions CF11 filtration is a cost-effective, scalable, one-step approach to remove human DNA from P. falciparum-infected whole blood samples.

  12. Direct diagnosis ofMycobacterium tuberculosis in blood samples of HIV infected patients by polymerase chain reaction

    OpenAIRE

    Kamatchiammal, Senthilkumar; Saravanakumar, Dhashinamoorthy; Kumarasamy, Nagalingeswaran; Solomon, Sunithi; Sritharan, Manjula; Sritharan, Venkataraman

    2000-01-01

    We have developed a simple, economical and reproducible method for processing blood samples from HIV infected patients for diagnosis of tuberculosis. The procedure was validated on 55 samples selected for tuberculosis based on clinical criteria. 52 patients had radiological changes indicative of pulmonary tuberculosis of which only 28 were positive for AFB in sputum (sensitivity 54%) and 27 for tuberculin (sensitivity 52%). 26 HIV positive patients who showed positive X-ray did not react to t...

  13. State of the art of diagnosis of rickettsial diseases: the use of blood specimens for diagnosis of scrub typhus, spotted fever group rickettsiosis, and murine typhus

    Science.gov (United States)

    Paris, Daniel H.; Dumler, J. Stephen

    2016-01-01

    Purpose of review With improved malaria control, acute undifferentiated febrile illness studies in tropical regions reveal a startling proportion of rickettsial illnesses, especially scrub typhus, murine typhus, and spotted fever group rickettsioses. Laboratory diagnosis of these infections evolved little over the past 40 years, but combinations of technologies like PCR and loop-mediated isothermal amplification, with refined rapid diagnostic tests and/or ELISA, are promising for guidance for early antirickettsial treatment. Recent findings The long-term reliance on serological tests – useful only late in rickettsial infections – has led to underdiagnosis, inappropriate therapies, and undocumented morbidity and mortality. Recent approaches integrate nucleic acid amplification and recombinant protein-based serological tests for diagnosing scrub typhus. Optimized using Bayesian latent class analyses, this strategy increases diagnostic confidence and enables early accurate diagnosis and treatment – a model to follow for lagging progress in murine typhus and spotted fever. Summary A laboratory diagnostic paradigm shift in rickettsial infections is evolving, with replacement of indirect immunofluorescence assay by the more objective ELISA coupled with nucleic acid amplification assays to expand the diagnostic window toward early infection intervals. This approach supports targeted antirickettsial therapy, reduces morbidity and mortality, and provides a robust evidence base for further development of diagnostics and vaccines. PMID:27429138

  14. 7 CFR 28.423 - Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody...

  15. The impact of lymphocyte isolation on induced DNA damage in human blood samples measured by the comet assay.

    Science.gov (United States)

    Bausinger, Julia; Speit, Günter

    2016-09-01

    The comet assay is frequently used in human biomonitoring for the detection of exposure to genotoxic agents. Peripheral blood samples are most frequently used and tested either as whole blood or after isolation of lymphocytes (i.e. peripheral blood mononuclear cells, PBMC). To investigate a potential impact of lymphocyte isolation on induced DNA damage in human blood samples, we exposed blood ex vivo to mutagens with different modes of genotoxic action. The comet assay was performed either directly with whole blood at the end of the exposure period or with lymphocytes isolated directly after exposure. In addition to the recommended standard protocol for lymphocyte isolation, a shortened protocol was established to optimise the isolation procedure. The results indicate that the effects of induced DNA strand breaks and alkali-labile sites induced by ionising radiation and alkylants, respectively, are significantly reduced in isolated lymphocytes. In contrast, oxidative DNA base damage (induced by potassium bromate) and stable bulky adducts (induced by benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide; BPDE) seem to be less affected. Our findings suggest that in vivo-induced DNA damage might also be reduced in isolated lymphocytes in comparison with the whole blood depending of the types of DNA damage induced. Because only small genotoxic effects can generally be expected in human biomonitoring studies with the comet assay after occupational and environmental exposure to genotoxic agents, any loss might be relevant and should be avoided. The possibility of such effects and their potential impact on variability of comet assay results in human biomonitoring should be considered when performing or evaluating such kind of studies. PMID:27154923

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

    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

  18. Thyroxine (T4) radioimmunoassay using filter paper dried blood sample: an attempt for screening of neonates for hypothyroidism

    International Nuclear Information System (INIS)

    This paper describes a sensitive but simple and less expensive method suitable for estimation of thyroxine (T4) level. Deficiency of iodine during fetal life results in neonatal hypothyroidism and critinism. Frequency of neonatal hypothyroidism is 1 in 5000 to 7000 in countries having iodine deficiency. It is therefore important to diagnose the neonatal hypothyroidism as soon as possible after birth. The estimation of thyroxine has been found to the a reliable index for diagnosis of hypothyroidism and has long been used for screening of neonatal hypothyroidism. In the present study, instead of serum sample, a 6 mm disc of filter paper containing dried blood sample was used. The test was carried out in the laboratory with 40 samples. As compared to the sensitivity of serum sample technique which is 15.19 n mol/L, the filter paper technique has the sensitivity of 17.23 n mol/L. The work revealed that the T4 concentration do not depend upon the amount of blood on the filter paper. Effect of temperature on filter paper disc was evaluated at 4o c, at 25o c and at 37o c. Results obtained showed significant variation and the best result was obtained for the sample kept at 4o c. The method is simple, rapid, less expensive and needs a small amount of blood and is, therefore, a useful technique for mass screening of neonatal hypothyroidism. 6 refs., 4 tables (author)

  19. Evaluation of biomarkers in plasma, blood, and urine samples from coke oven workers: significance of exposure to polycyclic aromatic hydrocarbons.

    OpenAIRE

    Ovrebø, S; Haugen, A; Farmer, P B; Anderson, D.(California Institute of Technology, Pasadena, USA)

    1995-01-01

    OBJECTIVE--The aim was to assess the significance of two biomarkers; antibody to benzo(a)pyrene DNA adducts and concentration of hydroxyethylvaline haemoglobin adducts in samples from a well studied group of coke oven workers. As a measure of exposure we have used 1-hydroxypyrene in urine. METHODS--Urine and blood samples were collected from coke oven workers and a control group. Samples from coke oven plant workers were collected in January and June. 1-Hydroxypyrene was measured in urine by ...

  20. [Use of C-arm CT for improving the hit rate for selective blood sampling from adrenal veins].

    Science.gov (United States)

    Georgiades, C; Kharlip, J; Valdeig, S; Wacker, F K; Hong, K

    2009-09-01

    Primary hyperaldosteronism is the most common curable cause of hypertension with a prevalence of up to 12% among patients with hypertension. Selective blood sampling from adrenal veins is considered the diagnostic gold standard. However, it is underutilized due to the high technical failure rate. The use of C-arm CT during the sampling procedure can reduce or even eliminate this failure rate. If adrenal vein sampling is augmented by native C-arm CT to check for the correct catheter position, the technical success rate increases substantially. General use of this technique will result in correct diagnosis and treatment for patients with primary hyperaldosteronism.

  1. Reagent deposition for rapid multiplex pathogen identification in human blood culture samples

    DEFF Research Database (Denmark)

    Mogensen, Klaus Bo; Machado, Ana Manuel; Dufva, Martin

    2014-01-01

    -situ hybridization assay is used (PNA-FISH) for diagnosis of hospital acquired bacteria, such as staphylococcus aureus, enterococcus faecalis, E. coli, candida albicans etc. The test covers 90-95 % of the species by prevalence. It is based on a microscope slide format with inspection of the fluorescing bacteria...... viewed in a dual-color microscope configuration. The test takes 20-30 min to perform. In order to lower the cost of the test, rapid automated reagent deposition is needed. Here, ultrasonic spray coating of polyvinyl alcohol/PNA-probes on microscope glass slides is presented. Different wetting regimes...... are explored in order to control the spot profile from convex to concave. The spray coated test are compared with manually prepared tests to ensure that the same performance is obtained....

  2. Strong Correlation Between Concentrations of Tenofovir (TFV) Emtricitabine (FTC) in Hair and TFV Diphosphate and FTC Triphosphate in Dried Blood Spots in the iPrEx Open Label Extension: Implications for Pre-exposure Prophylaxis Adherence Monitoring.

    Science.gov (United States)

    Gandhi, Monica; Glidden, David V; Liu, Albert; Anderson, Peter L; Horng, Howard; Defechereux, Patricia; Guanira, Juan V; Grinsztejn, Beatriz; Chariyalertsak, Suwat; Bekker, Linda-Gail; Grant, Robert M

    2015-11-01

    Self-reported adherence to pre-exposure prophylaxis (PrEP) has limitations, raising interest in pharmacologic monitoring. Drug concentrations in hair and dried blood spots (DBS) are used to assess long-term-exposure; hair shipment/storage occurs at room temperature. The iPrEx Open Label Extension collected DBS routinely, with opt-in hair collection; concentrations were measured with liquid chromatography/tandem mass spectrometry. In 806 hair-DBS pairs, tenofovir (TFV) hair levels and TFV diphosphate (DP) in DBS were strongly correlated (Spearman coefficient r = 0.734; P hair TFV/DBS emtricitabine (FTC) triphosphate (TP) (r = 0.781; P hair FTC/DBS TFV-DP (r = 0.74; P hair FTC/DBS FTC-TP (r = 0.587; P Hair TFV/FTC concentrations correlate strongly with DBS levels, which are predictive of PrEP outcomes.

  3. Analysis of hemoglobin adducts from acrylamide, glycidamide, and ethylene oxide in paired mother/cord blood samples from Denmark.

    Science.gov (United States)

    von Stedingk, Hans; Vikström, Anna C; Rydberg, Per; Pedersen, Marie; Nielsen, Jeanette K S; Segerbäck, Dan; Knudsen, Lisbeth E; Törnqvist, Margareta

    2011-11-21

    The knowledge about fetal exposure to acrylamide/glycidamide from the maternal exposure through food is limited. Acrylamide, glycidamide, and ethylene oxide are electrophiles and form adducts with hemoglobin (Hb), which could be used for in vivo dose measurement. In this study, a method for analysis of Hb adducts by liquid chromatography-mass spectrometry, the adduct FIRE procedure, was applied to measurements of adducts from these compounds in maternal blood samples (n = 87) and umbilical cord blood samples (n = 219). The adduct levels from the three compounds, acrylamide, glycidamide, and ethylene oxide, were increased in tobacco smokers. Highly significant correlations were found between cord and maternal blood with regard to measured adduct levels of the three compounds. The mean cord/maternal hemoglobin adduct level ratios were 0.48 (range 0.27-0.86) for acrylamide, 0.38 (range 0.20-0.73) for glycidamide, and 0.43 (range 0.17-1.34) for ethylene oxide. In vitro studies with acrylamide and glycidamide showed a lower (0.38-0.48) rate of adduct formation with Hb in cord blood than with Hb in maternal blood, which is compatible with the structural differences in fetal and adult Hb. Together, these results indicate a similar life span of fetal and maternal erythrocytes. The results showed that the in vivo dose in fetal and maternal blood is about the same and that the placenta gives negligible protection of the fetus to exposure from the investigated compounds. A trend of higher levels of the measured adducts in cord blood with gestational age was observed, which may reflect the gestational age-related change of the cord blood Hb composition toward a higher content of adult Hb. The results suggest that the Hb adduct levels measured in cord blood reflect the exposure to the fetus during the third trimester. The evaluation of the new analytical method showed that it is suitable for monitoring of background exposures of the investigated electrophilic compounds in large

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

    DEFF Research Database (Denmark)

    Jørgensen, Pelle Morten Thomas; 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...

  5. Prevalence of Fragilysin Gene in Bacteroides fragilis Isolates from Blood and Other Extraintestinal Samples

    OpenAIRE

    Foulon, Ina; Piérard, Denis; Muyldermans, Gaëtan; Vandoorslaer, Kristof; Soetens, Oriane; Rosseel, Paul; Lauwers, Sabine

    2003-01-01

    Of 166 Bacteroides fragilis isolates, 26.2% of 103 isolates from blood and 20.6% of 63 extraintestinal isolates harbored the fragilysin gene (difference not statistically significant). Clinical characteristics and evolution were comparable in patients with B. fragilis bacteremia with or without this enterotoxin. Fragilysin seems not to be an important virulence factor in B. fragilis disease.

  6. Comparison of Chlorhexidine and Tincture of Iodine for Skin Antisepsis in Preparation for Blood Sample Collection

    OpenAIRE

    Barenfanger, Joan; Drake, Cheryl; Lawhorn, Jerry; Verhulst, Steven J.

    2004-01-01

    Rates of contamination of blood cultures obtained when skin was prepared with iodine tincture versus chlorhexidine were compared. For iodine tincture, the contamination rate was 2.7%; for chlorhexidine, it was 3.1%. The 0.41% difference is not statistically significant. Chlorhexidine has comparable effectiveness and is safer, cheaper, and preferred by staff, so it is an alternative to iodine tincture.

  7. Discrepancies between VEGF −1154 G>A Polymorphism Analysis Performed in Peripheral Blood Samples and FFPE Tissue

    Directory of Open Access Journals (Sweden)

    Giorgia Marisi

    2014-07-01

    Full Text Available Single nucleotide polymorphisms (SNPs may be associated with the response or toxicity to different types of treatment. Although SNP analysis is usually performed on DNA from peripheral blood, formalin fixed paraffin-embedded (FFPE tissue is often used for retrospective studies. We analyzed VEGF (−2578C>A, −1498C>T, −1154G>A, −634C>G, +936C>T and eNOS (+894G>T, −786T>C, VNTR (variable number of tandem repeats 27bp intron 4 polymorphisms by direct sequencing or Real Time PCR in 237 patients with advanced colorectal cancer. Peripheral blood was used for 153 patients, whereas only FFPE tumor tissue was available for 84 patients. All SNP frequencies were in Hardy-Weinberg Equilibrium (HWE, with the exception of VEGF −1154, which was only in HWE in peripheral blood specimens. We therefore analyzed this SNP in DNA extracted from FFPE tumor tissue compared to FFPE healthy tissue and peripheral blood from 20 patients. Numerous heterozygous patients in peripheral blood DNA were homozygous for the A-allele in both tumor and healthy FFPE tissues. Our findings indicate that, although FFPE tissue might be a suitable specimen for genotyping, VEGF −1154 does not give reliable results on this type of material. As other SNPs may also have this limitation, genotype concordance should first be confirmed by comparing results obtained from FFPE and fresh sample analyses.

  8. Evaluation of carbon monoxide in blood samples from the second health and nutrition survey. Progress report No. 1

    Energy Technology Data Exchange (ETDEWEB)

    Radford, E.P.

    1976-01-01

    This is a study of carbon monoxide (CO) in the blood of human subjects participating in the Second National Health and Nutrition Survey (HANES II), a detailed study of health indicators in sample populations of many communities throughout the U.S. The purpose of this aspect of the survey is to evaluate the levels of blood carboxyhemoglobin in normal individuals of all ages in typical U.S. communities, from whom accurate histories and clinical studies are available. This report gives results of the first of three years of analyses. A careful calibration of the analytical method has been completed, and more than 3000 blood samples have been analyzed. Although smoking histories are not yet available to permit evaluation of carboxyhemoglobin in non-smokers, in children under 12 years of age, blood COHb has been found to be consistently low, with less than 3% greater than 1.5% COHb. These preliminary results suggest that urban exposure to carbon monoxide among the general population is not now significant in the U.S., at least during the period of these early examinations.

  9. Inorganic elements determination in human and animal whole blood samples by X-ray fluorescence technique (EDXRF)

    International Nuclear Information System (INIS)

    Blood is a suspension of cells contained in a complex liquid called plasma. The term 'whole blood' refers to samples with both solid and liquid parts. Inorganic elements are responsible for essential functions, such as osmotic regulation, cardiac frequency and contractibility, blood clotting and neuromuscular excitability. The determination of inorganic elements in corporeal fluids such as blood, serum, plasma, tissue and urine is used as a monitor for a part or the whole organism. In this work, the X-Ray fluorescence technique (EDXRF) was used for the determination of inorganic elements in whole blood samples from humans and animals (golden hamsters, Mesocricetus auratus and crioula breed horses, Equus caballus). The reference intervals of Na (1788 - 1826 μg g'-1), Mg (63 - 75 μg g-1), P (602 - 676 μg g-1), S (1519 - 1718 μg g-1), Cl (2743 - 2867 μg g-1), K (1508 - 1630 μg g-1), Ca (214 - 228 μg g'-1), Cu (4 -6 μg g-1) e Zn (1 - 3 μg g'-1) were determined for human blood. The reference intervals, for golden hamster blood were found to be: Na (1714 - 1819 μg g-1), Mg (51 - 79 μg g-1), P (970 - 1080 μg g-1), S (1231 - 1739 μg g-1), Cl (2775 - 2865 μg g-1), K (1968 - 2248 μg g-1), Ca (209 - 257 μg g-1), Cu (4 - 6 μg g-1) e Zn (3 - 5 μg g-1). The reference intervals, for crioula breed horse blood, showed to be: Na (1955 - 2013 μg g-1), Mg (51 - 75 μg g-1), P (443 - 476 μg g-1), S (1038 - 1140 μg g-'1), Cl (2388 - 2574 μg g-1), K (1678 - 1753 μg g-1), Ca (202 - 213 μg g-1), Cu (4,1 - 4,5 μg g-1) e Zn (2,0 - 2,2 μg g-1). Comparative study between NAA and EDXRF, both techniques showed the same performance for the analyses of biological matrices. The results contribute for the establishment of reference intervals for the Brazilian healthy population and the referred animal species. (author)

  10. Systemic Metabolomic Changes in Blood Samples of Lung Cancer Patients Identified by Gas Chromatography Time-of-Flight Mass Spectrometry

    Directory of Open Access Journals (Sweden)

    Suzanne Miyamoto

    2015-04-01

    Full Text Available Lung cancer is a leading cause of cancer deaths worldwide. Metabolic alterations in tumor cells coupled with systemic indicators of the host response to tumor development have the potential to yield blood profiles with clinical utility for diagnosis and monitoring of treatment. We report results from two separate studies using gas chromatography time-of-flight mass spectrometry (GC-TOF MS to profile metabolites in human blood samples that significantly differ from non-small cell lung cancer (NSCLC adenocarcinoma and other lung cancer cases. Metabolomic analysis of blood samples from the two studies yielded a total of 437 metabolites, of which 148 were identified as known compounds and 289 identified as unknown compounds. Differential analysis identified 15 known metabolites in one study and 18 in a second study that were statistically different (p-values <0.05. Levels of maltose, palmitic acid, glycerol, ethanolamine, glutamic acid, and lactic acid were increased in cancer samples while amino acids tryptophan, lysine and histidine decreased. Many of the metabolites were found to be significantly different in both studies, suggesting that metabolomics appears to be robust enough to find systemic changes from lung cancer, thus showing the potential of this type of analysis for lung cancer detection.

  11. Vector-borne pathogens in ticks and EDTA-blood samples collected from client-owned dogs, Kiev, Ukraine.

    Science.gov (United States)

    Hamel, Dietmar; Silaghi, Cornelia; Zapadynska, Svitlana; Kudrin, Anton; Pfister, Kurt

    2013-02-01

    Due to the availability of adequate habitats in urban environments, e.g. city parks and recreational green areas, ticks from such settings may also carry pathogens of veterinary and public health concern. Thus, tick-borne infections may readily be identified in companion animals residing in urbanised areas. To investigate the presence of vector-borne pathogens in Kiev, Ukraine, 52 engorged adult ticks, 33 Dermacentor reticulatus and 19 Ixodes ricinus, were collected from 15 dogs in the spring of 2010, and further 23 canine EDTA-blood samples were obtained in the spring of 2011 from client-owned patients presented in a veterinary clinic in Kiev. DNA of 9 pathogens was detected by PCR in ticks and canine EDTA-blood samples: Babesia canis canis, Anaplasma phagocytophilum, Rickettsia helvetica, Ri. monacensis, Ri. raoultii, and Dirofilaria repens (by proxy) were identified in engorged ticks and B. c. canis, Hepatozoon canis, Di. immitis, Di. repens, and Mycoplasma haemocanis in canine EDTA-blood samples. This is the first description of Ri. raoultii in the Ukraine. This study adds information on the occurrence of vector-borne pathogens of veterinary and public health importance in Kiev, Ukraine. PMID:23069260

  12. Nucleic Acid, Antibody, and Virus Culture Methods to Detect Xenotropic MLV-Related Virus in Human Blood Samples

    Directory of Open Access Journals (Sweden)

    M. F. Kearney

    2011-01-01

    Full Text Available The MLV-related retrovirus, XMRV, was recently identified and reported to be associated with both prostate cancer and chronic fatigue syndrome. At the National Cancer Institute-Frederick, MD (NCI-Frederick, we developed highly sensitive methods to detect XMRV nucleic acids, antibodies, and replication competent virus. Analysis of XMRV-spiked samples and/or specimens from two pigtail macaques experimentally inoculated with 22Rv1 cell-derived XMRV confirmed the ability of the assays used to detect XMRV RNA and DNA, and culture isolatable virus when present, along with XMRV reactive antibody responses. Using these assays, we did not detect evidence of XMRV in blood samples ( or prostate specimens ( from two independent cohorts of patients with prostate cancer. Previous studies detected XMRV in prostate tissues. In the present study, we primarily investigated the levels of XMRV in blood plasma samples collected from patients with prostate cancer. These results demonstrate that while XMRV-related assays developed at the NCI-Frederick can readily measure XMRV nucleic acids, antibodies, and replication competent virus, no evidence of XMRV was found in the blood of patients with prostate cancer.

  13. Phosphatidylethanol (PEth) in blood samples from "driving under the influence" cases as indicator for prolonged excessive alcohol consumption.

    Science.gov (United States)

    Schröck, Alexandra; Hernández Redondo, Ana; Martin Fabritius, Marie; König, Stefan; Weinmann, Wolfgang

    2016-03-01

    Phosphatidylethanol (PEth) is considered as specific biomarker of alcohol consumption. Due to accumulation after repeated drinking, PEth is suitable to monitor long-term drinking behavior. To examine the applicability of PEth in "driving under the influence of alcohol" cases, 142 blood samples with blood alcohol concentrations (BAC) ranging from 0.0-3.12‰ were analyzed for the presence of PEth homologues 16:0/18:1 (889 ± 878 ng/mL; range analysis, PEth thresholds were evaluated to differentiate moderate and excessive alcohol consumption with acceptable sensitivity and specificity in accordance with the 1.6‰ BAC limit. With a threshold of 700 ng/mL for PEth 16:0/18:1, prolonged excessive alcohol consumption was detected in 65.9% of drunk drivers with a BAC ≥ 1.6‰ and in 31.6% of the samples with a BAC habits in 88.7% of blood samples. These results show the possibility to detect prolonged excessive alcohol consumption, even if the BAC is below the legal threshold of 1.6‰ for driving aptitude assessment. As a consequence, concentrations of PEth 16:0/18:1 ≥ 700 ng/mL and of PEth 16:0/18:2 ≥ 300 ng/mL may be considered as indicators for the necessity of driving aptitude assessment in addition to BAC. PMID:26671597

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

    Science.gov (United States)

    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). PMID:25930204

  15. Cervical dilatation and grade of doctor affects the interval between decision and result of fetal scalp blood sampling in labour.

    Science.gov (United States)

    Rimmer, Stephanie; Roberts, Stephen A; Heazell, Alexander E P

    2016-08-01

    Fetal scalp blood sampling (FSBS) is used to provide information regarding fetal acid-base status during labour. This study assessed the interval between the decision to perform the procedure and obtaining the result and evaluated whether it is affected by cervical dilatation or the experience of the doctor. The median time for FSBS was 10 min. When cervical dilatation was ≤4 cm samples took approximately 30% longer to obtain. After adjustment for dilation, there were no significant differences between different grades of doctors. FSBS is shorter than previously reported; clinicians should be aware that procedures in early labour take longer to complete. PMID:26399279

  16. Hepatitis B Virus DNA in Blood Samples Positive for Antibodies to Core Antigen and Negative for Surface Antigen

    Science.gov (United States)

    Gutiérrez, C.; León, G.; Loureiro, C. L.; Uzcátegui, N.; Liprandi, F.; Pujol, F. H.

    1999-01-01

    Anti-hepatitis B core antigen (HBcAg)-positive hepatitis B surface antigen (HBsAg)-negative plasma samples from blood donors were tested by nested PCR. DNA positivity was more significantly associated with high levels of anti-HBcAg than with low levels of anti-HBsAg antibodies. Analysis of a dilution of anti-HBcAg antibodies might result in a more rational exclusion of anti-HBcAg-positive HBsAg-negative samples, reducing the number of donations discarded and enabling more countries to incorporate anti-HBcAg testing. PMID:10473534

  17. Foetal scalp blood sampling during labour for pH and lactate measurements.

    Science.gov (United States)

    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. PMID:26253238

  18. Analysis of tumor template from multiple compartments in a blood sample provides complementary access to peripheral tumor biomarkers.

    Science.gov (United States)

    Strauss, William M; Carter, Chris; Simmons, Jill; Klem, Erich; Goodman, Nathan; Vahidi, Behrad; Romero, Juan; Masterman-Smith, Michael; O'Regan, Ruth; Gogineni, Keerthi; Schwartzberg, Lee; Austin, Laura K; Dempsey, Paul W; Cristofanilli, Massimo

    2016-05-01

    Targeted cancer therapeutics are promised to have a major impact on cancer treatment and survival. Successful application of these novel treatments requires a molecular definition of a patient's disease typically achieved through the use of tissue biopsies. Alternatively, allowing longitudinal monitoring, biomarkers derived from blood, isolated either from circulating tumor cell derived DNA (ctcDNA) or circulating cell-free tumor DNA (ccfDNA) may be evaluated. In order to use blood derived templates for mutational profiling in clinical decisions, it is essential to understand the different template qualities and how they compare to biopsy derived template DNA as both blood-based templates are rare and distinct from the gold-standard. Using a next generation re-sequencing strategy, concordance of the mutational spectrum was evaluated in 32 patient-matched ctcDNA and ccfDNA templates with comparison to tissue biopsy derived DNA template. Different CTC antibody capture systems for DNA isolation from patient blood samples were also compared. Significant overlap was observed between ctcDNA, ccfDNA and tissue derived templates. Interestingly, if the results of ctcDNA and ccfDNA template sequencing were combined, productive samples showed similar detection frequency (56% vs 58%), were temporally flexible, and were complementary both to each other and the gold standard. These observations justify the use of a multiple template approach to the liquid biopsy, where germline, ctcDNA, and ccfDNA templates are employed for clinical diagnostic purposes and open a path to comprehensive blood derived biomarker access. PMID:27049831

  19. Immunoelectrophoresis - blood

    Science.gov (United States)

    IEP - serum; Immunoglobulin electrophoresis - blood; Gamma globulin electrophoresis; Serum immunoglobulin electrophoresis ... A blood sample is needed. For information on how this is done, see: Venipuncture

  20. 32P-postlabeling assay for carcinogen-DNA adducts: description of beta shielding apparatus and semi-automatic spotting and washing devices that facilitate the handling of multiple samples

    International Nuclear Information System (INIS)

    The utilization of the 32P-postlabeling assay in combination with TLC for the sensitive detection and estimation of aromatic DNA adducts has been increasing. The procedure consists of 32P-labeling of carcinogen-adducted 3'-nucleotides in the DNA digests using γ-32P ATP and polynucleotide kinase, separation of 32P-labeled adducts by TLC, and their detection by autoradiography. During both 32P-labeling and initial phases of TLC, a relatively high amount of γ-32P ATP is handled when 30 samples are processed simultaneously. We describe the design of acrylic shielding apparatus, semi-automatic TLC spotting devices, and devices for development and washing of multiple TLC plates, which not only provide substantial protection from exposure to 32P beta radiation, but also allow quick and easy handling of a large number of samples. Specifically, the equipment includes: (i) a multi-tube carousel rack having 15 wells to hold capless Eppendorf tubes and a rotatable lid with an aperture to access individual tubes; (ii) a pipette shielder; (iii) two semi-automatic spotting devices to apply radioactive solutions to TLC plates; (iv) a multi-plate holder for TLC plates; and (v) a mechanical device for washing multiple TLC plates. Item (i) is small enough to be held in one-hand, vortexed, and centrifuged to mix the solutions in each tube while beta radiation is shielded. Items (iii) to (iv) aid in the automation of the assay. (author)

  1. iRSpot-GAEnsC: identifing recombination spots via ensemble classifier and extending the concept of Chou's PseAAC to formulate DNA samples.

    Science.gov (United States)

    Kabir, Muhammad; Hayat, Maqsood

    2016-02-01

    Meiotic recombination is vital for maintaining the sequence diversity in human genome. Meiosis and recombination are considered the essential phases of cell division. In meiosis, the genome is divided into equal parts for sexual reproduction whereas in recombination, the diverse genomes are combined to form new combination of genetic variations. Recombination process does not occur randomly across the genomes, it targets specific areas called recombination "hotspots" and "coldspots". Owing to huge exploration of polygenetic sequences in data banks, it is impossible to recognize the sequences through conventional methods. Looking at the significance of recombination spots, it is indispensable to develop an accurate, fast, robust, and high-throughput automated computational model. In this model, the numerical descriptors are extracted using two sequence representation schemes namely: dinucleotide composition and trinucleotide composition. The performances of seven classification algorithms were investigated. Finally, the predicted outcomes of individual classifiers are fused to form ensemble classification, which is formed through majority voting and genetic algorithm (GA). The performance of GA-based ensemble model is quite promising compared to individual classifiers and majority voting-based ensemble model. iRSpot-GAEnsC has achieved 84.46 % accuracy. The empirical results revealed that the performance of iRSpot-GAEnsC is not only higher than the examined algorithms but also better than existing methods in the literature developed so far. It is anticipated that the proposed model might be helpful for research community, academia and for drug discovery. PMID:26319782

  2. Comprehensive kinetics of triiodothyronine production, distribution, and metabolism in blood and tissue pools of the rat using optimized blood-sampling protocols.

