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.
Full Text Available [ Credits ] BLAST Search Image Search Home About Archive Update History Contact us DGBY Tran...scriptome data - Air-drying stress Data detail Data name Transcriptome data - Air-drying stress Des...suggested that the genes involved in protein folding were transiently upregulated at early stages, and that ... License Update History of This Database Site Policy | Contact Us Transcriptome data - Air-drying stress - DGBY | LSDB Archive ...
Remco A. Koster
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.
A sensitive, semi-quantitative radioimmunoassay method to screen for elevations of TSH concentration in blood is described. The method requires two 0.32 cm dots of dried blood-impregnated filter paper (equivalent to 3 μl plasma) and a 3-day incubation. Separation of bound and free is obtained using polyethylene glycol. The method can recognize TSH concentrations as low as 22 μU/ml using a highly sensitive antiserum developed by one of us (AFP). TSH in dried cord and newborn blood from four infants with congenital hypothyroidism was clearly higher than in normal infants. The method is suitable for use in a newborn screening program to confirm the suspicion of primary hypothyroidism in specimens with low T4 concentrations
Wihokhoen, Benchawan; Dondorp, Arjen M.; Turner, Paul; Woodrow, Charles J.; Imwong, Mallika
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. PMID:26711525
Wihokhoen, Benchawan; Dondorp, Arjen M; Turner, Paul; Woodrow, Charles J; Imwong, Mallika
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. PMID:26711525
Full Text Available functions and in connection with vacuolar H + -ATPase, which plays a role in vacuolar acidification. To dete...rmine the role of vacuolar acidification in air-drying stress tolerance, we monit...ored intracellular pH. The results showed that intracellular acidification was induced during air-drying and that this acidification
The authors have undertaken a bacteriological study of irradiated albumin obtained from the blood of cattle. The albumin was obtained by drying blood collected in bulk at a slaughterhouse of the Argentine Meat Producers Corporation. The blood was separated by centrifuging into two fractions, white and red, the white fraction containing plasma, leukocytes and platelets, and the red fraction consisting of erythrocytes. The albumin is contaminated by microbes due to: (a) the impossibility of extracting the amount of blood involved (100 000 litres daily) under aseptic conditions; (b) transport of microbes in the blood during slaughter; (c) possible contamination by the blood of sick animals (veterinary examinations are carried out after extraction of the blood); (d) contamination caused by actual carcass processing. At present this material is used as a fertilizer, and for other industrial applications, but only in a minor degree for human nutrition. The reaction of the bacterial flora to gamma radiation was studied with the aid of a Gamma cell 220 apparatus operating in the dose range-0.5-4 Mrad. In the first test the whole material was irradiated, and total and differential counts performed for the purpose of identifying the different strains. The isolated bacterial flora was then studied together with the particular response, in each case, to different doses. The authors describe changes in the biochemical behaviour of some strains after irradiation. (author)
Chambers, Andrew G.; Percy, Andrew J.; Hardie, Darryl B.; Borchers, Christoph H.
Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach.
Tvedegaard, Kristine C.; Parner, Erik; Hooper, Craig W.;
Multiplex SNP analysis on whole genome amplified DNA from archived dried bloodspots, a validation study Kristine C. Tvedegaard,1 Erik Parner,1 Craig W. Hooper,2 Jørn Atterman,1 Niels Gregersen3, Poul Thorsen,1 1Institute of Public Health, NANEA at Department of Epidemiology, University of Aarhus...... further development of allele specific primer extension (ASPE) for multiplex SNP analysis based on the Luminex 100 IS platform. It uses isobases (isoC and isoG) and the software MultiCode-PLx platform for data analysis and data handling. We validate the EraGen multicode system in two 6-plex assays used on.......3-100%, repeatability ranged from 99.2-99.7% and robustness ranged from 94.1-99.3%. CONCLUSION: The Multi-Code System is a highly sensitive and specific method for multiplex SNP analysis on WGA DNA from archived dried bloodspots....
Wagner, Michel; Tonoli, David; Varesio, Emmanuel; Hopfgartner, Gérard
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
de Las Hazas, María Carmen López; Motilva, Maria José; Piñol, Carme; Macià, Alba
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
Batterman, Stuart A.; Chernyak, Sergey; Su, Feng-Chiao
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
The goals of the research were to determine the biochemical differences between freeze dried bovine and porcine blood relative to their nutritional value to Glossina palpalis palpalis and Stomoxys calcitrans and to develop an artificial diet for mass rearing these flies. Freeze dried bovine and porcine blood were found to differ in their amino acid content; total dietary lipids did not significantly differ, but some notable exceptions were found in fatty acid content. Both sonication and addition of foetal bovine serum to freeze dried bovine blood improved its nutritional value for G. p. palpalis. A two component, semi-defined artificial diet was developed for G. p. palpalis and S. calcitrans. The College Station diet consisted of lipid contaminated bovine haemoglobin (BHb) and bovine serum albumin (BSA). To conduct dietary deletion tests, a process was developed for preparing large quantities of ultrapure lipid free bovine haemoglobin. S. calcitrans fed on lipid free BHb plus BSA had zero fecundity. Lipids were re-added to the protein diet in three forms: (1) lipid contaminated BHb, (2) pure erythrocyte ghosts, and (3) pure lipids. It was found that membrane lipid from the erythrocyte is required by S. calcitrans. A defined artificial diet consisting of lipid free BHb, BSA, sphingomyelin, phosphatidyl ethanolamine, phosphatidyl serine and cholesterol gave normal adult survival, as well as near normal fecundity and percentage egg hatch for S. calcitrans. Knowing the identity of the lipids, it is now possible to prepare dietary formulations to alleviate dependency on the blood proteins BHb and BSA. (author). 34 refs, 1 fig., 15 tabs
McCleary, John M.
This Records and Archives Management Programme (RAMP) study covers the conservation of archival documents and the application of freeze-drying to the salvage of documents damaged by flood. Following an introductory discussion of the hazards of water, the study presents a broad summary of data on freeze-drying, including the behavior of…
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.
Sikarwar, Basant S; Roy, Mukesh; Ranjan, Priya; Goyal, Ayush
This paper examines programmed automatic recognition of infection from samples of dried stains of micro-scale drops of patient blood. This technique has the upside of being low-cost and less-intrusive and not requiring puncturing the patient with a needle for drawing blood, which is especially critical for infants and the matured. It also does not require expensive pathological blood test laboratory equipment. The method is shown in this work to be successful for ailment identification in patients suffering from tuberculosis and anaemia. Illness affects the physical properties of blood, which thus influence the samples of dried micro-scale blood drop stains. For instance, if a patient has a severe drop in platelet count, which is often the case of dengue or malaria patients, the blood's physical property of viscosity drops substantially, i.e. the blood is thinner. Thus, the blood micro-scale drop stain samples can be utilised for diagnosing maladies. This paper presents programmed automatic examination of the dried micro-scale drop blood stain designs utilising an algorithm based on pattern recognition. The samples of micro-scale blood drop stains of ordinary non-infected people are clearly recognisable as well as the samples of micro-scale blood drop stains of sick people, due to key distinguishing features. As a contextual analysis, the micro-scale blood drop stains of patients infected with tuberculosis have been contrasted with the micro-scale blood drop stains of typical normal healthy people. The paper dives into the fundamental flow mechanics behind how the samples of the dried micro-scale blood drop stain is shaped. What has been found is a thick ring like feature in the dried micro-scale blood drop stains of non-ailing people and thin shape like lines in the dried micro-scale blood drop stains of patients with anaemia or tuberculosis disease. The ring like feature at the periphery is caused by an outward stream conveying suspended particles to the edge
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
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...
Skogstrand, K.; Thorsen, P.; Vogel, I.;
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...
Full Text Available BACKGROUND: Hemoglobin (Hb is the iron-containing oxygen transport protein present in the red blood cells of vertebrates. Ancient DNA and forensic scientists are particularly interested in Hb reactions in the dry state because both regularly encounter aged, dried bloodstains. The DNA in such stains may be oxidatively damaged and, in theory, may be deteriorated by the presence of Hb. To understand the nature of the oxidative systems potentially available to degrade DNA in the presence of dried Hb, we need to determine what molecular species Hb forms over time. These species will determine what type of iron (i.e. Fe(2+/Fe(3+/Fe(4+ is available to participate in further chemical reactions. The availability of "free" iron will affect the ability of the system to undergo Fenton-type reactions which generate the highly reactive hydroxyl radical (OH*. The OH* can directly damage DNA. METHODOLOGY/PRINCIPAL FINDINGS: Oxygenated Hb (oxyHb converts over time to oxidized Hb (metHb, but this happens more quickly in the dry state than in the hydrated state, as shown by monitoring stabilized oxyHb. In addition, dry state oxyHb converts into at least one other unknown species other than metHb. Although "free" iron was detectable as both Fe(2+ and Fe(3+ in dry and hydrated oxyHb and metHb, the amount of ions detected did not increase over time. There was no evidence that Hb becomes more prone to generating OH* as it ages in either the hydrated or dry states. CONCLUSIONS: The Hb molecule in the dried state undergoes oxidative changes and releases reactive Fe(II cations. These changes, however, do not appear to increase the ability of Hb to act as a more aggressive Fenton reagent over time. Nevertheless, the presence of Hb in the vicinity of DNA in dried bloodstains creates the opportunity for OH*-induced oxidative damage to the deoxyribose sugar and the DNA nucleobases.
Aabye, Martine G; Eugen-Olsen, Jesper; Werlinrud, Anne Marie;
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...
Poulsen, Jesper Buchhave; Lescai, Francesco; Grove, Jakob; Bækvad-Hansen, Marie; Christiansen, Michael; Hagen, Christian Munch; Maller, Julian; Stevens, Christine; Li, Shenting; Li, Qibin; Sun, Jihua; Wang, Jun; Nordentoft, Merete; Werge, Thomas Mears; Mortensen, Preben Bo; Børglum, Anders Dupont; Daly, Mark; Hougaard, David Michael; Bybjerg-Grauholm, Jonas; Hollegaard, Mads Vilhelm
Stored neonatal dried blood spot (DBS) samples from neonatal screening programmes are a valuable diagnostic and research resource. Combined with information from national health registries they can be used in population-based studies of genetic diseases. DNA extracted from neonatal DBSs can be...
Thilsing-Hansen, T; Jørgensen, R J; Enemark, J M;
treated experimental cows according to expected date of calving and parity. The experimental cows were fed between 0.5 and 1.0 kg of zeolite A per day during the last 2 to 4 weeks of the dry period. Blood samples were drawn on the day of calving and day one and two after calving (all experiments), three...
A highly sensitive and specific polymerase chain reaction (PCR) assay for the detection of Trypanosoma evansi present in the dried blood on FTA Paper was developed. A simple lysis method was used to remove of the red blood cells on a 1.2 x 1.2 mm paper in 0.2 mL PCR tube. The dried paper sample was placed directly into a 25 μL PCR reaction as a DNA template. The primer set was designed and synthesized to amplify a single band of 257 bp PCR product. The sensitivity limit of test was 2.2 x 105 parasites/mL that was less than DNA templates derived from whole blood at 6.0 x 10-2 parasites/mL. The use of dried blood on FTA paper was sensitive equivalent to the determination by using the conventional wet blood film (WBF) and less sensitive than microcentrifuge heamatocrit test (MHCT) as well as mouse inoculation test (MIT). This application is not only beneficial for detection of the parasite but also useful for epidemiological study and designing Trypanosomiasis control programme. (author)
Sirdah, Mahmoud Mohammed
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).
Motohara, K.; Endo, F; Matsuda, I
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...
Ghantous, Akram; Saffery, Richard; Cros, Marie-Pierre; Ponsonby, Anne-Louise; Hirschfeld, Steven; Kasten, Carol; Dwyer, Terence; Herceg, Zdenko; Hernandez-Vargas, Hector
Background Neonatal dried blood spots (DBS) represent an inexpensive method for long-term biobanking worldwide and are considered gold mines for research for several human diseases, including those of metabolic, infectious, genetic and epigenetic origin. However, the utility of DBS is restricted by the limited amount and quality of extractable biomolecules (including DNA), especially for genome wide profiling. Degradation of DNA in DBS often occurs during storage and extraction. Moreover, amp...
Zanet, DeAnna L.; Sara Saberi; Laura Oliveira; Beheroze Sattha; Izabella Gadawski; Côté, Hélène C. F.
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...
A method of processing blood meal in which nutritive value of the protein is preserved is described, since appreciable losses occur in the nutritive value of the protein when prepared by heat sterilization with drying at atmospheric pressure in steam jacketed vessels. Blood was spray dried and irradiated at an intensity of 10 kGy. Collectively the heat of spray drying and irradiation was effective in killing both the virus plaque-forming units and the bacteria, thus producing a commercially acceptable sterile product of higher nutritive value. The relative nutritive values (RNV) of 50:50 protein were 0,56 for whey protein concentrate plus heat-sterilized blood meal and 0.90 for whey protein concentrate plus irradiated spray-dried blood powder. Whey protein concentrate used as a control has a RNV of 1,0
Full Text Available Background. FISH is a molecular cytogenetic technique enabling rapid detection of genetic abnormalities. Facilities that can run fresh/wet samples for molecular diagnosis and monitoring of neoplastic disorders are not readily available in Ghana and other neighbouring countries. This study aims to demonstrate that interphase FISH can successfully be applied to archival methanol-fixed bone marrow and peripheral blood smear slides transported to a more equipped facility for molecular diagnosis of CML. Methods. Interphase FISH was performed on 22 archival methanol-fixed marrow (BM and 3 peripheral blood (PB smear slides obtained at diagnosis. The BM smears included 20 CML and 2 CMML cases diagnosed by morphology; the 3 PB smears were from 3 of the CML patients at the time of diagnosis. Six cases had known BCR-ABL fusion results at diagnosis by RQ-PCR. Full blood count reports at diagnosis were also retrieved. Result. 19 (95% of the CML marrow smears demonstrated the BCR-ABL translocation. There was a significant correlation between the BCR-ABL transcript detected at diagnosis by RQ-PCR and that retrospectively detected by FISH from the aged BM smears at diagnosis (r=0.870; P=0.035. Conclusion. Archival methanol-fixed marrow and peripheral blood smears can be used to detect the BCR-ABL transcript for CML diagnosis.
