Diet of spotted bats (Euderma maculatum) in Arizona as indicated by fecal analysis and stable isotopes

EPA Science Inventory

We assessed diet of spotted bats (Euderma maculatum (J.A. Allen, 1891)) by visual analysis of bat feces and stable carbon (δ13C) and nitrogen (δ15N) isotope analysis of bat feces, wing, hair, and insect prey. We collected 33 fecal samples from spotted bats and trapped 3755 insect...

Evaluation of dried blood spot as an alternative sample collection method for hepatitis C virus RNA quantitation and genotyping using a commercial system.

PubMed

Mahajan, Supriya; Choudhary, Manish Chandra; Kumar, Guresh; Gupta, Ekta

2018-06-01

Dried blood spot (DBS) is a minimally invasive sampling method suitable for sample collection, storage and transportation in resource limited areas. Aim of this study was to compare the diagnostic utility of DBS with plasma sample for HCV RNA quantitation and genotyping using commercial systems. Plasma and DBS card spotted samples were collected from 95 HCV seropositive patients. Both types of samples were subjected to HCV RNA by real-time PCR (Abbott m2000rt, USA). Genotyping was performed using Abbott HCV genotype II kit (Abbott diagnostics, USA) in samples with viral load > 3 log 10  IU/ml. In both plasma and DBS, 14 (14.7%) samples were negative and 81 (85.3%) were positive for HCV RNA. Median viral load in plasma (3.78; range 0-7.43) log 10  IU/ml was comparable to DBS (3.93; range 0-7.24) log 10  IU/ml. DBS demonstrated sensitivity and specificity of 97.5 and 85.7% respectively, with positive predictive value (PPV) of 97.5% and negative predictive value (NPV) of 85.7%. DBS showed good correlation (r 2  = 0.866) and agreement (93.5%) with plasma. Genotyping in 20 patients showed 100% concordance between DBS and plasma samples. DBS showed good sensitivity and specificity as a sampling method for HCV RNA quantitation and genotyping.

Two-dimensional polyacrylamide gel electrophoresis of equine seminal plasma proteins and their relation with semen freezability.

PubMed

Jobim, M I M; Trein, C; Zirkler, H; Gregory, R M; Sieme, H; Mattos, R C

2011-09-01

The objective was to evaluate protein profiles of equine seminal plasma using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and to determine whether any of these proteins were related to semen freezability. Seminal plasma was collected from 10 stallions, of high and low semen freezability, housed at the State Stud of Lower Saxony, and routinely used in AI programs. Twenty-five protein spots were identified from the two-dimensional gel (12%), seven of which were present in all samples (all proteins were identified by MALDI-MS). Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been used to generate ion images of samples in one or more mass-to-charge (m/z) values, providing the capability of mapping specific molecules to two-dimensional coordinates of the original sample. Of the 25 proteins identified, two spots had greater relative content (P < 0.05) in seminal plasma samples collected from stallions with high semen freezability: spot 5 (80-85 kDa, isoelectric point [pI] 7.54), identified as CRISP-3; and spot 45 (18.2 kDa, pI 5.0-5.2), identified as HSP-2. Conversely, protein content was greater (P < 0.05) in seminal plasma samples from stallions with low semen freezability: spot 7 (75.4 kDa, pI 6.9-7.4), identified as lactoferrin; spot 15 (26.7 kDa, pI 5.51), identified as kallikrein; spot 25 (25 kDa, pI 7.54), identified as CRISP-3; and spot 35 (13.9 kDa, pI 3.8-4.2), identified as HSP-1. In conclusion, there were differences in the seminal plasma protein profile from stallions with high and low semen freezability. Furthermore, CRISP-3 and HSP-2 were potential seminal plasma markers of high semen freezability. Copyright © 2011 Elsevier Inc. All rights reserved.

Quantitative determination of atenolol in dried blood spot samples by LC-HRMS: a potential method for assessing medication adherence.

PubMed

Lawson, Graham; Cocks, Elizabeth; Tanna, Sangeeta

2012-05-15

The use of blood spot collection cards was investigated as a means of obtaining small volume samples for the quantification of therapeutic drugs for assessing medication adherence. A liquid chromatography-high resolution TOF mass spectrometry (LC-HRMS) method, based on the measurement at the accurate mass to charge ratio of the target analyte, was used to ensure specificity for atenolol in the dried blood spot (DBS) samples. A working method was developed and validated. For the preparation of DBS samples whole blood spiked with analyte was used to produce 30 μl blood spots on specimen collection cards. A 5mm disc was cut from the dried blood spot and extracted using methanol:water (60:40, v/v) containing the internal standard, atenolol-d(7). Extracts were vortexed, sonicated and then centrifuged. Gradient chromatographic elution was achieved using an Ascentis Express C18 100mm×2.1mm column and a mobile phase flow rate of 0.2 ml/min and the column oven temperature at 30 °C. MS detection was carried out in electrospray positive ion mode for target ions at accurate mass m/z 267.1703 for atenolol and 274.2143 for the IS. Drug extraction efficiency from spiked blood spots was demonstrated to be 96±5% and the drug was stable in DBS for at least 10 weeks. The developed LC-HRMS method was linear within the tested calibration range of 25-1500 ng/ml and validation showed the accuracy (relative error) and precision (coefficient of variation) values were within the pre-defined limits of ≤ 5% at all concentrations with a limit of quantification of 25 ng/ml. Factors with potential to affect drug quantification measurements such as the matrix effects, volume of blood applied onto the collection card and effect of different sampling cards were investigated. The developed LC-HRMS method was applied to blood spots on sampling card taken from adult healthy volunteers previously administered a 50mg atenolol tablet and a DBS concentration-time profile was obtained for atenolol. Requiring only a micro volume (30 μl) blood sample for analysis, the developed DBS based assay has the potential to assess patient adherence to atenolol. Copyright © 2012 Elsevier B.V. All rights reserved.