    Science.gov (United States)

    DiStefano, J J; Jang, M; Malone, T K; Broutman, M

    1982-01-01

    We have determined estimates for 24 physiological parameters of production, interpool transport, distribution, and metabolism of T3 in the major T3 pools of the unanesthetized male Sprague-Dawley rat, from blood-borne data and a comprehensive model and analysis of this system. Most of these indices have previously been unavailable. Whereas only 3% (2 ng/100 g BW) of the total body T3 pool (74 ng/100 g BW) is in plasma, the composite of slowly equilibrating (slow) tissue pools (e.g. muscle, skin, and brain) appears to contain most of the T3, 76% (57 ng/100 g BW) of the total. The composite of rapidly equilibrating (fast) tissue pools (e.g. liver and kidney) contains the remaining 19% (16 ng/100 g BW). The total body T3 production rate is 0.12 ng/100 g BW . min, and we estimate that about half of this emanates directly from T4 in the slow pools, whereas the remainder is derived from both thyroidal secretion and T4 to T3 conversion in the fast pools. Our results also indicate that T3 molecules spend an average of only 0.5 min in transit each time through plasma, whereas the single pass mean transit times in fast and slow tissue pools (the times available for hormone action) are 10 times and 200 times greater. In contrast, the mean residence time for T3 in the entire system is greater than 12 h despite the extremely rapid early disappearance of injected T3 from plasma. To obtain the required accuracy, we used a novel optimization approach for choosing blood-sampling schedules (1, 4, 44, 202, and 600 min), a remarkably small number of sample times, and each was adjustable by about +/- 20% without effect on optimized parameter accuracies. PMID:7053984

  3. Assessment of the levels of polybrominated diphenyl ethers in blood samples from Guadalajara, Jalisco, Mexico.

    Science.gov (United States)

    Orta-Garcia, Sandra Teresa; León-Moreno, Lilia Carolina; González-Vega, Carolina; Dominguez-Cortinas, Gabriela; Espinosa-Reyes, Guillermo; Pérez-Maldonado, Iván N

    2012-10-01

    The purpose of this study was to measure levels of polybrominated diphenyl ethers (PBDEs) in the blood of children (50 individuals) living in Guadalajara, Jalisco, Mexico. We analyzed six PBDE congeners by gas chromatography-mass spectrometry. Total PBDE levels ranged from not detectable (nd) to 15.2 μg/L on a whole-weight basis and from nd to 6,435 ng/g lipid on a lipid-weight basis. The dominant congener in our study was BDE-153, followed by BDE-154, BDE-99, BDE-100, and BDE-47. Levels of BDE-209 were below the detection limit. Our data indicate that children living in the areas studied in this work are exposed to high levels of PBDEs.

  4. Rocky Mountain spotted fever

    Science.gov (United States)

    ... Mountain spotted fever is caused by the bacteria Rickettsia rickettsii (R. Rickettsii) , which is carried by ticks. ... Saunders; 2014:chap 212. Walker DH, Blaton LS. Rickettsia rickettsii and other spotted fever group rickettsiae (Rocky ...

  5. Sequencing CYP2D6 for the detection of poor-metabolizers in post-mortem blood samples with tramadol.

    Science.gov (United States)

    Fonseca, Suzana; Amorim, António; Costa, Heloísa Afonso; Franco, João; Porto, Maria João; Santos, Jorge Costa; Dias, Mário

    2016-08-01

    Tramadol concentrations and analgesic effect are dependent on the CYP2D6 enzymatic activity. It is well known that some genetic polymorphisms are responsible for the variability in the expression of this enzyme and in the individual drug response. The detection of allelic variants described as non-functional can be useful to explain some circumstances of death in the study of post-mortem cases with tramadol. A Sanger sequencing methodology was developed for the detection of genetic variants that cause absent or reduced CYP2D6 activity, such as *3, *4, *6, *8, *10 and *12 alleles. This methodology, as well as the GC/MS method for the detection and quantification of tramadol and its main metabolites in blood samples was fully validated in accordance with international guidelines. Both methodologies were successfully applied to 100 post-mortem blood samples and the relation between toxicological and genetic results evaluated. Tramadol metabolism, expressed as its metabolites concentration ratio (N-desmethyltramadol/O-desmethyltramadol), has been shown to be correlated with the poor-metabolizer phenotype based on genetic characterization. It was also demonstrated the importance of enzyme inhibitors identification in toxicological analysis. According to our knowledge, this is the first study where a CYP2D6 sequencing methodology is validated and applied to post-mortem samples, in Portugal. The developed methodology allows the data collection of post-mortem cases, which is of primordial importance to enhance the application of these genetic tools to forensic toxicology and pathology.

  6. Determination of gamma-hydroxybutyrate (GHB) and its precursors in blood and urine samples: a salting-out approach.

    Science.gov (United States)

    Kankaanpää, Aino; Liukkonen, Raija; Ariniemi, Kari

    2007-08-01

    Gamma-hydroxybutyrate (GHB) is an increasingly popular drug of abuse that causes stimulation, euphoria, anxiolysis or hypnosis, depending on the dose used. Low doses of the drug are used recreationally, and also implicated in drug-facilitated sexual assaults. Because of the unusually steep dose-response curves, accidental GHB overdosing, leading to coma, seizures or death can occur. Being a controlled substance, GHB is often substituted with its non-scheduled precursors gamma-butyrolactone (GBL) and 1,4-butanediol (BD), which are rapidly metabolized into GHB in the body. Here we describe an assay for GHB, GBL and BD in blood and/or urine samples. GHB and BD were extracted from diluted 200 microL aliquots of samples with t-butylmethylether (plus internal standard benzyl alcohol) in test tubes preloaded with NaCl. After acidification and centrifugation the solvent phase was transferred to a test tube preloaded with Na(2)SO(4), incubated for 30 min, centrifuged again, and evaporated in vacuum. The residue was mixed with N-methyl-N-trimethylsilyl-trifluoroacetamide (MSTFA) in acetonitrile, and injected into a GC-MS. When analyzing GBL, the salting-out step was omitted, and analysis was performed with a GC-FID apparatus. As revealed by the validation data this procedure is suitable for quantitative determination of GHB and its precursors in blood and/or urine samples. PMID:17658710

  7. Blood feeding by the Rocky Mountain spotted fever vector, Dermacentor andersoni, induces interleukin-4 expression by cognate antigen responding CD4+ T cells

    Directory of Open Access Journals (Sweden)

    Wikel Stephen K

    2009-10-01

    Full Text Available Abstract Background Tick modulation of host defenses facilitates both blood feeding and pathogen transmission. Several tick species deviate host T cell responses toward a Th2 cytokine profile. The majority of studies of modulation of T cell cytokine expression by ticks were performed with lymphocytes from infested mice stimulated in vitro with polyclonal T cell activators. Those reports did not examine tick modulation of antigen specific responses. We report use of a transgenic T cell receptor (TCR adoptive transfer model reactive with influenza hemagglutinin peptide (110-120 to examine CD4+ T cell intracellular cytokine responses during infestation with the metastriate tick, Dermacentor andersoni, or exposure to salivary gland extracts. Results Infestation with pathogen-free D. andersoni nymphs or administration of an intradermal injection of female or male tick salivary gland extract induced significant increases of IL-4 transcripts in skin and draining lymph nodes of BALB/c mice as measured by quantitative real-time RT-PCR. Furthermore, IL-10 transcripts were significantly increased in skin while IL-2 and IFN-γ transcripts were not significantly changed by tick feeding or intradermal injection of salivary gland proteins, suggesting a superimposed Th2 response. Infestation induced TCR transgenic CD4+ T cells to divide more frequently as measured by CFSE dilution, but more notably these CD4+ T cells also gained the capacity to express IL-4. Intracellular levels of IL-4 were significantly increased. A second infestation administered 14 days after a primary exposure to ticks resulted in partially reduced CFSE dilution with no change in IL-4 expression when compared to one exposure to ticks. Intradermal inoculation of salivary gland extracts from both male and female ticks also induced IL-4 expression. Conclusion This is the first report of the influence of a metastriate tick on the cytokine profile of antigen specific CD4+ T cells. Blood feeding

  8. Catecholamine blood test

    Science.gov (United States)

    Norepinephrine -- blood; Epinephrine -- blood; Adrenalin -- blood; Dopamine -- blood ... A blood sample is needed. ... the test. This is especially true if both blood and urine catecholamines are to be measured. You ...

  9. Isotope dilution analysis for the determination of zinc in blood samples of diabetic patients

    International Nuclear Information System (INIS)

    Isotope dilution analysis (IDA) based on solvent extraction has been developed for the determination of zinc in the blood of diabetic patients and healthy adults as controls. The method using 65Zn as a tracer is based on the formation of a red colored complex with dithizone in chloroform which is measured by counting of the 1115 keV γ-rays by gamma-ray spectrometry. Various extraction parameters such as pH, nature of solvent and amount of reagent were optimized. Zinc concentration in diabetic patients (n = 10) was found in a much wider range (1.5-157 μg/ml) compared to those in healthy adults (3.1-95.9 μg/ml for n = 5). t-Test of data shows 80-90% confidence limits. A comparison of mean values, 28.5 ± 48.5 μg/ml for diabetics and 33.1 ± 34.5 μg/ml for controls shows 13.9% lower zinc concentration in diabetics. No correlation was found with eating (vegetarian/nonvegetarian)/drinking or smoking habits, but in general, females showed somewhat lower concentration compared to those in males though population size in each case was very small. (author)

  10. Sensitivity of laser light depolarization analysis for detection of malaria in blood samples.

    Science.gov (United States)

    Padial, Manuel Martínez; Subirats, Mercedes; Puente, Sabino; Lago, Mar; Crespo, Santiago; Palacios, Gonzalo; Baquero, Margarita

    2005-05-01

    Automated light depolarization analysis could be a useful tool for diagnosing malarial infections. This work discusses the results of a diagnostic efficacy study on 411 samples from patients with suspected malaria infection performed with a Cell-Dyn 4000 analyser. Light dispersed at 90 degrees and depolarized can be used for identifying and counting eosinophils. However, other cell populations with depolarizing capacity occur in malarial samples; these result from leukocytes ingesting haemozoin that is derived from the degradation of the haem group of haemoglobin performed by the parasite. A sensitivity of 72 % and specificity of 98 % were recorded, with positive and negative predictive values of 78 % and 97 %, respectively. Although the sensitivity level of the automated light depolarization analysis is not adequate to replace the existing methods for the diagnosis of parasitic diseases, it could alert clinicians to unsuspected infections by parasites, particularly those from the genus Plasmodium. PMID:15824421

  11. Highly Effective DNA Extraction Method from Fresh, Frozen, Dried and Clotted Blood Samples

    OpenAIRE

    Jaleh Barar; Sina Atashpaz; Abolfazl Barzegari; Vala Kafil; Sepideh Zununi Vahed; Farzaneh Soltanzad; Sara Samadi Shams

    2011-01-01

    Introduction: Today, with the tremendous potential of genomics and other recent advances in science, the role of science to improve reliable DNA extraction methods is more relevant than ever before. The ideal process for genomic DNA extraction demands high quantities of pure, integral and intact genomic DNA (gDNA) from the sample with minimal co-extraction of inhibitors of downstream processes. Here, we report the development of a very rapid, less-hazardous, and high throughput protocol for e...

  12. Measurement of Nitrite in Blood Samples Using the Ferricyanide-Based Hemoglobin Oxidation Assay

    OpenAIRE

    Piknova, Barbora; Schechter, Alan N.

    2011-01-01

    Nitrite is currently recognized as a biomarker of the state of nitric oxide metabolism. Therefore, assessing nitrite levels in various organs and compartments is an important issue. As nitrite levels in most organs and tissues are low (in high nanomolar or low micromolar range) several new sensitive methods for quantifying nitrite in various biological samples have been developed. Chemiluminescence, combined with tri-iodide reducing solution, is currently considered the most sensitive method,...

  13. Interpreting spotted dolphin age distributions

    OpenAIRE

    Barlow, Jay; Hohn, Aleta A.

    1984-01-01

    Previous work has determined the age distribution from a sample of spotted dolphins (Stenella attenuata) killed in the eastern Pacific tuna purse-seine fishery. In this paper we examine the usefulness of this age distribution for estimating natural mortality rates. The observed age distribution has a deficiency of individuals from 5-15 years and cannot represent a stable age distribution. Sampling bias and errors in age interpretation are examined as possible causes of the "dip" in the obs...

  14. Sampling

    CERN Document Server

    Thompson, Steven K

    2012-01-01

    Praise for the Second Edition "This book has never had a competitor. It is the only book that takes a broad approach to sampling . . . any good personal statistics library should include a copy of this book." —Technometrics "Well-written . . . an excellent book on an important subject. Highly recommended." —Choice "An ideal reference for scientific researchers and other professionals who use sampling." —Zentralblatt Math Features new developments in the field combined with all aspects of obtaining, interpreting, and using sample data Sampling provides an up-to-date treat

  15. International study to evaluate PCR methods for detection of Trypanosoma cruzi DNA in blood samples from Chagas disease patients.

    Directory of Open Access Journals (Sweden)

    Alejandro G Schijman

    Full Text Available 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

  16. Mercury in human hair and blood samples from people living in Wanshan mercury mine area, Guizhou, China: an XAS study.

    Science.gov (United States)

    Li, Yu-Feng; Chen, Chunying; Li, Bai; Li, Wei; Qu, Liya; Dong, Zeqin; Nomura, Masaharu; Gao, Yuxi; Zhao, Jinxuan; Hu, Wei; Zhao, Yuliang; Chai, Zhifang

    2008-03-01

    Human hair and blood samples from persons living in the town of Wanshan, a mercury mine area in Guizhou Province of China, were collected and the quantitative speciation and structural information of Hg and S in hair samples and of Hg in erythrocyte and serum samples were studied using X-ray absorption spectroscopy. Least-squares fitting of the X-ray absorption near-edge spectra found that inorganic mercury is the major mercury species in hair samples (91.74%), while inorganic and methyl mercury are both about 50% of total mercury in RBC and serum samples, which is in agreement with the data obtained by acidic extraction, fractionation of Hg(2+) and CH(3)Hg(+) and quantification by ICP-MS. Curve-fitting analysis revealed that the Hg-S bond length and coordination number in hair were 0.248+/-0.002 nm and 3.10, respectively, while the S-Hg bond length and coordination number in hair were 0.236+/-0.002 nm and 4.05. The Hg-S bond length and coordination number in RBC were 0.251+/-0.003 nm and 4.09, respectively, while they were 0.228+/-0.002 nm and 4.08 in serum, respectively. The techniques for speciation, structural and binding information described in this study will find the potential application in similar studies of other elements.

  17. Measurement of carboxyhemoglobin in forensic blood samples using UV-visible spectrometry and improved principal component regression

    Energy Technology Data Exchange (ETDEWEB)

    Egan, William; Morgan, Stephen L. [Department of Chemistry and Biochemistry, The University of South Carolina, Columbia, South Carolina 29208 (United States)] Brewer, William E. [Toxicology Department, South Carolina Law Enforcement Division, 4416 Broad River Road, Columbia, South Carolina 29210 (United States)

    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}

  18. An experimental model for simultaneous chronic sampling of portal and systemic blood and gastrointestinal lymph via cannulae in conscious swine.

    Science.gov (United States)

    Manolas, K J; Farmer, H M; Cussen, M; Welbourn, R B

    1983-10-01

    Surgical techniques are described whereby safe chronic cannulations of the portal vein, the external iliac artery and vein and the cisterna chyli of pigs were performed. The pigs tolerated the operations well and there was a short recovery period. They were unrestrained during the subsequent feeding experiments, when large sequential blood and lymph samples were withdrawn readily. The experimental periods varied from 3 to 46 days (mean : 13.4 days, SE: 2.0). All of 22 arterial cannulae remained patent (mean : 16 days, SE : 2.2), nineteen of 22 portal cannulae (mean : 15 days, SE : 1.8) and eighteen of 22 venous cannulae (mean : 14 days, SE : 1.9). The lymph cannula patency varied from 2 to 7 days, but lymph samples were easily obtained through all but one of them during the third postoperative day. PMID:6627950

  19. Spot test analysis of microbial contents during composting of kitchen- and garden biowaste: sampling procedures, bacterial reductions, time-temperature relationships, and their relevance for EU-regulations concerning animal by-products.

    Science.gov (United States)

    Bijlsma, P B; de Wit, D H; Duindam, J W; Elsinga, G J; Elsinga, W

    2013-01-30

    This study was aimed to collect data and develop methodologies to determine if and how Dutch biowaste composting plants can meet the microbiological requirements set out in EU-Regulations (EC) 1774/2002 and (EC) 1069/2009, and to provide the European Food and Safety Authority (EFSA) with data and analysis for evaluation of these regulations. We examined twenty plant locations and four types of composting technologies, all with forced aeration and without an anaerobic digestion phase. Raw biowaste, material after sanitation and compost were sampled by spot test analysis according to a standard protocol, and according to an additional protocol with enhanced hygienic precautions. Samples were analyzed for Escherichia coli, Enterococcaceae and Salmonella content. The latter protocol resulted in improved bacterial reductions after sanitation, whereas in compost Enterococcus levels but not E. coli levels increased substantially with both protocols, due to more thermo-resistant regrowth. Salmonella presence in compost coincided with low temperatures and increased levels of E. coli and Enterococcus, absence of Salmonella was associated with absence of E. coli (74%), but not with absence of Enterococcus (17%). In compost, E. coli and Salmonella showed a comparable time-temperature inactivation pattern. A pilot study with co-composting of biowaste and poultry manure indicated a similar inactivation pattern for ESBL-containing bacteria. We conclude that the abundance of Enterococcus in compost is caused by regrowth and not by (re)contamination, and that E. coli is a more reliable indicator species for the absence/presence of Salmonella in compost. Compliance with current EU-regulations concerning biowaste composting can be shown by spot test analysis at all examined plants, provided that adequate hygienic precautions are taken during sampling. PMID:23262408

  20. Accurate measurement of circulating mitochondrial DNA content from human blood samples using real-time quantitative PCR.

    Science.gov (United States)

    Ajaz, Saima; Czajka, Anna; Malik, Afshan

    2015-01-01

    We describe a protocol to accurately measure the amount of human mitochondrial DNA (MtDNA) in peripheral blood samples which can be modified to quantify MtDNA from other body fluids, human cells, and tissues. This protocol is based on the use of real-time quantitative PCR (qPCR) to quantify the amount of MtDNA relative to nuclear DNA (designated the Mt/N ratio). In the last decade, there have been increasing numbers of studies describing altered MtDNA or Mt/N in circulation in common nongenetic diseases where mitochondrial dysfunction may play a role (for review see Malik and Czajka, Mitochondrion 13:481-492, 2013). These studies are distinct from those looking at genetic mitochondrial disease and are attempting to identify acquired changes in circulating MtDNA content as an indicator of mitochondrial function. However, the methodology being used is not always specific and reproducible. As more than 95 % of the human mitochondrial genome is duplicated in the human nuclear genome, it is important to avoid co-amplification of nuclear pseudogenes. Furthermore, template preparation protocols can also affect the results because of the size and structural differences between the mitochondrial and nuclear genomes. Here we describe how to (1) prepare DNA from blood samples; (2) pretreat the DNA to prevent dilution bias; (3) prepare dilution standards for absolute quantification using the unique primers human mitochondrial genome forward primer (hMitoF3) and human mitochondrial genome reverse primer(hMitoR3) for the mitochondrial genome, and human nuclear genome forward primer (hB2MF1) and human nuclear genome reverse primer (hB2MR1) primers for the human nuclear genome; (4) carry out qPCR for either relative or absolute quantification from test samples; (5) analyze qPCR data; and (6) calculate the sample size to adequately power studies. The protocol presented here is suitable for high-throughput use.

  1. Bio-monitoring of persistent organochlorines in human milk and blood samples from sub-Himalayan region of India.

    Science.gov (United States)

    Rai, Swapnil; Dua, Virendra K; Chopra, A K

    2012-09-01

    In the present study, concentrations of organochlorine pesticide residues viz. Dichlorodiphenyltrichloroethane and its metabolites (DDTs) and Hexachlorocyclohexane isomers (HCHs) in human breast milk and human blood samples, collected from several high altitude regions of Garhwal Himalaya in Uttarakhand, India viz. Devprayag, Chamoli, Uttarkashi, Joshimath, Bhatwari and Gangnani (altitude ranging from 472 to 1,982 m above sea level) were determined. Mean concentrations of HCH and DDT in human milk samples ranged from 4.53 to 34.32 mg/kg and 6.09 to 12.98 mg/kg, respectively. While the human blood showed mean values ranging from 6.64 to 281.7 μg/L and 12.37 to 104.10 μg/L for HCH and DDT, respectively. The study showed much higher concentrations of organochlorine residue contamination in the Garhwal region as compared to other parts of India. Risk assessments for infants were also calculated and were found within WHO limits. PMID:22885541

  2. Optimization of loop-mediated isothermal amplification (LAMP) assays for the detection of Leishmania DNA in human blood samples.

    Science.gov (United States)

    Abbasi, Ibrahim; Kirstein, Oscar D; Hailu, Asrat; Warburg, Alon

    2016-10-01

    Visceral leishmaniasis (VL), one of the most important neglected tropical diseases, is caused by Leishmania donovani eukaryotic protozoan parasite of the genus Leishmania, the disease is prevalent mainly in the Indian sub-continent, East Africa and Brazil. VL can be diagnosed by PCR amplifying ITS1 and/or kDNA genes. The current study involved the optimization of Loop-mediated isothermal amplification (LAMP) for the detection of Leishmania DNA in human blood or tissue samples. Three LAMP systems were developed; in two of those the primers were designed based on shared regions of the ITS1 gene among different Leishmania species, while the primers for the third LAMP system were derived from a newly identified repeated region in the Leishmania genome. The LAMP tests were shown to be sufficiently sensitive to detect 0.1pg of DNA from most Leishmania species. The green nucleic acid stain SYTO16, was used here for the first time to allow real-time monitoring of LAMP amplification. The advantage of real time-LAMP using SYTO 16 over end-point LAMP product detection is discussed. The efficacy of the real time-LAMP tests for detecting Leishmania DNA in dried blood samples from volunteers living in endemic areas, was compared with that of qRT-kDNA PCR. PMID:27288706

  3. Loop-mediated isothermal amplification assay for the detection of Ehrlichia canis DNA in blood samples from dogs

    Directory of Open Access Journals (Sweden)

    SA Faggion

    2013-01-01

    Full Text Available The rickettsial bacterium Ehrlichia canis is the etiological agent of canine monocytic ehrlichiosis, one of the most important canine tick-borne diseases in the world. In this study, a loop-mediated isothermal amplification (LAMP assay was developed for detection of E. canis DNA using LAMP primers targeting the groESL operon. Reactions were performed at 60°C for 60 min and the results were visualized by gel electrophoresis. Successful amplification was obtained using plasmid DNA containing a fragment of the groESL operon and DNA extracted from blood samples that tested positive for E. canis by real-time PCR. The specificity of amplification was confirmed by EcoRI restriction of internal sites in the LAMP primers and no cross-reactivity with blood samples positive for Babesia spp., another common tick-borne pathogen, was observed. The high cost of nucleic acid tests (NAT is one of the disadvantages for their large-scale use as routine diagnostic tests. The E. canis LAMP assay developed here is an interesting alternative to PCR since it does not require a thermocycler, thus reducing costs for the veterinary clinical laboratory.

  4. Strong Correlation Between Concentrations of Tenofovir (TFV) Emtricitabine (FTC) in Hair and TFV Diphosphate and FTC Triphosphate in Dried Blood Spots in the iPrEx Open Label Extension: Implications for Pre-exposure Prophylaxis Adherence Monitoring

    Science.gov (United States)

    Gandhi, Monica; Glidden, David V.; Liu, Albert; Anderson, Peter L.; Horng, Howard; Defechereux, Patricia; Guanira, Juan V.; Grinsztejn, Beatriz; Chariyalertsak, Suwat; Bekker, Linda-Gail; Grant, Robert M.

    2015-01-01

    Self-reported adherence to pre-exposure prophylaxis (PrEP) has limitations, raising interest in pharmacologic monitoring. Drug concentrations in hair and dried blood spots (DBS) are used to assess long-term-exposure; hair shipment/storage occurs at room temperature. The iPrEx Open Label Extension collected DBS routinely, with opt-in hair collection; concentrations were measured with liquid chromatography/tandem mass spectrometry. In 806 hair-DBS pairs, tenofovir (TFV) hair levels and TFV diphosphate (DP) in DBS were strongly correlated (Spearman coefficient r = 0.734; P < .001), as were hair TFV/DBS emtricitabine (FTC) triphosphate (TP) (r = 0.781; P < .001); hair FTC/DBS TFV-DP (r = 0.74; P < .001); hair FTC/DBS FTC-TP (r = 0.587; P < .001). Drug detectability was generally concordant by matrix. Hair TFV/FTC concentrations correlate strongly with DBS levels, which are predictive of PrEP outcomes. PMID:25895984

  5. Electrooxidation of antihistamine drug methdilazine and its analysis in human urine and blood samples

    Directory of Open Access Journals (Sweden)

    Nagaraj P. Shetti

    2016-12-01

    Full Text Available The electrochemical oxidation of an antihistamine drug, methdilazine, was studied in 9.2 pH with 0.2 M phosphate buffer as supporting electrolyte at 25 ± 0.2°C. Glassy carbon electrode was used to perform the experiment at cyclic voltammetry, linear sweep voltammetry and differential pulse voltammetric techniques. The dependence of the current on pH, concentration and scan rate were investigated. Differential pulse voltammetric technique was adopted to know the linear relation between peak current and methdilazine concentration. The linear response was obtained in the range of 3.0 μM–1.0 mM with a detection limit of 0.1 μM. The proposed method was also applied for the quantitative determination of methdilazine in pharmaceuticals and biological samples.

  6. Sources of pre-analytical variations in yield of DNA extracted from blood samples: analysis of 50,000 DNA samples in EPIC.

    Directory of Open Access Journals (Sweden)

    Elodie Caboux

    Full Text Available The European Prospective Investigation into Cancer and nutrition (EPIC is a long-term, multi-centric prospective study in Europe investigating the relationships between cancer and nutrition. This study has served as a basis for a number of Genome-Wide Association Studies (GWAS and other types of genetic analyses. Over a period of 5 years, 52,256 EPIC DNA samples have been extracted using an automated DNA extraction platform. Here we have evaluated the pre-analytical factors affecting DNA yield, including anthropometric, epidemiological and technical factors such as center of subject recruitment, age, gender, body-mass index, disease case or control status, tobacco consumption, number of aliquots of buffy coat used for DNA extraction, extraction machine or procedure, DNA quantification method, degree of haemolysis and variations in the timing of sample processing. We show that the largest significant variations in DNA yield were observed with degree of haemolysis and with center of subject recruitment. Age, gender, body-mass index, cancer case or control status and tobacco consumption also significantly impacted DNA yield. Feedback from laboratories which have analyzed DNA with different SNP genotyping technologies demonstrate that the vast majority of samples (approximately 88% performed adequately in different types of assays. To our knowledge this study is the largest to date to evaluate the sources of pre-analytical variations in DNA extracted from peripheral leucocytes. The results provide a strong evidence-based rationale for standardized recommendations on blood collection and processing protocols for large-scale genetic studies.

  7. Seroepidemiological study of human cysticercosis with blood samples collected on filter paper, in Lages, State of Santa Catarina, Brazil, 2004-2005

    Directory of Open Access Journals (Sweden)

    Maria Márcia Imenes Ishida

    2011-06-01

    Full Text Available INTRODUCTION: Human serofrequency of antibodies against Taenia solium antigens was determined and risk factors for cysticercosis transmission were identified. METHODS: Individuals (n=878 from periurban and rural locations of Lages, SC, were interviewed to gather demographic, sanitary and health information. Interviews and blood sample collections by finger prick on Whatman filter paper were performed from August 2004 to May 2005. Observation determined that 850 samples were suitable for analysis and were tested by ELISA using vesicular fluid of Taenia crassiceps heterologous antigen. To ensure the reliability of the results, 77 samples of the dried blood were matched with sera. The reactive samples were submitted to a serum confirmatory immunoblot (IB test using purified Taenia crassiceps glycoproteins. RESULTS: The ELISA results for the dried blood and serum samples were statistically consistent. ELISA was positive in 186 (21.9% out of 850 individuals. A group of 213 individuals were asked to collect vein blood for IB (186 with positive result in ELISA and 27 with inappropriate whole blood samples and 130 attended the request. The IB was positive in 29 (3.4% out of 850 individuals. A significant correlation (p = 0.0364 was determined among individuals who tested positive in the IB assay who practiced both pig rearing and kitchen gardening. CONCLUSIONS: ELISA with dried blood eluted from filter paper was suitable for cysticercosis population surveys. In Lages, human infection was associated with pig rearing and kitchen gardening. The prevalence index was compatible with other Latin American endemic areas.

  8. Facile synthesis of copper(II)-decorated magnetic particles for selective removal of hemoglobin from blood samples.

    Science.gov (United States)

    Ding, Chun; Ma, Xiangdong; Yao, Xin; Jia, Li

    2015-12-11

    In this report, the Cu(2+)-immobilized magnetic particles were prepared by a facile route and they were used as adsorbents for removal of high abundance of hemoglobin in blood based on immobilized metal affinity chromatography. Ethylenediaminetetraacetic acid modified magnetic particles (EDTA-Fe3O4) were first synthesized through a one-pot solvothermal method and then charged with copper ions. The as-prepared Cu(2+)-EDTA-Fe3O4 particles were characterized by Fourier transform infrared spectrometry, scanning electron microscopy, transmission electron microscopy, energy dispersive X-ray spectroscopy, vibrating sample magnetometry and zeta potential. Factors affecting the adsorption of bovine hemoglobin on Cu(2+)-EDTA-Fe3O4 particles (including contact time, solution pH, ionic strength and initial concentration of protein) were investigated. The adsorption process followed a pseudo-second-order kinetic model and the adsorption equilibrium could be achieved in 60min. The adsorption isotherm data could be well described by a Langmuir model and the maximum adsorption capacity was 1250mgg(-1). The as-prepared particles showed high efficiency and excellent selectivity for removal of hemoglobin from bovine and human blood. The removal process integrated the selectivity of immobilized metal affinity chromatography and the convenience of magnetic separation. The results demonstrated that Cu(2+)-EDTA-Fe3O4 particles had potential application in removal of abundant histidine-rich proteins in biomedical diagnosis analysis.