Alice Charwudzi; Olayemi, Edeghonghon E; Ivy Ekem; Olufunmilayo Olopade; Mariann Coyle; Amma Anima Benneh; Emmanuel Alote Allotey
Background. FISH is a molecular cytogenetic technique enabling rapid detection of genetic abnormalities. Facilities that can run fresh/wet samples for molecular diagnosis and monitoring of neoplastic disorders are not readily available in Ghana and other neighbouring countries. This study aims to demonstrate that interphase FISH can successfully be applied to archival methanol-fixed bone marrow and peripheral blood smear slides transported to a more equipped facility for molecular diagnosis o...
Lewis, Michelle Huckaby; Goldenberg, Aaron J
There may be compelling reasons to return to parents a limited subset of results from research conducted using residual newborn screening dried blood samples (DBS). This article explores the circumstances under which research results might be returned, as well as the mechanisms by which state newborn screening programs might facilitate the return of research results. The scope of any responsibility to return results of research conducted using DBS should be assessed in light of the potential impact on the primary mission of state newborn screening programs. PMID:26479566
The demonstration of very high levels of immunoreactive trypsin in the blood of newborn infants with cystic fibrosis has provided a new way of detecting the disease soon after birth. A radioimmunoassay of trypsin in the eluate of blood dried on filter paper has now been developed. The sensitivity and accuracy of the method, as well as the good correlation observed between the values obtained and those of the conventional plasma assay, indicate that it is reliable and well adapted to the newborn. The new assay can easily be inserted into the present system of neonatal disease detection. A preliminary assessment of more than 5000 tests enables the authors to report an early diagnosis of proven cystic fibrosis and to discuss an essential aspect of mass-detection methods: the indicence of false-positive results
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%.
Sadones, Nele; Archer, John R H; Ingels, Ann-Sofie M E; Dargan, Paul I; Wood, David M; Wood, Michelle; Neels, Hugo; Lambert, Willy E; Stove, Christophe P
Gamma-hydroxybutyric acid (GHB) is a well-known illicit club and date-rape drug. Dried blood spot (DBS) sampling is a promising alternative for classical venous sampling in cases of (suspected) GHB intoxication since it allows rapid sampling, which is of interest for the extensively metabolized GHB. However, there is limited data if -and how- capillary DBS concentrations correlate with venous concentrations. We conducted a comparative study in 50 patients with suspected GHB intoxication, to determine and to correlate GHB concentrations in venous DBS (vDBS) and capillary DBS (cDBS). This is the first study that evaluates in a large cohort the correlation between capillary and venous concentrations of an illicit drug in real-life samples. Of the 50 paired samples, 7 were excluded: the vDBS concentration was below the LLOQ of 2 µg/mL in 3 cases and 4 samples were excluded after visual inspection of the DBS. Bland-Altman analysis revealed a mean % difference of -2.8% between cDBS and vDBS concentrations, with the zero value included in the 95% confidence interval of the mean difference in GHB concentration. A paired sample t-test confirmed this observation (p = 0.17). Also the requirement for incurred sample reproducibility was fulfilled: for more than two-thirds of the samples the concentrations obtained in cDBS and those in vDBS were within 20% of their mean. Since equivalent concentrations were observed in cDBS and vDBS, blood obtained by fingerprick can be considered a valid alternative for venous blood for GHB determination. PMID:25565078
Yourno, J; Conroy, J.
A method for detection of proviral human immunodeficiency virus DNA in dried blood spots on filter paper by direct polymerase chain reaction (PCR) has been developed. To develop the method, a standard system was used which was prepared from cells each containing a single integrated provirus and titrated with normal donor blood. This rapid procedure provides virtually quantitative yields of nuclear DNA and exploits most of the standard methodology described for blood specimens. A nested PCR us...
Full Text Available Mucopolysaccharidosis IVA (MPS IVA; Morquio A disease is an autosomal recessive disease caused and characterized by a decreased activity of N-acetylgalactosamine-6-sulfate sulfatase (GALNS, resulting in accumulation of keratan sulfate and chondroitin-6-sulfate in tissues and secondary organ damage. Recently approved enzyme replacement therapy renders the easy and early identification of MPS IVA of out-most importance.We propose a completely new assay for the stable and reproducible detection of GALNS deficiency in dry blood spots (DBS. For the validation blood samples were taken from 59 healthy individuals and 24 randomly selected genetically confirmed MPS IVA patients. The material extracted from DBS was incubated with a 4-methylumbelliferyl-β-D-galactopyranoside-6-sulfate as a specific substrate. Final enzymatic product, 4-methylumbelliferone, obtained after adding exogenous beta-galactosidase, was quantified by LC/MRM-MS (liquid-chromatography/multiple-reaction-monitoring mass-spectrometry. 4-propyl-5-hydroxy-7-methyl-2h-chromen-2-one was used as internal standard, a compound with a similar molecular structure and fragmentation pattern in negative ion mode as 4-methylumbelliferone.The enzymatic assay yielded a positive and negative predictive value of 1.0 for genetically confirmed MPS IVA patients (GALNS activity of 0.35 ± 0.21 μmol/L/h and for controls with normal GALNS activity (23.1 ± 5.3 μmol/L /h. With present enzymatic conditions, the reaction yield in dried blood spots is at least 20 fold higher than any previously reported data with other assays.The present LC/MRM-MS based assay for MPS IVA diagnosis provides an easy, highly-standardized, accurate and innovative quantification of the enzymatic product in vitro and distinguishes perfectly between MPS IVA affected patients and normal controls. This technique will significantly simplify the early detection of MPS IVA patients.
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
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.
Huan Huang; Ying Bu; Guo-Hua Zhou
AIM: To demonstrate an inexpensive method for typing gastric cancer related single nucleotide polymorphisms (SNPs) using whole blood or paper-dried blood as starting materials.METHODS: PCR amplification is directly carried out from the whole blood or paper-dried blood sample without any DNA extraction step. Before PCR, a blood sample, four primers, and all of biological reagents necessary for PCR were added at a time; After PCR, the amplified products were directly separated by slab gel electrophoresis or microchip CE without any purification. SNP typing was performed by tetra-primer PCR with two inner primers specific to each allele and two outer primers defining the length of allele-specific amplicons. Genotypes were directly discriminated by the size of amplicons specific to each allele, thereby avoiding any post-PCR process.RESULTS: Using a special PCR buffer, inhibitory substances in blood (including the anticoagulant in blood) and filter paper were effectively suppressed;a "true" single-tube-genotyping is thus realized. We successfully determined genotypes IL-1B-511 and IL-1B-31 polymorphisms at the gene IL-1B by using whole-blood and paper-dried blood samples as starting materials respectively. The method is so sensitive that 0.5-1.0 μL of blood sample is enough to give a satisfactory typing results. The genotyping results were confirmed by RFLP-PCR using purified genome DNA,indicating that amplification specificity was not affected by inhibitory components (including coagulants) in blood or filter paper.CONCLUSION: Compared with SNP typing methods based on purified DNA, the proposed method is laborsaving, simple, inexpensive, and less cross-contaminated.It is promising to use this method to type other SNPs.
Hamelin, Elizabeth I.; Blake, Thomas A.; Perez, Jonas W.; Crow, Brian S.; Shaner, Rebecca L.; Coleman, Rebecca M.; Johnson, Rudolph C.
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.
Full Text Available Abstract Background Testing for viral DNA in neonatal blood dried on paper (DBS has proved a valid means of diagnosing congenital CMV infection with both clinical and epidemiological relevance. To assess the quality of the detection of CMV-DNA on DBS in laboratories performing this test a proficiency panel consisting of nine samples with two blood spots on each filter paper was produced and distributed. Six samples were derived from whole blood, negative for CMV DNA and antibody, and spiked with cell-grown CMV Towne in various concentrations (7.3 × 102 – 9.6 × 105 copies/ml, one was a CMV positive clinical specimen (3.9 × 106 copies/ml, and two samples were CMV-negative whole blood. Results The 27 responding laboratories from 14 countries submitted 33 datasets obtained by means of conventional PCR (n = 5 or real-time PCR (n = 28 technologies. A correct positive result was reported in at least 91% of datasets in samples with a viral load of 8.8 × 104 copies/ml or higher. However only 59% and 12% identified the 9.4 × 103 and 7.3 × 102 copies/ml samples, respectively, correctly as positive. False positive results were reported by 9% of laboratories and in 11% of datasets. Conclusion These results indicate a clear need for improvement of methods as sensitivity and false-positivity still appear to be a major problem in a considerable number of laboratories.
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.
A routine and automatized methodology for thyroxine (T4) and thyrotropin (TSH) radioimmunoassay (RIA) using dried blood samples on filter paper is described. Five mm diameter dots were prepared. One eluted dot, corresponding to 4 μl of plasma, was used for T4-RIA while two were necessary for TSH-RIA. Reference filter papers were introduced in each assay for quality control. In a preliminary study on 1903 newborns, samples were obtained, generally between the 5th-7th day. Mean dot T4 was 7.38 +- 2.5 μg/dl. Mean dot TSH was 11.83 +- 9.1 μU/ml, the equation of the regression line between dot TSH (y) and serum TSH (x) being Y = 10.29 + 0.623x. (orig.)
De Kesel, Pieter M M; Lambert, Willy E; Stove, Christophe P
Volumetric absorptive microsampling (VAMS) is a novel sampling technique that allows the straightforward collection of an accurate volume of blood (approximately 10μL) from a drop or pool of blood by dipping an absorbent polymeric tip into it. The resulting blood microsample is dried and analyzed as a whole. The aim of this study was to evaluate the potential of VAMS to overcome the hematocrit bias, an important issue in the analysis of dried blood microsamples. An LC-MS/MS method for analysis of the model compounds caffeine and paraxanthine in VAMS samples was fully validated and fulfilled all pre-established criteria. In conjunction with previously validated procedures for dried blood spots (DBS) and blood, this allowed us to set up a meticulous comparative study in which both compounds were determined in over 80 corresponding VAMS, DBS and liquid whole blood samples. These originated from authentic human patient samples, covering a wide hematocrit range (0.21-0.50). By calculating the differences with reference whole blood concentrations, we found that analyte concentrations in VAMS samples were not affected by a bias that changed over the evaluated hematocrit range, in contrast to DBS results. However, VAMS concentrations tend to overestimate whole blood concentrations, as a consistent positive bias was observed. A different behavior of VAMS samples prepared from incurred and spiked blood, combined with a somewhat reduced recovery of caffeine and paraxanthine from VAMS tips at high hematocrit values, an effect that was not observed for DBS using a very similar extraction procedure, was found to be at the basis of the observed VAMS-whole blood deviations. Based on this study, being the first in which the validity and robustness of VAMS is evaluated by analyzing incurred human samples, it can be concluded that VAMS effectively assists in eliminating the effect of hematocrit. PMID:26041521
Vnučec Popov, Tanja; Cvitkovič-Maričič, Lea; Prosen, Helena; Brodnjak-Vončina, Darinka
An LC-MS/MS method for determination of the anti-epileptic drug topiramate (TPM) in dried blood spot (DBS) samples was developed and validated. DBS samples were prepared by spotting 30 ŽL of spiked whole blood onto FTATM DMPK-C cards and drying for at least 3 h. Six-millimetre punched spots were then extracted by using a mixture of methanol and water (90:10, v/v) with deuterated internal standard (topiramate-d12). The extracted samples were injected into a liquid chromatograph equipped with a...
Tretzel, Laura; Thomas, Andreas; Geyer, Hans; Gmeiner, Günter; Forsdahl, Guro; Pop, Valentin; Schänzer, Wilhelm; Thevis, Mario
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
Hendrix, Kristin S.; Meslin, Eric M.; Carroll, Aaron E.; Downs, Stephen M.
Objective Identify the relative importance of factors that impact parents' attitudes towards use of their child's dried newborn blood spots for research purposes. Methods Respondents were parents aged 18 and older with at least one child aged 17 or younger born in Indiana visiting an urban pediatrics clinic. They were asked to rate the acceptability of hypothetical scenarios involving the research use of blood spots. Three pieces of information varied between the scenarios: 1) who would be conducting the research; 2) whether the child's identity would be linked to the spots; and 3) whether and how often the parents' consent would be sought before the research began. Results 506 predominantly Black and low-income parents completed the survey. The conjoint analysis model showed good fit (Pearson's R=0.998, p<0.001). The rank order of factors affecting parents' attitudes was: 1) consent (importance score=64.9), 2) whether the child's identity was linked to the spot (importance score=19.4), 3) affiliation of the researcher using the spots (importance score=14.6). Respondents preferred being asked for their consent each time their children's spots would be used. They preferred that the children's identity not be linked to the spots and that the research is conducted by university researchers, though these issues had less impact on attitudes than consent. Conclusions Parents strongly prefer that consent be sought for each use of their children's spots. These findings have implications for future research and policymaking decisions. PMID:24011748
Foivas, Anargyros; Malenović, Anđelija; Kostić, Nađa; Božić, Marija; Knežević, Miroslav; Loukas, Yannis L; Dotsikas, Yannis
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
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.
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Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.
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
Winkel, Bo G; Hollegaard, Mads Vilhelm; Olesen, Morten S; Svendsen, Jesper H; Haunsø, Stig; Hougaard, David M; Tfelt-Hansen, Jacob
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...
Skogstrand, Kristin; Thorsen, Poul; Nørgaard-Pedersen, Bent;
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...
Klamer, A; Skogstrand, Kristin; Hougaard, D M;
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...