Resurrection of New Caledonian maskray Neotrygon trigonoides (Myliobatoidei: Dasyatidae) from synonymy with N. kuhlii, based on cytochrome-oxidase I gene sequences and spotting patterns.

PubMed

Borsa, Philippe; Arlyza, Irma S; Chen, Wei-Jen; Durand, Jean-Dominique; Meekan, Mark G; Shen, Kang-Ning

2013-04-01

The maskray from New Caledonia, Neotrygon trigonoides Castelnau, 1873, has been recently synonymized with the blue-spotted maskray, N. kuhlii (Müller and Henle, 1841), a species with wide Indo-West Pacific distribution, but the reasons for this are unclear. Blue-spotted maskray specimens were collected from the Indian Ocean (Tanzania, Sumatra) and the Coral Triangle (Indonesia, Taiwan, and West Papua), and N. trigonoides specimens were collected from New Caledonia (Coral-Sea). Their partial COI gene sequences were generated to expand the available DNA-barcode database on this species, which currently comprises homologous sequences from Ningaloo Reef, the Coral Triangle and the Great Barrier Reef (Coral-Sea). Spotting patterns were also compared across regions. Haplotypes from the Coral-Sea formed a haplogroup phylogenetically distinct from all other haplotypes sampled in the Indo-West Pacific. No clear-cut geographic composition relative to DNA-barcodes or spotting patterns was apparent in N. kuhlii samples across the Indian Ocean and the Coral Triangle. The New Caledonian maskray had spotting patterns markedly different from all the other samples. This, added to a substantial level of net nucleotide divergence (2.6%) with typical N. kuhlii justifies considering the New Caledonian maskray as a separate species, for which we propose to resurrect the name Neotrygon trigonoides. Copyright © 2013. Published by Elsevier SAS.

Spot urine and 24-h diet recall estimates of dietary sodium intake from the 2008/09 New Zealand Adult Nutrition Survey: a comparison.

PubMed

McLean, Rachael M; Williams, Sheila M; Te Morenga, Lisa A; Mann, Jim I

2018-05-22

We aimed to test the difference between estimates of dietary sodium intake using 24-h diet recall and spot urine collection in a large sample of New Zealand adults. We analysed spot urine results, 24-h diet recall, dietary habits questionnaire and anthropometry from a representative sample of 3312 adults aged 15 years and older who participated in the 2008/09 New Zealand Adult Nutrition Survey. Estimates of adult population sodium intake were derived from 24-h diet recall and spot urine sodium using a formula derived from analysis of INTERSALT data. Correlations, limits of agreement and mean difference were calculated for the total sample, and for population subgroups. Estimated total population 24-h urinary sodium excretion (mean (95% CI)) from spot urine samples was 3035 mg (2990, 3079); 3612 mg (3549, 3674) for men and 2507 mg (2466, 2548) for women. Estimated mean usual daily sodium intake from 24-h diet recall data (excluding salt added at the table) was 2564 mg (2519, 2608); 2849 mg (2779, 2920) for men and 2304 mg (2258, 2350) for women. Correlations between estimates were poor, especially for men, and limits of agreement using Bland-Altman mean difference analysis were wide. There is a poor agreement between estimates of individual sodium intake from spot urine collection and those from 24-hour diet recall. Although, both 24-hour dietary recall and estimated urinary excretion based on spot urine indicate mean population sodium intake is greater than 2 g, significant differences in mean intake by method deserve further investigation in relation to the gold standard, 24-hour urinary sodium excretion.

Mean population salt intake estimated from 24-h urine samples and spot urine samples: a systematic review and meta-analysis.