  9. Robust and efficient direct multiplex amplification method for large-scale DNA detection of blood samples on FTA cards

    International Nuclear Information System (INIS)

    Deoxyribonucleic acid (DNA) damage arising from radiations widely occurred along with the development of nuclear weapons and clinically wide application of computed tomography (CT) scan and nuclear medicine. All ionizing radiations (X-rays, γ-rays, alpha particles, etc.) and ultraviolet (UV) radiation lead to the DNA damage. Polymerase chain reaction (PCR) is one of the most wildly used techniques for detecting DNA damage as the amplification stops at the site of the damage. Improvements to enhance the efficiency of PCR are always required and remain a great challenge. Here we establish a multiplex PCR assay system (MPAS) that is served as a robust and efficient method for direct detection of target DNA sequences in genomic DNA. The establishment of the system is performed by adding a combination of PCR enhancers to standard PCR buffer, The performance of MPAS was demonstrated by carrying out the direct PCR amplification on l.2 mm human blood punch using commercially available primer sets which include multiple primer pairs. The optimized PCR system resulted in high quality genotyping results without any inhibitory effect indicated and led to a full-profile success rate of 98.13%. Our studies demonstrate that the MPAS provides an efficient and robust method for obtaining sensitive, reliable and reproducible PCR results from human blood samples. (authors)

  10. Early detection of HIV infection with Dried Blood Spot testing among infants in Yunnan province%滤纸片干血斑HIV-1DNA检测技术在婴儿HIV早期诊断中的应用

    Institute of Scientific and Technical Information of China (English)

    杨朝军; 陈敏; 陈玲; 苏莹珍; 陈会超; 闫文云; 杨莉

    2012-01-01

    目的 探讨滤纸片干血斑技术在婴儿HIV早期诊断中的应用效果.方法 于2010-2011年在云南省昆明、大理、德宏和临沧市(州)的14个妇幼保健院中,对所有感染HIV的孕妇所生的6周至18个月的婴儿进行调查,共计286名.采用滤纸片干血斑采血与罗氏HIV-1 DNA检测技术对HIV感染产妇所生的婴儿进行HIV早期诊断研究,并与18个月时婴儿的HIV抗体结果进行比较.同时阶段性采集并检测滤纸片干血斑的HIV抗体,了解未感染HIV婴儿的抗体阴转时间.并对孕妇抗病毒治疗情况及婴儿母乳喂养情况进行调查.结果 在286名婴儿中,有148名男性、138名女性.对286名婴儿进行了HIV-1 DNA检测,有8名婴儿HIV-1 DNA检测结果为阳性,HIV感染率为2.8%(8/286),与18个月时婴儿的HIV抗体检测结果完全一致;其余278名DNA检测结果为阴性的婴儿,其抗体也均为阴性.对143名HIV-1 DNA阴性的婴儿进行随访,其在出生后6、9、12和18个月时的累计抗体阴转率分别是14.0%(20/143)、61.5%(88/143)、88.1% (126/143)和100.0%( 143/143).286例感染HIV的孕妇中,抗病毒治疗组孕妇所生婴儿的HIV感染率为2.14%(6/280),未抗病毒治疗孕妇组婴儿HIV感染率为33.33% (2/6),差异有统计学意义(P<0.01).人工喂养组婴儿的HIV感染率为2.55% (7/274),纯母乳喂养组婴儿HIV感染率为8.33%(1/12).结论 滤纸片干血斑HIV-1 DNA检测方法可以较好地应用于6周至18个月龄婴儿HIV感染的早期诊断.%Objective To explore the application od Dried Blood Spot (DBS) testing for early detection of HIV infection among infants.Methods All of the infants aged between 6 weeks and 18 months and born by HIV positive mothers from 14 Maternity and Child Health Care Hospitals in Kunming,Dali,Dehong,Lincang of Yunnan province were investigated from 2010 to 2011.By using DBS and Roche HIV-1 DNA test techniques,286 infants were tested for HIV early diagnosis and

  11. An integrative pharmacological approach to radio telemetry and blood sampling in pharmaceutical drug discovery and safety assessment

    Directory of Open Access Journals (Sweden)

    Kamendi Harriet W

    2011-01-01

    Full Text Available Abstract Background A successful integration of the automated blood sampling (ABS and telemetry (ABST system is described. The new ABST system facilitates concomitant collection of physiological variables with blood and urine samples for determination of drug concentrations and other biochemical measures in the same rat without handling artifact. Method Integration was achieved by designing a 13 inch circular receiving antenna that operates as a plug-in replacement for the existing pair of DSI's orthogonal antennas which is compatible with the rotating cage and open floor design of the BASi Culex® ABS system. The circular receiving antenna's electrical configuration consists of a pair of electrically orthogonal half-toroids that reinforce reception of a dipole transmitter operating within the coil's interior while reducing both external noise pickup and interference from other adjacent dipole transmitters. Results For validation, measured baclofen concentration (ABST vs. satellite (μM: 69.6 ± 23.8 vs. 76.6 ± 19.5, p = NS and mean arterial pressure (ABST vs. traditional DSI telemetry (mm Hg: 150 ± 5 vs.147 ± 4, p = NS variables were quantitatively and qualitatively similar between rats housed in the ABST system and traditional home cage approaches. Conclusion The ABST system offers unique advantages over traditional between-group study paradigms that include improved data quality and significantly reduced animal use. The superior within-group model facilitates assessment of multiple physiological and biochemical responses to test compounds in the same animal. The ABST also provides opportunities to evaluate temporal relations between parameters and to investigate anomalous outlier events because drug concentrations, physiological and biochemical measures for each animal are available for comparisons.

  12. Detection of African swine fever virus DNA in blood samples stored on FTA cards from asymptomatic pigs in Mbeya region, Tanzania.

    Science.gov (United States)

    Braae, U C; Johansen, M V; Ngowi, H A; Rasmussen, T B; Nielsen, J; Uttenthal, Å

    2015-02-01

    The aim of the study was to assess whether blood samples collected onto FTA(®) cards could be used in combination with real-time PCR for the detection of African swine fever virus (ASFV) DNA in samples from resource-poor settings under the assumption that asymptomatically (sub-clinically) infected pigs may be present. Blood samples were collected from clinically healthy pigs from Mbeya Region, Tanzania. The blood samples were stored on FTA(®) cards and analysed by real-time PCR assays in duplicate; three pigs had high levels of viral DNA (Ct values of 27-29), and three pigs had a low level of viral DNA (Ct 36-45). Four pigs were positive in one of the duplicate samples only, but clear products of the expected size were obtained when the reactions were analysed by gel electrophoresis. For comparison, blood samples from pigs experimentally infected with either a pathogenic (OURT T88/1) or a non-pathogenic (OURT T88/3) isolate of ASFV were collected, stored on FTA(®) cards and analysed in the same way. The blood from pigs infected with the OURT T88/1 isolate showed high levels of viral DNA (Ct 22-33), whereas infection with non-pathogenic OURT T88/3 isolate resulted in only low levels of viral DNA (Ct 39) in samples collected at 10-14 days after inoculation.

  13. Identification of pyrimethamine- and chloroquine-resistant Plasmodium falciparum in Africa between 1984 and 1998: genotyping of archive blood samples

    Directory of Open Access Journals (Sweden)

    Saito-Nakano Yumiko

    2011-12-01

    Full Text Available Abstract Background Understanding the geographical distribution of drug resistance of Plasmodium falciparum is important for the effective treatment of malaria. Drug resistance has previously been inferred mainly from records of clinical resistance. However, clinical resistance is not always consistent with the parasite's genetic resistance. Thus, molecular identification of the parasite's drug resistance is required. In Africa, clinical resistance to pyrimethamine (Pyr and chloroquine (CQ was evident before 1980 but few studies investigating the genetic resistance to these drugs were conducted before the late 1990s. In this study, genotyping of genes involved in resistance to Pyr and CQ was performed using archive blood samples from Africa between 1984 and 1998. Methods Parasite DNA was extracted from P. falciparum-infected blood smears collected from travellers returning to Japan from Africa between 1984 and 1998. Genotypes of the dihydrofolate reductase gene (dhfr and CQ-resistance transporter gene (pfcrt were determined by polymerase chain reaction amplification and sequencing. Results Genotyping of dhfr and pfcrt was successful in 59 and 80 samples, respectively. One wild-type and seven mutant dhfr genotypes were identified. Three dhfr genotypes lacking the S108N mutation (NRSI, ICSI, IRSI; amino acids at positions 51, 59, 108, and 164 with mutations underlined were highly prevalent before 1994 but reduced after 1995, accompanied by an increase in genotypes with the S108N mutation. The dhfr IRNI genotype was first identified in Nigeria in 1991 in the present samples, and its frequency gradually increased. However, two double mutants (ICNI and NRNI, the latter of which was exclusively found in West Africa, were more frequent than the IRNI genotype. Only two pfcrt genotypes were found, the wild-type and a Southeast Asian type (CVIET; amino acids at positions 72-76 with mutations underlined. The CVIET genotype was already present as early as

  14. Perfluoroalkyl substances in the blood of wild rats and mice from 47 prefectures in Japan: use of samples from nationwide specimen bank.

    Science.gov (United States)

    Taniyasu, Sachi; Senthilkumar, Kurunthachalam; Yamazaki, Eriko; Yeung, Leo W Y; Guruge, Keerthi S; Kannan, Kurunthachalam; Yamashita, Nobuyoshi

    2013-07-01

    Numerous studies have reported on the global distribution, persistence, fate, and toxicity of perfluoroalkyl and polyfluoroalkyl substances (PFASs). However, studies on PFASs in terrestrial mammals are scarce. Rats can be good sentinels of human exposure to toxicants because of their habitat, which is in close proximity to humans. Furthermore, exposure data measured for rats can be directly applied for risk assessment because many toxicological studies use rodent models. In this study, a nationwide survey of PFASs in the blood of wild rats as well as surface water samples collected from rats' habitats from 47 prefectures in Japan was conducted. In addition to known PFASs, combustion ion chromatography technique was used for analysis of total fluorine concentrations in the blood of rats. In total, 216 blood samples representing three species of wild rats (house rat, Norway rats, and field mice) were analyzed for 23 PFASs. Perfluorooctanesulfonate (PFOS; concentration range 80 % of the blood samples. Concentrations of several PFASs in rat blood were similar to those reported for humans. PFSAs (mainly PFOS) accounted for 45 % of total PFASs, whereas perfluoroalkyl carboxylates (PFCAs), especially PFUnDA and PFNA, accounted for 20 and 10 % of total PFASs, respectively. In water samples, PFCAs were the predominant compounds with PFOA and PFNA found in >90 % of the samples. There were strong correlations (p < 0.001 to p < 0.05) between human population density and levels of PFOS, PFNA, PFOA, and PFOSA in wild rat blood. PMID:23494483

  15. Prevalence of Mycobacterium avium subsp. paratuberculosis and Escherichia coli in blood samples from patients with inflammatory bowel disease.

    Science.gov (United States)

    Nazareth, Nair; Magro, Fernando; Machado, Elisabete; Ribeiro, Teresa Gonçalves; Martinho, António; Rodrigues, Pedro; Alves, Rita; Macedo, Gonçalo Nuno; Gracio, Daniela; Coelho, Rosa; Abreu, Candida; Appelberg, Rui; Dias, Camila; Macedo, Guilherme; Bull, Tim; Sarmento, Amélia

    2015-12-01

    Mycobacterium avium subsp. paratuberculosis (MAP) and adherent-invasive Escherichia coli (AIEC) have been implicated as primary triggers in Crohn's disease (CD). In this study, we evaluated the prevalence of MAP and E. coli (EC) DNA in peripheral blood from 202 inflammatory bowel disease (IBD) patients at various disease periods and compared against 24 cirrhotic patients with ascites (CIR) (non-IBD controls) and 29 healthy controls (HC). MAP DNA was detected by IS900-specific nested PCR, EC DNA by malB-specific nested PCR and AIEC identity, in selected samples, by sequencing of fimH gene. CD patients with active disease showed the highest MAP DNA prevalence among IBD patients (68 %). Infliximab treatment resulted in decreased MAP detection. CIR patients had high individual and coinfection rates (75 % MAP, 88 % EC and 67 % MAP and EC), whilst HC controls had lower MAP prevalence (38 %) and EC was undetectable in this control group. EC DNA prevalence in IBD patients was highly associated with CD, and 80 % of EC from the selected samples of CD patients analyzed carried the fimH30 allele, with a mutation strongly associated with AIEC. Our results show that coinfection with MAP and AIEC is common and persistent in CD, although the high MAP and EC detection in CIR patients suggested that colonization is, at least, partially dependent on increased gut permeability. Nevertheless, facilitative mechanisms between a susceptible host and these two potential human pathogens may allow their implication in CD pathogenesis.

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

    Science.gov (United States)

    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 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.

  17. Blood cholesterol screening in several environments using a portable, dry-chemistry analyzer and fingerstick blood samples. Lipid Research Clinics Cholesterol Screening Study Group.

    Science.gov (United States)

    Bradford, R H; Bachorik, P S; Roberts, K; Williams, O D; Gotto, A M

    1990-01-01

    A multicenter study of blood cholesterol screening was performed in several typical environments, such as community sites (shopping malls and a supermarket), health care sites, work sites, a blood bank and a school. Cholesterol was measured with a portable, dry-chemistry analyzer using capillary blood obtained by fingerstick. Data are reported from a total of 13,824 participants, spanning the entire age spectrum. Overall, 25% of screened subjects had blood cholesterol levels above the age-specific cutpoints used in the current study. Although in the aggregate this screening experience very closely approximates the expected level of referrals, the proportion of referred screened subjects differed significantly among the 5 types of screening environments and by gender. Follow-up telephone interviews indicated that 53% of referrals had initiated a physician contact. More than 75% of those who had seen a physician reported that the diagnosis of hypercholesterolemia had been confirmed, and almost 72% had been prescribed a diet. A large proportion of referred screened subjects reported having modified their diet, particularly when recommended to do so by a physician. This study has yielded encouraging evidence that physicians gave referred screened subjects appropriate initial advice for managing hypercholesterolemia. The new technology for blood cholesterol measurement evaluated in the current study has proven to be a feasible and reliable means for measuring blood cholesterol in typical screening settings.

  18. Supported liquid membrane extraction coupled in-line to commercial capillary electrophoresis for rapid determination of formate in undiluted blood samples.

    Science.gov (United States)

    Pantůčková, Pavla; Kubáň, Pavel; Boček, Petr

    2013-07-19

    A cheap, disposable sample pretreatment device with planar supported liquid membrane (SLM) was proposed, assembled and placed into an autosampler carousel of a commercial capillary electrophoresis (CE) instrument for automated pretreatment and analysis of formate in undiluted whole blood and serum samples. All analytical procedures except for filling the pretreatment device with donor and acceptor solutions, i.e., extraction across SLM, injection of the extracted sample and CE-UV determination of formate, were performed fully automatically. The pretreatment device required only μL volumes of blood sample and organic solvent per extraction and was disposed off after each extraction. Good repeatability of peak areas (≤7.7%) and migration times (≤1.5%), linear relationship (r(2)=0.998-0.999) and limits of detection (≤35μM) were achieved. The overall analytical process including blood withdrawal, filling the SLM device with respective solutions, extraction of blood sample, injection into separation capillary and CE separation of formate from other anions took less than 4min. The method was proved useful by direct determination of elevated formate concentrations in undiluted serum samples of a methanol intoxicated patient. Due to its compatibility with currently commercially available CE instrumentation, disposability of extraction devices, minimum sample handling/consumption, and short extraction/analysis times, the developed method might be attractive for rapid diagnosis of methanol poisoning in clinical and toxicological laboratories. PMID:23777836

  19. Hematocrit, Anemia, and Arm Preference for Blood Sample Collection: A Cross-Sectional Study of Pregnant Women in Enugu, South-Eastern, Nigeria

    OpenAIRE

    Dim, CC; Ugwu, EO; Dim, NR; Anyaehie, UB

    2015-01-01

    Background: Anemia in pregnancy is a common cause of maternal morbidity and mortality in developing countries. Regular review of hematocrit (HCT) and anemia patterns in pregnancy is necessary in our environment. Aim: The aim was to determine the average HCT, prevalence, and pattern of anemia, as well the arm preferences for blood sample collection among pregnant women in Enugu, South East Nigeria. Subjects and Methods: HCT was determined using venous blood of 200 antenatal women at the Univer...

  20. Selection of suitable reference genes for normalization of quantitative RT-PCR in peripheral blood samples of bottlenose dolphins (Tursiops truncatus)

    OpenAIRE

    I-Hua Chen; Lien-Siang Chou; Shih-Jen Chou; Jiann-Hsiung Wang; Jeffrey Stott; Myra Blanchard; I-Fan Jen; Wei-Cheng Yang

    2015-01-01

    Quantitative RT-PCR is often used as a research tool directed at gene transcription. Selection of optimal housekeeping genes (HKGs) as reference genes is critical to establishing sensitive and reproducible qRT-PCR-based assays. The current study was designed to identify the appropriate reference genes in blood leukocytes of bottlenose dolphins (Tursiops truncatus) for gene transcription research. Seventy-five blood samples collected from 7 bottlenose dolphins were used to analyze 15 candidate...

  1. Untargeted metabolomics applied retrospectively to UPLC-HR-TOFMS data of whole blood samples from Danish drivers exposed to 3,4-Methylenedioxymethamphetamine (MDMA, Ecstasy)

    DEFF Research Database (Denmark)

    Nielsen, Kirstine Lykke; Telving, Rasmus; Andreasen, Mette Findal;

    to evaluate the drug metabolism of 3,4-methylenedioxymethamphetamine (MDMA, “Ecstasy”). Despite of the untraditional experimental setup, and a very heterogeneous population with different concentrations of MDMA/kg blood weight, as well as unknown information about amount and time of administration in relation...... to blood sampling, it was possible to extract meaningful information. Various statistical methods were tested and their predictability was validated by the positive identification of MDMA blood metabolites. In addition, endogenous metabolites that may be related to energy metabolism, the serotonergic...

  2. Paper-based assay for red blood cell antigen typing by the indirect antiglobulin test.

    Science.gov (United States)

    Yeow, Natasha; McLiesh, Heather; Guan, Liyun; Shen, Wei; Garnier, Gil

    2016-07-01

    A rapid and simple paper-based elution assay for red blood cell antigen typing by the indirect antiglobulin test (IAT) was established. This allows to type blood using IgG antibodies for the important blood groups in which IgM antibodies do not exist. Red blood cells incubated with IgG anti-D were washed with saline and spotted onto the paper assay pre-treated with anti-IgG. The blood spot was eluted with an elution buffer solution in a chromatography tank. Positive samples were identified by the agglutinated and fixed red blood cells on the original spotting area, while red blood cells from negative samples completely eluted away from the spot of origin. Optimum concentrations for both anti-IgG and anti-D were identified to eliminate the washing step after the incubation phase. Based on the no-washing procedure, the critical variables were investigated to establish the optimal conditions for the paper-based assay. Two hundred ten donor blood samples were tested in optimal conditions for the paper test with anti-D and anti-Kell. Positive and negative samples were clearly distinguished. This assay opens up new applications of the IAT on paper including antibody detection and blood donor-recipient crossmatching and extends its uses into non-blood typing applications with IgG antibody-based diagnostics. Graphical abstract A rapid and simple paper-based assay for red blood cell antigen typing by the indirect antiglobulin test. PMID:27185543

  3. Evaluating the effect of sample type on American alligator (Alligator mississippiensis) analyte values in a point-of-care blood analyser.

    Science.gov (United States)

    Hamilton, Matthew T; Finger, John W; Winzeler, Megan E; Tuberville, Tracey D

    2016-01-01

    The assessment of wildlife health has been enhanced by the ability of point-of-care (POC) blood analysers to provide biochemical analyses of non-domesticated animals in the field. However, environmental limitations (e.g. temperature, atmospheric humidity and rain) and lack of reference values may inhibit researchers from using such a device with certain wildlife species. Evaluating the use of alternative sample types, such as plasma, in a POC device may afford researchers the opportunity to delay sample analysis and the ability to use banked samples. In this study, we examined fresh whole blood, fresh plasma and frozen plasma (sample type) pH, partial pressure of carbon dioxide (PCO2), bicarbonate (HCO3 (-)), total carbon dioxide (TCO2), base excess (BE), partial pressure of oxygen (PO2), oxygen saturation (sO2) and lactate concentrations in 23 juvenile American alligators (Alligator mississippiensis) using an i-STAT CG4+ cartridge. Our results indicate that sample type had no effect on lactate concentration values (F 2,65 = 0.37, P = 0.963), suggesting that the i-STAT analyser can be used reliably to quantify lactate concentrations in fresh and frozen plasma samples. In contrast, the other seven blood parameters measured by the CG4+ cartridge were significantly affected by sample type. Lastly, we were able to collect blood samples from all alligators within 2 min of capture to establish preliminary reference ranges for juvenile alligators based on values obtained using fresh whole blood. PMID:27382469

  4. Correlation of antigen-specific IFN-γ responses of fresh blood samples from Mycobacterium avium subsp. paratuberculosis infected heifers with responses of day-old samples co-cultured with IL-12 or anti-IL-10 antibodies

    DEFF Research Database (Denmark)

    Mikkelsen, Heidi; Aagaard, Claus; Nielsen, Søren Saxmose;

    2012-01-01

    Paratuberculosis is a chronic infection of the intestine of ruminants caused by Mycobacterium avium subsp. paratuberculosis (MAP). Early stage MAP infection can be detected by measuring cell-mediated immune responses using the interferon gamma (IFN-γ) assay. Whole blood samples are cultured...... to enhance IFN-γ responses of cultures stimulated with Johnin purified protein derivative (PPDj). Here we examined the correlation of IFN-γ production in response to PPDj and 15 recombinant antigens in day-old blood samples from heifers 10–21 months of age from a MAP infected herd with addition of either...

  5. Association between plasma leptin and blood pressure in two population-based samples of children and adolescents

    DEFF Research Database (Denmark)

    Grøntved, Anders; Steene-Johannessen, Jostein; Kynde, Iben;

    2011-01-01

    In this study we examined the association between leptin and blood pressure in a population-based study of Danish and Norwegian children and adolescents. Because of the putative bidirectional relationship between leptin and adiposity we formally tested (i) the mediating effect of body mass index in...... the association between leptin and blood pressure, and (ii) the mediating effect of leptin in the association between body mass index and blood pressure....

  6. A Rapid and Sensitive UPLC-MS/MS-Method for the Separation and Quantification of Branched-Chain Amino Acids from Dried Blood Samples of Patients with Maple Syrup Urine Disease (MSUD

    Directory of Open Access Journals (Sweden)

    Ralph Fingerhut

    2016-06-01

    Full Text Available Newborn screening for MSUD is a special challenge since patients with MSUD can metabolically decompensate rapidly without adequate treatment within the first two weeks of life. However, the screening method does not detect the actual marker metabolite (alloisoleucine specifically, but only as part of the group of the other isobaric amino acids leucine, isoleucine and hydroxyproline. We describe a sensitive and rapid second-tier UPLC-MS/MS method to determine branched-chain amino acids from the initial extraction of the screening sample. Quantification is based on a seven-point calibration curve. Reference ranges (mean ± SD in µmol/L were determined from 179 normal, not pre-selected samples from the newborn screening: leucine: 72 ± 27; isoleucine: 37 ± 19; valine: 98 ± 46; hydroxyproline: 23 ± 13. The concentration of alloisoleucine was below the detection limit in about 55% of the cases, and the highest concentration was 1.9 µmol/L. In all 30 retrospectively studied screening samples from patients with confirmed MSUD the concentration of alloisoleucine was significantly increased. In 238 samples with false-positive newborn screening due to a significant increase in the combined concentration of leucine + isoleucine + alloisoleucine + hydroxyproline (400 to >4000 µmol/L, alloisoleucine was below 6.5 µmol/L (n = 57 or not detectable (n = 181. The application of this assay markedly reduces the false-positive rate and the associated anxiety and costs. It is also suitable for routinely monitoring blood spots of patients with MSUD.

  7. Mean hemoglobin levels in venous blood samples and prevalence of anemia in Japanese elementary and junior high school students

    International Nuclear Information System (INIS)

    Screening for anemia has been performed in schools in Japan for over 30 years. The long-term effect of the nuclear power plant disaster on the prevalence of anemia in school age children is unknown. This research was performed to evaluate the prevalence of anemia in school age children and to determine grade-level and gender-related reference hemoglobin (Hb) levels prior to the nuclear disaster. Data for this research were obtained from results of screening for anemia obtained by venous blood sampling in schools in 2002. Mean Hb levels were calculated for each grade level (elementary school grades 1-6 and junior high school years 1-3) and according to gender, and the prevalence of anemia was determined. In our research, Tokyo Health Service Association guidelines were used to determine reference Hb levels for anemia. We demonstrated that Hb levels in boys increased with age during childhood and adolescence (from 13.1±0.7 g/dL in 7 year olds to 14.9±1.1 g/dL in 15 year olds); in girls, Hb levels peaked at menarche (13.7±0.8 g/dL in 12 year olds), decreasing slightly thereafter (13.4±1.1 g/dL in 15 year olds). The prevalence of anemia was 0.26% in elementary school boys, 0.27% in elementary school girls, and 1.21% in junior high school boys. The prevalence of anemia in second- and third-year junior high school girls was lower than that in first-year junior high school girls. Among all junior high school girls, 5.73% had mild anemia. Iron-deficiency anemia is the commonest type of anemia in high school girls, secondary to the relative lack of iron due to menstruation, the growth spurt and exercise. Appropriate dietary therapy and treatment of anemia, together with education about the dietary prevention of anemia, are important to reduce the prevalence of anemia in high school students. When complete blood counts are performed in regions thought to be affected by the Fukushima nuclear power plant disaster, our report can serve as a reference during evaluation of Hb

  8. Spotted Seal Distribution Map

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This dataset contains GIS layers that depict the known spatial distributions (i.e., ranges) and reported breeding areas of spotted seals (Phoca largha). It was...

  9. Mononucleosis spot test

    Science.gov (United States)

    Monospot test; Heterophile antibody test; Heterophile agglutination test; Paul-Bunnell test; Forssman antibody test ... The mononucleosis spot test is done when symptoms of mononucleosis are ... Fatigue Fever Large spleen (possibly) Sore throat Tender ...

  10. Expression and methylation data from SLE patient and healthy control blood samples subdivided with respect to ARID3a levels.

    Science.gov (United States)

    Ward, Julie M; Ratliff, Michelle L; Dozmorov, Mikhail G; Wiley, Graham; Guthridge, Joel M; Gaffney, Patrick M; James, Judith A; Webb, Carol F

    2016-12-01

    Previously published studies revealed that variation in expression of the DNA-binding protein ARID3a in B lymphocytes from patients with systemic lupus erythematosus (SLE) correlated with levels of disease activity ("Disease activity in systemic lupus erythematosus correlates with expression of the transcription factor AT-rich-interactive domain 3A" (J.M. Ward, K. Rose, C. Montgomery, I. Adrianto, J.A. James, J.T. Merrill et al., 2014) [1]). The data presented here compare DNA methylation patterns from SLE peripheral blood mononuclear cells obtained from samples with high numbers of ARID3a expressing B cells (ARID3a(H)) versus SLE samples with normal numbers of ARID3a(+) B cells (ARID3a(N)). The methylation data is available at the gene expression omnibus (GEO) repository, "Gene Expression Omnibus: NCBI gene expression and hybridization array data repository" (R. Edgar, M. Domrachev, A.E. Lash, 2002) [2]. Isolated B cells from SLE ARID3a(H) and ARID3a(N) B samples were also evaluated via qRT-PCR for Type I interferon (IFN) signature and pathway gene expression levels by qRT-PCR. Similarly, healthy control B cells and B cells stimulated to express ARID3a with the TLR agonist, CpG, were also compared via qRT-PCR. Primers designed to detect 6 IFNa subtype mRNAs were tested in 4 IFNa, Epstein-Barr Virus-transformed B cell lines ("Reduced interferon-alpha production by Epstein-Barr virus transformed B-lymphoblastoid cell lines and lectin-stimulated lymphocytes in congenital dyserythropoietic anemia type I" (S.H. Wickramasinghe, R. Hasan, J. Smythe, 1997) [3]). The data in this article support the publication, "Human effector B lymphocytes express ARID3a and secrete interferon alpha" (J.M. Ward, M.L. Ratliff, M.G. Dozmorov, G. Wiley, J.M. Guthridge, P.M. Gaffney, J.A. James, C.F. Webb, 2016) [4]. PMID:27656675

  11. Cloud point extraction for determination of lead in blood samples of children, using different ligands prior to analysis by flame atomic absorption spectrometry: A multivariate study

    Energy Technology Data Exchange (ETDEWEB)

    Shah, Faheem, E-mail: shah_ceac@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080 (Pakistan); Kazi, Tasneem Gul, E-mail: tgkazi@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080 (Pakistan); Afridi, Hassan Imran, E-mail: hassanimranafridi@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080 (Pakistan); Naeemullah, E-mail: khannaeemullah@ymail.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080 (Pakistan); Arain, Muhammad Balal, E-mail: bilal_ku2004@yahoo.com [Department of Chemistry, University of Science and Technology, Bannu, KPK (Pakistan); Baig, Jameel Ahmed, E-mail: jab_mughal@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080 (Pakistan)

    2011-09-15

    Highlights: {yields} Trace levels of lead in blood samples of healthy children and with different kidney disorders {yields} Pre-concentration of Pb{sup +2} in acid digested blood samples after chelating with two complexing reagents. {yields} Multivariate technique was used for screening of significant factors that influence the CPE of Pb{sup +2} {yields} The level of Pb{sup +2} in diseased children was significantly higher than referents of same age group. - Abstract: The phase-separation phenomenon of non-ionic surfactants occurring in aqueous solution was used for the extraction of lead (Pb{sup 2+}) from digested blood samples after simultaneous complexation with ammonium pyrrolidinedithiocarbamate (APDC) and diethyldithiocarbamate (DDTC) separately. The complexed analyte was quantitatively extracted with octylphenoxypolyethoxyethanol (Triton X-114). The multivariate strategy was applied to estimate the optimum values of experimental factors. Acidic ethanol was added to the surfactant-rich phase prior to its analysis by flame atomic absorption spectrometer (FAAS). The detection limit value of Pb{sup 2+} for the preconcentration of 10 mL of acid digested blood sample was 1.14 {mu}g L{sup -1}. The accuracy of the proposed methods was assessed by analyzing certified reference material (whole blood). Under the optimized conditions of both CPE methods, 10 mL of Pb{sup 2+} standards (10 {mu}g L{sup -1}) complexed with APDC and DDTC, permitted the enhancement factors of 56 and 42, respectively. The proposed method was used for determination of Pb{sup 2+} in blood samples of children with kidney disorders and healthy controls.

  12. Application of mRNA Expression Analysis to Human Blood Identification in Degenerated Samples that were False-negative by Immunochromatography(,) (.).