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)
Lemonnier, F; Masson, J; Laroche, D; Travert, J; Travert, G
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
Bosilkovska, M; Samer, C F; Déglon, J; Rebsamen, M; Staub, C; Dayer, P; Walder, B; Desmeules, J A; Daali, Y
The suitability of the capillary dried blood spot (DBS) sampling method was assessed for simultaneous phenotyping of cytochrome P450 (CYP) enzymes and P-glycoprotein (P-gp) using a cocktail approach. Ten volunteers received an oral cocktail capsule containing low doses of the probes bupropion (CYP2B6), flurbiprofen (CYP2C9), omeprazole (CYP2C19), dextromethorphan (CYP2D6), midazolam (CYP3A), and fexofenadine (P-gp) with coffee/Coke (CYP1A2) on four occasions. They received the cocktail alone (session 1), and with the CYP inhibitors fluvoxamine and voriconazole (session 2) and quinidine (session 3). In session 4, subjects received the cocktail after a 7-day pretreatment with the inducer rifampicin. The concentrations of probes/metabolites were determined in DBS and plasma using a single liquid chromatography-tandem mass spectrometry method. The pharmacokinetic profiles of the drugs were comparable in DBS and plasma. Important modulation of CYP and P-gp activities was observed in the presence of inhibitors and the inducer. Minimally invasive one- and three-point (at 2, 3, and 6 h) DBS-sampling methods were found to reliably reflect CYP and P-gp activities at each session. PMID:24722393
Hoshi, F; Satho, M; Koyama, S; Nakadaka, K; Chiba, M; Ikeda, N; Hakamada, R; Higuchi, S; Kawamura, S
The usefulness of a dry-chemistry blood analyzer, Spotchem SP-4410 (SP-4410) in a veterinary clinic for analysis of bovine and equine blood chemistry was studied. We quantitated total protein (TP), albumin (Alb), total bilirubin (T-Bil), blood urea nitrogen (BUN), total cholesterol (T-Cho), glucose (Glu), calcium (Ca), aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), creatinine phosphokinase (CPK), and alkaline phosphatase (ALP) in bovine sera. Each sample was assayed with both the SP-4410 and an automated blood analyzer which served as a wet-chemistry reference system, and the data were analyzed with regression analysis. The correlation coefficient for AST was 0.997 being the highest for all the parameters, and all the correlation coefficients were 0.93 or higher. The coefficients of variation were lower than 5.0 except in the case of bovine T-Bil where it was 5,756. The ranges of normal reference values measured by SP-4410 were the same as those reported by other investigators in most cases, but those for GGT and CPK were slightly higher. The strongest interference was observed with hemoglobin. It seems that dry-chemical-analysis of blood serum using the SP-4410 is useful for analysis of bovine and equine blood. PMID:8085395
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.
Archives of blood stations are formed directly to engage in the work of this unit, it has a reference value of various kinds of different forms such as text, graphics, audio and video historical records in the blood stations , including blood stations business archives is an important part of blood stations archives. Strengthen the management of blood sta-tions archives, can effectively safeguard the rights and interests of blood donors and recipients, and blood services, regu-late professional behavior of medical institutions to provide important basis. Combining the reality of this unit, by improv-ing the archives management of blood stations a recognition of the importance of comprehensive construction, to strength-en the standardized management of archives to speed up the pace of blood stations modernization construction, to promote the development of career of blood donation without compensation.%血站档案是血站从事各项工作中直接形成的对本单位有参考利用价值的各种文字、图表、声像等不同形式的历史记录，其中血站业务档案是血站档案的重要组成部分。加强对血站档案资料的管理，能够有效维护献血者及受血者的权益，并为采供血机构、医疗机构规范执业行为提供重要依据。结合本单位实际，通过提高档案管理对血站全面建设重要性的认识，加强档案规范化管理，加快血站现代化建设步伐，促进无偿献血事业发展。
Smit Pieter W
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.
Randriamparany, T; Kouakou, K V; Michaud, V; Fernández-Pinero, J; Gallardo, C; Le Potier, M-F; Rabenarivahiny, R; Couacy-Hymann, E; Raherimandimby, M; Albina, E
The performance of Whatman 3-MM filter papers for the collection, drying, shipment and long-term storage of blood at ambient temperature, and for the detection of African swine fever virus and antibodies was assessed. Conventional and real-time PCR, viral isolation and antibody detection by ELISA were performed on paired samples (blood/tissue versus dried-blood 3-MM filter papers) collected from experimentally infected pigs and from farm pigs in Madagascar and Côte d'Ivoire. 3-MM filter papers were used directly in the conventional and real-time PCR without previous extraction of nucleic acids. Tests that performed better with 3-MM filter papers were in descending order: virus isolation, real-time UPL PCR and conventional PCR. The analytical sensitivity of real-time UPL PCR on filter papers was similar to conventional testing (virus isolation or conventional PCR) on organs or blood. In addition, blood-dried filter papers were tested in ELISA for antibody detection and the observed sensitivity was very close to conventional detection on serum samples and gave comparable results. Filter papers were stored up to 9 months at 20-25°C and for 2 months at 37°C without significant loss of sensitivity for virus genome detection. All tests on 3-MM filter papers had 100% specificity compared to the gold standards. Whatman 3-MM filter papers have the advantage of being cheap and of preserving virus viability for future virus isolation and characterization. In this study, Whatman 3-MM filter papers proved to be a suitable support for the collection, storage and use of blood in remote areas of tropical countries without the need for a cold chain and thus provide new possibilities for antibody testing and virus isolation. PMID:25430732
Soetens, Oriane; Vauloup-Fellous, Christelle; Foulon, Ina; Dubreuil, Pascal; De Saeger, Ben; Grangeot-Keros, Liliane; Naessens, Anne
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...
Skogstrand, Kristin; Thorsen, Poul; Nørgaard-Pedersen, Bent;
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...... residual DBSS constitute a valuable resource for research into the etiology of these diseases. The small amount of blood available, however, limits the number of analytes that can be determined by traditional immunoassay methodologies. METHODS: We used new multiplexed sandwich immunoassays based on...
Natan, Dity; Nagler, Arnon; Arav, Amir
Background We recently showed that freeze-dried cells stored for 3 years at room temperature can direct embryonic development following cloning. However, viability, as evaluated by membrane integrity of the cells after freeze-drying, was very low; and it was mainly the DNA integrity that was preserved. In the present study, we improved the cells' viability and functionality after freeze-drying. Methodology/Principal Findings We optimized the conditions of directional freezing, i.e. interface ...
Chaillet, P; Zachariah, R.; Harries, K; Rusanganwa, E; Harries, A. D.
A study was conducted in two primary health facilities in Kigali, Rwanda, to determine whether dried blood spots (DBS) used for quality control of HIV testing would give comparable results with serum after being stored for a period of 14 days and 30 days at ambient temperature. DBS and serum specimens were collected from patients undergoing HIV testing. ELISA performed on serum at baseline (gold standard) was compared with DBS results. The study included a total of 491 patients, comprising 92...
Aarif, Ovais; Aggarwal, Anjali
This study aimed to evaluate the impact of evaporative cooling during late gestation on physiological responses, blood gas and acid base balance and subsequent milk production of Murrah buffaloes. To investigate this study sixteen healthy pregnant dry Murrah buffaloes (second to fourth parity) at sixty days prepartum were selected in the months of May to June and divided into two groups of eight animals each. One group of buffaloes (Cooled/CL) was managed under fan and mist cooling system during dry period. Group second buffaloes (Noncooled/NCL) remained as control without provision of cooling during dry period. The physiological responses viz. Rectal temperature (RT), Respiratory rate (RR) and Pulse rate were significantly ( P group 2, with the provision of cooling. Skin surface temperature at thorax was significantly lower in cooled group relative to noncooled group. Blood pH and pO2 were significantly ( P group as compared to the cooled group. pCO2, TCO2, HCO3, SBC, base excess in extracellular fluid (BEecf), base excess in blood (BEb), PCV and Hb were significantly ( P group as compared to noncooled group. DMI was significantly ( P group of Murrah buffaloes.
Aarif, Ovais; Aggarwal, Anjali
This study aimed to evaluate the impact of evaporative cooling during late gestation on physiological responses, blood gas and acid base balance and subsequent milk production of Murrah buffaloes. To investigate this study sixteen healthy pregnant dry Murrah buffaloes (second to fourth parity) at sixty days prepartum were selected in the months of May to June and divided into two groups of eight animals each. One group of buffaloes (Cooled/CL) was managed under fan and mist cooling system during dry period. Group second buffaloes (Noncooled/NCL) remained as control without provision of cooling during dry period. The physiological responses viz. Rectal temperature (RT), Respiratory rate (RR) and Pulse rate were significantly ( P < 0.05) lower in group 2, with the provision of cooling. Skin surface temperature at thorax was significantly lower in cooled group relative to noncooled group. Blood pH and pO2 were significantly ( P < 0.05) higher in heat stressed group as compared to the cooled group. pCO2, TCO2, HCO3, SBC, base excess in extracellular fluid (BEecf), base excess in blood (BEb), PCV and Hb were significantly ( P < 0.05) higher in cooled group as compared to noncooled group. DMI was significantly ( P < 0.05) higher in cooled relative to noncooled animals. Milk yield, FCM, fat yield, lactose yield and total solid yield was significantly higher ( P < 0.05) in cooled group of Murrah buffaloes.
Laura Russo Zillmer
Full Text Available OBJECTIVE: To validate and develop an immunonephelometric assay for the determination of alpha-1 antitrypsin (AAT levels in dried blood spots from COPD patients in Brazil. METHODS: We determined AAT levels in serum samples and dried blood spots from 192 COPD patients. For the preparation of dried blood spots, a disk (diameter, 6 mm was placed into a tube, eluted with 200 µL of PBS, and stored overnight at 4ºC. All of the samples were analyzed by immunonephelometry in duplicate. We used the bootstrap resampling method in order to determine a cut-off point for AAT levels in dried blood spots. RESULTS: The correlation coefficient between the AAT levels in serum samples and those in dried blood spots was r = 0.45. For dried blood spots, the cut-off value was 2.02 mg/dL (97% CI: 1.45-2.64 mg/dL, with a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 95.7%, 27.2%, and 100%, respectively. CONCLUSIONS: This method for the determination of AAT levels in dried blood spots appears to be a reliable screening tool for patients with AAT deficiency.
Full Text Available Due to the widely supported theory of bovine spongiform encephalopathy (BSE spread in cattle by contaminated animal feeds, screening of feed products has become essential. For many years, manufacturers have used blood and plasma proteins as high quality ingredients of foods for both pets and farm animals. However, in Europe, the Commission Regulation 1234/2003/EC temporally bans the use of processed animal proteins, including blood-derivative products, in feedstuffs for all farm animals which are fattened or bred for the production of food. This regulation has some exceptions, such as the use of non ruminant blood products into the feed of farm fish. Authorization of the re-introduction of these proteins into animal feed formulations, especially non ruminant proteins into the feed for non ruminant farm animals, is expected when adequate control methods to discriminate ruminant proteins exist. Currently, the number of validated methods to differentiate the species of origin for most of the animal by-products is limited. Here we report the development of a rapid and sensitive polymerase chain reaction (PCR-based assay, which allows detection of bovine or porcine specific mitochondrial DNAfrom spray-dried blood derivate products (plasma, whole blood and red cells, as a marker for bovine contamination in porcine products. Sample extracts, suitable for PCR, were easily and quickly obtained with the commercial PrepManTM Ultra reagent (Applied Biosystems. To confirm the porcine origin of the samples, primers targeting a specific region of 134 bp of the porcine cytochrome b coding sequence were designed (cytbporc1-F and cytbporc2-R. Previously published PCR primers (L8129 and H8357, specific for a 271 bp fragment of the bovine mitochondrial ATPase 8-ATPase 6 genes, were chosen to accomplish amplification of bovine DNA. The limit of detection (LOD of the bovine PCR assay was at least of 0.05% (v/v of bovine inclusion in spray-dried porcine plasma or red
Gulay, M S; Hayen, M J; Head, H H; Bachman, K C
The objective was to determine the effect of supplemental estrogen (estradiol cypionate, ECP) at dry-off on temporal changes in concentrations of lactose in blood plasma of Holstein cows as an indicator of rate of mammary involution. Thirty-two Holstein cows (8/group) were assigned randomly to 4 treatment groups: 30-d dry, 30-d dry + ECP, 60-d dry, and 60-d dry + ECP. A single injection (7.5 mL) of cottonseed oil (30- and 60-d dry) or ECP (15 mg) in oil (30- and 60-d dry + ECP) was administered intramuscularly at dry-off. Blood samples were collected from the coccygeal vein of all cows 24 h before dry-off and at dry-off, and then 8 samples were collected throughout the subsequent 48 h to monitor concentrations of lactose in blood plasma. No significant effects of ECP on the overall mean concentrations of lactose were detected. Concentrations of lactose increased and were greatest in blood collected 20 h (520.4 +/- 54.1, 268.1 +/- 48.2, 345.0 +/- 52.3, 418.4 +/- 49.8 microM, for the 4 treatment groups respective to the order listed above) after supplemental ECP and final milk removal. At 40 h, concentrations approached those observed 24 h before dry-off (140.5 +/- 52.1, 57.6 +/- 47.1, 90.1 +/- 51.4, 61.2 +/- 48.4 microM, respectively). Concentrations of lactose at 20 h were positively correlated with milk yield of cows at dry-off. Similar temporal profiles of lactose in blood plasma of cows supplemented or not with ECP indicated that ECP at dry-off did not markedly alter the course of tight junction leakage that typically occurs in mammary epithelial tissue during progressive early involution when milk removal is discontinued. PMID:19620664
Jamroz, D; Wiliczkiewicz, A; Orda, J; Skorupińska, J; Słupczyńska, M; Kuryszko, J
The chemical composition of spray dried porcine blood by-products is characterised by wide variation in crude protein contents. In spray dried porcine blood plasma (SDBP) it varied between 670-780 g/kg, in spray dried blood cells (SDBC) between 830-930 g/kg, and in bone protein hydrolysate (BPH) in a range of 740-780 g/kg. Compared with fish meal, these feeds are poor in Met and Lys. Moreover, in BPH deep deficits of Met, Cys, Thr and other amino acids were found. The experiment comprised 7 dietary treatments: SDBP, SDBC, and BPH, each at an inclusion rate of 20 or 40 g/kg diet, plus a control. The addition of 20 or 40 g/kg of the analysed meals into feeds for very young chickens (1-28 d post hatch) significantly decreased the body weight (BW) of birds. Only the treatments with 40 g/kg of SDBP and SDBC showed no significant difference in BW as compared with the control. There were no significant differences between treatments and type of meal for feed intake, haematocrit and haemoglobin concentrations in blood. Addition of bone protein and blood cell meals to feed decreased the IgG concentration in blood and caused shortening of the femur and tibia bones. However, changes in the mineral composition of bones were not significantly affected by the type of meal used. The blood by-products, which are rich in microelements, improved retention of Ca and Cu only. In comparison to control chickens, significantly better accretion of these minerals was found in treatments containing 20 g/kg of SDBP or 40 g/kg of SDBC. Great variability in apparent ileal amino acid digestibility in chickens was determined. In this respect, some significant differences related to the type of meal fed were confirmed for Asp, Pro, Val, Tyr and His. In general, the apparent ileal digestibility of amino acids was about 2-3 percentage units better in chickens fed on diets containing the animal by products than in control birds. PMID:22029787
M. A. Gordukova
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.
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
Anirudh J. Ullal
Full Text Available Mucopolysaccharidosis type VI or Maroteaux–Lamy syndrome is an autosomal recessive lysosomal storage disorder caused by deficiency of arylsulfatase B (ARS-B enzyme activity. It results in mild to severe multi-organ system failure from accumulation of undigested glycosaminoglycans (GAGs; dermatan sulfate and chondroitin-4-sulfate. We have developed a single-step enzyme assay using a fluorescent substrate and dried blood spots to measure ARS-B activity to identify disease patients. This assay is robust, reproducible, specific and convenient to perform.