    Science.gov (United States)

    Matsumura, Shusaku; Matsusue, Aya; Waters, Brian; Kashiwagi, Masayuki; Hara, Kenji; Kubo, Shin-Ichi

    2016-07-01

    Forensic laboratories are often faced with cases in which methamphetamine hydrochloride-mixed blood is unable to be identified as human blood by immunochromatography against human hemoglobin A0. The application of mRNA expression analysis to samples that showed a false-negative with immunochromatography was investigated as an alternative approach that did not depend on the antigen-antibody reaction. Real-time PCR was used to examine the expression levels of blood markers such as glycophorin A, spectrin beta, and hemoglobin beta. Hemoglobin beta was the only marker that was specifically detected in blood, while glycophorin A was useful for determining human specificity. Hemoglobin beta showed good detection sensitivity and was detectable in 37-year-old blood stains. Hemoglobin beta was exclusively detectable in methamphetamine hydrochloride-mixed blood stains. Detergents and disinfectants did not significantly influence mRNA markers. The proposed mRNA expression analysis was suitable for human blood identification as an alternative method to immunochromatography. PMID:27364269

  13. 血液标本存放时间及存放方式对血糖检测结果的影响%Influence of storage time and storage way of blood sample on blood glucose testing result

    Institute of Scientific and Technical Information of China (English)

    周正国

    2015-01-01

    Objective To research and observe the influence of storage time and storage way of blood sample on blood glucose testing result, for providing reference for clinical detection in clinical practice. Methods 20 cases were selected randomly, and the patients were collected blood samples with empty belly. 14mL blood was collected for each patient, and 2mL once. The blood samples were stored in vacuum tubes which were marked as control(1), indoor temperature(2), 37℃thermostatic waterbath(2) and 0℃fridge(2). And the influence of storage time and storage way of blood sample on blood glucose testing result was analyzed. Results The influence of storage time of blood on blood glucose test is very evident. After 6h, the blood glucose level was 60%of the initial value. After 18h, he blood glucose level was 27%of initial value.And the blood glucose level after 24 h is 22%of initial value. The longer blood glucose was stored, the faster the blood glucose level reduced. After blood samples were stored in indoor temperature and 37℃thermostatic waterbath for 2.5h and 5h, the blood glucose testing result had evident difference from the initial value, which had statistical significance. Blood samples are stored in sodium fluoride potassium oxalate tube and separation gel coagulant tube, and the influence of it on blood glucose detection had no evident difference, which had no statistical significance. Conclusion In order to ensure that the result of blood glucose detection is accurate, the blood samples should be stored in the environment with low temperature, and the blood samples should be detected timely, to reduce the error of detection results and ensure that the result is reliable and accurate.%目的:研究和探讨血液标本存放时间和存放方式对血糖检测结果的影响,为日后临床检验提供参考价值。方法随机抽取20例患者,空腹采集血液标本,每位患者共采集14mL,每次2mL,存放于真空

  14. Chromosome aberrations induced in human lymphocytes by U-235 fission neutrons: I. Irradiation of human blood samples in the "dry cell" of the TRIGA Mark II nuclear reactor.

    Science.gov (United States)

    Fajgelj, A; Lakoski, A; Horvat, D; Remec, I; Skrk, J; Stegnar, P

    1991-11-01

    A set-up for irradiation of biological samples in the TRIGA Mark II research reactor in Ljubljana is described. Threshold activation detectors were used for characterisation of the neutron flux, and the accompanying gamma dose was measured by TLDs. Human peripheral blood samples were irradiated "in vitro" and biological effects evaluated according to the unstable chromosomal aberrations induced. Biological effects of two types of cultivation of irradiated blood samples, the first immediately after irradiation and the second after 96 h storage, were studied. A significant difference in the incidence of chromosomal aberrations between these two types of samples was obtained, while our dose-response curve fitting coefficients alpha 1 = (7.71 +/- 0.09) x 10(-2) Gy-1 (immediate cultivation) and alpha 2 = (11.03 +/- 0.08) x 10(-2) Gy-1 (96 h delayed cultivation) are in both cases lower than could be found in the literature.

  15. Chromosome aberrations induced in human lymphocytes by U-235 fission neutrons: I. Irradiation of human blood samples in the "dry cell" of the TRIGA Mark II nuclear reactor.

    Science.gov (United States)

    Fajgelj, A; Lakoski, A; Horvat, D; Remec, I; Skrk, J; Stegnar, P

    1991-11-01

    A set-up for irradiation of biological samples in the TRIGA Mark II research reactor in Ljubljana is described. Threshold activation detectors were used for characterisation of the neutron flux, and the accompanying gamma dose was measured by TLDs. Human peripheral blood samples were irradiated "in vitro" and biological effects evaluated according to the unstable chromosomal aberrations induced. Biological effects of two types of cultivation of irradiated blood samples, the first immediately after irradiation and the second after 96 h storage, were studied. A significant difference in the incidence of chromosomal aberrations between these two types of samples was obtained, while our dose-response curve fitting coefficients alpha 1 = (7.71 +/- 0.09) x 10(-2) Gy-1 (immediate cultivation) and alpha 2 = (11.03 +/- 0.08) x 10(-2) Gy-1 (96 h delayed cultivation) are in both cases lower than could be found in the literature. PMID:1962281

  16. Star spot location estimation using Kalman filter for star tracker.

    Science.gov (United States)

    Liu, Hai-bo; Yang, Jian-kun; Wang, Jiong-qi; Tan, Ji-chun; Li, Xiu-jian

    2011-04-20

    Star pattern recognition and attitude determination accuracy is highly dependent on star spot location accuracy for the star tracker. A star spot location estimation approach with the Kalman filter for a star tracker has been proposed, which consists of three steps. In the proposed approach, the approximate locations of the star spots in successive frames are predicted first; then the measurement star spot locations are achieved by defining a series of small windows around each predictive star spot location. Finally, the star spot locations are updated by the designed Kalman filter. To confirm the proposed star spot location estimation approach, the simulations based on the orbit data of the CHAMP satellite and the real guide star catalog are performed. The simulation results indicate that the proposed approach can filter out noises from the measurements remarkably if the sampling frequency is sufficient. PMID:21509065

  17. Correlation of omega-3 levels in serum phospholipid from 2053 human blood samples with key fatty acid ratios

    Directory of Open Access Journals (Sweden)

    Rowe William

    2009-12-01

    Full Text Available Abstract Background This research was conducted to explore the relationships between the levels of omega-3 fatty acids in serum phospholipid and key fatty acid ratios including potential cut-offs for risk factor assessment with respect to coronary heart disease and fatal ischemic heart disease. Methods Blood samples (n = 2053 were obtained from free-living subjects in North America and processed for determining the levels of total fatty acids in serum phospholipid as omega-3 fatty acids including EPA (eicosapentaenoic acid, 20:5 n-3 and DHA (docosahexaenoic acid, 22:6 n-3 by combined thin-layer and gas-liquid chromatographic analyses. The omega-3 levels were correlated with selected omega-6: omega-3 ratios including AA (arachidonic acid, 20:4n-6: EPA and AA:(EPA+DHA. Based on previously-published levels of omega-3 fatty acids considered to be in a 'lower risk' category for heart disease and related fatality, 'lower risk' categories for selected fatty acid ratios were estimated. Results Strong inverse correlations between the summed total of omega-3 fatty acids in serum phospholipid and all four ratios (omega-6:omega-3 (n-6:n-3, AA:EPA, AA:DHA, and AA:(EPA+DHA were found with the most potent correlation being with the omega-6:omega-3 ratio (R2 = 0.96. The strongest inverse relation for the EPA+DHA levels in serum phospholipid was found with the omega-6: omega-3 ratio (R2 = 0.94 followed closely by the AA:(EPA+DHA ratio at R2 = 0.88. It was estimated that 95% of the subjects would be in the 'lower risk' category for coronary heart disease (based on total omega-3 ≥ 7.2% with omega-6:omega-3 ratios Conclusions Strong inverse correlations between the levels of omega-3 fatty acids in serum (or plasma phospholipid and omega-6: omega-3 ratios are apparent based on this large database of 2053 samples. Certain fatty acid ratios may aid in cardiovascular disease-related risk assessment if/when complete profiles are not available.

  18. A high incidence of polymorphic CYP2C19 variants in archival blood samples from Papua New Guinea

    Directory of Open Access Journals (Sweden)

    Hsu Huai-Ling

    2008-09-01

    Full Text Available Abstract There is considerable inter-ethnic variability in the incidence of CYP2C19 genetic poor metabolisers (var/var. About 3 per cent of Caucasians are CYP2C19 var/var. By contrast, an extremely high incidence (70 per cent is observed in the Melanesian island of Vanuatu. The colonisation of the Pacific Islands is believed to have involved migration through Papua New Guinea (PNG, and hence a high incidence may also be expected in this population. The reported incidence in PNG was only 36 per cent, however. PNG is a country of extensive ethnic diversity, and the incidence of the CYP2C19 var/var in other regional populations of PNG is currently not established. In this study, restriction fragment length polymorphism-polymerase chain reaction analysis of archival blood serum samples was used to determine the prevalence of the CYP2C19*2 and *3 variant alleles in three different ethnic and geographically isolated populations of PNG. In the largest population studied (Iruna, the frequency of both variant CYP2C19 alleles was high (0.37 and 0.34, respectively. Specifically, the frequency of the CYP2C19*3 allele was significantly higher than in the PNG (East Sepik population reported previously (0.34 vs 0.16; p 0.0001. In the Iruna population, 48.9 per cent of the samples were homozygous variants for CYP2C19*2 or *3, which although higher was not statistically different from the East Sepik population (36 per cent. The results of this study indicated that other regional populations of PNG also have a relatively high incidence of the CYP2C19 genetic polymorphism compared with Caucasian populations. The high incidence reported in Vanuatu, however, may be due to genetic drift rather than a PNG founder population, as the Vanuatu population is dominated by the CYP2C19*2 allele, with a lower contribution from the *3 allelic variant.

  19. Kinetics of Dengue Non-Structural Protein 1 Antigen and IgM and IgA Antibodies in Capillary Blood Samples from Confirmed Dengue Patients

    OpenAIRE

    Matheus, Séverine; Pham, Thai Binh; Labeau, Bhetty; Huong, Vu Thi Que; Lacoste, Vincent; Deparis, Xavier; Marechal, Vincent

    2014-01-01

    Large-scale epidemiological surveillance of dengue in the field and dengue patient management require simple methods for sample collection, storage, and transportation as well as effective diagnostic tools. We evaluated the kinetics of three biological markers of dengue infection—non-structural protein 1 (NS1) antigen, immunoglobulin M (IgM), and IgA—in sequential capillary blood samples collected from fingertips of confirmed dengue patients. The overall sensitivities and specificities of the...

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

    OpenAIRE

    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 collec...

  1. Development and validation of an indirect Enzyme-linked Immunosorbent Assay for the detection of antibodies against Schmallenberg virus in blood samples from ruminants

    NARCIS (Netherlands)

    Heijden, van der H.M.J.F.; Bouwstra, R.J.; Mars, M.H.; Poel, van der W.H.M.; Wellenberg, G.J.; Maanen, van C.

    2013-01-01

    To detect Schmallenberg virus (SBV) infections in ruminants and to perform SBV epidemiological studies a cost-effective serological test is required. For these purposes an indirect whole virus Enzyme-linked Immunosorbent Assay (ELISA) for detection of SBV specific antibodies in ruminant blood sample

  2. Diagnosis of visceral leishmaniasis by the polymerase chain reaction using blood, bone marrow and lymph node samples from patients from the Sudan

    DEFF Research Database (Denmark)

    Andresen, K; Gasim, S; Elhassan, A M;

    1997-01-01

    We have evaluated the sensitivity of the polymerase chain reaction (PCR) as a diagnostic tool for Leishmania donovani using blood, bone marrow and lymph node samples from Sudanese patients with a confirmed infection. Forty patients were diagnosed by microscopic examination of bone marrow or lymph...

  3. Evaluation of Trapper-Collected Nobuto Filter-Paper Blood Samples for Distemper and Parvovirus Antibody Detection in Coyotes (Canis latrans) and Raccoons (Procyon lotor).

    Science.gov (United States)

    Kamps, Amanda J; Dubay, Shelli A; Langenberg, Julie; Maes, Roger K

    2015-07-01

    Blood samples are often collected from free-ranging wildlife for antibody detection. However, filter-paper (FP) strips are more cost efficient and easy to collect and store. We evaluated trapper-collected FP strips and body-cavity blood for canine distemper (CDV) and parvovirus (CPV-2) antibody detection in raccoons (Procyon lotor) and coyotes (Canis latrans). From 2008 to 2010, licensed trappers near Madison and Milwaukee, Wisconsin, US collected paired samples from harvested animals. Canine distemper antibodies were detected using virus neutralization and parvovirus antibodies were detected using hemagglutination inhibition. Titers ≥ 1:32 for CDV and ≥ 1:25 for CPV-2 were considered evidence of exposure. Using Cohen's kappa test of agreement, FP strip titers agreed with sera for CDV in coyotes (n = 28, K = 0.772) and raccoons (n = 29, K = 0.858) and for CPV-2 in coyotes (n = 40, K = 0.775) and raccoons (n = 70, K = 0.646). However, raccoons determined to be exposed to CPV-2 from sera were unexposed by FP strips in 35% of the samples. Titer results may be affected by quality and volume of blood samples, interval between collection and processing, small sample sizes, and diagnostic testing procedures. Filter-paper strips can be useful for detecting CDV and CPV-2 exposure in coyotes and raccoons with correct field sample collection and appropriate diagnostic testing procedures. PMID:25973631

  4. Ultra-fast local-haplotype variant calling using paired-end DNA-sequencing data reveals somatic mosaicism in tumor and normal blood samples.

    Science.gov (United States)

    Sengupta, Subhajit; Gulukota, Kamalakar; Zhu, Yitan; Ober, Carole; Naughton, Katherine; Wentworth-Sheilds, William; Ji, Yuan

    2016-02-18

    Somatic mosaicism refers to the existence of somatic mutations in a fraction of somatic cells in a single biological sample. Its importance has mainly been discussed in theory although experimental work has started to emerge linking somatic mosaicism to disease diagnosis. Through novel statistical modeling of paired-end DNA-sequencing data using blood-derived DNA from healthy donors as well as DNA from tumor samples, we present an ultra-fast computational pipeline, LocHap that searches for multiple single nucleotide variants (SNVs) that are scaffolded by the same reads. We refer to scaffolded SNVs as local haplotypes (LH). When an LH exhibits more than two genotypes, we call it a local haplotype variant (LHV). The presence of LHVs is considered evidence of somatic mosaicism because a genetically homogeneous cell population will not harbor LHVs. Applying LocHap to whole-genome and whole-exome sequence data in DNA from normal blood and tumor samples, we find wide-spread LHVs across the genome. Importantly, we find more LHVs in tumor samples than in normal samples, and more in older adults than in younger ones. We confirm the existence of LHVs and somatic mosaicism by validation studies in normal blood samples. LocHap is publicly available at http://www.compgenome.org/lochap. PMID:26420835

  5. Systolic Blood Pressure, Socioeconomic Status, and Biobehavioral Risk Factors in a Nationally Representative U.S Young Adult Sample

    OpenAIRE

    Brummett, Beverly H.; Babyak, Michael A; Siegler, Ilene C.; Shanahan, Michael; Harris, Kathleen Mullan; Elder, Glen H.; Williams, Redford B.

    2011-01-01

    In the National Longitudinal Study of Adolescent Health, a US longitudinal study of over 15,000 young adults, we examined the extent to which socioeconomic status is linked to systolic blood pressure, and whether biobehavioral risk factors mediate the association. Over 62% of the participants had systolic blood pressure >120 mmHg and 12% with systolic blood pressure >140 mmHg. Over 66% were classified as at least overweight (Body Mass Index>25 kg/m2), with over 36% meeting criteria for at lea...

  6. Novel system using microliter order sample volume for measuring arterial radioactivity concentrations in whole blood and plasma for mouse PET dynamic study.

    Science.gov (United States)

    Kimura, Yuichi; Seki, Chie; Hashizume, Nobuya; Yamada, Takashi; Wakizaka, Hidekatsu; Nishimoto, Takahiro; Hatano, Kentaro; Kitamura, Keishi; Toyama, Hiroshi; Kanno, Iwao

    2013-11-21

    This study aimed to develop a new system, named CD-Well, for mouse PET dynamic study. CD-Well allows the determination of time-activity curves (TACs) for arterial whole blood and plasma using 2-3 µL of blood per sample; the minute sample size is ideal for studies in small animals. The system has the following merits: (1) measures volume and radioactivity of whole blood and plasma separately; (2) allows measurements at 10 s intervals to capture initial rapid changes in the TAC; and (3) is compact and easy to handle, minimizes blood loss from sampling, and delay and dispersion of the TAC. CD-Well has 36 U-shaped channels. A drop of blood is sampled into the opening of the channel and stored there. After serial sampling is completed, CD-Well is centrifuged and scanned using a flatbed scanner to define the regions of plasma and blood cells. The length measured is converted to volume because the channels have a precise and uniform cross section. Then, CD-Well is exposed to an imaging plate to measure radioactivity. Finally, radioactivity concentrations are computed. We evaluated the performance of CD-Well in in vitro measurement and in vivo (18)F-fluorodeoxyglucose and [(11)C]2-carbomethoxy-3β-(4-fluorophenyl) tropane studies. In in vitro evaluation, per cent differences (mean±SE) from manual measurement were 4.4±3.6% for whole blood and 4.0±3.5% for plasma across the typical range of radioactivity measured in mouse dynamic study. In in vivo studies, reasonable TACs were obtained. The peaks were captured well, and the time courses coincided well with the TAC derived from PET imaging of the heart chamber. The total blood loss was less than 200 µL, which had no physiological effect on the mice. CD-Well demonstrates satisfactory performance, and is useful for mouse PET dynamic study.

  7. Comparison of real-time PCR and conventional PCR with two DNA targets for detection of Leishmania (Leishmania) infantum infection in human and dog blood samples.

    Science.gov (United States)

    Mohammadiha, A; Mohebali, M; Haghighi, A; Mahdian, R; Abadi, A R; Zarei, Z; Yeganeh, F; Kazemi, B; Taghipour, N; Akhoundi, B

    2013-01-01

    Zoonotic visceral leishmaniasis (VL) is endemic in northwestern Iran. Real-time PCR, conventional PCR, and the direct agglutination test (DAT) were used to diagnose Leishmania infantum infection in blood samples from 100 domestic dogs and 100 humans. Based on clinical evaluation, 82 humans and 72 dogs from the endemic area were categorized as having asymptomatic infection, DAT positive with no clinical signs of VL, or symptomatic infection, DAT positive with at least one sign of VL. Eighteen human samples containing no Leishmania antibodies (DAT(-)) and 28 dog DAT(-) sera from non-endemic areas with no history of VL constituted negative controls. All 46 DAT(-) samples were also negative by Dipstick rK39. Bone marrow material was used for parasitological examinations in symptomatic VL, and peripheral blood samples were used for detection of L. infantum infection using conventional PCR and real-time PCR in non-symptomatic subjects. Two DNA targets (ITS1 kDNA) were used for conventional PCR. L. infantum antibodies in sera were detected by DAT. Parasitemia was measured by real-time PCR targeting kDNA using Taqman Assay. All 72 (100%) symptomatic (38/38) and asymptomatic (34/34) dog DAT(+)samples, 45 of 48 (93.8%) symptomatic human DAT(+) samples, and 32 of 34 (94.1%) human asymptomatic cases were identified by real-time PCR. The mean (59.19 vs 12.38 parasite equivalents/mL of blood) and median (16.15 vs 1 parasite equivalents/mL of blood) ranges of parasitemia were higher in dogs than in humans (PPCR and DAT (99% in dogs and 95% in humans). Sensitivity of 100% and 93.9%, specificity of 96.4% and 100%, positive predictive values of 98.6% and 100%, and negative predictive values of 100% and 78.3% were found by real-time PCR for dog and human samples, respectively.

  8. Time Resolved X-Ray Spot Size Diagnostic

    CERN Document Server

    Richardson, Roger; Falabella, Steven; Guethlein, Gary; Raymond, Brett; Weir, John

    2005-01-01

    A diagnostic was developed for the determination of temporal history of an X-ray spot. A pair of thin (0.5 mm) slits image the x-ray spot to a fast scintillator which is coupled to a fast detector, thus sampling a slice of the X-Ray spot. Two other scintillator/detectors are used to determine the position of the spot and total forward dose. The slit signal is normalized to the dose and the resulting signal is analyzed to get the spot size. The position information is used to compensate for small changes due to spot motion and misalignment. The time resolution of the diagnostic is about 1 ns and measures spots from 0.5 mm to over 3 mm. The theory and equations used to calculate spot size and position are presented, as well as data. The calculations assume a symmetric, Gaussian spot. The spot data is generated by the ETA II accelerator, a 2kA, 5.5 MeV, 60ns electron beam focused on a Tantalum target. The spot generated is typically about 1 mm FWHM. Comparisons are made to an X-ray pinhole camera which images th...

  9. SPIDIA-RNA: second external quality assessment for the pre-analytical phase of blood samples used for RNA based analyses.

    Directory of Open Access Journals (Sweden)

    Francesca Malentacchi

    Full Text Available One purpose of the EC funded project, SPIDIA, is to develop evidence-based quality guidelines for the pre-analytical handling of blood samples for RNA molecular testing. To this end, two pan-European External Quality Assessments (EQAs were implemented. Here we report the results of the second SPIDIA-RNA EQA. This second study included modifications in the protocol related to the blood collection process, the shipping conditions and pre-analytical specimen handling for participants. Participating laboratories received two identical proficiency blood specimens collected in tubes with or without an RNA stabilizer. For pre-defined specimen storage times and temperatures, laboratories were asked to perform RNA extraction from whole blood according to their usual procedure and to return extracted RNA to the SPIDIA facility for further analysis. These RNA samples were evaluated for purity, yield, integrity, stability, presence of interfering substances, and gene expression levels for the validated markers of RNA stability: FOS, IL1B, IL8, GAPDH, FOSB and TNFRSF10c. Analysis of the gene expression results of FOS, IL8, FOSB, and TNFRSF10c, however, indicated that the levels of these transcripts were significantly affected by blood collection tube type and storage temperature. These results demonstrated that only blood collection tubes containing a cellular RNA stabilizer allowed reliable gene expression analysis within 48 h from blood collection for all the genes investigated. The results of these two EQAs have been proposed for use in the development of a Technical Specification by the European Committee for Standardization.

  10. Lead levels - blood

    Science.gov (United States)

    Blood lead levels ... A blood sample is needed. Most of the time blood is drawn from a vein located on the inside ... may be used to puncture the skin. The blood collects in a small glass tube called a ...

  11. Quantitative analysis of human herpesvirus-6 genome in blood and bone marrow samples from Tunisian patients with acute leukemia: a follow-up study

    Directory of Open Access Journals (Sweden)

    Faten Nefzi

    2012-11-01

    Full Text Available Abstract Background Infectious etiology in lymphoproliferative diseases has always been suspected. The pathogenic roles of human herpesvirus-6 (HHV-6 in acute leukemia have been of great interest. Discordant results to establish a link between HHV-6 activation and the genesis of acute leukemia have been observed. The objective of this study was to evaluate a possible association between HHV-6 infection and acute leukemia in children and adults, with a longitudinal follow-up at diagnosis, aplasia, remission and relapse. Methods HHV-6 load was quantified by a quantitative real-time PCR in the blood and bone marrow samples from 37 children and 36 adults with acute leukemia: 33 B acute lymphoblastic leukemia (B-ALL, 6 T acute lymphoblastic leukemia (T-ALL, 34 acute myeloid leukemia (AML. Results HHV-6 was detected in 15%, 8%, 30% and 28% of the blood samples at diagnosis, aplasia, remission and relapse, respectively. The median viral loads were 138, 244, 112 and 78 copies/million cells at diagnosis, aplasia, remission and relapse, respectively. In the bone marrow samples, HHV-6 was detected in 5%, 20% and 23% of the samples at diagnosis, remission and relapse, respectively. The median viral loads were 34, 109 and 32 copies/million cells at diagnosis, remission and relapse, respectively. According to the type of leukemia at diagnosis, HHV-6 was detected in 19% of the blood samples and in 7% of the bone marrow samples (with median viral loads at 206 and 79 copies/million cells, respectively from patients with B-ALL. For patients with AML, HHV-6 was present in 8% of the blood samples and in 4% of the bone marrow samples (with median viral loads at 68 and 12 copies/million cells, respectively. HHV-6 was more prevalent in the blood samples from children than from adults (25% and 9%, respectively and for the bone marrow (11% and 0%, respectively. All typable HHV-6 were HHV-6B species. No link was shown between neither the clinical symptoms nor the

  12. Incidence of Isolation of Mycobacterium Tuberculosis from Blood Samples in Tuberculosis Patients in Imam Khomeini Hospital, Tehran, Iran

    Directory of Open Access Journals (Sweden)

    Mehrnaz Rasoolinejad

    2011-08-01

    Full Text Available It is estimated that one third of the world's population is latently infected with tuberculosis (TB. The HIV epidemic fuels the TB epidemic by increasing the risk of reactivation of latent TB infection and by facilitating a more rapid progression of TB disease. Although the incidence of TB is constant or decreasing in many regions of the world, rates remain high in developing countries as a consequence of the HIV epidemic. This study was conducted as a collaboration of the Infectious Diseases department of Imam Khomeini Hospital with the Microbiology department of Tehran University of Medical Sciences. The hospital dataset of 94 patients admitted with TB during 2003-2005 was reviewed. We aimed to study factors correlating with positive blood culture including age, sex, immune deficiency status, HIV serology and SIRS (Systemic Inflammatory Response Syndrome status. In this study, we found that positive blood cultures are more frequent in patients less than 45 years old. Positive blood cultures were also more frequent in HIV infected patients and there was a significant correlation between blood culture and SIRS status. Therefore, we recommend that we obtain blood cultures from these high-risk groups in order to increase early detection of TB.

  13. Distribution of bacteria and yeasts within the 10-ml Isolator during the processing of seeded blood samples.

    Science.gov (United States)

    Kellogg, J A; Levisky, J S

    1986-02-01

    Forty-five organisms consisting of stock cultures and clinical isolates of bacteria and yeast were separately inoculated into outdated blood bank blood to achieve a concentration of approximately 100 CFU/ml. Blood with each organism was introduced into groups of four Isolators (E. I. du Pont de Nemours & Co., Inc., Wilmington, Del.), which were then processed according to the Isostat instructions of the manufacturer. The supernatant, sediment, and wash (material removed from the surface of the slanted stopper after sediment removal) were inoculated onto 5% sheep blood agar plates. Cultures were incubated aerobically (5 to 10% CO2) at 35 degrees C for 48 to 72 h. From the 180 Isolators, the mean recovery was 6% (range, 0 to 48%) for the supernatant, 87% (range, 47 to 98%) for the sediment, and 8% (range, 3 to 23%) for the wash. Neither variation among technologists nor intentional misalignment of additional Isolators in the centrifuge could explain all of the losses of microorganisms from the sediment. The manual nature of the Isolator procedure, which led to the loss of significant amounts of organisms from the sediment, may help to explain false-negative Isolator results obtained from blood of patients, particularly when small numbers of pathogens are present. PMID:3084546

  14. Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan

    Directory of Open Access Journals (Sweden)

    A Rasul

    2013-06-01

    Full Text Available Background: The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp. in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.Methods: Reverse line blot (RLB assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.Results: Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.Conclusion: Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001. Risk factor analysis revealed that male (P = 0.03, ani­mals infested by ticks (P = 0.03 and herd composed of sheep only (P = 0.001 were more infected by blood parasites.

  15. SpotADAPT

    DEFF Research Database (Denmark)

    Kaulakiene, Dalia; Thomsen, Christian; Pedersen, Torben Bach;

    2015-01-01

    Having constantly increasing amounts of data, the analysis of it is often entrusted for a MapReduce framework. The execution of an analytical workload can be cheapened by adopting cloud computing resources, and in particular by using spot instances (cheap, fluctuating price instances) offered by ...

  16. Feasibility of Using the Mosquito Blood Meal for Rapid and Efficient Human and Animal Virus Surveillance and Discovery.

    Science.gov (United States)

    Yang, Yu; Garver, Lindsey S; Bingham, Karen M; Hang, Jun; Jochim, Ryan C; Davidson, Silas A; Richardson, Jason H; Jarman, Richard G

    2015-12-01

    Mosquito blood meals taken from humans and animals potentially represent a useful source of blood for the detection of blood-borne pathogens. In this feasibility study, Anopheles stephensi mosquitoes were fed with blood meals spiked with dengue virus type 2 (DENV-2) and harvested at serial time points. These mosquitoes are not competent vectors, and the virus is not expected to replicate. Ingested blood was spotted on Whatman FTA cards and stored at room temperature. Mosquito abdomens were removed and stored at -80°C. Control blood meal aliquots were stored in vials or applied onto FTA cards. After 4 weeks of storage, the samples were extracted using beadbeating and QIAamp Viral RNA kit (Qiagen Sciences, Germantown, MD). Recovered viral RNA was analyzed by DENV-2 TaqMan RT-PCR assay and next-generation sequencing (NGS). Overall viral RNA recovery efficiency was 15% from the directly applied dried blood spots and approximately 20% or higher for dried blood spots made by blotting mosquito midgut on FTA cards. Viral RNA in mosquito-ingested blood decreases over time, but remains detectable 24 hours after blood feeding. The viral sequences in FTA-stored specimens can be maintained at room temperature. The strategy has the potential utility in expedited zoonotic virus discovery and blood-borne pathogen surveillance. PMID:26416112

  17. Comparison of Performance Characteristics of Aspergillus PCR in Testing a Range of Blood-Based Samples in Accordance with International Methodological Recommendations.

    Science.gov (United States)

    Springer, Jan; White, P Lewis; Hamilton, Shanna; Michel, Denise; Barnes, Rosemary A; Einsele, Hermann; Löffler, Juergen

    2016-03-01

    Standardized methodologies for the molecular detection of invasive aspergillosis (IA) have been established by the European Aspergillus PCR Initiative for the testing of whole blood, serum, and plasma. While some comparison of the performance of Aspergillus PCR when testing these different sample types has been performed, no single study has evaluated all three using the recommended protocols. Standardized Aspergillus PCR was performed on 423 whole-blood pellets (WBP), 583 plasma samples, and 419 serum samples obtained from hematology patients according to the recommendations. This analysis formed a bicenter retrospective anonymous case-control study, with diagnosis according to the revised European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) consensus definitions (11 probable cases and 36 controls). Values for clinical performance using individual and combined samples were calculated. For all samples, PCR positivity was significantly associated with cases of IA (for plasma, P = 0.0019; for serum, P = 0.0049; and for WBP, P = 0.0089). Plasma PCR generated the highest sensitivity (91%); the sensitivities for serum and WBP PCR were 80% and 55%, respectively. The highest specificity was achieved when testing WBP (96%), which was significantly superior to the specificities achieved when testing serum (69%, P = 0.0238) and plasma (53%, P = 0.0002). No cases were PCR negative in all specimen types, and no controls were PCR positive in all specimens. This study confirms that Aspergillus PCR testing of plasma provides robust performance while utilizing commercial automated DNA extraction processes. Combining PCR testing of different blood fractions allows IA to be both confidently diagnosed and excluded. A requirement for multiple PCR-positive plasma samples provides similar diagnostic utility and is technically less demanding. Time

  18. An approach for addressing hard-to-detect hot spots.

    Science.gov (United States)

    Abelquist, Eric W; King, David A; Miller, Laurence F; Viars, James A

    2013-05-01

    The Multi-Agency Radiation Survey and Site Investigation Manual (MARSSIM) survey approach is comprised of systematic random sampling coupled with radiation scanning to assess acceptability of potential hot spots. Hot spot identification for some radionuclides may not be possible due to the very weak gamma or x-ray radiation they emit-these hard-to-detect nuclides are unlikely to be identified by field scans. Similarly, scanning technology is not yet available for chemical contamination. For both hard-to-detect nuclides and chemical contamination, hot spots are only identified via volumetric sampling. The remedial investigation and cleanup of sites under the Comprehensive Environmental Response, Compensation, and Liability Act typically includes the collection of samples over relatively large exposure units, and concentration limits are applied assuming the contamination is more or less uniformly distributed. However, data collected from contaminated sites demonstrate contamination is often highly localized. These highly localized areas, or hot spots, will only be identified if sample densities are high or if the environmental characterization program happens to sample directly from the hot spot footprint. This paper describes a Bayesian approach for addressing hard-to-detect nuclides and chemical hot spots. The approach begins using available data (e.g., as collected using the standard approach) to predict the probability that an unacceptable hot spot is present somewhere in the exposure unit. This Bayesian approach may even be coupled with the graded sampling approach to optimize hot spot characterization. Once the investigator concludes that the presence of hot spots is likely, then the surveyor should use the data quality objectives process to generate an appropriate sample campaign that optimizes the identification of risk-relevant hot spots. PMID:23528274

  19. A non-organic and non-enzymatic extraction method gives higher yields of genomic DNA from whole-blood samples than do nine other methods tested.

    Science.gov (United States)

    Lahiri, D K; Bye, S; Nurnberger, J I; Hodes, M E; Crisp, M

    1992-12-01

    We compared ten methods for extraction of DNA from whole blood. Nine methods require incubation with either enzymes or treatment of organic solvents or both. The 'Rapid Method' (RM) (Method 10) avoids the use of organic solvents (phenol/chloroform) and eliminates completely the use of proteinase K. Thus, the time and cost of DNA extraction are reduced significantly. This is accomplished by salting out and precipitation of the cellular proteins in saturated sodium chloride. This method takes less than an hour to completion, without compromising the yield or the quality of DNA. Using RM, we can make DNA from 0.1 ml of whole blood and as little as 0.5 ml of blood yields DNA sufficient to run a few Southern blots. The RM can also be applied to packed cells. The DNA is free of RNA, protein and degrading enzymes. The uncut DNA runs as a typical slow-migrating, high-molecular-weight and undegraded species in an agarose gel. The DNA is suitable for digestion by various restriction endonucleases. This procedure works equally well with fresh blood samples and with those that are stored at 4 degrees C and -70 degrees C. To our knowledge the RM reported here is the safest, fastest and most quantitative and economical method for preparation of DNA from whole blood and cells.

  20. Selection of suitable reference genes for normalization of quantitative RT-PCR in peripheral blood samples of bottlenose dolphins (Tursiops truncatus).

    Science.gov (United States)

    Chen, I-Hua; Chou, Lien-Siang; Chou, Shih-Jen; Wang, Jiann-Hsiung; Stott, Jeffrey; Blanchard, Myra; Jen, I-Fan; Yang, Wei-Cheng

    2015-01-01

    Quantitative RT-PCR is often used as a research tool directed at gene transcription. Selection of optimal housekeeping genes (HKGs) as reference genes is critical to establishing sensitive and reproducible qRT-PCR-based assays. The current study was designed to identify the appropriate reference genes in blood leukocytes of bottlenose dolphins (Tursiops truncatus) for gene transcription research. Seventy-five blood samples collected from 7 bottlenose dolphins were used to analyze 15 candidate HKGs (ACTB, B2M, GAPDH, HPRT1, LDHB, PGK1, RPL4, RPL8, RPL18, RPS9, RPS18, TFRC, YWHAZ, LDHA, SDHA). HKG stability in qRT-PCR was determined using geNorm, NormFinder, BestKeeper and comparative delta Ct algorithms. Utilization of RefFinder, which combined all 4 algorithms, suggested that PGK1, HPRT1 and RPL4 were the most stable HKGs in bottlenose dolphin blood. Gene transcription perturbations in blood can serve as an indication of health status in cetaceans as it occurs prior to alterations in hematology and chemistry. This study identified HKGs that could be used in gene transcript studies, which may contribute to further mRNA relative quantification research in the peripheral blood leukocytes in captive cetaceans. PMID:26486099

  1. Hot spots of mutualistic networks.

    Science.gov (United States)

    Gilarranz, Luis J; Sabatino, Malena; Aizen, Marcelo A; Bascompte, Jordi

    2015-03-01

    Incorporating interactions into a biogeographical framework may serve to understand how interactions and the services they provide are distributed in space. We begin by simulating the spatiotemporal dynamics of realistic mutualistic networks inhabiting spatial networks of habitat patches. We proceed by comparing the predicted patterns with the empirical results of a set of pollination networks in isolated hills of the Argentinian Pampas. We first find that one needs to sample up to five times as much area to record interactions as would be needed to sample the same proportion of species. Secondly, we find that peripheral patches have fewer interactions and harbour less nested networks - therefore potentially less resilient communities - compared to central patches. Our results highlight the important role played by the structure of dispersal routes on the spatial distribution of community patterns. This may help to understand the formation of biodiversity hot spots. PMID:25402941

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

    DEFF Research Database (Denmark)

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

    2011-01-01

    We have implemented and validated automated protocols for DNA extraction and PCR setup using a Tecan Freedom EVO liquid handler mounted with the Te-MagS magnetic separation device (Tecan, Männedorf, Switzerland). The protocols were validated for accredited forensic genetic work according to ISO...... 17025 using the Qiagen MagAttract DNA Mini M48 kit (Qiagen GmbH, Hilden, Germany) from fresh whole blood and blood from deceased individuals. The workflow was simplified by returning the DNA extracts to the original tubes minimizing the risk of misplacing samples. The tubes that originally contained...... the samples were washed with MilliQ water before the return of the DNA extracts. The PCR was setup in 96-well microtiter plates. The methods were validated for the kits: AmpFlSTR Identifiler, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, WI...

  3. PCB Concentrations and Dioxin-like Activity in Blood Samples from Danish School Children and Their Mothers living in Urban and Rural Areas

    DEFF Research Database (Denmark)

    Mørck, Thit A; Erdmann, Simon E; Long, Manhai;

    2014-01-01

    Human exposure to persistent organic pollutants (POPs) is of major concern due to a diversity of adverse effects from prolonged exposure and bioaccumulation. Manufacturing of polychlorinated biphenyls (PCBs), a subgroup of POPs, has been prohibited for many decades; however, human exposure still......R transactivity assay were analysed in blood samples from Danish schoolchildren and their mothers in the European framework of the DEMOCOPHES/COPHES projects. The participants were selected from an urban and a rural area, respectively. The PCB concentrations and the AhR-TEQ (TCDD toxic equivalent) were......, there was a positive association between age and PCB concentration. These results show that both PCBs and dioxin-like activity can be measured as biomarkers of exposure and effects in blood samples from children and women. The results indicate that people living in urban areas may be exposed to higher concentrations...

  4. 7 CFR 28.425 - Low Middling Spotted Color.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples...

  5. A randomised clinical trial on cardiotocography plus fetal blood sampling versus cardiotocography plus ST-analysis of the fetal electrocardiogram (STAN®) for intrapartum monitoring

    OpenAIRE

    Rijnders Robbert JP; Porath Martina M; Oei S Guid; Nijhuis Jan G; Mol Ben WJ; van Lith Jan MM; van Geijn Herman P; Drogtrop Addy P; Bijvoet Saskia M; van Beek Erik; Moons Karel GM; Westerhuis Michelle EMH; Schuitemaker Nico WE; van der Tweel Ingeborg; Visser Gerard HA

    2007-01-01

    Abstract Background Cardiotocography (CTG) is worldwide the method for fetal surveillance during labour. However, CTG alone shows many false positive test results and without fetal blood sampling (FBS), it results in an increase in operative deliveries without improvement of fetal outcome. FBS requires additional expertise, is invasive and has often to be repeated during labour. Two clinical trials have shown that a combination of CTG and ST-analysis of the fetal electrocardiogram (ECG) reduc...

  6. Determining the Diagnostic Value of Mycobacterium Tuberculosis DNA in the Differentiation of Blood Samples of Patients with Active Pulmonary Tuberculosis and Healthy Controls Using Polymerase Chain Reaction

    OpenAIRE

    Abasali Niazi; Nezarali Muolai; Mosayeb Shahriar; Reza Karimian; Farzaneh Peykfalak

    2013-01-01

    Background: Tuberculosis (TB) is now a major cause of mortality and morbidity in the world. Nowadays, different methods are used to diagnose tuberculosis. Although classical microbiological methods (such as sputum smear) are specific, they have little sensitivity and the culture is also time-consuming. Using Polymerase Chain Reaction (PCR) in blood samples in terms of Mycobacterium tuberculosis DNA, this study examines diagnostic power of this test in the diagnosis of pulmonary tuberculosis c...

  7. Biomarkers of polycyclic aromatic hydrocarbon-DNA damage and cigarette smoke exposures in paired maternal and newborn blood samples as a measure of differential susceptibility

    Energy Technology Data Exchange (ETDEWEB)

    Whyatt, R.M.; Jedrychowski, W.; Hemminki, K.; Santella, R.M.; Tsai WeiYann; Yang Ke; Perera, F.P. [Columbia University, New York, NY (US). Division of Environmental Health Sciences, Mailman School of Public Health

    2001-07-01

    In this study, we report on three biomarkers measured in paired blood samples collected at birth from 160 mother/newborn pairs from Poland: 70 pairs from Krakow (a city with high air pollution including PAHs) and 90 pairs from Limanowa (an area with lower ambient pollution but greater indoor coal use). Field studies were conducted during January-March 1992. Biomarkers were: WBC aromatic-DNA adducts by {sup 32}P-postlabeling and PAH-DNA adducts by ELISA and plasma cotinine. Correlations were assessed by Spearman's rank test, and differences in biomarker levels were assessed by the Wilcoxon signed-ranks test. A significant correlation between paired newborn/maternal samples was seen for aromatic-DNA adduct levels and plasma cotinine, but not PAH-DNA adduct levels. Among the total cohort, levels of the three biomarkers were higher in newborn samples compared with paired maternal samples. The difference was significant for aromatic-DNA adduct levels (16.6 plus or minus 12.5 versus 14.21 plus or minus 15.4/10{sup 8} nucleotides; P=0.002) and plasma cotinine, but not for PAH-DNA adduct levels. When analyses were restricted to the 80 mother/newborn pairs from whom the blood sample was drawn concurrently, levels of all of the three biomarkers were significantly higher in the newborn compared with paired maternal blood samples (P {lt} 0.05). These results suggest that the fetus has reduced detoxification capabilities and increased susceptibility to DNA damage, especially in light of experimental evidence that transplacental exposures to PAHs are 10-fold lower than paired maternal exposures. Also, these results have implications for risk assessment, which currently does not adequately account for sensitive subsets of the population. 64 refs.

  8. How well do blood folate concentrations predict dietary folate intakes in a sample of Canadian lactating women exposed to high levels of folate? An observational study

    Directory of Open Access Journals (Sweden)

    Sherwood Kelly L

    2007-10-01

    Full Text Available Abstract Background In 1998, mandatory folic acid fortification of white flour and select cereal grain products was implemented in Canada with the intention to increase dietary folate intakes of reproducing women. Folic acid fortification has produced a dramatic increase in blood folate concentrations among reproductive age women, and a reduction in neural tube defect (NTD-affected pregnancies. In response to improved blood folate concentrations, many health care professionals are asking whether a folic acid supplement is necessary for NTD prevention among women with high blood folate values, and how reliably high RBC folate concentrations predict folate intakes shown in randomized controlled trials to be protective against NTDs. The objective of this study was to determine how predictive blood folate concentrations and folate intakes are of each other in a sample of well-educated lactating Canadian women exposed to high levels of synthetic folate. Methods The relationship between blood folate concentrations and dietary folate intakes, determined by weighed food records, were assessed in a sample of predominantly university-educated lactating women (32 ± 4 yr at 4-(n = 53 and 16-wk postpartum (n = 55. Results Median blood folate concentrations of all participants were well above plasma and RBC folate cut-off levels indicative of deficiency (6.7 and 317 nmol/L, respectively and all, except for 2 subjects, were above the cut-off for NTD-risk reduction (>906 nmol/L. Only modest associations existed between total folate intakes and plasma (r = 0.46, P P nd quartile of intake did not differ from that of women consuming >410 μg/d (3rd and 4th quartile. Conclusion Folate intakes, estimated by food composition tables, and blood folate concentrations are not predictive of each other in Canadian lactating women exposed to high levels of folate. Synthetic intakes > 151–410 μg/d in these women produced little additional benefit in terms of maximizing

  9. Detection and quantification of Wuchereria bancrofti and Brugia malayi DNA in blood samples and mosquitoes using duplex droplet digital polymerase chain reaction.

    Science.gov (United States)

    Jongthawin, Jurairat; Intapan, Pewpan M; Lulitanond, Viraphong; Sanpool, Oranuch; Thanchomnang, Tongjit; Sadaow, Lakkhana; Maleewong, Wanchai

    2016-08-01

    Lymphatic filariasis, a mosquito-borne disease, is still a major public health problem in tropical and sub-tropical countries. Effective diagnostic tools are required for identification of infected individuals, for epidemiological assessment, and for monitoring of control programs. A duplex droplet digital polymerase chain reaction (ddPCR) was conducted to differentiate and quantify Wuchereria bancrofti DNA by targeting the long DNA repeat (LDR) element and Brugia malayi DNA by targeting the HhaI element in blood samples and mosquito vectors. The analytical sensitivity and specificity were evaluated. Our results indicated that the duplex ddPCR assay could differentiate and quantify W. bancrofti and B. malayi DNA from blood samples and mosquitoes. DNA from a single larva in 50 μl of a blood sample, or in one mosquito vector, could be detected. The analytical sensitivity and specificity for W. bancrofti are both 100 %. Corresponding values for B. malayi are 100 and 98.3 %, respectively. Therefore, duplex ddPCR is a potential tool for simultaneous diagnosis and monitoring of bancroftian and brugian filariasis in endemic areas. PMID:27085707

  10. Separation and preconcentration of trace level of lead in one drop of blood sample by using graphite furnace atomic absorption spectrometry.

    Science.gov (United States)

    Shrivas, Kamlesh; Patel, Devesh Kumar

    2010-04-15

    Drop-to-drop solvent microextraction (DDSME) assisted with ultrasonication is applied for the determination of lead in one drop (30 microL) of blood sample by using graphite furnace atomic absorption spectrometry (GF-AAS). The optimum extraction efficiency of lead was observed for 10 min extraction time at pH 5.0 with 2 microL of organic solvent that containing 0.5 M of Cyanex-302. The optimized methodology exhibited good linearity in the range of 0.3-30.0 ng mL(-1) lead with relative standard deviations (RSD) from 2.5 to 4.4%. The method is found to be simple and rapid for the analysis of lead in micro amount of blood sample with the limit of detection (LOD) of 0.08 ng mL(-1). The application of the proposed method has been successfully tested for the determination of lead in blood samples. The results showed that under the optimized experimental conditions, the method showed good sensitivity and recovery %, as well as advantages such as linearity, simplicity, low cost and high feasibility. PMID:20004520

  11. Separation and preconcentration of trace level of lead in one drop of blood sample by using graphite furnace atomic absorption spectrometry

    International Nuclear Information System (INIS)

    Drop-to-drop solvent microextraction (DDSME) assisted with ultrasonication is applied for the determination of lead in one drop (30 μL) of blood sample by using graphite furnace atomic absorption spectrometry (GF-AAS). The optimum extraction efficiency of lead was observed for 10 min extraction time at pH 5.0 with 2 μL of organic solvent that containing 0.5 M of Cyanex-302. The optimized methodology exhibited good linearity in the range of 0.3-30.0 ng mL-1 lead with relative standard deviations (RSD) from 2.5 to 4.4%. The method is found to be simple and rapid for the analysis of lead in micro amount of blood sample with the limit of detection (LOD) of 0.08 ng mL-1. The application of the proposed method has been successfully tested for the determination of lead in blood samples. The results showed that under the optimized experimental conditions, the method showed good sensitivity and recovery %, as well as advantages such as linearity, simplicity, low cost and high feasibility.

  12. Detection of Babesia bovis in blood samples and its effect on the hematological and serum biochemical profile in large ruminants from Southern Punjab

    Institute of Scientific and Technical Information of China (English)

    Samreen Zulfiqar; Ali Saeed; Furhan Iqbal; Sadia Shahnawaz; Muhammad Ali; Arif Mahmood Bhutta; Shahid Iqbal; Sikandar Hayat; Shazia Qadir; Muhammad Latif; Nazia Kiran

    2012-01-01

    Objective:To determine the presence of Babesia bovis (B. bovis) in large ruminants in southern Punjab and its effect on hematological and serum biochemical profile of host animals. Methods:Blood samples were collected from 144 large ruminants, including 105 cattle and 39 buffaloes, from six districts in southern Punjab including Multan, Layyah, Muzaffar Garh, Bhakar, Bahawalnagar and Vehari. Data on the characteristics of animals and herds were collected through questionnaires. Different blood (hemoglobin, glucose) and serum (ALT, AST, LDH, cholesterol) parameters of calves and cattle were measured and compared between parasite positive and negative samples to demonstrate the effect of B. bovis on the blood and serological profile of infected animals. Results:27 out of 144 animals, from 5 out of 6 sampling districts, produced the 541-bp fragment specific for B. bovis. Age of animals (P=0.02), presence of ticks on animals (P=0.04) and presence of ticks on dogs associated with herds (P=0.5) were among the major risk factors involved in the spread of bovine babesiosis in the study area. ALT concentrations were the only serum biochemical values that significantly varied between parasite positive and negative cattle. Conclusions:This study has reported for the first time the presence of B. bovis in large ruminant and the results can lead to the prevention of babesiosis in the region to increase the livestock output.

  13. Field-adapted sampling of whole blood to determine the levels of amodiaquine and its metabolite in children with uncomplicated malaria treated with amodiaquine plus artesunate combination

    Directory of Open Access Journals (Sweden)

    Gustafsson Lars L

    2009-03-01

    Full Text Available Abstract Background Artemisinin combination therapy (ACT has been widely adopted as first-line treatment for uncomplicated falciparum malaria. In Uganda, amodiaquine plus artesunate (AQ+AS, is the alternative first-line regimen to Coartem® (artemether + lumefantrine for the treatment of uncomplicated falciparum malaria. Currently, there are few field-adapted analytical techniques for monitoring amodiaquine utilization in patients. This study evaluates the field applicability of a new method to determine amodiaquine and its metabolite concentrations in whole blood dried on filter paper. Methods Twelve patients aged between 1.5 to 8 years with uncomplicated malaria received three standard oral doses of AQ+AS. Filter paper blood samples were collected before drug intake and at six different time points over 28 days period. A new field-adapted sampling procedure and liquid chromatographic method was used for quantitative determination of amodiaquine and its metabolite in whole blood. Results The sampling procedure was successively applied in the field. Amodiaquine could be quantified for at least three days and the metabolite up to 28 days. All parasites in all the 12 patients cleared within the first three days of treatment and no adverse drug effects were observed. Conclusion The methodology is suitable for field studies. The possibility to determine the concentration of the active metabolite of amodiaquine up to 28 days suggested that the method is sensitive enough to monitor amodiaquine utilization in patients. Amodiaquine plus artesunate seems effective for treatment of falciparum malaria.

  14. Analytical sample preparation strategies for the determination of antimalarial drugs in human whole blood, plasma and urine

    DEFF Research Database (Denmark)

    Casas, Monica Escolà; Hansen, Martin; Krogh, Kristine A;

    2014-01-01

    Antimalarial drugs commonly referred to as antimalarials, include a variety of compounds with different physicochemical properties. There is a lack of information on antimalarial distribution in the body over time after administration, e.g. the drug concentrations in whole blood, plasma, and urine...... summarized. Finally, the main problems that the researchers have dealt with are highlighted. This information will aid analytical chemists in the development of novel methods for determining existing antimalarials and upcoming new drugs....

  15. Extensive surface protein profiles of extracellular vesicles from cancer cells may provide diagnostic signatures from blood samples

    OpenAIRE

    Belov, Larissa; Matic, Kieran J.; Hallal, Susannah; Mulligan, Stephen P.; Best, O. Giles; Christopherson, Richard I

    2016-01-01

    Extracellular vesicles (EV) are membranous particles (30–1,000 nm in diameter) secreted by cells. Important biological functions have been attributed to 2 subsets of EV, the exosomes (bud from endosomal membranes) and the microvesicles (MV; bud from plasma membranes). Since both types of particles contain surface proteins derived from their cell of origin, their detection in blood may enable diagnosis and prognosis of disease. We have used an antibody microarray (DotScan) to compare the surfa...

  16. The post-occipital spinal venous sinus of the Nile crocodile (Crocodylus niloticus: Its anatomy and use for blood sample collection and intravenous infusions

    Directory of Open Access Journals (Sweden)

    Jan G. Myburgh

    2014-02-01

    Full Text Available The post-occipital sinus of the spinal vein is often used for the collection of blood samples from crocodilians. Although this sampling method has been reported for several crocodilian species, the technique and associated anatomy has not been described in detail in any crocodilian, including the Nile crocodile (Crocodylus niloticus. The anatomy of the cranial neck region was investigated macroscopically, microscopically, radiographically and by means of computed tomography. Latex was injected into the spinal vein and spinal venous sinus of crocodiles to visualise the regional vasculature. The spinal vein ran within the vertebral canal, dorsal to and closely associated with the spinal cord and changed into a venous sinus cranially in the post-occipital region. For blood collection, the spinal venous sinus was accessed through the interarcuate space between the atlas and axis (C1 and C2 by inserting a needle angled just off the perpendicular in the midline through the craniodorsal cervical skin, just cranial to the cranial borders of the first cervical osteoderms. The most convenient method of blood collection was with a syringe and hypodermic needle. In addition, the suitability of the spinal venous sinus for intravenous injections and infusions in live crocodiles was evaluated. The internal diameter of the commercial human epidural catheters used during these investigations was relatively small, resulting in very slow infusion rates. Care should be taken not to puncture the spinal cord or to lacerate the blood vessel wall using this route for blood collection or intravenous infusions.

  17. Poisson Spot with Magnetic Levitation

    Science.gov (United States)

    Hoover, Matthew; Everhart, Michael; D'Arruda, Jose

    2010-01-01

    In this paper we describe a unique method for obtaining the famous Poisson spot without adding obstacles to the light path, which could interfere with the effect. A Poisson spot is the interference effect from parallel rays of light diffracting around a solid spherical object, creating a bright spot in the center of the shadow.

  18. Spotting a fake

    International Nuclear Information System (INIS)

    Diamonds are highly prized for their dazzling appearance and hardness, but would you be able to spot one that had been created in the laboratory? Simon Lawson describes how physics-based techniques can distinguish between natural and synthetic stones. For the last 50 years or so we have been able to make synthetic diamonds that replicate the superlative physical and chemical properties of natural diamonds, and these are used largely for industrial applications. But in the mind of the consumer, there is far more to a diamond than its hardness or brilliance. Research commissioned by the Diamond Trading Company (DTC) has shown that 94% of women surveyed prefer natural diamonds over synthetic ones as a symbol of love, possibly as a result of the immense age of natural stones. One of the key research activities at the DTC is therefore to ensure that synthetic diamonds can be spotted easily. (U.K.)

  19. Flying spot scanner

    International Nuclear Information System (INIS)

    An improved flying spot x-ray scanning equipment is described which includes a grid controlled x-ray tube and associated collimators for producing a pencil beam of x-rays. It is possible to control the position of the scan field relative to the patient, to control the width of the scan field and also to independently achieve an arbitary variation in the longitudinal dimension of the scan field. (U.K.)

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

    Science.gov (United States)

    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. PMID:26320872

  1. Feasibility study on blood sample investigations from former Wismut employees with respect to possible biomarkers for arsenic or radiation exposure using proteomics and cDNA microarray technologies. Final report

    International Nuclear Information System (INIS)

    The final report on the feasibility of blood sample investigations from former Wismut employees with respect to possible biomarkers for arsenic or radiation exposure using proteomics and cDNA microarray technologies covers the following topics: blood samples; methodologies: 2D gel electrophoresis; protein identification using MALDI-MS; accomplishment and evaluation of the proteomics and cDNA microarray analysis.

  2. Spot- Zombie Filtering System

    Directory of Open Access Journals (Sweden)

    Arathy Rajagopal

    2014-01-01

    Full Text Available A major security challenge on the Internet is the existence of the large number of compromised machines. Such machines have been increasingly used to launch various security attacks including spamming and spreading malware, DDoS, and identity theft. These compromised machines are called “Zombies”. In general E-mail applications and providers uses spam filters to filter the spam messages. Spam filtering is a technique for discriminating the genuine message from the spam messages. The attackers send the spam messages to the targeted machine by exalting the filters, which causes the increase in false positives and false negatives. We develop an effective spam zombie detection system named SPOT by monitoring outgoing messages of a network. SPOT focuses on the number of outgoing messages that are originated or forwarded by each computer on a network to identify the presence of Zombies. SPOT is designed based on a powerful statistical tool called Sequential Probability Ratio Test, which has bounded false positive and false negative error rates.

  3. Spot- Zombie Filtering System

    Directory of Open Access Journals (Sweden)

    Arathy Rajagopal

    2015-10-01

    Full Text Available A major security challenge on the Internet is the existence of the large number of compromised machines. Such machines have been increasingly used to launch various security attacks including spamming and spreading malware, DDoS, and identity theft. These compromised machines are called "Zombies". In general E-mail applications and providers uses spam filters to filter the spam messages. Spam filtering is a technique for discriminating the genuine message from the spam messages. The attackers send the spam messages to the targeted machine by exalting the filters, which causes the increase in false positives and false negatives. We develop an effective spam zombie detection system named SPOT by monitoring outgoing messages of a network. SPOT focuses on the number of outgoing messages that are originated or forwarded by each computer on a network to identify the presence of Zombies. SPOT is designed based on a powerful statistical tool called Sequential Probability Ratio Test, which has bounded false positive and false negative error rates.

  4. Determination of nickel in blood and serum samples of oropharyngeal cancer patients consumed smokeless tobacco products by cloud point extraction coupled with flame atomic absorption spectrometry.

    Science.gov (United States)

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Arain, Jamshed Bashir; Afridi, Hassan Imran; Kazi, Atif Gul; Nasreen, Syeda; Brahman, Kapil Dev

    2014-10-01

    Oropharyngeal cancer is a significant public health issue in the world. The incidence of oropharyngeal cancer has been increased among people who have habit of chewing smokeless tobacco (SLT) in Pakistan. The aim of present study was to evaluate the concentration of nickel (Ni) in biological samples (whole blood, serum) of oral (n = 95) and pharyngeal (n = 84) male cancer patients. For comparison purposes, the biological samples of healthy age-matched referents (n = 150), who consumed and did not consumed SLT products, were also analyzed for Ni levels. As the Ni level is very low in biological samples, a preconcentration procedure has been developed, prior to analysis of analyte by flame atomic absorption spectrometry (FAAS). The Ni in acid-digested biological samples was complexed with ammonium pyrrolidinedithio carbamate (APDC), and a resulted complex was extracted in a surfactant Triton X-114. Acidic ethanol was added to the surfactant-rich phase prior to its analysis by FAAS. The chemical variables, such as pH, amounts of reagents (APDC, Triton X-114), temperature, incubation time, and sample volume were optimized. The resulted data indicated that concentration of Ni was higher in blood and serum samples of cancer patients as compared to that of referents who have or have not consumed different SLT products (p = 0.012-0.001). It was also observed that healthy referents who consumed SLT products have two to threefold higher levels of Ni in both biological samples as compared to those who were not chewing SLT products (p < 0.01).

  5. Determination of nickel in blood and serum samples of oropharyngeal cancer patients consumed smokeless tobacco products by cloud point extraction coupled with flame atomic absorption spectrometry.

    Science.gov (United States)

    Arain, Sadaf Sadia; Kazi, Tasneem Gul; Arain, Jamshed Bashir; Afridi, Hassan Imran; Kazi, Atif Gul; Nasreen, Syeda; Brahman, Kapil Dev

    2014-10-01

    Oropharyngeal cancer is a significant public health issue in the world. The incidence of oropharyngeal cancer has been increased among people who have habit of chewing smokeless tobacco (SLT) in Pakistan. The aim of present study was to evaluate the concentration of nickel (Ni) in biological samples (whole blood, serum) of oral (n = 95) and pharyngeal (n = 84) male cancer patients. For comparison purposes, the biological samples of healthy age-matched referents (n = 150), who consumed and did not consumed SLT products, were also analyzed for Ni levels. As the Ni level is very low in biological samples, a preconcentration procedure has been developed, prior to analysis of analyte by flame atomic absorption spectrometry (FAAS). The Ni in acid-digested biological samples was complexed with ammonium pyrrolidinedithio carbamate (APDC), and a resulted complex was extracted in a surfactant Triton X-114. Acidic ethanol was added to the surfactant-rich phase prior to its analysis by FAAS. The chemical variables, such as pH, amounts of reagents (APDC, Triton X-114), temperature, incubation time, and sample volume were optimized. The resulted data indicated that concentration of Ni was higher in blood and serum samples of cancer patients as compared to that of referents who have or have not consumed different SLT products (p = 0.012-0.001). It was also observed that healthy referents who consumed SLT products have two to threefold higher levels of Ni in both biological samples as compared to those who were not chewing SLT products (p < 0.01). PMID:24920259

  6. Development of CT-guided biopsy sampling for time-dependent postmortem redistribution investigations in blood and alternative matrices--proof of concept and application on two cases.