Lorenz, I; Aigner, M; Klee, W
The usefulness of the dry-chemistry blood analyzer, SPOTCHEM SP-4410, for analysis of bovine blood chemistry was studied in a veterinary clinic. The control serum Precipath-U, Boehringer-Mannheim, was used to measure precision within each run and between days. The coefficients of variation (CV) ranged between 1.54% and 4.86%, with the exception of albumin and creatine phosphokinase showing a CV of 6.3% and 10.03% for between-day precision. For methodological comparison bovine serum samples were assayed with both the SPOTCHEM SP-4410 and the automated blood analyzer HITACHI 705, which served as a wet-chemistry reference system. The following analytes were measured: glucose, urea, creatinine, total protein, albumin, total bilirubin and the enzymes AST, CPK and gamma-GT. For hemoglobin, which was measured in heparinized whole blood, the CO oximeter 855, CIBA-CORNING, was used as a reference system. The comparative analysis showed very good correlation in eight of ten parameters and their correlation coefficients (r) ranged between 0.962 and 0.998. Only the correlation coefficients of the analysis of total bilirubin (r = 0.903) and albumin (r = 0.771) were less satisfactory. The recovery test was carried out with the two parameters glucose and blood urea. The recovery of glucose was 93.7% and of urea 98.8%. The SPOTCHEM SP-4410 is easy to use and proved to be reliable and accurate, and therefore it seems to be useful for analysis of bovine blood samples. PMID:11225499
Yu, Chaowen; Zhang, Juan; Yuan, Zhaojian; Liu, Hao; Wang, Xingbin; Wang, Ming; Zou, Lin
Quantification of human hemoglobin (Hb) is essential for diagnosis of anemia, especially for screening for thalassemia and sickle cell disease. The main methods currently used for quantification of Hb, including spectrophotometry, fluorimetry, and electrochemical assays, are all based on the structural integrity of Hb, which could be affected by hemolysis and degradation. When used for disease screening, whole blood specimens cannot meet requirements for sample collecting, transport, and storage. Here, we report a novel MS-MS method for quantification of Hb from dried blood spots (DBS) by use of a triple-quadrupole mass spectrometer. Proteospecific peptides from α-globin chains were selected after tryptic digestion. The precursor → product ion transitions of representative peptides were studied to identify the best choice with regard to sensitivity and chromatographic properties. For quantification, stable isotope-labeled peptides were used as internal standards. The concentration of Hb in each sample was obtained by calculation on the basis of established equations. The precision of the method was within 15 % and accuracy was in the range -7 to 13.0 %. Compared with routine clinical results obtained by use of the automated hematology analyzer (AHA) assay, the correlation, r (2), was >0.993. When used for determination of anemia levels the sensitivity of the assay was 95.7 % and specificity 96.5 %. Our new approach for quantification of the concentration of Hb from DBS is feasible, and precision is acceptable. The method could be used for determination of anemia levels when screening for hemoglobin disorders. Graphical Abstract Quantification of human hemoglobin from digested dried blood spot samples using tandem mass spectrometry. PMID:26345440
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)
This document deals with the comparative evaluation of the analytical data on the trace elements in beef blood obtained by 38 laboratories in 23 countries. The evaluations were based on 799 laboratory mean values of concentration of 41 elements. It was one of the series of IAEA intercomparisons on the determination of trace elements in animal materials. It was organized for the purpose of assisting the participating laboratories to check the accuracy of their work and to prepare a new reference material
Kyriakou, Chrystalla; Marchei, Emilia; Scaravelli, Giulia; García-Algar, Oscar; Supervía, August; Graziano, Silvia
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
Bou Zeid, Wassim; Brutin, David
Newtonian and non-Newtonian fluids start spreading after coming into contact with a solid substrate till the anchoring of the triple line. Our experimental work aims to study the effect of the relative humidity on the spreading dynamics of drops of whole blood. Drops of blood of same volume (+/-4.8%) are injected using a digital micropipette and gently deposited onto microscope ultraclean glass substrates. Experiments are conducted in a glove box at ambient temperature and a range of investigated relative humidities between 13.0% and 80.0%. The recorded images are post-processed using ImageJ in which the position of the contact line is measured every 20 ms. We show that the spreading dynamics is, in a first time, governed by relative humidity and later no more influence by relative humidity. Two spreading regimes are observed and analyzed compared to classical viscous drops. In previous work, we show that relative humidity influences the contact angle and the initial wetting radius. In the first regime, we find a spreading power law exponent that decreases for an increasing relative humidity. In the second regime, all the data collapse on each other and the evolution of the dimensionless radius no more depend on relative humidity.
Cardiovascular Diseases (CVDs) have been a major cause for deaths in both men and women in United States. Cerebrovascular Diseases like Strokes are known to have origins in CVDs as well. Moreover, nearly 18 Million Americans have a history of myocardial infarction and are currently undergoing cardiac rehabilitation. Consequently, CVDs are the highest costing disease groups and cost more than all types of cancer combined. However, significant cost reduction is possible through the effective use of the vast advances in embedded and pervasive electronic devices for healthcare. These devices can automate and move a significant portion of disease management to the patient's home through cyber connectivity, a concept known as point-of-care (POC) diagnostics and healthcare services. POC can minimize hospital visits and potentially avoid admission altogether with prognostic tools that give advanced notice of any abnormalities or chronic illnesses so that the treatment can be planned in advance. The POC concept requires continuous remote health monitoring. Therefore, the various sensors needed for comprehensive monitoring need to be worn daily and throughout the day. Moreover, true "roaming" capability is necessary so that it does not restrict the user's travel or his/her quotidian activities. Two biomedical signals namely, Electrocardiogram (ECG) and Blood Pressure are important diagnostic tests in assessing the cardiac health of a person. To that end, the research presented in this thesis: First , describes the development of a remote monitoring solution based on Bluetooth(TM), smartphones and cyber infrastructure for cardiac care called e-nanoflex. Second, Sensors for ECG that are compatible with everyday life style namely, (a) dry, gel-less vertically aligned gold nanowire electrodes, (b) dry textile-based conductive sensor electrodes to address the need for this technology to monitor cardiovascular diseases in women are tested with e-nanoflex and discussed. Third, non
Full Text Available Abstract Background Cytomegalovirus (CMV is a prevalent herpesvirus with links to both stress and aging. This paper describes and validates a minimally invasive method for assessing antibodies against CMV in finger stick whole blood spot samples for use as an indirect marker of an aspect of cell-mediated immunity. Results Analysis of CMV in dried blood spot samples (DBS was based on modifications of a commercially available protocol for quantifying CMV antibodies in serum or plasma. The method was evaluated through analysis of precision, reliability, linearity, and correlation between matched serum and DBS samples collected from 75 volunteers. Correlation between DBS and plasma values was linear and high (Pearson correlation R = .96, and precision, reliability, and linearity of the DBS assay were within acceptable ranges. Conclusions The validity of a DBS assay for CMV antibodies will enable its inclusion in population-based surveys and other studies collecting DBS samples in non-clinical settings, increasing scientific understanding of the interaction of social and biological stress and immune function.
Enderle, Yeliz; Meid, Andreas D; Friedrich, Jörg; Grünig, Ekkehard; Wilkens, Heinrike; Haefeli, Walter E; Burhenne, Jürgen
Endothelin receptor antagonists (ERA) and phosphodiesterase 5 inhibitors (PDE5I) are long-term therapeutics for the treatment of pulmonary arterial hypertension (PAH). Their interindividual pharmacokinetic variability is remarkably large, and despite the seriousness of the disease, nonadherence is occurring. Therefore, methods to monitor sufficient circulating drug levels are essential. The objectives of this study were to develop and validate dried blood spot (DBS) assays for the quantification of ambrisentan, bosentan, sildenafil, tadalafil, and their main metabolites. We also quantified the influence of different hematocrit levels and assessed the correlation of simultaneously taken capillary whole blood (DBS) and venous plasma samples. The aliquot punches were extracted by liquid/liquid extraction followed by liquid chromatography/tandem mass spectrometry (LC/MS/MS) quantification methods. All assays fulfilled the requirements of the FDA and EMA guidelines for assay validation with a lower limit of quantification of 2.5 ng/mL for the ERAs, 5 ng/mL for sildenafil, and 10 ng/mL for tadalafil. All analytes were stable for at least 147 days when stored on DBS filter paper cards at room temperature in the dark. Due to poor distribution into erythrocytes, drug concentrations in DBS were always lower than in plasma, resulting in conversion factors of 1.58 for ambrisentan and sildenafil and 1.52 for bosentan and tadalafil. PMID:26583764
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
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
Pieter W Smit
Full Text Available BACKGROUND: Dried blood spots (DBS have been used as alternative specimens to plasma to increase access to HIV viral load (VL monitoring and early infant diagnosis (EID in remote settings. We systematically reviewed evidence on the performance of DBS compared to plasma for VL monitoring and EID. METHODS AND FINDINGS: Thirteen peer reviewed HIV VL publications and five HIV EID papers were included. Depending on the technology and the viral load distribution in the study population, the percentage of DBS samples that are within 0.5 log of VL in plasma ranged from 52-100%. Because the input sample volume is much smaller in a blood spot, there is a risk of false negatives with DBS. Sensitivity of DBS VL was found to be 78-100% compared to plasma at VL below 1000 copies/ml, but this increased to 100% at a threshold of 5000 copies/ml. Unlike a plasma VL test which measures only cell free HIV RNA, a DBS VL also measures proviral DNA as well as cell-associated RNA, potentially leading to false positive results when using DBS. The systematic review showed that specificity was close to 100% at DBS VL above 5000 copies/ml, and this threshold would be the most reliable for predicting true virologic failure using DBS. For early infant diagnosis, DBS has a sensitivity of 100% compared to fresh whole blood or plasma in all studies. CONCLUSIONS: Although limited data are available for EID, DBS offer a highly sensitive and specific sampling strategy to make viral load monitoring and early infant diagnosis more accessible in remote settings. A standardized approach for sampling, storing, and processing DBS samples would be essential to allow successful implementation. TRIAL REGISTRATION: PROSPERO Registration #: CRD42013003621.
Pushpa,; Raghunath; Nimisha
INTRODUCTION: Surveillance for risk factors of heart diseases, like increased blood sugar, cholesterol becomes difficult in places with inadequate lab facilities due to difficulty in sample collection, transportation and processing. Feasibility of using dried blood/seru m spots in such situations can be thought of as an alternative to fresh serum total cholesterol, as sample collection does not require much expertise. AIM: The aim of this study is to determine whether ...
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Koster, Remco A.; Alffenaar, Jan-Willem C.; Greijdanus, Ben; Uges, Donald R. A.
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
Metherel, Adam H; Hogg, Ryan C; Buzikievich, Lindy M; Stark, Ken D.
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 ...
This experiment was carried out under hot environmental conditions, where temperature-humidity index was equivalent to 86 - 90 and 78 - 80 during day and night, respectively. Twelve bovine Baladi calves of 8 - 10 months old and 112 kg average initial live body weight were used in this study. The calves were divided into two groups of 6 animals each to study the effect of supplementation of dried whey milk on some blood biochemical and immunological indices and growth performance of calves under hot weather conditions of Egypt. The results showed that supplementation of dried whey milk to the diet of heat-stressed calves at the level of 150 g / calf / day reduced significantly each of respiration rate and rectal temperature as well as serum lipid concentrations and their fractions e.g. total cholesterol and phospholipids. Also, dried whey milk supplement caused a significant decline in both AST and ALT activities and reduced significantly alpha globulin concentration, while non-significant changes were observed in each of beta globulin, gamma globulin and immunoglobulin G. However, supplementing dried whey milk to growing calves increased significantly serum concentrations of total protein, albumin, calcium, phosphorous, T3 and T4 . Moreover, dried whey milk improved significantly both feed efficiency and daily gain of growing calves. It could be concluded that addition of dried whey milk to the diet reduced rectal temperature and respiration rate and induced an improvement in most blood biochemical parameters and growth performance of heat-stressed bovine Baladi calves.
Objectives The purpose of this study was to investigate whether blood mercury concentrations associated with the presence of dry eye symptoms in a nationally representative Korean population. Methods Population-based prospective cross-sectional study using the heavy metal data set of the 2010–2012 Korean National Health and Nutrition Examination Survey (KNHANES). A total of 4761 adult Koreans were the eligible population in this study. Of the 7162 survey participants, 2401 were excluded because they were mercury levels were measured on the day the participants completed a questionnaire regarding the presence of dry eye symptoms (persistent dryness or eye irritation). The population was divided into low and high groups by median level (4.26 and 2.89 µg/L for males and females, respectively). Results Self-reported dry eye symptoms were present in 13.0% of the cohort. Participants with dry eye symptoms were significantly more likely to have blood mercury levels exceeding the median than those without dry eye symptoms (45.7% vs 51.7%, p=0.021). Logistic regression analysis showed that, after adjusting for age, gender, education, total household income, smoking status, heavy alcohol use, sleep time, perceived stress status, total cholesterol levels and atopy history, dry eye symptoms were significantly associated with blood mercury levels that exceeded the median (reference: lower mercury group; OR, 1.324; 95% CI 1.059 to 1.655; pmercury levels were associated with dry eye symptoms in a nationally representative Korean population. PMID:27121705
Douglas, Conor M.W.; van El, Carla G.; Faulkner, Alex; Cornel, Martina C.
A series of governance issues currently surrounds the multiple uses and multiple users of dried blood spots (DBS) for research purposes. Internationally there is a discussion on storing DBS resulting from newborn screening for public health and using them as the basis for large biobank-like collections to facilitate biomedical research. If such a transformation were to be formalized, then DBS would sit at the intersection of care (ie, public health) and research, with the mechanisms through which such a collection could be managed not totally self-evident. What is more, a DBS collection raises questions about the fuzzy boundaries between privacy and anonymity; how to control or define quality control uses of DBS; medical vs nonmedical uses; as well as benefit sharing and stakeholder involvement. Our goal here is to explore some of the key questions relating to DBS governance by way of the bio-objects and bio-objectification concepts. By embracing – rather than resisting to – the blurring of boundaries and problems in categorization that have come to characterize bio-objects and bio-objectification processes recently described in this journal, we attempt to highlight some issues that might not be currently considered, and to point to some possible directions to go (or avoid). Building from our knowledge of the current DBS situation in the Netherlands, we outline questions concerning the uses, management, collection, and storage of DBS. PMID:22911534
Zheng, Yajun; Wang, Qian; Wang, Xiaoting; Chen, Ying; Wang, Xuan; Zhang, Xiaoling; Bai, Zongquan; Han, Xiaoxiao; Zhang, Zhiping
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
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
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
Kong, Sing Teang; Lim, Shih-Hui; Lee, Wee Beng; Kumar, Pasikanthi Kishore; Wang, Hwee Yi Stella; Ng, Yan Lam Shannon; Wong, Pei Shieen; Ho, Paul C.