    Science.gov (United States)

    Staeheli, Sandra N; Gascho, Dominic; Fornaro, Juergen; Laberke, Patrick; Ebert, Lars C; Martinez, Rosa Maria; Thali, Michael J; Kraemer, Thomas; Steuer, Andrea E

    2016-02-01

    The postmortem redistribution (PMR) phenomenon complicates interpretation in forensic toxicology. Human data on time-dependent PMR are rare and only exist for blood so far. A new method for investigation of time-dependent PMR in blood as well as in alternative body fluids and tissues was developed and evaluated using automated biopsy sampling. At admission of the bodies, introducer needles were placed in liver, lung, kidney, muscle, spleen, adipose tissue, heart, femoral vein, and lumbar spine using a robotic arm guided by a computed tomography scanner (CT). Needle placement accuracy was analyzed and found to be acceptable for the study purpose. Tissue biopsies and small volume body fluid samples were collected in triplicate through the introducer needles. At autopsy (around 24 h after admission), samples from the same body regions were collected. After mastering of the technical challenges, two authentic cases were analyzed as a proof of concept. Drug concentrations of venlafaxine, O-desmethylvenlafaxine, bromazepam, flupentixol, paroxetine, and lorazepam were determined by LC-MS/MS, and the percentage concentration changes between the two time points were calculated. Concentration changes were observed with both increases and decreases depending on analyte and matrix. While venlafaxine, flupentixol, paroxetine, and lorazepam generally showed changes above 30% and more, O-desmethylvenlafaxine and bromazepam did not undergo extensive PMR. The presented study shows that CT-controlled biopsy collection provides a valuable tool for systematic time-dependent PMR investigation, demanding only minimal sample amount and causing minimal damage to the body. PMID:26677021

  7. Problems and Countermeasures of Animal Blood Samples Collection%动物血液样品的采集保管存在问题及对策

    Institute of Scientific and Technical Information of China (English)

    蒋金贵; 张迎锐

    2012-01-01

    血液样品的采集和保管,在动物抗体监测检验工作中是一个十分重要的环节,也是保证监测结果正确性的必要前提,如果忽视了这一点,就会直接影响血样的质量,进而造成监测结论不正确而对免疫程序和免疫效果错误评估,最终导致动物综合防疫措施达不到预期目标。因此,笔者结合动物抗体监测工作中出现的问题,就如何正确进行血液样品的采集和保管谈几点体会,仅供参考。%It is a critical collectiong and preserving blood samples link,also guarantee the correctness of monitoring results,if neglect this point,the blood quality will be affected,furthermore,it will cause uncorrect results and mistake immune program and mistake evaluation,as a result,it will lead animal comprehensive prevention anticipate object.Therefore,the author discussed some experiences in this article on how to collect and preserve blood samples combination with animal antibody monitoring problems.

  8. Artificial neural networks to evaluate the boron concentration decreasing profile in Blood-BPA samples of BNCT patients

    Energy Technology Data Exchange (ETDEWEB)

    Garcia-Reiriz, Alejandro, E-mail: garciareiriz@gmail.com [Department of Analytical Chemistry, Faculty of Biochemical and Pharmaceutical Sciences, National University of Rosario, Rosario Institute of Chemistry (IQUIR-CONICET), Suipacha 531, Rosario S2002LRK (Argentina); Magallanes, Jorge [Comision Nacional de Energia Atomica, Av. Gral. Paz 1499, San Martin, B1650KNA, Buenos Aires (Argentina); Zupan, Jure [National Institute of Chemistry, Hajdrihova 19, SLO-1000 Ljubljana, Eslovenia (Slovenia); Liberman, Sara [Comision Nacional de Energia Atomica, Av. Gral. Paz 1499, San Martin, B1650KNA, Buenos Aires (Argentina)

    2011-12-15

    For the prediction of decay concentration profiles of the p-boronophenylalanine (BPA) in blood during BNCT treatment, a method is suggested based on Kohonen neural networks. The results of a model trained with the concentration profiles from the literature are described. The prediction of the model was validated by the leave-one-out method. Its robustness shows that it is mostly independent on small variations. The ability to fit retrospective experimental data shows an uncertainty lower than the two compartment model used previously. - Highlights: Black-Right-Pointing-Pointer We predicted decaying concentration profiles of BPA in blood during BNCT therapy. Black-Right-Pointing-Pointer Is suggested a method based on Kohonen neural networks. Black-Right-Pointing-Pointer The results show that it is very robust and mostly independent of small variations. Black-Right-Pointing-Pointer It has a better ability to fit retrospective experimental data. Black-Right-Pointing-Pointer The model could be progressively improved by adding new data to the training matrix.

  9. Postmortem detection of hepatitis B, C, and human immunodeficiency virus genomes in blood samples from drug-related deaths in Denmark*

    DEFF Research Database (Denmark)

    Eriksen, Mette Brandt; Jakobsen, Marianne Antonius; Kringsholm, Birgitte;

    2009-01-01

    Blood-borne viral infections are widespread among injecting drug users; however, it is difficult to include these patients in serological surveys. Therefore, we developed a national surveillance program based on postmortem testing of persons whose deaths were drug related. Blood collected...... at autopsy was tested for anti-HBc, anti-HBs, anti-hepatitis C virus (HCV), or anti-human immunodeficiency virus (HIV) antibodies using commercial kits. Subsets of seropositive samples were screened for viral genomes using sensitive in-house and commercial polymerase chain reaction (PCR) assays. Hepatitis B....... Postmortem HIV RNA testing was less sensitive than antemortem testing. Thus, postmortem PCR analysis for HBV and HBC infection is feasible and relevant for demonstrating ongoing infections at death or for transmission analysis during outbreaks....

  10. Corrections of arterial input function for dynamic H215O PET to assess perfusion of pelvic tumours: arterial blood sampling versus image extraction

    Science.gov (United States)

    Lüdemann, L.; Sreenivasa, G.; Michel, R.; Rosner, C.; Plotkin, M.; Felix, R.; Wust, P.; Amthauer, H.

    2006-06-01

    Assessment of perfusion with 15O-labelled water (H215O) requires measurement of the arterial input function (AIF). The arterial time activity curve (TAC) measured using the peripheral sampling scheme requires corrections for delay and dispersion. In this study, parametrizations with and without arterial spillover correction for fitting of the tissue curve are evaluated. Additionally, a completely noninvasive method for generation of the AIF from a dynamic positron emission tomography (PET) acquisition is applied to assess perfusion of pelvic tumours. This method uses a volume of interest (VOI) to extract the TAC from the femoral artery. The VOI TAC is corrected for spillover using a separate tissue TAC and for recovery by determining the recovery coefficient on a coregistered CT data set. The techniques were applied in five patients with pelvic tumours who underwent a total of 11 examinations. Delay and dispersion correction of the blood TAC without arterial spillover correction yielded in seven examinations solutions inconsistent with physiology. Correction of arterial spillover increased the fitting accuracy and yielded consistent results in all patients. Generation of an AIF from PET image data was investigated as an alternative to arterial blood sampling and was shown to have an intrinsic potential to determine the AIF noninvasively and reproducibly. The AIF extracted from a VOI in a dynamic PET scan was similar in shape to the blood AIF but yielded significantly higher tissue perfusion values (mean of 104.0 ± 52.0%) and lower partition coefficients (-31.6 ± 24.2%). The perfusion values and partition coefficients determined with the VOI technique have to be corrected in order to compare the results with those of studies using a blood AIF.

  11. Quantitative detection of epstein-barr virus DNA in cerebrospinal fluid and blood samples of patients with relapsing-remitting multiple sclerosis.

    Directory of Open Access Journals (Sweden)

    Clementina E Cocuzza

    Full Text Available The presence of Epstein-Barr Virus (EBV DNA in cerebrospinal fluid (CSF and peripheral blood (PB samples collected from 55 patients with clinical and radiologically-active relapsing-remitting MS (RRMS and 51 subjects with other neurological diseases was determined using standardized commercially available kits for viral nucleic acid extraction and quantitative EBV DNA detection. Both cell-free and cell-associated CSF and PB fractions were analyzed, to distinguish latent from lytic EBV infection. EBV DNA was detected in 5.5% and 18.2% of cell-free and cell-associated CSF fractions of patients with RRMS as compared to 7.8% and 7.8% of controls; plasma and peripheral blood mononuclear cells (PBMC positivity rates were 7.3% and 47.3% versus 5.8% and 31.4%, respectively. No significant difference in median EBV viral loads of positive samples was found between RRMS and control patients in all tested samples. Absence of statistically significant differences in EBV positivity rates between RRMS and control patients, despite the use of highly sensitive standardized methods, points to the lack of association between EBV and MS disease activity.

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

    Science.gov (United States)

    Stangegaard, Michael; Frøslev, Tobias G; Frank-Hansen, Rune; Hansen, Anders J; Morling, Niels

    2011-04-01

    We have implemented and validated automated protocols for DNA extraction and PCR setup using a Tecan Freedom EVO liquid handler mounted with the Te-MagS magnetic separation device (Tecan, Männedorf, Switzerland). The protocols were validated for accredited forensic genetic work according to ISO 17025 using the Qiagen MagAttract DNA Mini M48 kit (Qiagen GmbH, Hilden, Germany) from fresh whole blood and blood from deceased individuals. The workflow was simplified by returning the DNA extracts to the original tubes minimizing the risk of misplacing samples. The tubes that originally contained the samples were washed with MilliQ water before the return of the DNA extracts. The PCR was setup in 96-well microtiter plates. The methods were validated for the kits: AmpFℓSTR Identifiler, SGM Plus and Yfiler (Applied Biosystems, Foster City, CA), GenePrint FFFL and PowerPlex Y (Promega, Madison, WI). The automated protocols allowed for extraction and addition of PCR master mix of 96 samples within 3.5h. In conclusion, we demonstrated that (1) DNA extraction with magnetic beads and (2) PCR setup for accredited, forensic genetic short tandem repeat typing can be implemented on a simple automated liquid handler leading to the reduction of manual work, and increased quality and throughput. PMID:21609694

  13. The Spotting Distribution of Wildfires

    Directory of Open Access Journals (Sweden)

    Jonathan Martin

    2016-06-01

    Full Text Available In wildfire science, spotting refers to non-local creation of new fires, due to downwind ignition of brands launched from a primary fire. Spotting is often mentioned as being one of the most difficult problems for wildfire management, because of its unpredictable nature. Since spotting is a stochastic process, it makes sense to talk about a probability distribution for spotting, which we call the spotting distribution. Given a location ahead of the fire front, we would like to know how likely is it to observe a spot fire at that location in the next few minutes. The aim of this paper is to introduce a detailed procedure to find the spotting distribution. Most prior modelling has focused on the maximum spotting distance, or on physical subprocesses. We will use mathematical modelling, which is based on detailed physical processes, to derive a spotting distribution. We discuss the use and measurement of this spotting distribution in fire spread, fire management and fire breaching. The appendix of this paper contains a comprehensive review of the relevant underlying physical sub-processes of fire plumes, launching fire brands, wind transport, falling and terminal velocity, combustion during transport, and ignition upon landing.

  14. Empirical Bayes accomodation of batch-effects in microarray data using identical replicate reference samples: application to RNA expression profiling of blood from Duchenne muscular dystrophy patients

    Directory of Open Access Journals (Sweden)

    McCulloch Charles E

    2008-10-01

    Full Text Available Abstract Background Non-biological experimental error routinely occurs in microarray data collected in different batches. It is often impossible to compare groups of samples from independent experiments because batch effects confound true gene expression differences. Existing methods can correct for batch effects only when samples from all biological groups are represented in every batch. Results In this report we describe a generalized empirical Bayes approach to correct for cross-experimental batch effects, allowing direct comparisons of gene expression between biological groups from independent experiments. The proposed experimental design uses identical reference samples in each batch in every experiment. These reference samples are from the same tissue as the experimental samples. This design with tissue matched reference samples allows a gene-by-gene correction to be performed using fewer arrays than currently available methods. We examine the effects of non-biological variation within a single experiment and between experiments. Conclusion Batch correction has a significant impact on which genes are identified as differentially regulated. Using this method, gene expression in the blood of patients with Duchenne Muscular Dystrophy is shown to differ for hundreds of genes when compared to controls. The numbers of specific genes differ depending upon whether between experiment and/or between batch corrections are performed.

  15. 门诊采血患者投诉原因分析及对策%Complaint reason analysis and countermeasures of blood sampling patients in outpatient department

    Institute of Scientific and Technical Information of China (English)

    吴志琴; 付艳

    2012-01-01

    Objectives: To analyze complaint reasons of blood sampling patients and to improve patients satisfaction and nursing service quality in blood sampling room of outpatient department. Methods: We analyzed patients' complaint reason according to the relevant data from January 2005 to January 2011 and proposed protective countermeasures. Results: There were 174 complaints and the main complaint reason were not receiving laboratory sheet in time, bad service attitude, complicated diagnostic steps and so on. Conclusion: Good communication and earnest nurse service could decrease complaint rate, increase patient's satisfaction, improve working flow in blood sampling room, and achieve win-win outcome.%目的:通过分析门诊采血患者投诉原因,提高门诊采血室护理服务质量,进而提高患者满意度.方法:回顾性分析自2005年1月至2011年1月共6年间的门诊采血患者投诉情况,分析投诉原因并提出防范对策.结果:6年间共发生174例患者投诉事件,投诉原因主要涉及未能及时领取化验单、服务态度、就诊流程等方面.结论:通过良好的沟通,认真的处理,可有效降低患者投诉率,提高患者满意度,改进采血室工作流程,使护患双方得到共赢.

  16. Characterization of multi-drug resistant ESBL producing nonfermenter bacteria isolated from patients blood samples using phenotypic methods in Shiraz (Iran

    Directory of Open Access Journals (Sweden)

    Maneli Amin Shahidi

    2015-10-01

    Full Text Available Background and Aim: The emergence of  nonfermenter bacteria that are resistant to multidrug resistant ESBL  are  nowadays a principal problem  for hospitalized patients. The present study aimed at surveying the emergence of nonfermenter bacteria resistant to multi-drug ESBL producing isolated from patients blood samples using BACTEC 9240 automatic system in Shiraz. Materials and Methods: In this cross-sectional study, 4825 blood specimens were collected from hospitalized patients in Shiraz (Iran, and positive samples were detected by means of  BACTEC 9240 automatic system. The isolates  containing nonfermenter bacteria were identified based on biochemical tests embedded in the API-20E system. Antibiotic sensitivity  test was performed  and identification of  ESBL producing strains were done  using phenotypic detection of extended spectrum beta-lactamase producing isolates(DDST according to CLSI(2013 guidelines.   Results: Out of 4825 blood samples, 1145 (24% specimen were gram-positive using BACTEC system. Among all isolated microorganisms, 206 isolates were non-fermenting gram- negative bacteria. The most common non-fermenter isolates were Pseudomonas spp. (48%, Acinetobacter spp. (41.7% ,and Stenotrophomonas spp. (8.2%. Seventy of them (81.4% were  Acinetobacter spp. which were ESBL positive. Among &beta-lactam antibiotics, Pseudomonas spp. showed  the best sensitivity to piperacillin-tazobactam (46.5%.  Conclusion: It was found that  &beta-lactam antibiotics are not effective against more than 40% of Pseudomonas spp. infections and 78% Acinetobacter infections. Emergence of multi-drug resistant strains that are resistant to most antibiotic classes is a major public health problem in Iran. To resolve this problem using of practical guidelines is critical.

  17. Rapid Detection and Identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in Mosquito Vectors and Blood Samples by High Resolution Melting Real-Time PCR

    OpenAIRE

    Thanchomnang, Tongjit; Intapan, Pewpan M.; Tantrawatpan, Chairat; Lulitanond, Viraphong; Chungpivat, Sudchit; Taweethavonsawat, Piyanan; Kaewkong, Worasak; Sanpool, Oranuch; Janwan, Penchom; Choochote, Wej; Maleewong, Wanchai

    2013-01-01

    A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. ban...

  18. 采血管中添加剂对血样中乙醇含量的影响%Effects of Additives in Blood Collection Tubes on Testing the Alcohol Concentra-tion in Blood Samples

    Institute of Scientific and Technical Information of China (English)

    刘冬娴; 贺江南

    2014-01-01

    Objective To discuss blood collection tubes with different additives and their effects on the testing results of alcohol concentration in blood sam ples. Methods Blood sam ples from 10 volunteers were collected 2 hours after drinking with seven different types of disposable vacuum blood collection tubes, including ordinary tube without anticoagulant, coagulant tube, separating gel-coagulant tube, sodi-um citrate (1∶4) tube, sodium citrate (1∶9) tube, sodium citrate (9∶1) tube and EDTA-K2 tube. The al-cohol concentrations in these blood sam ples were analyzed by headspace gas chrom atography. Results The concentration testing results of the sam e blood sam ples in different types of tubes were different from one to another. The sequence was as follows:separating gel-coagulant tube>coagulant tube>ordi-nary tube without anticoagulant>EDTA-K2 tube>sodium citrate (1∶9) tube>sodium citrate (1∶4) tube, whereas the results of the sam e blood sam ple in sodium citrate (1∶9) tube and sodium citrate (9∶1) tube showed no obvious difference. Conclusion It is better to collect a suspicious drunk driver’s blood sam-ple using a disposable vacuum blood collection tube, with the EDTA-K2 tube being preferred.%目的:探讨不同种类采血管对血样中乙醇含量检测结果的影响。方法分别用7种一次性真空采血管[无抗凝剂管、促凝剂管、分离胶-促凝剂管、枸橼酸钠(1∶4)管、枸橼酸钠(1∶9)管、柠檬酸钠(9∶1)管、EDTA-K2管]采集10名志愿者饮酒后2 h血液,用顶空气相色谱法检测血样中乙醇含量。结果相同血样用不同的采血管,其乙醇含量检测结果不同,依次为分离胶-促凝剂管>促凝剂管>无抗凝剂管>EDTA-K2管>枸橼酸钠(1∶9)管>枸橼酸钠(1∶4)管,柠檬酸钠(9∶1)管与枸橼酸钠(1∶9)管检测结果基本一致。结论采集涉嫌酒后驾驶的驾驶员血样,应选用一次性真空采血管,首选EDTA-K2管。

  19. 干血滤纸片和白细胞酸性α-葡萄糖苷酶活性测定平台的建立及临床应用%Establishment and clinical application of dried blood spots and mixed leukocytes for determination of acid α-glucosidase activity

    Institute of Scientific and Technical Information of China (English)

    邱文娟; 王霞; 王瑜; 叶军; 韩连书; 张惠文; 顾学范

    2010-01-01

    目的 建立干血滤纸片和门细胞酸性α-葡萄糖苷酶(GAA)活性测定方法 及正常参考值,用于糖原累积病Ⅱ型(GSD Ⅱ)患者的酶学诊断.方法 利用GAA特异性水解荧光底物4-MUG的原理,采用阿卡波糖抑制其同工酶,建立干血滤纸片(DBS)和白细胞测定GAA活性方法 .取700例DBS样本和100例白细胞样本作为正常对照建立DBS和白细胞测定GAA活性正常参考值,并对临床疑诊4例患者及其父母进行GAA活性测定.结果 DBS法具有良好的精密度,室温或低于室温保存至少4周时DBS GAA活性无明显影响.正常新生儿和儿童-成人DBS的GAA参考范围分别为8.92~60.03 pmol/(punch·h)和8.00~37.43 pmol/(punch·h);正常人白细胞GAA参考范围:12.56~50.26 nmol/(mg蛋白·h),共检出4例GSD Ⅱ型患者.结论 DBS法测定GAA活性具有敏感、快速、高通量等特点,适于GSD Ⅱ型高危人群筛查和诊断;白细胞法测定GAA活性也具有快速、特异性高等特点,适于患者的确诊.%Objective Glycogen storage disease type Ⅱ(GSD Ⅱ,Pompe disease)is caused by the deficiency of acid α-glucosidase(GAA)that leads to lysosomal glycogen accumulation.Early diagnosis and treatment of GSD Ⅱ are considered to be critical for maximum efficacy of the enzyme replacement therapy.The aim of this study was to introduce two reliable methods and to generate the reference range of GAA activitv.Method The assay of GAA activity was performed in dried blood spots(DBS)and mixed leukocytes with acarbose to eliminate isoenzyme interference and to generate the reference range.GAA activitv was assayed in 700 specimens for DBS from normal subjects and 100 specimens for mixed leukocytes from normal subjects to set up reference range.GAA activity in the samples of 4 patients who were clinically suspected of GSD Ⅱ and their parents were also assayed.Result The intra-run and inter-run precision of the DBS method wag less than 10%.GAA activity tested by DBS was

  20. CEA blood test

    Science.gov (United States)

    Carcinoembryonic antigen blood test ... A blood sample is needed . ... When the needle is inserted to draw blood, some people feel moderate pain. Others feel only a prick or stinging sensation. Afterward, there may be some throbbing or a slight bruise. ...

  1. Ketones blood test

    Science.gov (United States)

    ... Ketones - serum; Nitroprusside test; Ketone bodies - serum; Ketones - blood ... A blood sample is needed. ... When the needle is inserted to draw blood, some people feel slight ... there may be some throbbing or a slight bruise. This soon ...

  2. Magnesium blood test

    Science.gov (United States)

    Magnesium - blood ... A blood sample is needed. ... When the needle is inserted to draw blood, some people feel slight pain. Others feel a prick or stinging. Afterward, there may be some throbbing or a slight bruise. This soon ...

  3. Cord Blood

    Directory of Open Access Journals (Sweden)

    Saeed Abroun

    2014-05-01

    Full Text Available   Stem cells are naïve or master cells. This means they can transform into special 200 cell types as needed by body, and each of these cells has just one function. Stem cells are found in many parts of the human body, although some sources have richer concentrations than others. Some excellent sources of stem cells, such as bone marrow, peripheral blood, cord blood, other tissue stem cells and human embryos, which last one are controversial and their use can be illegal in some countries. Cord blood is a sample of blood taken from a newborn baby's umbilical cord. It is a rich source of stem cells, umbilical cord blood and tissue are collected from material that normally has no use following a child’s birth. Umbilical cord blood and tissue cells are rich sources of stem cells, which have been used in the treatment of over 80 diseases including leukemia, lymphoma and anemia as bone marrow stem cell potency.  The most common disease category has been leukemia. The next largest group is inherited diseases. Patients with lymphoma, myelodysplasia and severe aplastic anemia have also been successfully transplanted with cord blood. Cord blood is obtained by syringing out the placenta through the umbilical cord at the time of childbirth, after the cord has been detached from the newborn. Collecting stem cells from umbilical blood and tissue is ethical, pain-free, safe and simple. When they are needed to treat your child later in life, there will be no rejection or incompatibility issues, as the procedure will be using their own cells. In contrast, stem cells from donors do have these potential problems. By consider about cord blood potency, cord blood banks (familial or public were established. In IRAN, four cord blood banks has activity, Shariati BMT center cord blood bank, Royan familial cord blood banks, Royan public cord blood banks and Iranian Blood Transfusion Organ cord blood banks. Despite 50,000 sample which storage in these banks, but the

  4. Detection and serotyping of pneumococci in community acquired pneumonia patients without culture using blood and urine samples

    OpenAIRE

    Elberse, K. (Karin); Mens, S.; Cremers, A.J.; Meijvis, S.C.A.; Vlaminckx, B.; de Jonge, M. I.; Meis, J. F. G. M.; Blauwendraat, C.; Pol, I. van de; Schouls, L. M.

    2015-01-01

    Background Treatment of community acquired pneumonia (CAP) patients with antibiotics before laboratory-confirmed diagnosis leads to loss of knowledge on the causative bacterial pathogen. Therefore, an increasing number of pneumococcal infections is identified using non-culture based techniques. However, methods for serotyping directly on the clinical specimen remain scarce. Here we present three approaches for detection and serotyping of pneumococci using samples from patients with CAP. Metho...

  5. Essential and Toxic Elements in Blood Samples of Harbor Seals (Phoca vitulina) from the Islands Helgoland (North Sea) and Anholt (Baltic Sea): A Comparison Study with Urbanized Areas.

    Science.gov (United States)

    Kakuschke, Antje; Griesel, Simone

    2016-01-01

    The harbor seals (Phoca vitulina) from Helgoland (North Sea) and Anholt (Kattegat, Baltic Sea) are top predators within the marine food web and an indicator species of the environmental contamination. Furthermore, they are a main tourist attraction. Despite these important roles, little is known about the health and pollutant contamination of these seals. The objective of this study was therefore to investigate 18 essential and nonessential/toxic elements (Al, As, Be, Ca, Cr, Cu, Fe, K, Mn, Mo, Ni, P, Pb, Rb, S, Se, Sr, and Zn) in blood samples using inductively coupled plasma mass spectrometry and total X-ray-fluorescence spectrometry. Blood concentrations of mineral nutrients, such as Ca, K, P, and S, were within the reference ranges described for harbor seals. Likewise, for the trace elements, As, Be, Rb, Se, and Sr, no significant differences were observed compared with previous studies. Interestingly, blood concentrations of nine nonessential as well as essential trace metals (Al, Cr, Cu, Fe, Mn, Mo, Ni, Pb, Zn) measured significantly lower in the offshore living seals from Helgoland and Anholt compared with results obtained from animals living close to urbanized areas, such as the Wadden Sea and Elbe estuary. This suggests that industrial emissions, sewage deposition, shipping traffic and dredging tasks might be the cause of increased metal concentrations of inshore harbor seals.

  6. Geographic variation and genetic structure in Spotted Owls

    Science.gov (United States)

    Haig, Susan M.; Wagner, R.S.; Forsman, E.D.; Mullins, Thomas D.

    2001-01-01

    We examined genetic variation, population structure, and definition of conservation units in Spotted Owls (Strix occidentalis). Spotted Owls are mostly non-migratory, long-lived, socially monogamous birds that have decreased population viability due to their occupation of highly-fragmented late successional forests in western North America. To investigate potential effects of habitat fragmentation on population structure, we used random amplified polymorphic DNA (RAPD) to examine genetic variation hierarchically among local breeding areas, subregional groups, regional groups, and subspecies via sampling of 21 breeding areas (276 individuals) among the three subspecies of Spotted Owls. Data from 11 variable bands suggest a significant relationship between geographic distance among local breeding groups and genetic distance (Mantel r = 0.53, P genetic drift. Merging nuclear data with recent mitochondrial data provides support for designation of an Evolutionary Significant Unit for Mexican Spotted Owls and two overlapping Management Units for Northern and California Spotted Owls.

  7. Analysis of the HLA-A,-B allele polymorphism in 5844 umbilical cord blood samples taken from Han population of Shandong province

    Institute of Scientific and Technical Information of China (English)

    YUN PENG DAI; WEN YING YAN; BAI JUN SHEN; LI JUN CHEN; FEI GAO; HONG MEI WANG

    2006-01-01

    To investigate the HLA-A, -B allele polymorphism in Han population of Shandong province and to explore the possibility to find out the HLA-A,-B-matched cord blood donors for stem cell transplantation to be used in other area in China, 5844 umbilical cord blood samples were taken from Han population donors of Shandong province, and assayed with PCR-sequence-oligonucleotide (PCR-SSO) assay. In Shandong Han donors, 20 alleles at HLA-A locus and 46 alleles at HLA-B locus could be detected as revealed in the present study. Among the 20 alleles at HLA-A locus, the most prevalent five alleles included A * 02(0. 3041), A * 11 (0. 1443), A * 24(0. 1434), A * 30(0. 0975) and A * 33(0.0859), while, the alleles with lower gene frequencies included A * 34(0. 0006), A * 25 (0.0005), A * 66(0.0005), A * 74(0.0004) and A * (0.0001). Of the 46 HLA-B alleles detected, the most prevalent five alleles were B * 13(0. 1348), B * 51(0.0713), B * 62(0.0712), B * 61 (0.0676) and B * 60(0.0642); while alleles with lower gene frequencies included B * 77(0.0001),B * 76(0.0002), B * 47(0.0003), B * 42(0.0003) and B * 72(0.0004). In comparison with those of the other Han population in China, the HLA-A, -B gene frequencies in the umbilical cord blood of Shandong province possess unique distribution features among the investigated populations from various regions of the same race origin, and the differences in various regions of the same race were less than those among the different race. It is evident that the HLA-A,-B alleles of the umbilical cord blood taken in Shangdong province show high degree of polymorphism, and it might be part of those of Northern Han population in China. So, it is reasonable for patients of Northern Chinese to receive HLA class Ⅰ -match transplant of cord blood stem cells for tissue and organ transplantation from Shangdong umbilical cord blood bank.

  8. Detection of Bartonella henselae DNA in clinical samples including peripheral blood of immune competent and immune compromised patients by three nested amplifications

    Directory of Open Access Journals (Sweden)

    Karina Hatamoto Kawasato

    2013-02-01

    Full Text Available Bacteria of the genus Bartonella are emerging pathogens detected in lymph node biopsies and aspirates probably caused by increased concentration of bacteria. Twenty-three samples of 18 patients with clinical, laboratory and/or epidemiological data suggesting bartonellosis were subjected to three nested amplifications targeting a fragment of the 60-kDa heat shock protein (HSP, the internal transcribed spacer 16S-23S rRNA (ITS and the cell division (FtsZ of Bartonella henselae, in order to improve detection in clinical samples. In the first amplification 01, 04 and 05 samples, were positive by HSP (4.3%, FtsZ (17.4% and ITS (21.7%, respectively. After the second round six positive samples were identified by nested-HSP (26%, eight by nested-ITS (34.8% and 18 by nested-FtsZ (78.2%, corresponding to 10 peripheral blood samples, five lymph node biopsies, two skin biopsies and one lymph node aspirate. The nested-FtsZ was more sensitive than nested-HSP and nested-ITS (p < 0.0001, enabling the detection of Bartonella henselae DNA in 15 of 18 patients (83.3%. In this study, three nested-PCR that should be specific for Bartonella henselae amplification were developed, but only the nested-FtsZ did not amplify DNA from Bartonella quintana. We conclude that nested amplifications increased detection of B. henselae DNA, and that the nested-FtsZ was the most sensitive and the only specific to B. henselae in different biological samples. As all samples detected by nested-HSP and nested-ITS, were also by nested-FtsZ, we infer that in our series infections were caused by Bartonella henselae. The high number of positive blood samples draws attention to the use of this biological material in the investigation of bartonellosis, regardless of the immune status of patients. This fact is important in the case of critically ill patients and young children to avoid more invasive procedures such as lymph nodes biopsies and aspirates.

  9. Application of cloud point preconcentration and flame atomic absorption spectrometry for the determination of cadmium and zinc ions in urine, blood serum and water samples

    Directory of Open Access Journals (Sweden)

    Ardeshir Shokrollahi

    2013-01-01

    Full Text Available A simple, sensitive and selective cloud point extraction procedure is described for the preconcentration and atomic absorption spectrometric determination of Zn2+ and Cd2+ ions in water and biological samples, after complexation with 3,3',3",3'"-tetraindolyl (terephthaloyl dimethane (TTDM in basic medium, using Triton X-114 as nonionic surfactant. Detection limits of 3.0 and 2.0 µg L-1 and quantification limits 10.0 and 7.0 µg L-1were obtained for Zn2+ and Cd2+ ions, respectively. Relative standard deviation was 2.9 and 3.3, and enrichment factors 23.9 and 25.6, for Zn2+ and Cd2+ ions, respectively. The method enabled determination of low levels of Zn2+ and Cd2+ ions in urine, blood serum and water samples.

  10. A compact and high sensitivity positron detector using dual-layer thin GSO scintillators for a small animal PET blood sampling system

    International Nuclear Information System (INIS)

    For quantitative measurements of small animals such as mice or rats, a compact and high sensitivity continuous blood sampling detector is required because their blood sampling volume is limited. For this purpose we have developed and tested a new positron detector. The positron detector uses a pair of dual-layer thin gadolinium orthosilicate (GSO) scintillators with different decay times. The front layer detects the positron and the background gamma photons, and the back layer detects the background gamma photons. By subtracting the count rate of the latter from that of the former, the count rate of the positrons can be estimated. The GSO for the front layer has a Ce concentration of 1.5 mol% (decay time of 35 ns), and that for the back layer has a Ce concentration of 0.5 mol% (decay time of 60 ns). By using the pulse shape analysis, the count rate of these two GSOs can be discriminated. The thickness is 0.5 mm, which is thick enough to detect positrons while minimizing the detection of the background gamma photons. These two types of thin GSOs were optically coupled to each other and connected to a metal photomultiplier tube (PMT) through triangular light guides. The signal from the PMT was digitized by 100 MHz free-running A-D converters in the data acquisition system and digitally integrated at two different integration times for the pulse shape analysis. We obtained good separation of the pulse shape distributions of these two GSOs. The energy threshold level was decreased to 80 keV, increasing the sensitivity of the detector. The sensitivity of a small diameter plastic tube was 8.6% and 24% for the F-18 and C-11 positrons, respectively. The count rate performance was linear up to ∼50 kcps. The background counts from the gamma photons could be precisely corrected. The time-activity curve (TAC) of the rat artery blood was successfully obtained and showed a good correlation with that measured using a well counter. With these results, we confirmed that the

  11. Global distribution of polymorphisms associated with delayed Plasmodium falciparum parasite clearance following artemisinin treatment: genotyping of archive blood samples.

    Science.gov (United States)

    Murai, Kenji; Culleton, Richard; Hisaoka, Teruhiko; Endo, Hiroyoshi; Mita, Toshihiro

    2015-06-01

    The recent emergence and spread of artemisinin-resistant Plasmodium falciparum isolates is a growing concern for global malaria-control efforts. A recent genome-wide analysis study identified two SNPs at genomic positions MAL10-688956 and MAL13-1718319, which are linked to delayed clearance of parasites following artemisinin combination therapy (ACT). It is expected that continuous artemisinin pressure will affect the distribution of these SNPs. Here, we investigate the worldwide distribution of these SNPs using a large number of archived samples in order to generate baseline data from the period before the emergence of ACT resistance. The presence of SNPs in MAL10-688956 and MAL13-1718319 was assessed by nested PCR RFLP and direct DNA sequencing using 653 global P. falciparum samples obtained before the reported emergence of ACT resistance. SNPs at MAL10-688956 and MAL13-1718319 associated with delayed parasite clearance following ACT administration were observed in 8% and 3% of parasites, respectively, mostly in Cambodia and Thailand. Parasites harbouring both SNPs were found in only eight (1%) isolates, all of which were from Cambodia and Thailand. Linkage disequilibrium was detected between MAL10-688956 and MAL13-1718319, suggesting that this SNP combination may have been selected by ACT drug pressure. Neither of the SNPs associated with delayed parasite clearance were observed in samples from Africa or South America. Baseline information of the geographical difference of MAL10-688956 and MAL13-1718319 SNPs provides a solid basis for assessing whether these SNPs are selected by artemisinin-based combination therapies.

  12. Expression profiling of blood samples from an SU5416 Phase III metastatic colorectal cancer clinical trial: a novel strategy for biomarker identification

    Directory of Open Access Journals (Sweden)

    Smolich Beverly D

    2003-02-01

    Full Text Available Abstract Background Microarray-based gene expression profiling is a powerful approach for the identification of molecular biomarkers of disease, particularly in human cancers. Utility of this approach to measure responses to therapy is less well established, in part due to challenges in obtaining serial biopsies. Identification of suitable surrogate tissues will help minimize limitations imposed by those challenges. This study describes an approach used to identify gene expression changes that might serve as surrogate biomarkers of drug activity. Methods Expression profiling using microarrays was applied to peripheral blood mononuclear cell (PBMC samples obtained from patients with advanced colorectal cancer participating in a Phase III clinical trial. The PBMC samples were harvested pre-treatment and at the end of the first 6-week cycle from patients receiving standard of care chemotherapy or standard of care plus SU5416, a vascular endothelial growth factor (VEGF receptor tyrosine kinase (RTK inhibitor. Results from matched pairs of PBMC samples from 23 patients were queried for expression changes that consistently correlated with SU5416 administration. Results Thirteen transcripts met this selection criterion; six were further tested by quantitative RT-PCR analysis of 62 additional samples from this trial and a second SU5416 Phase III trial of similar design. This method confirmed four of these transcripts (CD24, lactoferrin, lipocalin 2, and MMP-9 as potential biomarkers of drug treatment. Discriminant analysis showed that expression profiles of these 4 transcripts could be used to classify patients by treatment arm in a predictive fashion. Conclusions These results establish a foundation for the further exploration of peripheral blood cells as a surrogate system for biomarker analyses in clinical oncology studies.

  13. Induction of PGC-1α expression can be detected in blood samples of patients with ST-segment elevation acute myocardial infarction.

    Directory of Open Access Journals (Sweden)

    Óscar Fabregat-Andrés

    Full Text Available Following acute myocardial infarction (MI, cardiomyocyte survival depends on its mitochondrial oxidative capacity. Cell death is normally followed by activation of the immune system. Peroxisome proliferator activated receptor γ-coactivator 1α (PGC-1α is a transcriptional coactivator and a master regulator of cardiac oxidative metabolism. PGC-1α is induced by hypoxia and facilitates the recovery of the contractile capacity of the cardiac muscle following an artery ligation procedure. We hypothesized that PGC-1α activity could serve as a good molecular marker of cardiac recovery after a coronary event. The objective of the present study was to monitor the levels of PGC-1α following an ST-segment elevation acute myocardial infarction (STEMI episode in blood samples of the affected patients. Analysis of blood mononuclear cells from human patients following an STEMI showed that PGC-1α expression was increased and the level of induction correlated with the infarct size. Infarct size was determined by LGE-CMR (late gadolinium enhancement on cardiac magnetic resonance, used to estimate the percentage of necrotic area. Cardiac markers, maximum creatine kinase (CK-MB and Troponin I (TnI levels, left ventricular ejection function (LVEF and regional wall motion abnormalities (RWMA as determined by echocardiography were also used to monitor cardiac injury. We also found that PGC-1α is present and active in mouse lymphocytes where its expression is induced upon activation and can be detected in the nuclear fraction of blood samples. These results support the notion that induction of PGC-1α expression can be part of the recovery response to an STEMI and could serve as a prognosis factor of cardiac recovery.

  14. Determining the Diagnostic Value of Mycobacterium Tuberculosis DNA in the Differentiation of Blood Samples of Patients with Active Pulmonary Tuberculosis and Healthy Controls Using Polymerase Chain Reaction

    Directory of Open Access Journals (Sweden)

    Abasali Niazi

    2013-10-01

    Full Text Available Background: Tuberculosis (TB is now a major cause of mortality and morbidity in the world. Nowadays, different methods are used to diagnose tuberculosis. Although classical microbiological methods (such as sputum smear are specific, they have little sensitivity and the culture is also time-consuming. Using Polymerase Chain Reaction (PCR in blood samples in terms of Mycobacterium tuberculosis DNA, this study examines diagnostic power of this test in the diagnosis of pulmonary tuberculosis compared with other standard methods. Materials and Methods: In a cross-sectional descriptive-analytic study, blood samples were taken from 40 TB patients and 40 non-TB cases. Following DNA extraction by the commercial kit QIAGEN, the PCR assay was performed using IS6110 primer.Results: In this study, there were 80 people in two groups of TB and non-TB cases. Each group composed of 14 men (35% and 26 women (65%. Sensitivity, specificity as well as positive and negative predictive values obtained 37.5, 100, 100 and 61.5%, respectively.Conclusion: Despite high costs of using PCR for TB diagnosis, sensitivity of this method is low due to various factors and cannot replace current standard methods for TB diagnosis such as smear and culture. It can only be used as a complementary method to confirm diagnosis in strongly suspected cases of tuberculosis.

  15. The SPOT satellite

    Science.gov (United States)

    Fouquet, J.-P.

    1981-03-01

    The background, objectives and data products of the French SPOT remote sensing satellite system are presented. The system, which was developed starting in 1978 with the subsequent participation of Sweden and Belgium, is based on a standard multimission platform with associated ground control station and a mission-specific payload, which includes two High-Resolution Visible range instruments allowing the acquisition of stereoscopic views from different orbits. Mission objectives include the definition of future remote sensing systems, the compilation of a cartographic and resources data base, the study of species discrimination and production forecasting based on frequent access and off-nadir viewing, the compilation of a stereoscopic data base, and platform and instrument qualification, for possible applications in cartography, geology and agriculture. Standard data products will be available at three levels of preprocessing: radiometric correction only, precision processing for vertical viewing, and cartographic quality processing.

  16. Development and validation of an indirect Enzyme-linked immunosorbent assay for the detection of antibodies against Schmallenberg virus in blood samples from ruminants.

    Science.gov (United States)

    van der Heijden, H M J F; Bouwstra, R J; Mars, M H; van der Poel, W H M; Wellenberg, G J; van Maanen, C

    2013-10-01

    To detect Schmallenberg virus (SBV) infections in ruminants and to perform SBV epidemiological studies a cost-effective serological test is required. For these purposes an indirect whole virus Enzyme-linked Immunosorbent Assay (ELISA) for detection of SBV specific antibodies in ruminant blood samples was developed. Schmallenberg virus antigen was produced by propagation on Vero cells, partly purified and coated onto ELISA plates. The indirect ELISA procedure included the subsequent incubation of diluted samples, protein-G-HRP conjugate and TMB substrate solution. Net Optical Densities (OD) values were calculated and expressed as a sample to positive percentage (S/P%) by comparison of the average net OD with the OD of the positive control. Validation of this assay was performed using 633 samples from SBV-free sheep, goats and cattle, and 141 samples from SBV suspect ruminants. The diagnostic specificity was 98.8%. Test results of 86 ruminant serum samples using both the SBV-ELISA and an SBV virus neutralization test (VNT), designated as the gold standard serological test for SBV, showed good correlation: at an S/P cut-off of 15% only one VNT positive sample tested negative in the SBV ELISA. The diagnostic sensitivity of the ELISA, relative to the VNT, was 98.8% (95% CI: 93.3-100.0%). The ELISA showed a high repeatability (cv=6.5%) and reproducibility (100% agreement). It was concluded that this ELISA is a suitable test method for the detection of SBV antibodies in sera from cows, sheep and, possibly, goats.

  17. Specific, sensitive and rapid detection of human plasmodium knowlesi infection by loop-mediated isothermal amplification (LAMP in blood samples

    Directory of Open Access Journals (Sweden)

    Anthony Claudia N

    2011-07-01

    Full Text Available Abstract Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plasmodium ssrRNA genes has been described as the most sensitive and specific method for Plasmodium detection. However, this method is costly and requires trained personnel for its implementation. Loop-mediated isothermal amplification (LAMP, a novel nucleic acid amplification method was developed for the clinical detection of P. knowlesi. The sensitivity and specificity of LAMP was evaluated in comparison to the results obtained via microscopic examination and nested PCR. Methods LAMP assay was developed based on P. knowlesi genetic material targeting the apical membrane antigen-1 (AMA-1 gene. The method uses six primers that recognize eight regions of the target DNA and it amplifies DNA within an hour under isothermal conditions (65°C in a water-bath. Results LAMP is highly sensitive with the detection limit as low as ten copies for AMA-1. LAMP detected malaria parasites in all confirm cases (n = 13 of P. knowlesi infection (sensitivity, 100% and none of the negative samples (specificity, 100% within an hour. LAMP demonstrated higher sensitivity compared to nested PCR by successfully detecting a sample with very low parasitaemia ( Conclusion With continuous efforts in the optimization of this assay, LAMP may provide a simple and reliable test for detecting P. knowlesi malaria parasites in areas where malaria is prevalent.

  18. zeta-, epsilon-, and gamma-Globin mRNA in blood samples and CD71(+) cell fractions from fetuses and from pregnant and nonpregnant women, with special attention to identification of fetal erythroblasts

    DEFF Research Database (Denmark)

    Høgh, A M; Hviid, T V; Christensen, B;

    2001-01-01

    erythroblasts. A specific marker is necessary for isolation and identification of fetal nucleated red blood cells from maternal blood samples for use in antenatal diagnosis of fetal genetic or chromosomal abnormalities. METHODS: We used a very sensitive reverse transcription-PCR (RT-PCR) method, coamplification...

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

    Science.gov (United States)

    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. PMID:24199057

  20. Lyme disease blood test

    Science.gov (United States)

    ... antibodies in the blood sample using the ELISA test . If the ELISA test is positive, it must be confirmed with another ... were seen in your blood sample. If the ELISA test is negative, usually no other testing is needed. ...

  1. Comparison of DNA Extraction Methods from Small Samples of Newborn Screening Cards Suitable for Retrospective Perinatal Viral Research

    OpenAIRE

    McMichael, Gai L.; Highet, Amanda R.; Gibson, Catherine S; Goldwater, Paul N; O'Callaghan, Michael E.; Alvino, Emily R.; MacLennan, Alastair H

    2011-01-01

    Reliable detection of viral DNA in stored newborn screening cards (NSC) would give important insight into possible silent infection during pregnancy and around birth. We sought a DNA extraction method with sufficient sensitivity to detect low copy numbers of viral DNA from small punch samples of NSC. Blank NSC were spotted with seronegative EDTA-blood and seropositive EBV EDTA-blood. DNA was extracted with commercial and noncommercial DNA extraction methods and quantified on a spectrofluorome...

  2. Promoter Region Hypermethylation and mRNA Expression of MGMT and p16 Genes in Tissue and Blood Samples of Human Premalignant Oral Lesions and Oral Squamous Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Vikram Bhatia

    2014-01-01

    Full Text Available Promoter methylation and relative gene expression of O6-methyguanine-DNA-methyltransferase (MGMT and p16 genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs and oral squamous cell carcinoma (OSCC. Methylation-specific PCR and reverse transcriptase PCR were performed in 146 tissue and blood samples from controls and patients with PMOLs and OSCC. In PMOL group, significant promoter methylation of MGMT and p16 genes was observed in 59% (P=0.0010 and 57% (P=0.0016 of tissue samples, respectively, and 39% (P=0.0135 and 33% (P=0.0074 of blood samples, respectively. Promoter methylation of both genes was more frequent in patients with OSCC, that is, 76% (P=0.0001 and 82% (P=0.0001 in tissue and 57% (P=0.0002 and 70% (P=0.0001 in blood, respectively. Significant downregulation of MGMT and p16 mRNA expression was observed in both tissue and blood samples from patients with PMOLs and OSCC. Hypermethylation-induced transcriptional silencing of MGMT and p16 genes in both precancer and cancer suggests important role of these changes in progression of premalignant state to malignancy. Results support use of blood as potential surrogate to tissue samples for screening or diagnosing PMOLs and early OSCC.

  3. Promoter Region Hypermethylation and mRNA Expression of MGMT and p16 Genes in Tissue and Blood Samples of Human Premalignant Oral Lesions and Oral Squamous Cell Carcinoma

    Science.gov (United States)

    Bhatia, Vikram; Makker, Annu; Tewari, Shikha; Yadu, Alka; Shilpi, Priyanka; Kumar, Sandeep; Agarwal, S. P.; Goel, Sudhir K.

    2014-01-01

    Promoter methylation and relative gene expression of O6-methyguanine-DNA-methyltransferase (MGMT) and p16 genes were examined in tissue and blood samples of patients with premalignant oral lesions (PMOLs) and oral squamous cell carcinoma (OSCC). Methylation-specific PCR and reverse transcriptase PCR were performed in 146 tissue and blood samples from controls and patients with PMOLs and OSCC. In PMOL group, significant promoter methylation of MGMT and p16 genes was observed in 59% (P = 0.0010) and 57% (P = 0.0016) of tissue samples, respectively, and 39% (P = 0.0135) and 33% (P = 0.0074) of blood samples, respectively. Promoter methylation of both genes was more frequent in patients with OSCC, that is, 76% (P = 0.0001) and 82% (P = 0.0001) in tissue and 57% (P = 0.0002) and 70% (P = 0.0001) in blood, respectively. Significant downregulation of MGMT and p16 mRNA expression was observed in both tissue and blood samples from patients with PMOLs and OSCC. Hypermethylation-induced transcriptional silencing of MGMT and p16 genes in both precancer and cancer suggests important role of these changes in progression of premalignant state to malignancy. Results support use of blood as potential surrogate to tissue samples for screening or diagnosing PMOLs and early OSCC. PMID:24991542

  4. Number of Persistent Organic Pollutants Detected at High Concentrations in Blood Samples of the United States Population

    Science.gov (United States)

    Gasull, Magda; Lee, Duk-Hee; López, Tomàs

    2016-01-01

    Human exposure to environmental chemicals as persistent organic pollutants (POPs) is usually assessed considering each pollutant individually, with little attention to concentrations of mixtures in individuals or social groups. Yet, it may be relatively common for humans to have low and high concentrations of numerous POPs. The study objectives were to analyze the number of POPs detected per person at high concentrations in the U.S. population, and the associations between such type of indicators and socioeconomic factors as gender, race / ethnicity, education, and poverty level. From 91 POPs analyzed in serum samples of 4,739 individuals in three subsamples of the National Health and Nutrition Examination Survey (NHANES) 2003–2004 (the last period with valid updated individual data for the compounds considered in the present study), we computed the number of POPs whose serum concentrations were above selected cutoff points. POPs included were 13 organochlorine compounds (OCs), 10 polybrominated diphenyl ethers (PBDEs), the polybrominated biphenyl (PBB) 153, 38 polychlorinated biphenyls (PCBs), 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/Fs), and 12 perfluorinated compounds (PFCs). Over 13% of participants had ≥10 of the 37 most detected POPs each at a concentration in the top decile (P90). Over 30% of subjects with total toxic equivalency (TEQ) ≥P75, had ≥10 of 24 POPs not included in TEQ calculations at concentrations ≥P90. Compared to non-Hispanic whites, the adjusted odds ratio of having ≥10 of the 37 POPs at P90 was 9.2 for non-Hispanic blacks and 0.18 for Mexican Americans. Poverty, body mass index, age, and gender were also independently associated with having ≥10 POPs in the top decile. More than one tenth of the US population may have ≥10 POPs each at concentrations in the top decile. Such pattern is nine times more frequent in Non-Hispanic blacks and four times less frequent in Mexican Americans than in non

  5. Number of Persistent Organic Pollutants Detected at High Concentrations in Blood Samples of the United States Population.

    Science.gov (United States)

    Pumarega, José; Gasull, Magda; Lee, Duk-Hee; López, Tomàs; Porta, Miquel

    2016-01-01

    Human exposure to environmental chemicals as persistent organic pollutants (POPs) is usually assessed considering each pollutant individually, with little attention to concentrations of mixtures in individuals or social groups. Yet, it may be relatively common for humans to have low and high concentrations of numerous POPs. The study objectives were to analyze the number of POPs detected per person at high concentrations in the U.S. population, and the associations between such type of indicators and socioeconomic factors as gender, race / ethnicity, education, and poverty level. From 91 POPs analyzed in serum samples of 4,739 individuals in three subsamples of the National Health and Nutrition Examination Survey (NHANES) 2003-2004 (the last period with valid updated individual data for the compounds considered in the present study), we computed the number of POPs whose serum concentrations were above selected cutoff points. POPs included were 13 organochlorine compounds (OCs), 10 polybrominated diphenyl ethers (PBDEs), the polybrominated biphenyl (PBB) 153, 38 polychlorinated biphenyls (PCBs), 17 polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/Fs), and 12 perfluorinated compounds (PFCs). Over 13% of participants had ≥10 of the 37 most detected POPs each at a concentration in the top decile (P90). Over 30% of subjects with total toxic equivalency (TEQ) ≥P75, had ≥10 of 24 POPs not included in TEQ calculations at concentrations ≥P90. Compared to non-Hispanic whites, the adjusted odds ratio of having ≥10 of the 37 POPs at P90 was 9.2 for non-Hispanic blacks and 0.18 for Mexican Americans. Poverty, body mass index, age, and gender were also independently associated with having ≥10 POPs in the top decile. More than one tenth of the US population may have ≥10 POPs each at concentrations in the top decile. Such pattern is nine times more frequent in Non-Hispanic blacks and four times less frequent in Mexican Americans than in non-Hispanic whites

  6. Image-derived and arterial blood sampled input functions for quantitative PET imaging of the angiotensin II subtype 1 receptor in the kidney

    Energy Technology Data Exchange (ETDEWEB)

    Feng, Tao; Tsui, Benjamin M. W.; Li, Xin; Vranesic, Melin; Lodge, Martin A.; Gulaldi, Nedim C. M.; Szabo, Zsolt, E-mail: zszabo@jhmi.edu [Russell H. Morgan Department of Radiology and Radiological Science, The Johns Hopkins School of Medicine, Baltimore, Maryland 21287 (United States)

    2015-11-15

    Purpose: The radioligand {sup 11}C-KR31173 has been introduced for positron emission tomography (PET) imaging of the angiotensin II subtype 1 receptor in the kidney in vivo. To study the biokinetics of {sup 11}C-KR31173 with a compartmental model, the input function is needed. Collection and analysis of arterial blood samples are the established approach to obtain the input function but they are not feasible in patients with renal diseases. The goal of this study was to develop a quantitative technique that can provide an accurate image-derived input function (ID-IF) to replace the conventional invasive arterial sampling and test the method in pigs with the goal of translation into human studies. Methods: The experimental animals were injected with [{sup 11}C]KR31173 and scanned up to 90 min with dynamic PET. Arterial blood samples were collected for the artery derived input function (AD-IF) and used as a gold standard for ID-IF. Before PET, magnetic resonance angiography of the kidneys was obtained to provide the anatomical information required for derivation of the recovery coefficients in the abdominal aorta, a requirement for partial volume correction of the ID-IF. Different image reconstruction methods, filtered back projection (FBP) and ordered subset expectation maximization (OS-EM), were investigated for the best trade-off between bias and variance of the ID-IF. The effects of kidney uptakes on the quantitative accuracy of ID-IF were also studied. Biological variables such as red blood cell binding and radioligand metabolism were also taken into consideration. A single blood sample was used for calibration in the later phase of the input function. Results: In the first 2 min after injection, the OS-EM based ID-IF was found to be biased, and the bias was found to be induced by the kidney uptake. No such bias was found with the FBP based image reconstruction method. However, the OS-EM based image reconstruction was found to reduce variance in the subsequent

  7. Proteomic analysis of blood level of proteins before and after operation in patients with esophageal squamous cell carcinoma at high-incidence area in Henan Province

    Institute of Scientific and Technical Information of China (English)

    Ji-Ye An; Zong-Min Fan; Ze-Hao Zhuang; Yan-Ru Qin; Shan-Shan Gao; Ji-Lin Li; Li-Dong Wang

    2004-01-01

    AIM: To characterize the protein files in blood from same patients with esophageal squamous cell carcinoma (ESCC)before and after operation at the high-incidence area for ESCC in Henan Province, China.METHODS: Two-dimensional electrophoresis, silver staining and ImageMaster 2-DE analysis software were applied to the determination of protein files in the blood obtained from normal controls and ESCC patients before and after operation.RESULTS: A total of 655, 662 and 677 protein spots were identified, respectively, from the normal controls and ESCC patients before and after operation. No significant difference in the number of protein spots was observed between the normal group and ESCC patients. A total of seven protein spots were identified with a dramatic difference among the samples before and after operation. Six protein spots were up-regulated and one protein spot was down-regulated in the group after operation compared with those in normal and before operation. Three protein spots were further characterized by matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS). The proteins from these three spots were identified as serum amyloid A(SAA), amyloid related serum protein and haptoglobin.CONCLUSION: Serum amyloid A, amyloid related serum protein and haptoglobin may be related with ESCC and/or surgery. The significance of these proteins needs to be further characterized. The present study provides informative data for the establishment of serum protein profiles related with ESCC.

  8. Analytical performance of a multiplex Real-Time PCR assay using TaqMan probes for quantification of Trypanosoma cruzi satellite DNA in blood samples.

    Directory of Open Access Journals (Sweden)

    Tomas Duffy

    Full Text Available BACKGROUND: The analytical validation of sensitive, accurate and standardized Real-Time PCR methods for Trypanosoma cruzi quantification is crucial to provide a reliable laboratory tool for diagnosis of recent infections as well as for monitoring treatment efficacy. METHODS/PRINCIPAL FINDINGS: We have standardized and validated a multiplex Real-Time quantitative PCR assay (qPCR based on TaqMan technology, aiming to quantify T. cruzi satellite DNA as well as an internal amplification control (IAC in a single-tube reaction. IAC amplification allows rule out false negative PCR results due to inhibitory substances or loss of DNA during sample processing. The assay has a limit of detection (LOD of 0.70 parasite equivalents/mL and a limit of quantification (LOQ of 1.53 parasite equivalents/mL starting from non-boiled Guanidine EDTA blood spiked with T. cruzi CL-Brener stock. The method was evaluated with blood samples collected from Chagas disease patients experiencing different clinical stages and epidemiological scenarios: 1- Sixteen Venezuelan patients from an outbreak of oral transmission, 2- Sixty three Bolivian patients suffering chronic Chagas disease, 3- Thirty four Argentinean cases with chronic Chagas disease, 4- Twenty seven newborns to seropositive mothers, 5- A seronegative receptor who got infected after transplantation with a cadaveric kidney explanted from an infected subject. CONCLUSIONS/SIGNIFICANCE: The performing parameters of this assay encourage its application to early assessment of T. cruzi infection in cases in which serological methods are not informative, such as recent infections by oral contamination or congenital transmission or after transplantation with organs from seropositive donors, as well as for monitoring Chagas disease patients under etiological treatment.

  9. Preservation in 70% ethanol solution does not affect δ13C and δ15N values of reindeer blood samples – relevance for stable isotope studies of diet

    Directory of Open Access Journals (Sweden)

    Duncan J. Halley

    2008-04-01

    Full Text Available We compared duplicate samples of whole blood samples from 18 reindeer that were preserved either by immediate freezing or by immersion in 70 % ethanol. All samples were dried at 60 °C, powdered, treated with 1:1 chloroform: methanol, and dried again before isotope analysis. There were no differences in the values of δ13C and δ15N between the methods of preservation. Isotopic differences were absolutely small (δ13C = 0.1±0.10/00; δ15N=0.2±0.20/00, random in direction, and within the limits of analytical precision for the mass spectrometer. Preservation in ethanol thus appears to be an effective and efficient method for preserving blood samples for stable isotope analysis under field conditions. Abstract in Norwegian / Sammendrag:Konservering av blodprøver fra rein i 70% etanolløsning påvirker ikke verdiene av δ13C and δ15N–verdiene og er en fullgod metode for analyse av stabile isotoperVi sammenlignet to og to prøver av blodprøver fra 18 reinsdyr. Prøvene var enten konservert ved umiddelbar frysing eller ved bruk av 70% etanol. Alle prøver ble tørket ved 60 °C, pulverisert og behandlet med kloroform:metanol i forholdet 1:1. Til slutt ble de tørket på nytt før gjennomføring av isotopanalysen. Vi fant ingen forskjell i verdiene av δ13C and δ15N mellom de to konserveringsmetodene. I absolutte verdier var isotopforskjellene små (δ13C = 0.1±0.1 0/00; δ15N=0.2±0.2 0/00. Forskjellene var tilfeldige og innenfor grensene for massespektrometerets presisjon. Bruk av etanol framstår som en effektiv og fullgod metode til konservering av blodprøver for analyse av stabile isotoper under feltforhold.

  10. AhR- and ER-mediated activities in human blood samples collected from PCB-contaminated and background region in Slovakia

    Energy Technology Data Exchange (ETDEWEB)

    Pliskova, M. [Veterinary Researcch Institute, Brno (Czech Republic); Canton, R.F.; Duursen, M.B.M. van [Utrecht Univ. (NL). Institute for Risk Assessment Sciences (IRAS)] (and others)

    2004-09-15

    Endocrine disruption mediated through activation of aryl hydrocarbon receptor (AhR) and estrogen receptor (ER) by polychlorinated biphenyls (PCBs) and other persistent organic pollutants (POPs) has been studied extensively both in vivo and in vitro. Non-ortho- and mono-ortho-substituted polychlorinated biphenyls (PCBs) are potent AhR agonists therefore, increased dioxin-like activity of complex blood samples might reflect an increased exposure to PCBs. The induction of expression of CYP1A1 and CYP1B1 in different tissues, including lymphocytes, also depends on activation of AhR and it could be useful as a potential biomarker of exposure to dioxin-like compounds. Using various in vivo and in vitro models, the exposure to PCBs or hydroxy-PCBs has been reported to lead to either induction of ER-mediated activity or to an antiestrogenic effect associated with a suppression of estradiol-induced ER-dependent gene expression. Nevertheless, relative (anti)estrogenic potencies of a large set of prevalent environmental PCBs have not been yet compared in a single bioassay. A cross-talk between AhR and ER has been suggested to lead to a suppression of ER-mediated gene expression. Therefore, presence of dioxin-like compounds in blood could potentially suppress the ER-mediated activity. Additionally, AhR-dependent induction of CYP1A1 and especially CYP1B1, two enzymes involved in oxidative metabolism of estradiol and other estrogens, might enhance the metabolism of estradiol and it has been suggested to cause a potential depression of estrogen levels in the body. The aim of the present study was to determine dioxin-like, estrogenic and antiestrogenic activities in human blood samples collected in two Eastern Slovakia regions differently polluted with PCBs using established in vitro bioassays. We also studied mRNA expression of CYP1A1 and 1B1 in lymphocytes and the genotypes of CYP1B1 as possible biomarkers of exposure for PCBs and related compounds. The biological data obtained

  11. Avaliação de multielementos em amostras de sangue humano usando SR-TXRF Evaluation of multielements in human blood samples using synchrotron radiation

    Directory of Open Access Journals (Sweden)

    Nivia Graciele V. Pinto

    2010-01-01

    Full Text Available A técnica de fluorescência de raios X por reflexão total usando radiação síncrotron (SR-TXRF é uma poderosa ferramenta utilizada para a determinação das concentrações elementares presentes em amostras biológicas. O objetivo deste estudo é avaliar as possíveis alterações causadas por processos de irradiação na concentração de elementos-traço em amostras de sangue humano. As amostras de sangue foram coletadas no Laboratório de Análises Clínicas Dr. Elilel Figueiredo, Rio de Janeiro, e divididas em dois grupos. O primeiro grupo foi irradiado com doses de 1.500, 2.500 e 3.000 cGy, utilizando o irradiador Gammacell 220 Excel, e o segundo foi irradiado com doses que variaram de 2 cGy a 100 cGy, utilizando uma bomba de cobalto Theratron 780 C do Inca, Rio de Janeiro. Todas as amostras de sangue total, plasma e matriz celular foram então liofilizadas e, em seguida, passaram pelo procedimento padrão de digestão. Todas as medidas foram realizadas na linha de fluorescência de raios X do Laboratório Nacional de Luz Síncrotron (LNLS, em Campinas, Brasil. Não se verificou variação significativa na concentração de Ca e, em contrapartida, o K foi o único elemento que sofreu alterações significativas para todas as amostras analisadas em função da dose. A concentração de Fe diminuiu apenas para as amostras de sangue total e plasma. A concentração de Zn apresentou uma diminuição significativa somente para as amostras de sangue total.Total-reflection X-ray fluorescence using synchrotron radiation (SR-TXRF is a powerful analytical technique to study trace elements in biomedical samples. The aim of this study was to investigate possible changes in essential trace element concentrations caused by irradiation procedures. Fresh blood samples were obtained from the Dr. Eliel Figueiredo Laboratory, Rio de Janeiro. The samples were separated in two groups. The first was irradiated with doses of 1500, 2500 and 3000cGy, using a

  12. Coulomb explosion of "hot spot"

    CERN Document Server

    Oreshkin, V I; Chaikovsky, S A; Artyomov, A P

    2016-01-01

    The study presented in this paper has shown that the generation of hard x rays and high-energy ions, which are detected in pinch implosion experiments, may be associated with the Coulomb explosion of the hot spot that is formed due to the outflow of the material from the pinch cross point. During the process of material outflow, the temperature of the hot spot plasma increases, and conditions arise for the plasma electrons to become continuously accelerated. The runaway of electrons from the hot spot region results in the buildup of positive space charge in this region followed by a Coulomb explosion. The conditions for the hot spot plasma electrons to become continuously accelerated have been revealed and estimates have been obtained for the kinetic energy of the ions generated by the Coulomb explosion.