To facilitate therapeutic monitoring of antiepileptic drugs (AEDs) by healthcare professionals for patients with epilepsy (PWE), we applied a GC-MS assay to measure three AEDs: carbamazepine (CBZ), phenytoin (PHT) and valproic acid (VPA) levels concurrently in one dried blood spot (DBS), and validated the DBS-measured levels to their plasma levels. 169 PWE on either mono- or polytherapy of CBZ, PHT or/and VPA were included. One DBS, containing ∼15 µL of blood, was acquired for the simultaneous measurement of the drug levels using GC-MS. Simple Deming regressions were performed to correlate the DBS levels with the plasma levels determined by the conventional immunoturbimetric assay in clinical practice. Statistical analyses of the results were done using MedCalc Version 22.214.171.124 and SPSS 21. DBS concentrations (Cdbs) were well-correlated to the plasma concentrations (Cplasma): r = 0.8381, 0.9305 and 0.8531 for CBZ, PHT and VPA respectively, The conversion formulas from Cdbs to plasma concentrations were [0.89×CdbsCBZ+1.00]µg/mL, [1.11×CdbsPHT−1.00]µg/mL and [0.92×CdbsVPA+12.48]µg/mL respectively. Inclusion of the red blood cells (RBC)/plasma partition ratio (K) and the individual hematocrit levels in the estimation of the theoretical Cplasma from Cdbs of PHT and VPA further improved the identity between the observed and the estimated theoretical Cplasma. Bland-Altman plots indicated that the theoretical and observed Cplasma of PHT and VPA agreed well, and >93.0% of concentrations was within 95% CI (±2SD); and similar agreement (1∶1) was also found between the observed Cdbs and Cplasma of CBZ. As the Cplasma of CBZ, PHT and VPA can be accurately estimated from their Cdbs, DBS can therefore be used for drug monitoring in PWE on any of these AEDs. PMID:25255292
In France neonatal screening for hypothyroidism is performed by mean of radioimmunoassay for TSH in dried blood spots, collected on the 5th day of life. In order to be able to get more data from the day 5 blood sample and thus confirm congenital hypothyroidism in all cases when elevated TSH levels have been found, we developed a simple radioimmunoassay for free T4 in eluates of dried blood spots on filter paper. We used reagents from the Amerlex FT4 RIA kit. The ready for use supplied anti-T4 antibody suspension and 125I-T4 derivative solution are respectively 2-fold and 3-fold diluted in 0,154 M phosphate saline buffer-pH 7,4. Whole blood free T4 standards are prepared by adding serum standards in the kit to an equal volume of washed and packed red blood cells. Free T4 concentrations in these dried standards are expressed as picomoles per liter of whole blood. One blood spot 4.25 mm in diameter, equivalent to about 5-6 μl test blood or standard is incubated for 1 hour at room temperature in 500 μl of antibody suspension. The assay sensitivity (smallest detectable FT4 level) is 1.4 pmol.l-1. Within assay and between assays coefficients of variation are respectively 9,5 and 13%. Free T4 levels measured in eluates of dried blood closely correlate (r=0,946) with plasma levels in the same subjects. Free T4 in 690 five day old neonates shows a non gaussian distribution, with a mean value: 13,6 pmol.l-1 and standard deviation: 4,3 (range 3,6-30 pmol.l-1). In 13 newborns with congenital hypothyroidism, day 5 free T4 levels were found within the range: 0 to 3,5 pmol.l-1, and thus did not overlap with free T4 measured in age-matched euthyroid neonates
Travert, G.; Lemonnier, F.; Fernandez, Y. (C.H.R.U. de Caen, Service de Biophysique-Radioisotopes, 14 - Caen (France))
In France neonatal screening for hypothyroidism is performed by mean of radioimmunoassay for TSH in dried blood spots, collected on the 5th day of life. In order to be able to get more data from the day 5 blood sample and thus confirm congenital hypothyroidism in all cases when elevated TSH levels have been found, we developed a simple radioimmunoassay for free T4 in eluates of dried blood spots on filter paper. We used reagents from the Amerlex FT4 RIA kit. The ready for use supplied anti-T4 antibody suspension and /sup 125/I-T4 derivative solution are respectively 2-fold and 3-fold diluted in 0,154 M phosphate saline buffer-pH 7,4. Whole blood free T4 standards are prepared by adding serum standards in the kit to an equal volume of washed and packed red blood cells. Free T4 concentrations in these dried standards are expressed as picomoles per liter of whole blood. One blood spot 4.25 mm in diameter, equivalent to about 5-6 ..mu..l test blood or standard is incubated for 1 hour at room temperature in 500 ..mu..l of antibody suspension. The assay sensitivity (smallest detectable FT4 level) is 1.4 pmol.l/sup -1/. Within assay and between assays coefficients of variation are respectively 9,5 and 13%. Free T4 levels measured in eluates of dried blood closely correlate (r=0,946) with plasma levels in the same subjects. Free T4 in 690 five day old neonates shows a non gaussian distribution, with a mean value: 13,6 pmol.l/sup -1/ and standard deviation: 4,3 (range 3,6-30 pmol.l/sup -1/). In 13 newborns with congenital hypothyroidism, day 5 free T4 levels were found within the range: 0 to 3,5 pmol.l/sup -1/, and thus did not overlap with free T4 measured in age-matched euthyroid neonates.
During July 1986-June 1987, blood samples were collected onto filter papers (Type 8402) from neonates born in 21 hospitals in Tianjin City. The TSH concentration in umbilical cord blood and T4 concentration in heel blood during the period of 4∼7 postnatal days were measured. Dried blood on discs of 9.5 mm and 6.2 mm in diameters were used for TSH, T4 RIA respectively. The neonates with low T4 (20 mU/L) were recalled for further investigation. A total of 46425 neonates were screened in one year, the screening rate being 90.8%. Congenital hypothyroidism (CH) was confirmed in 6 neonates, with an incidence of 1 in 7843.5 of the 6 were primary CH while the other one was secondary CH. Thyroid tablets administration showed significant improvement
Full Text Available Aim: The aim was to determine the chemical composition of solar dried blood and rumen content (DBRC and further ascertain the concentration at which DBRC could be included in Japanese quail diets without any adverse effect on its performance. Materials and Methods: Feeding trial on the effect of DBRC on performance of Japanese quails was studied up to 5 weeks. 252 numbers of day old (Nandanam Type III breed Japanese quails were purchased from Poultry Research Station, Madhavaram and divided into 7 batches (control+ six treatments each consisting of 36 birds. The DBRC was included at 0%, 5%, 10%, 15%, 20%, 25% and 30% in diets as control, treatment-1 (T1, treatment-2 (T2, treatment-3 (T3, treatment-4 (T4, treatment-5 (T5 and treatment-6 (T6 respectively in a completely randomized design to replace soybean meal in Japanese quail feed. The birds were provided with ad-labidum feed and drinking water ad-libitum during the entire experimental period. Results: The crude protein (CP, crude fiber (CF, ether extract (EE and ash contents of DBRC were 35.87%, 17.40%, 3.6% and 12.6%, respectively. The amount of essential amino acids and non-essential amino acid content were found to be 12.98 and 4.87 (g/100 g of feed respectively in DBRC feed. Result showed that all birds fed DBRC diets performed better than the control group. Mortality was unaffected by dietary treatments. There was a significant difference (p<0.01 observed in weight gain in treatment groups compared to the control. Conclusion: Up to 30% DBRC could be incorporated in the diets of Japanese quails without any adverse effects on its performance.
Soetens, Oriane; Vauloup-Fellous, Christelle; Foulon, Ina; Dubreuil, Pascal; De Saeger, Ben; Grangeot-Keros, Liliane; Naessens, Anne
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
Mitchell, Caroline; Kraft, Kelli; Peterson, Do; Frenkel, Lisa
Dried blood spot (DBS) samples are a convenient way to collect infant blood for HIV-1 diagnostic testing. Minimizing the risk of false positives is critical for diagnostic tests. A protocol for processing and testing DBS for infant HIV-1 diagnosis was evaluated to identify the rate and source of false positive results. DBS were created on Flinders Technology Associates (FTA) filter paper with 500 copies/punch (high) or 5000 copies/punch (very high) concentrations of HIV-1 DNA. Blank discs of ...
Bank, Steffen; Nexø, Bjørn Andersen; Andersen, Vibeke;
Background: The recovery of biological samples for genetic epidemiological studies can be cumbersome. Blood clots are routinely collected for serological examinations. However, the extraction of DNA from blood clots can be difficult and often results in low yields. Aim: The aim was to compare the...
Wondafrash, Beyene; Hiko, Desta
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...
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.
The experiment was designed to study the effect of supplemented dried live yeast (DLY) + vitamin E to the diet of growing calves under hot summer conditions in Egypt. Six bovine Baladi calves with 115 kg initial body weight and 8-10 months old were used during two periods. In the first period, the calves were offered the concentrated basal diet only for one month and considered as a control period. In the second period, the calves were fed the same basal diet which supplemented with 15 g dried live yeast (Saccharomyces cerevisiae) + 600 IU vitamin E (alpha- tocopherol) per calf daily for one month and considered as a treated period. Body weight was recorded at the beginning and the end of each period, and daily gain was calculated for each animal. Blood samples were collected from each animal at the end of each period to determine some blood biochemical parameters and T3 and T4 concentrations as well as some immunological indices.The results showed that supplementation of DLY + 600 IU vitamin E to the diet of calves reduced significantly (P3 and T4 levels and improved feed efficiency and daily gain. It is concluded that supplementation of growing calves with 15 g DLY + 600 IU vitamin E / calf / day under Egyptian hot summer conditions reduced the effect of heat stress as shown by a decline in RT and modified most blood constituents and thyroid function which leads to an improvement in growing calves
Teaching slides on: What is a digital archive? Parties involved in digital archiving Archiving challenges organization of data coherence and persistency access and safety Language archiving software Different users, different needs For: Saami Language Documentation and Revitalization. Winter school, Bodø, Norway
Sørensen, Karina M; Agergaard, Peter; Olesen, Charlotte; Andersen, Paal S; Larsen, Lars A; Ostergaard, John R; Schouten, Jan P; Christiansen, Michael
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...
Murai, Kenji; Culleton, Richard; Hisaoka, Teruhiko; Endo, Hiroyoshi; Mita, Toshihiro
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. PMID:25449286
G.P. Hooff (Gero); R.J.W. Meesters (Roland); J.J.A. van Kampen (Jeroen); N.A. van Huizen (Nick); B. Koch (Birgit); A.F.Y. Al Hadithy (Asmar); T. van Gelder (Teun); A.D.M.E. Osterhaus (Albert); R.A. Gruters (Rob); T.M. Luider (Theo)
textabstractThe neuraminidase inhibitor oseltamivir (Tamiflu®) is currently the first-line therapy for patients with influenza virus infection. Common analysis of the prodrug and its active metabolite oseltamivircarboxylate is determined via extraction from plasma. Compared with these assays, dried
Full Text Available The aim of this study was to evaluate the glucogenic property of glycerol supplementation in the dairy cow’s diet. Sixty primiparous cows (control, n=30, and glycerol supplemented, n=30 were used to measure milk yield and components, blood hormone and metabolite profiles, and body condition score. Feed intake and apparent total-tract digestibility were also measured using 10 primiparous cows (control, n=5, and glycerol supplemented, n=5. Dry glycerol was top dressed at 250 g/day/cow from parturition to 21 days postpartum. Average feed intake, milk yield and components were not affected by glycerol supplementation. Apparent total–tract digestibility of organic matter and neutral detergent fibre were not influenced by dry glycerol supplementation, but lipid digestibility was greater (p=0.01 in cows fed glycerol. The serum concentration of glucose and insulin tended to be higher in dry glycerol-supplemented cows (p=0.1; p=0.06, respectively. While, serum concentrations of nonesterified fatty acids and β-hydroxybutyrate were not affected. Supplemented cows had lower body condition loss during weeks 1 to 5 after calving (p=0.09. The glucogenic effect of glycerol did not affect milk yield during the first 3 weeks of lactation. However, daily milk yield during the 13 weeks recording period was higher in the glycerol-supplemented cows (28.5 vs. 30.3 kg, p<0.001. Percentages of cows cycling at the planned breeding date was greater (p=0.01 for cows fed dry glycerol. The results demonstrated that feeding dry glycerol as a glucogenic supply could be useful in saving body reserves and improving energy balance of primiparous Holstein dairy cows during the early postpartum period.
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.
Lepczynski, A; Herosimczyk, A; Ozgo, M; Skomial, J; Taciak, M; Barszcz, M; Berezecka, N
The aim of the study was to characterize the systemic immune and metabolic alterations in the blood serum of growing pigs in response to a dietary supplementation with 4% of dried chicory roots. This was achieved by examining the influence of the experimental diet on serum protein changes especially these related with immunology and lipid metabolism. Serum proteins with the isoelectric point ranging from pH 3.0 to 10.0 were separated using high resolution two-dimensional electrophoresis. As a result, we found that experimental diet triggered significant changes in 37 protein spots. Of these, 14 were up-regulated, whereas 23 showed down-regulation. Of 37 significantly altered protein spots, 24 were successfully identified, representing 14 distinct gene products. Implementation of the dried chicory roots into the diet of growing pigs caused a significant down-regulation of apolipoprotein C-II complement component C6, C-reactive protein, CD14 antigen, C4b binding protein α and β chains, and fibrinogen. Piglets fed experimental diet had similar IgA, IgG and IgM concentrations, although the level of IgM tended to be lower compared to the control group. It is concluded that diet supplemented with 4% of dried chicory root may exert anti-inflammatory properties and affect lipid metabolism in growing pigs. PMID:25716964
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...
la Marca, Giancarlo; Malvagia, Sabrina; Materazzi, Serena; Della Bona, Maria Luisa; Boenzi, Sara; Martinelli, Diego; Dionisi-Vici, Carlo
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
van Thiel, D.; Marschner, I.; Wood, W.G.; Habermann, J.; Scriba, P.C.
The blood-spot thyrotropin RIA for detection of congenital hypothyroidism has been established as a screening programme. This article describes the problems in hand, namely optimisation of the method and a comparison of the performance in 7 commercial kits on the West German market. The following factors have been investigated: a. Shelf-life of standards and samples, b. Effect of filter paper quality and weight, c. Size of paper disc, d. Blood sampling time, e. Time of removal of paper disc, f. Effect on the results of washing the precipitate. The commercial kits at present on the market differ widely in method and price. Apart from this, there was no agreement between the results obtained, and the difference between the standard concentration given by the firm compared with the reference standard MRC 68/38 varied from 22-185%. Methodological guidelines and quality control measures, both internal and external, are suggested.
Brindle, Eleanor; FUJITA, MASAKO; Shofer, Jane; O’Connor, Kathleen A.
C-reactive protein (CRP) is used as a biomarker of morbidity and mortality risk in studies of population health, and is essential to interpretation of several micronutrient biomarkers. There is thus need for a robust high sensitivity CRP (hsCRP) measurement method for large-scale, non-clinical studies. We developed an efficient, inexpensive assay suitable for quantifying CRP across the physiological range using any blood specimen type. The ELISA uses readily available monoclonal antibodies to...
Zhou, Zhenming; Zhou, Bo; Ren, Liping; Meng, Qingxiang
Fifty-one Simmental crossbred steers (357.0 ± 16.5 kg) were used to compare a standard total mix ration (TMR) with variants on animal performance, ruminal fermentation, blood biochemical parameters, and carcass characteristics. Corn grain and cotton seed meal were partially replaced by ensiled mulberry leaves (EML) or sun-dried mulberry fruit pomace (SMFP). Experimental diets had similar amounts of crude protein (CP), acid detergent fiber (ADF), and metabolizable energy (ME). Animals were divided into three groups: control group (CONT), 8% EML group, and 6.3% SMFP group. Performance, including average daily weight gain (ADG), and dry matter intake (DMI), was measured. Blood and rumen samples were collected at the end of the experiment (16 weeks). There were no differences in final body weight (P = 0.743), ADG (P = 0.425), DMI (P = 0.642), or ADG/DMI (P = 0.236) between the groups. There were no differences (P = 0.2024) in rumen pH values; ammonia N was lower (P = 0.0076) in SMFP than in the EML and CONT groups. There were differences in the concentrations of total and individual volatile fatty acids, while no differences were determined in blood biochemical parameters (i.e., plasma glucose, urea concentrations, triglycerides, total protein, insulin, IgG, alanine transaminase, and aspartate aminotransferase, P ≥ 0.098). No differences were observed in carcass characteristics (P ≥ 0.513), tenderness (P = 0.844), adipose and lean color values (P ≥ 0.149), and chemical composition (P ≥ 0.400); however, intramuscular fat was lower in the EML and SMFP groups compared to the CONT animals (P = 0.034). In conclusion, diets supplemented with these two mulberry products in an isocaloric and isonitrogenous manner have similar effects to corn grain and cotton seed meals on steer performance, blood biochemical parameters and carcass characteristics, with the exception of ruminal VFA concentrations and lower intramuscular fat content. PMID:24427304
Full Text Available Fifty-one Simmental crossbred steers (357.0 ± 16.5 kg were used to compare a standard total mix ration (TMR with variants on animal performance, ruminal fermentation, blood biochemical parameters, and carcass characteristics. Corn grain and cotton seed meal were partially replaced by ensiled mulberry leaves (EML or sun-dried mulberry fruit pomace (SMFP. Experimental diets had similar amounts of crude protein (CP, acid detergent fiber (ADF, and metabolizable energy (ME. Animals were divided into three groups: control group (CONT, 8% EML group, and 6.3% SMFP group. Performance, including average daily weight gain (ADG, and dry matter intake (DMI, was measured. Blood and rumen samples were collected at the end of the experiment (16 weeks. There were no differences in final body weight (P = 0.743, ADG (P = 0.425, DMI (P = 0.642, or ADG/DMI (P = 0.236 between the groups. There were no differences (P = 0.2024 in rumen pH values; ammonia N was lower (P = 0.0076 in SMFP than in the EML and CONT groups. There were differences in the concentrations of total and individual volatile fatty acids, while no differences were determined in blood biochemical parameters (i.e., plasma glucose, urea concentrations, triglycerides, total protein, insulin, IgG, alanine transaminase, and aspartate aminotransferase, P ≥ 0.098. No differences were observed in carcass characteristics (P ≥ 0.513, tenderness (P = 0.844, adipose and lean color values (P ≥ 0.149, and chemical composition (P ≥ 0.400; however, intramuscular fat was lower in the EML and SMFP groups compared to the CONT animals (P = 0.034. In conclusion, diets supplemented with these two mulberry products in an isocaloric and isonitrogenous manner have similar effects to corn grain and cotton seed meals on steer performance, blood biochemical parameters and carcass characteristics, with the exception of ruminal VFA concentrations and lower intramuscular fat content.
The Gaia archive is being designed and implemented by the DPAC Consortium. The purpose of the archive is to maximize the scientific exploitation of the Gaia data by the astronomical community. Thus, it is crucial to gather and discuss with the community the features of the Gaia archive as much as possible. It is especially important from the point of view of the GENIUS project to gather the feedback and potential use cases for the archive. This paper presents very briefly the general ideas behind the Gaia archive and presents which tools are already provided to the community.
Ditrich, H; Splechtna, H
Microvascular corrosion casts of chicken embryos between four and 19 days after fertilization have been prepared. The developing kidney was investigated with scanning electron microscopy (SEM). The injection technique and resin composition were modified in order to facilitate the complete replication of native blood vascular systems of specimens as small as 15 mm body length. The development of the opisthonephros was followed from near the beginning of its function until a vascular development comparable to the adult situation was reached. Critical point dried glomeruli show the differentiation of the glomerular visceral epithelium (podocytes) from initially epithelioid to highly branched forms. The embryonic kidney (cranial part of the opisthonephros-mesonephros) shows a construction-principle resembling amphibians that is entirely different from the definitive excretory organ (caudal part of the opisthonephros-metanephros). PMID:2814402
This book treats the micro archiving of websites, i.e. archiving by researchers, students or others without special technical knowledge who, using a standard computer, wish to save a website for further study. The phenomenon is discussed from the standpoint that Internet research must be able...... is reached. Firstly, unlike other well-known media, the Internet does not simply exist in a form suited to being archived, but rather is first formed as an object of study in the archiving, and it is formed differently depending on who does the archiving, when, and for what purpose. Secondly, this means...... that there is an element of subjective creation in the archived material, so that methodical deliberations are necessary — in other words, the answers to why and how the archived material has been created. These conclusions form the starting point for the last section of the book, which, based on comprehensive tests...
Meister, Isabel; Leonidova, Anna; Kovač, Jana; Duthaler, Urs; Keiser, Jennifer; Huwyler, Jörg
Praziquantel (PZQ) is the treatment of choice against various trematode and cestode infections. To study the pharmacokinetics of PZQ in patients infected with the liver fluke Opisthorchis viverrini, we developed and validated an enantioselective liquid chromatography coupled to tandem mass spectrometry method for the analysis of R - and S -PZQ and its R -trans-4-OH-PZQ metabolite in human plasma, blood and dried blood spots (DBS). The analytes were detected in the positive mode using selected reaction monitoring (R- and S-PZQ: m/z 312.2 → 202.2; R-trans -4-OH-PZQ: m/z 328.0 → 202.0). Prior to the chiral separation with a cellulose tris(3-chloro-4-methylphenylcarbamate) column, the analytes were purified from matrix contaminants and concentrated on a C-18 trapping column. The analytical range for each PZQ enantiomer was 0.01-2.5 μg/mL, and 0.1-25 μg/mL for the metabolite. The method met the requirements regarding precision (± 15%, ± 20% at the lower limit of quantification-LLOQ), intra- and inter-assay accuracy (85-115%, 80-120% at LLOQ), and linearity (R(2) ≥ 0.998). The analytes were stable in stock solutions as well as in plasma, blood and DBS. For DBS, the influences of hematocrit and blood spot size were considered as minor. Our validation results show that the method presented here is precise, accurate and selective, and can be used for pharmacokinetic studies. Moreover, the enantioselective separation was achieved with a run time of 11.5 min and a simple sample processing method. PMID:26517852
National Archives and Records Administration — Archival Descriptions from the National Archives Catalog data set provides archival descriptions of the permanent holdings of the federal government in the custody...
Full Text Available Background: HIV has claimed millions of lives with the Sub-Saharan Africa being the most affected. There is a significant increase in access to antiretroviral drugs which also demands frequent monitoring to determine the drug effectiveness and efficacy. Thus there is a great need to evaluate simplified methods to monitor treatment with such antiretroviral drugs. Use of dried blood spots (DBS can be ideal if evaluated in resource limited countries such as Malawi since they are easy to collect, store and convenient. The main objective of this study was to evaluate the accuracy of a dry blood spot sample in the quantification of viral particles in HIV reactive patients using the Abbott m2000rt assay. Methods: 87 participants were recruited from the ART clinic at Queen Elizabeth Central Hospital using convenience sampling method. 29 were on antiretroviral therapy and 58 had not started the therapy. HIV-1 RNA extraction and quantification was performed from DBS and plasma using Abbott m2000sp and m2000rt systems respectively. The results were statistically analyzed by Bland-Altman method using medcalc software version 12.6.1. Results: 66 paired samples with detectable viral loads were analysed. These gave a correlation of 0.98. The mean difference was 0.05 log10 copies/ml with a standard deviation of 0.17 at 95% confidence interval.The Bland-Altman plots showed limits of agreement which ranged from -0.38 to 0.28 log10 copies/ml at 95% confidence interval. Conclusion: Results showed strong agreement between the plasma and DBS samples. A slight and clinically insignificant difference was observed between the two methods. A larger sample size can give support to the study findings. Since samples were less than a week old, it is not known if the results would be different if they were to be stored for a longer period. [Int J Res Med Sci 2013; 1(4.000: 338-342
Lee, Kyunghoon; Jun, Sun Hee; Han, Minje; Song, Sang Hoon; Park, Jong Sun; Lee, Jae Ho; Park, Kyoung Un; Song, Junghan
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
Meister, Isabel; Kovac, Jana; Duthaler, Urs; Odermatt, Peter; Huwyler, Jörg; Vanobberghen, Fiona; Sayasone, Somphou; Keiser, Jennifer
Background Praziquantel (PZQ) is the treatment of choice for infections with the liver fluke Opisthorchis viverrini, a major health problem in Southeast Asia. However, pharmacokinetic (PK) studies investigating the disposition of PZQ enantiomers (R- and S-PZQ) and its main metabolite, R-trans-4-OH-PZQ, in diseased patients are lacking. The implementation of a dried blood spot (DBS) sampling technique would ease the performance of PK studies in remote areas without clinical facilities. The aim of the present study is to provide data on the disposition of PZQ enantiomers and R-trans-4-OH-PZQ in opisthorchiasis patients and to validate the use of DBS compared to plasma and blood sampling. Methodology/Principal Findings PZQ was administered to nine O. viverrini-infected patients at 3 oral doses of 25 mg/kg in 4 h intervals. Plasma, blood and DBS were simultaneously collected at selected time points from 0 to 24 h post-treatment. PK parameters were determined using non-compartmental analysis. Drug concentrations and areas under the curve (AUC0–24h) measured in the 3 matrices were compared using Bland-Altman analysis. We observed plasma AUC0–24hs of 1.1, 9.0 and 188.7 μg/ml*h and half-lives of 1.1, 3.3 and 6.4 h for R-PZQ, S-PZQ and R-trans-4-OH, respectively. Maximal plasma concentrations (Cmax) of 0.2, 0.9 and 13.9 μg/ml for R-PZQ, S-PQZ and R-trans-4-OH peaked at 7 h for PZQ enantiomers and at 8.7 h for the metabolite. Individual drug concentration measurements and patient AUC0–24hs displayed ratios of blood or DBS versus plasma between 79–94% for R- and S-PZQ, and between 108–122% for R-trans-4-OH. Conclusions/Significance Pharmacodynamic (PD) in vitro studies on PZQ enantiomers and R-trans-4-OH-PZQ are necessary to be able to correlate PK parameters with efficacy. DBS appears to be a valid alternative to conventional venous sampling for PK studies in PZQ-treated patients. PMID:27152952
Full Text Available Praziquantel (PZQ is the treatment of choice for infections with the liver fluke Opisthorchis viverrini, a major health problem in Southeast Asia. However, pharmacokinetic (PK studies investigating the disposition of PZQ enantiomers (R- and S-PZQ and its main metabolite, R-trans-4-OH-PZQ, in diseased patients are lacking. The implementation of a dried blood spot (DBS sampling technique would ease the performance of PK studies in remote areas without clinical facilities. The aim of the present study is to provide data on the disposition of PZQ enantiomers and R-trans-4-OH-PZQ in opisthorchiasis patients and to validate the use of DBS compared to plasma and blood sampling.PZQ was administered to nine O. viverrini-infected patients at 3 oral doses of 25 mg/kg in 4 h intervals. Plasma, blood and DBS were simultaneously collected at selected time points from 0 to 24 h post-treatment. PK parameters were determined using non-compartmental analysis. Drug concentrations and areas under the curve (AUC0-24h measured in the 3 matrices were compared using Bland-Altman analysis. We observed plasma AUC0-24hs of 1.1, 9.0 and 188.7 μg/ml*h and half-lives of 1.1, 3.3 and 6.4 h for R-PZQ, S-PZQ and R-trans-4-OH, respectively. Maximal plasma concentrations (Cmax of 0.2, 0.9 and 13.9 μg/ml for R-PZQ, S-PQZ and R-trans-4-OH peaked at 7 h for PZQ enantiomers and at 8.7 h for the metabolite. Individual drug concentration measurements and patient AUC0-24hs displayed ratios of blood or DBS versus plasma between 79-94% for R- and S-PZQ, and between 108-122% for R-trans-4-OH.Pharmacodynamic (PD in vitro studies on PZQ enantiomers and R-trans-4-OH-PZQ are necessary to be able to correlate PK parameters with efficacy. DBS appears to be a valid alternative to conventional venous sampling for PK studies in PZQ-treated patients.