  13. Establishment of loop-mediated isothermal amplification (LAMP) for rapid detection of Brucella spp. and application to milk and blood samples.

    Science.gov (United States)

    Song, Liuyan; Li, Juntao; Hou, Shuiping; Li, Xunde; Chen, Shouyi

    2012-09-01

    Brucella spp. are facultative intracellular bacteria that infect humans and animals. In this study, the loop-mediated isothermal amplification (LAMP) was used to detect the Brucella-specific gene omp25. Reaction conditions were optimized as temperature 65°C, reaction time 60 min, Mg(2+) concentration 8.0 mmol/L, polymerase content Bst DNA, 0.5 μL, deoxyribonucleotide concentration 1.6 mmol/L, and inner/outer primer ratio 1:8. The LAMP method was evaluated with 4 Brucella species and 29 non-Brucella bacteria species. Positive reactions were observed on all the 4 Brucella species but not on any non-Brucella species. The limit of detection of the LAMP method was 3.81 CFU Brucella spp. Using the LAMP method, 7 of 110 raw milk samples and 5 of 59 sheep blood samples were detected positive of Brucella spp. Results indicated that LAMP is a fast, specific, sensitive, inexpensive, and suitable method for diagnosis of Brucella spp. infection.

  14. A novel surface molecularly imprinted polymer as the solid-phase extraction adsorbent for the selective determination of ampicillin sodium in milk and blood samples$

    Institute of Scientific and Technical Information of China (English)

    Ningli Wu; Qiang Fu; Zhimin Luo; Yanhui Ge; Pengqi Guo; Kangli Du; Weili Tang; Wei Du; Aiguo Zeng; Chun Chang

    2016-01-01

    Surface molecularly imprinted polymers (SMIPs) for selective adsorption of ampicillin sodium were synthesized using surface molecular imprinting technique with silica gel as a support. The physical and morphological characteristics of the polymers were investigated by scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FT-IR), thermogravimetric analysis (TGA), elemental analysis and nitrogen adsorption–desorption test. The obtained results showed that the SMIPs displayed great adsorption capacity (13.5μg/mg), high recognition ability (the imprinted factor is 3.2) and good binding kinetics for ampicillin sodium. Finally, as solid phase extraction adsorbents, the SMIPs coupled with HPLC method were validated and applied for the enrichment, purification and determination of ampicillin sodium in real milk and blood samples. The averages of spiked accuracy ranged from 92.1%to 107.6%. The relative standard deviations of intra-and inter-day precisions were less than 4.6%. This study provides a new and promising method for enriching, extracting and determining ampicillin sodium in complex biological samples.

  15. Detection of IgM antibodies against Toxoplasma gondii in blood samples absorbed onto filter paper, in order to assess a baseline for the occurence of congenital toxoplasma infections

    NARCIS (Netherlands)

    Conyn-van Spaendonck MAE; van Knapen F

    1990-01-01

    This report deals in the first place with some investigations on the utility of blood samples absorbed onto filter paper for determination of antibodies to Toxoplasma gondii. Secondly, the results of specific IgM detection in such samples of a cohort of neonates born in 1986/1987 are reported. The

  16. Spot14/Spot14R expression may be involved in MSC adipogenic differentiation in patients with adolescent idiopathic scoliosis

    Science.gov (United States)

    WANG, QIFEI; YANG, JUNLIN; LIN, XIANG; HUANG, ZIFANG; XIE, CHAOFAN; FAN, HENGWEI

    2016-01-01

    The aim of the present study was to evaluate the different expression levels of thyroid hormone responsive (THRSP; Spot14)/S14 related, Mig12 (S14R) during bone marrow mesenchymal stem cell (BM-MSC) adipogenesis in adolescent idiopathic scoliosis (AIS) patients. MSCs were retrospectively isolated from AIS patients and controls, and adipogenic differentiation was induced. Total RNA was extracted for Affymetrix 3′-IVT expression profiling microarrays and compared with the results from healthy controls. The results were confirmed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) validation and the protein expression levels of Spot14 and its paralogous gene S14R by western blotting and immunohistochemistry. A total of 300 significantly altered mRNAs were detected (111 upregulated and 189 downregulated) and confirmed by RT-qPCR. The mRNA expression levels of seven genes, including Spot14, were altered by >2-fold in AIS patients. Spot14/S14R was selected for further investigation. The results of the western blotting demonstrated that mRNA and protein expression levels of Spot14/S14R were significantly higher in AIS patients than the controls (P<0.05). Immunohistochemistry demonstrated Spot14 was expressed in 85% (17/20 cases) in adipose tissue samples from AIS patients and 23.1% (3/13 cases) of adipose tissue samples from controls. The positive ratio of Spot14 in adipose tissue samples from AIS was significantly higher than the controls (P<0.001). The results of the present study indicated that Spot14/S14R were differently expressed in MSC adipogenesis in AIS patients, and they may be important in the abnormal adipogenic differentiation in AIS. PMID:27082501

  17. Blood Pressure Problems During Pregnancy, Heart Trouble Later?

    Science.gov (United States)

    ... page: https://medlineplus.gov/news/fullstory_159580.html Blood Pressure Problems During Pregnancy, Heart Trouble Later? Spotting risk ... 27, 2016 (HealthDay News) -- Pregnant women who have blood pressure in the high-normal range may have an ...

  18. Hot spots in fiber pinches

    International Nuclear Information System (INIS)

    Results are presented on the behavior of ''bright spots'' in carbon and deuterium fiber Z-pinches using the MAGPIE generator at the I MA current level. The experimental diagnosis was carried out using optical and X-ray framing and streak images, along with Schlieren and shadowgraphy laser probing. After a short (∼4 ns) duration formation phase, these bright spots exhibit highly dynamic behavior. Bifurcation of the bright spots gives rise to rapid axial motion at 1-3x105 m/sec. The post-bifurcation bright spots persist for up to 40 ns. The important features of bright spot evolution can be reproduced using a 2-D MHD code incorporating LTE ionization dynamics and cold start conditions. Construction of ''artificial'' diagnostic images from the simulation data allows direct comparison to experiment. From the close agreement between experiment and 2-D simulation we infer that the observed bright spot behavior can be explained entirely in terms of the non-linear evolution of the m=0 instability

  19. Blood Clots

    Science.gov (United States)

    ... Index A-Z Blood Clots Blood clots are semi-solid masses of blood that can be stationary (thrombosis) ... treated? What are blood clots? Blood clots are semi-solid masses of blood. Normally, blood flows freely through ...

  20. BUN - blood test

    Science.gov (United States)

    Blood urea nitrogen ... A blood sample is needed. Most of the time blood is drawn from a vein located on the inside ... Many medicines can interfere with blood test results. Your health ... if you need to stop taking any medicines before you have this ...

  1. 同卵双生子外周血DNA甲基化谱的差异%Differences of DNA Methylation Profiles in Monozygotic Twins' Blood Samples

    Institute of Scientific and Technical Information of China (English)

    赵书民; 张素华; 陈金中; 李士林; 李成涛

    2011-01-01

    Objective To evaluate the potential usefulness of DNA methylation in individual discrimination of monozygotic twins by investigating the differences of DNA methylation profiles in monozygotic twins' blood samples. Methods Blood samples from 22 pairs of monozygotic twins were obtained with informed consent. Genomic DNA extracts were bisulfite treated followed by detection with Infinium (R) HumanMethyla-tion27 BeadChip Assays(Hlumina, USA). Epigenetic distances between each pair of monozygotic twins and each pair of unrelated individuals of same gender were calculated with Euclidean distance algorithms. Distribution of epigenetic distance in monozygotic twin group was statistically compared with that in unrelated individuals. Results Difference of epigenetic distance between male and female pairs was not statistically significant in unrelated individual group or in monozygotic twin group(P=0.0695 and 0.4825, respectively). Epigenetic distance of monozygotic twins was significantly lower than that of unrelated individual pair of same gender (Median: 6.02 vs 7.20, P=0.0002). However, all the epigenetic distance in monozygotic twin group or in unrelated individuals were significantly higher than 4.00(P<0.000 1). Conclusion DNA methylation profiles of monozygotic twin's blood samples were significandy different with each other, which was similar to that in unrelated individuals of same gender. These results indicated that DNA methylation was a useful biomarker in individual discrimination of monozygotic twins.%目的 通过比较不同个体外周血DNA甲基化谱的差异,评估DNA甲基化在同卵双生子个体甄别中的应用价值.方法 在知情同意基础上获得22对同卵双生子外周血样.抽提基因组DNA后进行重亚硫酸盐转化,采用Illumina公司的人27k甲基化微珠芯片检测基因组27578个CpG位点的甲基化程度(β值).依据常染色体CpG住点的β值,采用欧氏距离计算方法计算同卵双生子间以及同性别的

  2. Cold spots in neonatal incubators are hot spots for microbial contamination.

    Science.gov (United States)

    de Goffau, Marcus C; Bergman, Klasien A; de Vries, Hendrik J; Meessen, Nico E L; Degener, John E; van Dijl, Jan Maarten; Harmsen, Hermie J M

    2011-12-01

    Thermal stability is essential for the survival and well-being of preterm neonates. This is achieved in neonatal incubators by raising the ambient temperature and humidity to sufficiently high levels. However, potentially pathogenic microorganisms also can thrive in such warm and humid environments. We therefore investigated whether the level of microbial contamination (i.e., the bacterial load) inside neonatal incubators can be predicted on the basis of their average temperature and relative humidity settings, paying special attention to local temperature differences. Swab samples were taken from the warmest and coldest spots found within Caleo incubators, and these were plated to determine the number of microbial CFU per location. In incubators with high average temperature (≥ 34°C) and relative humidity (≥ 60%) values, the level of microbial contamination was significantly higher at cold spots than at hot spots. This relates to the fact that the local equilibrium relative humidity at cold spots is sufficiently high to sustain microbial growth. The abundance of staphylococci, which are the main causative agents of late-onset sepsis in preterm neonates, was found to be elevated significantly in cold areas. These findings can be used to improve basic incubator hygiene.

  3. Clinical application and experience of continuous improvement of heel blood sampling methods%持续性改进足跟采血方法在临床中的应用与体会

    Institute of Scientific and Technical Information of China (English)

    孙丽娟; 王志敏; 胡国丽

    2015-01-01

    Objective:to discuss the sustainability of improved heel blood sampling methods in clinical applications. Methods:Comprehensive introduction to improve neonatal heel blood after collection. Blood sampling timing, separately from the posture in neonates, needle position, selection of blood collection needles, blood collection, blood smear drying room environment, wound treatment, and many other improvements, summarized comparison. Results:using the improved heel blood collection, blood and wound treatment, improves blood quality and patient satisfaction. Conclusions:continuity improved heel blood collection meth-ods to improve the quality and efficiency of work, worth.%目的:讨论持续性改进足跟采血方法在临床中的应用。全面介绍改进后新生儿足跟血采集。方法:分别从采血时机、新生儿体位、针刺部位、采血针选择、采血室环境、血片的晾晒、创口的处理等多方面进行改进、总结对比。结果:运用改进后足跟血采集方法、血片及创口的处理,提高了血片质量和患者满意度。结论:持续性改进足跟采血方法提高了工作质量和效率,值得推广应用。

  4. Membrane materials for storing biological samples intended for comparative nanotoxicological testing

    Science.gov (United States)

    Metelkin, A.; Kuznetsov, D.; Kolesnikov, E.; Chuprunov, K.; Kondakov, S.; Osipov, A.; Samsonova, J.

    2015-11-01

    The study is aimed at identifying the samples of most promising membrane materials for storing dry specimens of biological fluids (Dried Blood Spots, DBS technology). Existing sampling systems using cellulose fiber filter paper have a number of drawbacks such as uneven distribution of the sample spot, dependence of the spot spreading area on the individual biosample properties, incomplete washing-off of the sample due to partially inconvertible sorption of blood components on cellulose fibers, etc. Samples of membrane materials based on cellulose, polymers and glass fiber with applied biosamples were studied using methods of scanning electron microscopy, FT-IR spectroscopy and surface-wetting measurement. It was discovered that cellulose-based membrane materials sorb components of biological fluids inside their structure, while membranes based on glass fiber display almost no interaction with the samples and biological fluid components dry to films in the membrane pores between the structural fibers. This characteristic, together with the fact that membrane materials based on glass fiber possess sufficient strength, high wetting properties and good storage capacity, attests them as promising material for dry samples of biological fluids storage systems.

  5. The influence of tube voltage and phantom size in computed tomography on the dose-response relationship of dicentrics in human blood samples

    International Nuclear Information System (INIS)

    The aim of this study was to investigate the dose response relationship of dicentrics in human lymphocytes after CT scans at tube voltages of 80 and 140 kV. Blood samples from a healthy donor placed in tissue equivalent abdomen phantoms of standard, pediatric and adipose sizes were exposed at dose levels up to 0.1 Gy using a 64-slice CT scanner. It was found that both the tube voltage and the phantom size significantly influenced the CT scan-induced linear dose-response relationship of dicentrics in human lymphocytes. Using the same phantom (standard abdomen), 80 kV CT x-rays were biologically more effective than 140 kV CT x-rays. However, it could also be determined that the applied phantom size had much more influence on the biological effectiveness. Obviously, the increasing slopes of the CT scan-induced dose response relationships of dicentrics in human lymphocytes obtained in a pediatric, a standard and an adipose abdomen have been induced by scattering effects of photons, which strongly increase with increasing phantom size.

  6. Rapid detection and identification of Wuchereria bancrofti, Brugia malayi, B. pahangi, and Dirofilaria immitis in mosquito vectors and blood samples by high resolution melting real-time PCR.

    Science.gov (United States)

    Thanchomnang, Tongjit; Intapan, Pewpan M; Tantrawatpan, Chairat; Lulitanond, Viraphong; Chungpivat, Sudchit; Taweethavonsawat, Piyanan; Kaewkong, Worasak; Sanpool, Oranuch; Janwan, Penchom; Choochote, Wej; Maleewong, Wanchai

    2013-12-01

    A simple, rapid, and high-throughput method for detection and identification of Wuchereria bancrofti, Brugia malayi, Brugia pahangi, and Dirofilaria immitis in mosquito vectors and blood samples was developed using a real-time PCR combined with high-resolution melting (HRM) analysis. Amplicons of the 4 filarial species were generated from 5S rRNA and spliced leader sequences by the real-time PCR and their melting temperatures were determined by the HRM method. Melting of amplicons from W. bancrofti, B. malayi, D. immitis, and B. pahangi peaked at 81.5±0.2℃, 79.0±0.3℃, 76.8±0.1℃, and 79.9±0.1℃, respectively. This assay is relatively cheap since it does not require synthesis of hybridization probes. Its sensitivity and specificity were 100%. It is a rapid and technically simple approach, and an important tool for population surveys as well as molecular xenomonitoring of parasites in vectors. PMID:24516268

  7. Highly sensitive tITP-CZE determination of l-histidine and creatinine in human blood plasma using field-amplified sample injection with mobility-boost effect.

    Science.gov (United States)

    Hattori, Takanari; Fukushi, Keiichi

    2016-01-01

    2D computer simulation revealed that amino acids and weak electrolytes were cationized because of the migration of counter-ion from a BGE zone to a sample zone, which encouraged electrokinetic injection (EKI) of these analytes (by the mobility-boost (MB) effect). To investigate the effects of kinds and concentrations of counter-ions on the MB effect and the analyte amount injected into the capillary, experiments, and 1D computer simulations were performed. When acetate was used as the counter-ion, the LODs (S/N = 3) of l-histidine and creatinine, respectively, reached 0.10 and 0.25 nM because of the concentration effect by transient ITP (tITP). The concentrations of l-histidine and creatinine in human blood plasma obtained using the proposed method were agreed with those obtained using the conventional methods. The proposed method can be applied to the analysis of amino acids and weak bases that have similar pI and pKa to l-histidine and creatinine. PMID:26454141

  8. Cross hedging jet fuel on the Singapore spot market.

    OpenAIRE

    Clark, Ephraim A.; Tan, Mark; Tunaru, Radu

    2003-01-01

    In this paper we test for the most effective cross hedging instrument for the Singapore spot market in jet fuel over the period February 4, 1997 to August 21, 2001. Our results are mixed. We find that the heating oil contract is the best in-sample cross-hedging instrument. It has the highest correlation with the spot price and gives the best regression results. However, after correcting for serial correlation, the goodness of fit measured by R2 is rather low. Out of sample results are weak fo...

  9. Genetics Blood Card Use

    Data.gov (United States)

    US Fish and Wildlife Service, Department of the Interior — SOP guiding collection of blood for genetics analysis. Provides stepwise instructions and guidance on how to collect DNA sample using a whole blood blot card

  10. Analysis of factors affecting pre-analytical quality of blood samples%影响检验分析前血标本质量的因素分析

    Institute of Scientific and Technical Information of China (English)

    赖琼凤

    2014-01-01

    目的:对影响检验分析前血标本质量的因素进行探讨。方法随机抽取我院在2011年1月-2013年12月检验的500例检验分析前质量出现问题的血标本,通过回顾性分析法对影响检验分析前血标本质量的因素进行探讨与分析。结果影响检验分析前血标本质量的主要因素有以下三种:饮食、用药状况、患者心理,分别占17.2%、9.6%、12.4%,其次是医生、送检人员以及护理人员等。结论在血标本检验分析前的采集、送检以及保存等操作中,医护人员要严格按照规定中的要求进行,以降低血标本质量检验分析前受到多种因素的影响,提高血标本检验结果的准确性与可靠性。%Objective To investigate the factors affecting the pre-analytical quality of blood samples. Methods Five hundred clinical blood samples with pre-analytical quality defects in our hospital from January 2011 to December 2013 were randomly selected and evaluated. Factors affecting the pre-analytical quality of blood samples were retrospectively analyzed. Results Food, medication, and psychological problems, which resulted in 17.2%, 9.6%, and 12.4% of defective blood samples, respectively, were three main factors influencing the pre-analytical quality of blood samples. There were also some secondary factors including doctors, laboratory technicians, and nurses. Conclusion To reduce the effects of multiple factors influencing the pre-analytical quality of blood samples and improve the accuracy and reliability of blood sample test, the medical staff should collect, transfer, and store pre-analytical blood samples in strict accordance with the provisions.

  11. Laser based spot weld characterization

    Science.gov (United States)

    Jonietz, Florian; Myrach, Philipp; Rethmeier, Michael; Suwala, Hubert; Ziegler, Mathias

    2016-02-01

    Spot welding is one of the most important joining technologies, especially in the automotive industry. Hitherto, the quality of spot welded joints is tested mainly by random destructive tests. A nondestructive testing technique offers the benefit of cost reduction of the testing procedure and optimization of the fabrication process, because every joint could be examined. This would lead to a reduced number of spot welded joints, as redundancies could be avoided. In the procedure described here, the spot welded joint between two zinc-coated steel sheets (HX340LAD+Z100MB or HC340LA+ZE 50/50) is heated optically on one side. Laser radiation and flash light are used as heat sources. The melted zone, the so called "weld nugget" provides the mechanical stability of the connection, but also constitutes a thermal bridge between the sheets. Due to the better thermal contact, the spot welded joint reveals a thermal behavior different from the surrounding material, where the heat transfer between the two sheets is much lower. The difference in the transient thermal behavior is measured with time resolved thermography. Hence, the size of the thermal contact between the two sheets is determined, which is directly correlated to the size of the weld nugget, indicating the quality of the spot weld. The method performs well in transmission with laser radiation and flash light. With laser radiation, it works even in reflection geometry, thus offering the possibility of testing with just one-sided accessibility. By using heating with collimated laser radiation, not only contact-free, but also remote testing is feasible. A further convenience compared to similar thermographic approaches is the applicability on bare steel sheets without any optical coating for emissivity correction. For this purpose, a proper way of emissivity correction was established.

  12. Color tunable LED spot lighting

    Science.gov (United States)

    Hoelen, C.; Ansems, J.; Deurenberg, P.; van Duijneveldt, W.; Peeters, M.; Steenbruggen, G.; Treurniet, T.; Valster, A.; ter Weeme, J. W.

    2006-08-01

    A new trend in illumination is to use dynamic light to set or dynamically vary the ambience of a room or office. For this we need color tunable spots that can reliably vary over at least a wide range of color temperatures, and preferably also more saturated colors. LEDs are in principle ideally suited for this application thanks to their nature of emitting light in a relatively narrow band. For color tunable spot lighting based on the concept of mixing RGB LED colors, the key results have been presented before. Limitations of these 3-intrinsic-color mixing systems with high color rendering properties are found in a limited operating temperature range due to wavelength shifts, a limited color temperature range, and a low maximum operating temperature due to a strong flux decrease with increasing temperature. To overcome these limitations, a 3-color R pcGB system with phosphor-converted red (R pc) and a 4-color RAGB system have been investigated. With both systems, a CRI of at least 80 can be maintained over the relevant color temperature range of approximately 2700 K to 6500 K. In this paper we compare these concepts on overall system aspects and report on the performance of prototype spot lamps. The main features of the RAGB and R pcGB spot lamp concepts can be summarized as: 1) The RAGB spot overcomes CRI and gamut shortcomings of RGB light sources and gives much freedom in wavelength selection, but suffers from temperature sensitivity and complex controls; 2) The R pcGB spot overcomes shortcomings concerning CRI and thermal dependence of RGB sources and enables relatively simple controls, but needs an improved overall red efficacy. With both color concepts, prototype spot lamps have been built. The amber to red emitting nitridosilicate-based phosphors can be wavelength-tuned for optimal performance, which is found at a peak emission around 610 nm for high color quality systems. This results in a simple and very robust system with good color consistency. For the

  13. Is this Red Spot the Blue Spot (locus ceruleum)?

    Energy Technology Data Exchange (ETDEWEB)

    Choe, Won Sick; Lee, Yu Kyung; Lee, Min Kyung; Hwang, Kyung Hoon [Gachon University Gil Hospital, Incheon (Korea, Republic of)

    2010-06-15

    The authors report brain images of 18F-FDG-PET in a case of schizophrenia. The images showed strikingly increased bilateral uptake in the locus ceruleum. The locus ceruleum is called the blue spot and known to be a center of the norepinephrinergic system.

  14. Determine the prevalence of Brucella spp. and Leptospira spp. in blood samples by multiplex polymerase chain reaction collected from cattle, sheep and goats in herds located in provinces of Iran

    Directory of Open Access Journals (Sweden)

    Faham Khamesipour

    2014-05-01

    Full Text Available Leptospirosis and brucellosis are common zoonosis that affect many species of mammals mostly causing economical losses. Further, very important fact is huge danger for human and animal health around the world. The purpose of the study is to determine the prevalence of Brucella spp. and Leptospira spp. using multiplex polymerase chain reaction (mPCR method, in blood samples collected from cattle, sheep and goats. In this study, a total number of 250 blood samples (5 cc of blood with ethilen diamin tetra asetic acid were collected randomly from 100 cattle, 80 sheep and 70 goats located on 6 herds in Chaharmahal Va Bakhtiari and Esfahan provinces, Iran. After DNA extraction and setting of mPCR for Brucella spp. and Leptospira spp. mPCR products were screened. The DNA of these microorganisms was detected by multiplex PCR from 31 and 21 out of 100 cattle, respectively. Four of 70 goat’s blood samples from goat breeding farms were positive for Leptospira spp. and 11 were positive for Brucella spp. Out of 80 sheep blood samples 23 were positive for Brucella spp. and 14 for Leptospira spp. The results of the present study show ruminant as an important reservoir for transmission of these zoonotic diseases to humans in Iran. mPCR has the ability to concurrently detect both Brucella and Leptospira species from blood samples of ruminants. The convenience and the possibility of detection of both bacteria at a time, strongly support the use of this mPCR for routine diagnostics.

  15. 一例A3血型样本的分子机制%Study of molecular mechanism for a blood sample with A3 phenotype

    Institute of Scientific and Technical Information of China (English)

    梁伟; 杨亮; 梅传亮; 许德义; 邓刚; 贺芸蕾; 刘奕宇; 张哲

    2015-01-01

    Objective To explore the molecular mechanism for a blood sample with mixed-field hemagglutination upon determination of ABO blood group.Methods Serological techniques were employed to identify the erythrocyte phenotype.The A and B antigens were detected by flow cytometry.The preliminary genotype of ABO gene was assayed with sequence-specific primer-polymerase chain reaction (PCR-SSP).Exons 6 and 7 of the ABO gene were amplified with PCR and analyzed by direct sequencing.Haplotypes of the ABO gene were analyzed by cloning sequencing as well.Results The serological reaction pattern has supported an O phenotype when all the tubes were centrifuged for the first time.However, a mixed-field hemagglutination of red blood cells (RBCs) with anti-A antibodies was present after the tube was centrifuged five times later.A antigens were detected on the surface of partial red blood cells of the sample by flow cytometry.PCR-SSP results have shown that the preliminary ABO genotype was A/O.Analysis of the fragments of exons 6 and 7 of the ABO gene has indicated that heterozygosis lied as follows:261G/A, 425T/T, 467C/T, 646A/T, 681A/G, 745C/T, 771C/T, 829A/G, conjecturing the genotype to be A307/O02, which was confirmed by haplotype sequence analysis.Compared with A101 allele, A307 allele has two missense mutations, 467C> T and 745C> T, which have resulted in substitutions Pro156Leu and Arg249Trp in the A glycosyltransferase polypeptide chain.Conclusion A variant allele (A307) has been identified for the first time in mainland China, which is responsible for the formation of A3 phenotype.%目的 研究1例ABO定型时出现混合外观凝集特征个体的分子遗传机制.方法 应用血清学方法和流式细胞术鉴定其ABO红细胞表型,序列特异性引物聚合酶链反应(sequence-specific primerpolymerase chain reaction,PCR-SSP)方法进行ABO基因型的初步检测,对ABO基因第6、7外显子进行聚合酶链反应和DNA序列分析,并进一步通过克隆

  16. Valores glicêmicos oferecidos pelo glicosímetro portátil, utilizando sangue de diferentes vias de coleta: estudo de validade Portable blood glucose meter values using different sampling ways: a validity study

    Directory of Open Access Journals (Sweden)

    Ana Paula Bello Argollo

    2010-12-01

    Full Text Available OBJETIVO: Identificar se há diferença significativa entre os resultados glicêmicos oferecidos pelo glicosímetro portátil, utilizando sangue obtido de diferentes vias de coleta e pela análise laboratorial. MÉTODOS: Estudo quantitativo de validade. Foram coletadas diariamente amostras de sangue capilar, do cateter venoso central e do catéter arterial para verificação da glicemia pelo glicosímetro, e feita análise da glicemia laboratorial. Para análise, foi utilizado o programa Statistical Package for the Social Sciences - SPSS. RESULTADOS: Observou-se que glicemia processada no glicosímetro portátil, com amostra de sangue do cateter venoso central, foi a que apresentou maior correlação com o valor oriundo do laboratório, considerando-se este o "padrão-ouro". CONCLUSÃO: Coletas por via capilar, em pacientes graves e instáveis hemodinamicamente, podem trazer resultados glicêmicos falsos, levando a alterações indevidas da solução de insulina É preciso atentar para a via de coleta ideal da glicemia em pacientes graves, afim de que não haja erro no manuseio da solução de insulina.OBJECTIVE: To identify the relevant differences between portable blood glucose meter readings in different sampling accesses blood and laboratory analysis. METHODS: Quantitative validity study. Daily samples were collected from capillary blood, central venous access catheter and arterial catheter and the blood glucose values checked using portable blood glucose meter and laboratory analysis. The findings were analyzed with the Statistical Package for the Social Sciences - SPSS software. RESULTS: Central venous catheter samples blood glucose meter readings were found to have the best correlation with the laboratory analysis results, considered as the gold-standard. CONCLUSION: Hemodynamically unstable patients' capillary blood samples may provide false blood glucose results, and lead to inappropriate insulin solution management. Therefore, ideal

  17. Association of Socioeconomic Status with Anthropometric Measures and Blood Pressure in a Representative Sample of Iranian Children and Adoles-cents: The CASPIAN-IV Study

    Directory of Open Access Journals (Sweden)

    Ramin HESHMAT

    2015-10-01

    Full Text Available Background: The aim of this study was to evaluate the association of Socioeconomic Status (SES with anthropometric measures and BP in Iranian children and adolescents.Methods: This nationwide study was conducted in 2011-2012 among 14,880 students, aged 6-18 years, selected by multistage, cluster-sampling method from rural and urban areas of 30 provinces of Iran. Anthropometric indexes and BP were measured by standard protocols and with calibrated instruments. SES was estimated based on family assets and parents’ job and education using principle component analysis method. SES was considered as “low”, “intermediate” and “high” in the statistical analysis.Results: Overall, 13486 children and adolescents out of 14,880 invited students (response rate 90.6% participated in this study. They consisted of 50.8% boys, 75.6% urban residents, with a mean age of 12.47 ±3.36 years. The prevalence of overweight, obesity and abdominal obesity and also mean of all anthropometric measures increased linearly with increasing SES. Inversely, underweight decreased linearly with increasing SE