Vidya, Madhavan; Saravanan, Shanmugam; Rifkin, Samara; Solomon, Sunil S; Waldrop, Greer; Mayer, Kenneth H; Solomon, Suniti; Balakrishnan, Pachamuthu
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
Simonavice, Emily; Liu, Pei-Yang; Ilich, Jasminka Z; Kim, Jeong-Su; Arjmandi, Bahram; Panton, Lynn B
The purpose of this study was to examine the effects of resistance training (RT) and dried plum (DP) consumption on strength, body composition, blood markers of bone, and inflammation in breast cancer survivors (BCS). Twenty-three BCS (RT, n = 12; RT+DP, n = 11), aged 64 ± 7 years, were evaluated at baseline and after 6 months of intervention on the following: muscular strength (chest press and leg extension) via 1-repetition maximums (1RMs); body composition, specifically bone mineral density (BMD) by dual energy X-ray absorptiometry; biochemical markers of bone turnover (bone-specific alkaline phosphatase (BAP), tartrate resistant acid phosphatase (TRAP-5b)); and inflammation (C-reactive protein (CRP)). Target RT prescription was 2 days/week of 10 exercises, including 2 sets of 8-12 repetitions at ∼60%-80% of 1RM. RT+DP also consumed 90 g of DP daily. There were no baseline differences between groups or any group-by-time interactions for any of the variables. BCS increased upper (p effective for improving biochemical markers of bone turnover and muscular strength in BCS. A longer and higher intensity intervention may be needed to reveal the true effects of RT and DP on body composition and biochemical markers of inflammation. PMID:24869977
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.
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
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
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
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.
Full Text Available ... Meeting Abstracts 2016 Call for Abstracts 2016 Abstract Review Categories Abstracts Archive View all Education ASH Academy ... Current Issue First Edition Abstracts Blood Advances A peer-reviewed, online only, open access journal with a ...
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.
Full Text Available Background: Poor quality dried blood spot (DBS specimens are usually rejected by virology laboratories, affecting early infant diagnosis of HIV. The practice of combining two incompletely-filled DBS in one specimen preparation tube during pre-analytical specimen processing (i.e., the two-spot method has been implemented to reduce the number of specimens being rejected for insufficient volume.Objectives: This study analysed laboratory data to describe the quality of DBS specimens and the use of the two-spot method over a one-year period, then validated the two-spot method against the standard (one-spot method.Methods: Data on HIV-1 PCR test requests submitted in 2014 to the Department of Virology at Inkosi Albert Luthuli Central Hospital in KwaZulu-Natal province, South Africa were analysed to describe reasons for specimen rejection, as well as results of the two-spot method. The accuracy, lower limit of detection and precision of the two-spot method were assessed.Results: Of the 88 481 specimens received, 3.7% were rejected for pre-analytical problems. Of those, 48.9% were rejected as a result of insufficient specimen volume. Two health facilities had significantly more specimen rejections than other facilities. The two-spot method prevented 10 504 specimen rejections. The Pearson correlation coefficient comparing the standard to the two-spot method was 0.997.Conclusions: The two-spot method was comparable with the standard method of pre-analytical specimen processing. Two health facilities were identified for targeted retraining on specimen quality. The two-spot method of DBS specimen processing can be used as an adjunct to retraining, to reduce the number of specimens rejected and improve linkage to care.
Van Berkel, Gary J [ORNL; Kertesz, Vilmos [ORNL
The utility of a liquid extraction based sealing surface sampling probe (SSSP) for the direct mass spectrometric analysis of targeted drugs and metabolites in dried blood spots (DBSs) and whole mouse thin tissue sections was demonstrated. The accuracy and precision for the quantitative analysis of a minimum of 50 ng/mL sitamaquine or acetaminophen in DBSs on paper were well within the required 15% dictated by internationally recognized acceptance criteria for assay validations. Analysis of whole-body mouse thin tissue sections from animals dosed with propranolol, adhered to an adhesive tape substrate, provided semi-quantitative information for propranolol and its hydroxyproranolol glucuronide metabolite within specific organs of the tissue. The relative abundances recorded for the two compounds in the brain, lung, kidney and liver were in nominal agreement with previously reported amounts based on analysis using a liquid microjunction surface sampling probe (LMJ-SSP), and whole-body autoradiography (WBA) and HPLC-MS analysis. The ability to sample and analyze from tape-adhered tissue samples, which are generally employed in WBA analysis, presents the possibility of consecutive WBA and SSSP-MS analysis of the same tissue section. This would facilitate assignment, and possibly quantitation, of the different molecular forms of total drug related material detected in the WBA analysis. The flexibility to sample larger or smaller spot sizes, alternative probe sealing mechanisms, and a reduction in internal volumes and associated sample carryover issues will be among the first simple improvements necessary to make the SSSP-MS method a practical DBS and/or thin tissue section analysis tool or to expand its use to other surface sampling applications.
Full Text Available Background: There is a paucity of data on the prevalence of hepatitis C virus (HCV in children, particularly in sub-Saharan Africa. A major obstacle in resource-limited settings for polymerase chain reaction (PCR testing is the necessity for specimen transportation and storage at low temperatures. There are numerous recent studies of using real-time HCV PCR for diagnosis and screening of plasma and serum, but few have looked at using dried blood spot (DBS specimens.Objectives: The aim of this study was to optimise a real-time HCV PCR method to detect HCV RNA from infant DBS specimens for use as a tool for HCV surveillance in KwaZulu-Natal, South Africa.Method: The LightCycler® 2.0 instrument was used for the HCV PCR using the LightCycler® RNA Master SYBR Green I kit. Template volume, primer concentration and primer annealing temperatures were optimised and the method was used on 179 DBS specimens from HIV-exposed infants in KwaZulu-Natal.Results: Primer concentrations adjusted to 0.25 µM and a template volume of 10 µL improved the PCR amplification. Primer annealing temperatures lowered from 65 °C to 58 °C resulted in higher quantities of amplified PCR product. The limit of detection of the optimised HCV PCR assay was between 1200 IU/mL and 3580 IU/mL of HCV RNA. HCV was not detected in any of the 179 DBS specimens.Conclusion: The optimised real-time HCV PCR on infant DBS specimens performed well, but HCV was not found in this surveillance study. HIV infection may have little impact on the vertical transmission of HCV in this region.
The 2014/2015 academic year was the first to be taught the Master in Archives, Records Management and Digital Continuity at the University Carlos III Madrid, based on a blended learning model of attendance. This article describes the implementation of a visualization project based on geolocation of Spanish archives and its justification. The project arose from an optional practice proposed to the students of the subject “Re-use of data and documents” of this master course. This subject is opt...
National Archives and Records Administration — The Archival Research Catalog (ARC) data set provides archival descriptions of the permanent holdings of the federal government in the custody of the National...
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
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
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
Mizuta, H.; Miyai, K.; Ichihara, K.; Amino, N.; Harada, T.; Nose, O.; Tanizawa, O.
In this sensitive, simple method for measuring ''free thyroxin'' (FT/sub 4/) in eluates of dried blood spots on filter paper by use of a radioimmunoassay kit (Amerlex Free T/sub 4/ RIA), the measurable range of FT/sub 4/ 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. FT/sub 4/ in blood spotted on filter paper is stable for at least a month when dried and kept at either -20/sup 0/C, 4/sup 0/C, room temperature (about 25/sup 0/C), or 37/sup 0/C. The results for FT/sub 4/ in dried blood spots correlated closely with the free-T/sub 4/ 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.
Heat stress (HT) during the dry period compromises mammary gland development, decreases future milk production, and impairs immune status of dairy cows. Our objective was to evaluate the effect of cooling HT cows during the dry period on gene expression of the mammary gland and lymphocytes. Cows wer...
Dry mouth is the feeling that there is not enough saliva in your mouth. Everyone has a dry mouth once in a while - if they are nervous, ... under stress. But if you have a dry mouth all or most of the time, it can ...
Performing Archives/Archives of Performance contributes to the ongoing critical discussions of performance and its disappearance, of the ephemeral and its reproduction, of archives and mediatized recordings of liveness. The many contributions by excellent scholars and artists from a broad range...... of interdisciplinary fields as well as from various locations in research geographies demonstrate that despite the extensive discourse on the relationship between performance and the archive, inquiry into the productive tensions between ephemerality and permanence is by no means outdated or exhausted. New ways...... of understanding archives, history, and memory emerge and address theories of enactment and intervention, while concepts of performance constantly proliferate and enable a critical focus on archival residue. The contributions in Performing Archives/Archives of Performance cover philosophical inquiries as well...
Kumar, D; Dhiman, S; Rabha, B; Goswami, D; Yadav, K; Deka, M; Veer, V; Baruah, I
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
Effects of feeding a spray-dried multivalent polyclonal antibody preparation on feedlot performance, feeding behavior, carcass characteristics, rumenitis, and blood gas profile of Brangus and Nellore yearling bulls.
Millen, D D; Pacheco, R D L; DiLorenzo, N; Martins, C L; Marino, C T; Bastos, J P S T; Mariani, T M; Barducci, R S; Sarti, L M N; DiCostanzo, A; Rodrigues, P H M; Arrigoni, M D B
The objective of this study was to evaluate the effects of replacing monensin (MON) with a spray-dried multivalent polyclonal antibody preparation (PAP) against several ruminal microorganisms on feedlot performance, carcass characteristics, feeding behavior, blood gas profile, and the rumenitis incidence of Brangus and Nellore yearling bulls. The study was designed as a completely randomized design with a 2 × 2 factorial arrangement, replicated 6 times (4 bulls per pen and a total of 24 pens), in which bulls ( = 48) of each biotype were fed diets containing either MON fed at 300 mg/d or PAP fed at 3 g/d. No significant feed additive main effects were observed for ADG ( = 0.27), G:F ( = 0.28), HCW ( = 0.99), or dressing percentage ( = 0.80). However, bulls receiving PAP had greater DMI ( = 0.02) and larger ( = 0.02) final LM area as well as greater ( Brangus bulls had greater ( Brangus bulls. In addition, Brangus bulls had greater ( Brangus bulls, MON led to greater ( Brangus bulls fed PAP. Feeding a spray-dried PAP led to similar feedlot performance compared with that when feeding MON. Spray-dried PAP might provide a new technology alternative to ionophores. PMID:26440339
Whalen, Lucille; And Others
This 16-article issue focuses on history, policy, services, users, organization, evaluation, and automation of the archival reference process. Collections at academic research libraries, a technical university, Board of Education, business archives, a bank, labor and urban archives, a manuscript repository, religious archives, and regional history…
Cook, Michael Garnet
Archives and the Computer deals with the use of the computer and its systems and programs in archiving data and other related materials. The book covers topics such as the scope of automated systems in archives; systems for records management, archival description, and retrieval; and machine-readable archives. The book also features examples of systems for records management from different institutions such as theTyne and Wear Archive Department, Dyfed Record Office, and the University of Liverpool. Included in the last part are appendices. Appendix A is a directory of archival systems, Appen
Cook, Michael Garnet
Archives and the Computer deals with the use of the computer and its systems and programs in archiving data and other related materials. The book covers topics such as the scope of automated systems in archives; systems for records management, archival description, and retrieval; and machine-readable archives. The selection also features examples of archives from different institutions such as the University of Liverpool, Berkshire County Record Office, and the National Maritime Museum.The text is recommended for archivists who would like to know more about the use of computers in archiving of
档案的管理是一种社会现象,它以一定的物质形式存在,其中一部分需要我们永久保存下去.随着我国献血事业的发展,血站档案管理的重要性及独特性也逐渐显现出来.本文就如何做好血站档案管理工作,吸收和借鉴优秀的档案管理工作研究成果,以及血站档案管理工作中存在的问题作一剖析,并提出相应的解决措施.%File management is a social phenomenon; il exists in the form of material, some of which we need to keep permanently. With the development of our blood career, it emerges that the importance and specialty of file management. The article analyzes the problems in file management, how to do well file management, absorb and learn from excellent file management, and proposes appropriate solutions.
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.
Fajgelj, A; Lakoski, A; Horvat, D; Remec, I; Skrk, J; Stegnar, P
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
Behets, F.; Kashamuka, M; Pappaioanou, M; Green, T A; Ryder, R W; Batter, V; George, J R; Hannon, W H; Quinn, T C
The use of whole-blood spots on filter paper for the detection of antibody to human immunodeficiency virus type 1 (HIV-1) was evaluated during a 20-week period under a variety of storage environments simulating the harsh tropical field conditions in Kinshasa, Zaire. During the first 6 weeks of storage, all replicates of high- and low-titer HIV-1-positive reference samples remained positive by enzyme immunoassay and Western blotting (immunoblotting), and all replicates of HIV-1-negative sample...
Plichart, Catherine; Lemoine, Aurore
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...
National Archives and Records Administration — The National Archives Catalog is the online catalog of NARA's nationwide holdings in the Washington, DC area, Regional Archives, and Presidential Libraries.
Dauner, Allison L.; Gilliland, Theron C; Mitra, Indrani; Pal, Subhamoy; Morrison, Amy C; Hontz, Robert D.; Wu, Shuenn-Jue L.
Loss of sample integrity during specimen transport can lead to false-negative diagnostic results. In an effort to improve upon the status quo, we used dengue as a model RNA virus to evaluate the stabilization of RNA and antibodies in three commercially available sample stabilization products: Whatman FTA Micro Cards (GE Healthcare Life Sciences, Pittsburgh, PA), DNAstāble Blood tubes (Biomātrica, San Diego, CA), and ViveST tubes (ViveBio, Alpharetta, GA). Both contrived and clinical dengue-po...
Janzen, N; Sander, S.; Terhardt, M.; Das, A.M; Sass, J.O.; Kraetzner, R.; Rosewich, H.; Peter, M.; Sander, J.
The aim of the study was to develop a method for fast and reliable diagnosis of peroxisomal diseases and to facilitate differential diagnosis of cholestatic hepatopathy. For the quantification of bile acids and their conjugates as well as C27 precursors di- and trihydroxycholestanoic acid (DHCA, THCA), in small pediatric blood samples we combined HPLC separation on a reverse-phase C18 column with ESI-MS/MS analysis in the negative ion mode. Analysis was done with good precision (CV 3,7%–11.1%...
Dauner, Allison L; Gilliland, Theron C; Mitra, Indrani; Pal, Subhamoy; Morrison, Amy C; Hontz, Robert D; Wu, Shuenn-Jue L
Loss of sample integrity during specimen transport can lead to false-negative diagnostic results. In an effort to improve upon the status quo, we used dengue as a model RNA virus to evaluate the stabilization of RNA and antibodies in three commercially available sample stabilization products: Whatman FTA Micro Cards (GE Healthcare Life Sciences, Pittsburgh, PA), DNAstāble Blood tubes (Biomātrica, San Diego, CA), and ViveST tubes (ViveBio, Alpharetta, GA). Both contrived and clinical dengue-positive specimens were stored on these products at ambient temperature or 37°C for up to 1 month. Antibody and viral RNA levels were measured by enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) assays, respectively, and compared with frozen unloaded controls. We observed reduced RNA and antibody levels between stabilized contrived samples and frozen controls at our earliest time point, and this was particularly pronounced for the FTA cards. However, despite some time and temperature dependent loss, a 94.6-97.3% agreement was observed between stabilized clinical specimens and their frozen controls for all products. Additional considerations such as cost, sample volume, matrix, and ease of use should inform any decision to incorporate sample stabilization products into a diagnostic testing workflow. We conclude that DNAstāble Blood and ViveST tubes are useful alternatives to traditional filter paper for ambient temperature shipment of clinical specimens for downstream molecular and serological testing. PMID:25940193
Holm, Isak Winkel; Bjering, Jens Christian Borrebye
The first season of Nic Pizzolatto's True Detective (2014) is not only a show about a murder, but also a show about how to gather and organize information in an archive. Having identified two archival problems—its temporal and topical extension—the article turns to Jacques Derrida's Archive Fever......: a Freudian Impression in order to explain how the archiving problems of the crime investigation are, in fact, intrinsic to any archiving practice. Lastly, the article addresses the political significance of the show's archiving problems by help of Derrida's text on the American Constitution and of Hardt...
The National Ecological Observatory Network (NEON) is a 30-year National Science Foundation-funded facility for understanding and forecasting the impacts of climate change, land use change, and invasive species on aspects of continental-scale ecology such as biodiversity, biogeochemistry, infectious diseases, and ecohydrology. NEON will measure a wide range of properties at 60 terrestrial and 36 aquatic sites throughout the US using in situ sensors, sample collection/lab analysis, and remote sensing, and all data will be made freely available. The Observatory is currently under construction and will be fully operational by 2017, however, limited data collection and release will begin in 2013. In addition, NEON is archiving large numbers of samples, including surface soils (top ~30 cm) collected from locations across each site, and soils collected by horizon to 2 m deep from a single soil pit at each site. Here I present information about the latter, focusing on sampling and processing, metadata, and currently available samples. At each terrestrial site the soil pit is dug in the locally dominant soil type and soil is collected by horizon, mixed, and ~4-8 liters soil is sent for processing. Soil samples are air-dried and sieved (mineral soil) or air-dried (organic soil) and 1.2 kg is split between 4 glass jars for archiving (protocol available upon request). To date 15 soil pits have been sampled, representing 7 soil orders, and soils from 110 horizons have been archived or are being processed. Metadata associated with each archive sample include a soil profile description, photos, and soil properties (total C, N, S, Al, Ca, Fe, K, Mg, Mn, Na, P, Si, Sr, Ti, Zr, bulk density, pH, and texture). The procedure for requesting samples from the archive is under development and I encourage scientists to use the archive in their future research. Collecting and processing samples for the NEON Soil Archive
Full Text Available ... Submit to Blood View all Annual Meeting Abstracts 2016 Call for Abstracts 2016 Abstract Review Categories Abstracts Archive View all Education ... ASH Workshop on Genome Editing July 14-15, 2016 Learn new techniques in the clinical application of ...
... in thyroid disease when the eye protrudes forward; cosmetic surgery, if the eyelids are opened too widely. Frequently ... disease when the eye protrudes forward or after cosmetic surgery if the eyelids are opened too widely, dry ...
National Archives and Records Administration — ALIC is an online library catalog of books, periodicals, and other materials contained in Archives I and II and book collections located in other facilities.
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
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
Cook, Benjamin I.; Smerdon, Jason E.; Seager, Richard; Coats, Sloan
Global warming is expected to increase the frequency and intensity of droughts in the twenty-first century, but the relative contributions from changes in moisture supply (precipitation) versus evaporative demand (potential evapotranspiration; PET) have not been comprehensively assessed. Using output from a suite of general circulation model (GCM) simulations from phase 5 of the Coupled Model Intercomparison Project, projected twenty-first century drying and wetting trends are investigated using two offline indices of surface moisture balance: the Palmer Drought Severity Index (PDSI) and the Standardized Precipitation Evapotranspiration Index (SPEI). PDSI and SPEI projections using precipitation and Penman-Monteith based PET changes from the GCMs generally agree, showing robust cross-model drying in western North America, Central America, the Mediterranean, southern Africa, and the Amazon and robust wetting occurring in the Northern Hemisphere high latitudes and east Africa (PDSI only). The SPEI is more sensitive to PET changes than the PDSI, especially in arid regions such as the Sahara and Middle East. Regional drying and wetting patterns largely mirror the spatially heterogeneous response of precipitation in the models, although drying in the PDSI and SPEI calculations extends beyond the regions of reduced precipitation. This expansion of drying areas is attributed to globally widespread increases in PET, caused by increases in surface net radiation and the vapor pressure deficit. Increased PET not only intensifies drying in areas where precipitation is already reduced, it also drives areas into drought that would otherwise experience little drying or even wetting from precipitation trends alone. This PET amplification effect is largest in the Northern Hemisphere mid-latitudes, and is especially pronounced in western North America, Europe, and southeast China. Compared to PDSI projections using precipitation changes only, the projections incorporating both
In this article, the author shares his experience as he traveled from island to island with a single objective--to reach the archives. He found out that not all archives are the same. In recent months, his daydreaming in various facilities has yielded a recurrent question on what would constitute the Ideal Archive. What follows, in no particular…
Huffman, David; /Fermilab
These steps were taken to maintain weekly archives: (1) Friday morning you stop the archiver and wait for it to finish writing data (the lock file will be removed from the directory); (2) move the current archive information to a PC via FTP; (3) remove all previous archive information in the previous directory; (4) move the current archive into the previous directory; (5) start a new archive; (6) burn a CDROM of the archive; and (7) copy the current archive to a specific directory. There are 2 ways to check if the Calorimeter Archiver is running, either through the WEB based front end or directly from a command line. Once the archiver is running it can be monitored from a WEB page. This only works with a browser launched from the online machine running the archiver. Each time the browser is reloaded there should be an update reported in the last write check field. You might have to wait a few minutes to see the update. Calorimetry currently takes readings every (300 sec.) 5 minutes. The second method to verify the archiver is running is to issue a command from a Linux cluster machine.
Batteries - dry cell ... Acidic dry cell batteries contain: Manganese dioxide Ammonium chloride Alkaline dry cell batteries contain: Sodium hydroxide Potassium hydroxide Lithium dioxide dry cell batteries ...
Scientific Opinion on the substantiation of a health claim related to the combination of artichoke leaf dry extract standardised in caffeoylquinic acids, monacolin K in red yeast rice, sugar-cane derived policosanols, OPC from French maritime pine bark, garlic dry extract standardised in allicin, d-α-tocopheryl hydrogen succinate, riboflavin and inositol hexanicotinate in Limicol® and reduction of blood LDL-cholesterol concentrations pursuant to Article 14 of Regulation (EC No 1924/2006
EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA
Full Text Available Following an application from Laboratoire Lescuyer, submitted pursuant to Article 14 of Regulation (EC No 1924/2006 via the Competent Authority of France, the Panel on Dietetic Products, Nutrition and Allergies (NDA was asked to deliver an opinion on the scientific substantiation of a health claim related to the combination of artichoke leaf dry extract standardised in caffeoylquinic acids, monacolin K in red yeast rice, sugar-cane derived policosanols, OPC from French maritime pine bark, garlic dry extract standardised in allicin, d-α-tocopheryl hydrogen succinate, riboflavin and inositol hexanicotinate in Limicol® and reduction of blood LDL-cholesterol concentrations. The Panel considers that the food which is the subject of the claim is sufficiently characterised. The Panel considers that reduction of blood LDL-cholesterol concentrations is a beneficial physiological effect. High LDL-cholesterol is a risk factor in the development of coronary heart disease. In weighing the evidence, the Panel took into account that, although no evidence was provided for an LDL-cholesterol lowering effect of any of the single food constituents in Limicol® at the proposed conditions of use or as to how the ingredients individually or in any combination could contribute to the claimed effect and despite the lack of a dose-response relationship observed in one human intervention study, three human intervention studies conducted by two independent research groups showed an effect of the combination of food ingredients in Limicol® on blood LDL-cholesterol concentrations. The Panel concludes that a cause and effect relationship has been established between the consumption of the combination of artichoke leaf dry extract standardised in caffeoylquinic acids, monacolin K in red yeast rice, sugar-cane derived policosanols, OPC from French maritime pine bark, garlic dry extract standardised in allicin, d-α-tocopheryl hydrogen succinate, riboflavin and inositol
In recent years, historians of archives have paid increasingly careful attention to the development of state, colonial, religious, and corporate archives in the early modern period, arguing that power (of various kinds) was mediated and extended through material writing practices in and around archives. The history of early modern science, likewise, has tracked the production of scientific knowledge through the inscription and circulation of written records within and between laboratories, libraries, homes, and public spaces, such as coffeehouses and bookshops. This Focus section interrogates these two bodies of scholarship against each other. The contributors ask how archival digitization is transforming historical practice; how awareness of archival histories can help us to reconceptualize our work as historians of science; how an archive's layered purposes, built up over centuries of record keeping, can shape the historical narratives we write; and how scientific knowledge emerging from archives gained authority and authenticity. PMID:27197412
Thomas, Carla; Amanuel, Benhur; Finlayson, Jill; Grieu-Iacopetta, Fabienne; Spagnolo, Dominic V; Erber, Wendy N
Hairy cell leukaemia (HCL) is a rare, indolent chronic B-cell leukaemia accounting for approximately 2% of all adult leukaemias. The recent association of the BRAF p.Val600Glu (V600E) mutation in HCL makes it a valuable molecular diagnostic marker. We compared the ability of Sanger sequencing, fluorescent single-strand conformational polymorphism (F-SSCP) and high resolution melting (HRM) analysis to detect BRAF mutations in 20 cases of HCL consisting of four archival Romanowsky stained air-dried peripheral blood and bone marrow aspirate smears, 12 mercury fixed decalcified bone marrow trephine biopsies, three formalin fixed, paraffin embedded (FFPE) splenectomy samples and one fresh peripheral blood sample. DNA was amplified and BRAF mutation status determined by the three methods above. V600E mutation was identified in 94%, 89% and 72% of HCL cases by F-SSCP, HRM and Sanger sequencing, respectively. In one case, in addition to the p.Val600Glu mutation, a p.Lys601Thr (K601T) mutation was identified. DNA from archival slide scrapings, mercury-fixed and FFPE tissue can be used to identify BRAF mutations with high sensitivity, especially using HRM/F-SSCP. The V600E mutation can be used as a supplementary molecular marker to aid in the diagnosis of HCL and the presence of the mutation may provide a target for therapy. PMID:25938346
Lakio, Satu; Heinämäki, Jyrki; Yliruusi, Jouko
Drying is one of the standard unit operations in the pharmaceutical industry and it is important to become aware of the circumstances that dominate during the process. The purpose of this study was to test microcapsulated thermochromic pigments as heat indicators in a fluid bed drying process. The indicator powders were manually granulated with alpha-lactose monohydrate resulting in three particle-size groups. Also, pellets were coated with the indicator powders. The granules and pellets were fluidized in fluid bed dryer to observe the progress of the heat flow in the material and to study the heat indicator properties of the indicator materials. A tristimulus colorimeter was used to measure CIELAB color values. Color indicator for heat detection can be utilized to test if the heat-sensitive API would go through physical changes during the pharmaceutical drying process. Both the prepared granules and pellets can be used as heat indicator in fluid bed drying process. The colored heat indicators give an opportunity to learn new aspects of the process at real time and could be exploded, for example, for scaling-up studies. PMID:20039220
Shirley, Lindsey; Weller, Chanae
Despite its name, commercial dry cleaning is not actually a “dry” process. Clothes are immersed in a solvent, most commonly perchlorethylene (perc), instead of in water. Perc or other similar solvents are effective in the removal of oil and grease-based stains without damaging or shrinking sensitive fabrics, unlike a regular detergents and fabric softeners.
... 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 ...
In the modern world, digital data archiving and accessing is an issue that needs tailored solutions. This thesis gives one example of designing and implementing a data archiving solution using time tested technologies. Helsinki Metropolia University of Applied Sciences had a need to archive their contracts in digital format instead of using old file cabinets. Specifications were created together with the customer at the start of the project. The project was to produce a complete product fo...
Xie, Zhiwu; Nayyar, Krati; Fox, Edward A
In this paper, we propose a modified approach to realtime transactional web archiving. It leverages the web caching infrastructure that is already prevalent on web servers. Instead of archiving web content at HTTP transaction time, in our approach the archiving happens when the cached copy expires and is about to be expunged. Before the deletion, all expired cache copies are combined and then sent to the web archive in small batches. Since the cache is purged at much lower frequency than HTT...
Alacid, J. Manuel; Solano, Enrique
The Gran Telescopio Canarias (GTC) archive is operational since November 2011. The archive, maintained by the Data Archive Unit at CAB in the framework of the Spanish Virtual Observatory project, provides access to both raw and science ready data and has been designed in compliance with the standards defined by the International Virtual Observatory Alliance (IVOA) to guarantee a high level of data accessibility and handling. In this presentation I will describe the main capabilities the GTC archive offers to the community, in terms of functionalities and data collections, to carry out an efficient scientific exploitation of GTC data.
Eskildsen, Kasper Risbjerg
This article investigates the emergence of the archive as the primary venue for the production of historical knowledge in the 19th century. The turn to archival research, the article argues, may be considered as a response to the discussions about the problems of testimony that dominated 18th......- and early 19th-century German writings on the methodology and epistemology of historical research. These discussions, especially regarding the epistemic virtues of witnesses, also helped create the particular culture of knowledge-making within German historical scholarship that enabled the archival turn...... of the 19th-century archival turn....
Lund, Bendik; Najmi, Laeya A; Wesolowska-Andersen, Agata;
AB Archival samples represent a significant potential for genetic studies, particularly in severe diseases with risk of lethal outcome, such as in cancer. In this pilot study, we aimed to evaluate the usability of archival bone marrow smears and biopsies for DNA extraction and purification, whole...... with samples stored for 4 to 10 years. Acceptable call rates for SNPs were detected for 7 of 42 archival samples. In conclusion, archival bone marrow samples are suitable for DNA extraction and multiple marker analysis, but WGA was less successful, especially when longer fragments were analyzed. Multiple SNP...