PubMed

Huang, Liping; Crino, Michelle; Wu, Jason H Y; Woodward, Mark; Barzi, Federica; Land, Mary-Anne; McLean, Rachael; Webster, Jacqui; Enkhtungalag, Batsaikhan; Neal, Bruce

2016-02-01

Estimating equations based on spot urine samples have been identified as a possible alternative approach to 24-h urine collections for determining mean population salt intake. This review compares estimates of mean population salt intake based upon spot and 24-h urine samples. We systematically searched for all studies that reported estimates of daily salt intake based upon both spot and 24-h urine samples for the same population. The associations between the two were quantified and compared overall and in subsets of studies. A total of 538 records were identified, 108 were assessed as full text and 29 were included. The included studies involved 10,414 participants from 34 countries and made 71 comparisons available for the primary analysis. Overall average population salt intake estimated from 24-h urine samples was 9.3 g/day compared with 9.0 g/day estimated from the spot urine samples. Estimates based upon spot urine samples had excellent sensitivity (97%) and specificity (100%) at classifying mean population salt intake as above or below the World Health Organization maximum target of 5 g/day. Compared with the 24-h samples, estimates based upon spot urine overestimated intake at lower levels of consumption and underestimated intake at higher levels of consumption. Estimates of mean population salt intake based upon spot urine samples can provide countries with a good indication of mean population salt intake and whether action on salt consumption is required. Published by Oxford University Press on behalf of the International Epidemiological Association 2015. This work is written by US Government employees and is in the public domain in the US.

Dried blood spots as a source of anti-malarial antibodies for epidemiological studies

PubMed Central

Corran, Patrick H; Cook, Jackie; Lynch, Caroline; Leendertse, Heleen; Manjurano, Alphaxard; Griffin, Jamie; Cox, Jonathan; Abeku, Tarekegn; Bousema, Teun; Ghani, Azra C; Drakeley, Chris; Riley, Eleanor

2008-01-01

Background Blood spots collected onto filter paper are an established and convenient source of antibodies for serological diagnosis and epidemiological surveys. Although recommendations for the storage and analysis of small molecule analytes in blood spots exist, there are no published systematic studies of the stability of antibodies under different storage conditions. Methods Blood spots, on filter paper or glass fibre mats and containing malaria-endemic plasma, were desiccated and stored at various temperatures for different times. Eluates of these spots were assayed for antibodies against two Plasmodium falciparum antigens, MSP-119 and MSP2, and calculated titres used to fit an exponential (first order kinetic) decay model. The first order rate constants (k) for each spot storage temperature were used to fit an Arrhenius equation, in order to estimate the thermal and temporal stability of antibodies in dried blood spots. The utility of blood spots for serological assays was confirmed by comparing antibodies eluted from blood spots with the equivalent plasma values in a series of samples from North Eastern Tanzania and by using blood spot-derived antibodies to estimate malaria transmission intensity in this site and for two localities in Uganda. Results Antibodies in spots on filter paper and glass fibre paper had similar stabilities but blood was more easily absorbed onto filter papers than glass fibre, spots were more regular and spot size was more closely correlated with blood volume for filter paper spots. Desiccated spots could be stored at or below 4°C for extended periods, but were stable for only very limited periods at ambient temperature. When desiccated, recoveries of antibodies that are predominantly of IgG1 or IgG3 subclasses were similar. Recoveries of antibodies from paired samples of serum and of blood spots from Tanzania which had been suitably stored showed similar recoveries of antibodies, but spots which had been stored for extended periods at ambient humidity and temperature showed severe loss of recoveries. Estimates of malaria transmission intensity obtained from serum and from blood spots were similar, and values obtained using blood spots agreed well with entomologically determined values. Conclusion This study has demonstrated the suitability of filter paper blood spots paper for collection of serum antibodies, and provided clear guidelines for the treatment and storage of filter papers which emphasize the importance of desiccation and minimisation of time spent at ambient temperatures. A recommended protocol for collecting, storing and assaying blood spots is provided. PMID:18826573

Relationship between timed and spot urine collections for measuring phosphate excretion.

PubMed

Tan, Sven-Jean; Smith, Edward R; Cai, Michael M X; Holt, Stephen G; Hewitson, Tim D; Toussaint, Nigel D

2016-01-01

Twenty-four hour urinary phosphate excretion (UPE) reflects intestinal phosphate absorption in steady state and may be more informative than serum phosphate (sPi) when assessing phosphate homoeostasis clinically. Timed urine collections are cumbersome and prone to collection errors. Spot urine phosphate/creatinine ratio (uPiCr) may be a useful, simple surrogate for 24-h UPE, but requires further validation. This study aimed to determine the relationship between uPiCr and 24-h UPE. This single-centre cross-sectional study examined contemporaneous serum, spot urine and 24-h urine. Serum biochemistry was analysed. Urine phosphate concentration (uPi) and creatinine concentration (uCr) were measured in spot and 24-h urine collections. Spearman's rank correlation coefficients and Bland-Altman plots were used to assess agreement between spot uPiCr and UPE. Backward multivariate analysis was undertaken for UPE prediction. One hundred and sixteen participants (77 with kidney disease and 39 healthy volunteers) were studied. Seventy-four (63.8 %) were male. Median (IQR) age was 61(49-71) years. Median (IQR) spot uPiCr and total UPE were 1.7 (1.3-2.2) mmol/mmol and 25.8 (19.9-35.0) mmol/d, respectively. There was no correlation between spot uPiCr and 24-h UPE (R = 0.064, P = 0.51) but spot uPi significantly correlated with 24-h UPE (R = 0.385, P < 0.001). Although spot and 24-h measures of phosphate handling correlated (all P < 0.001), Bland-Altman analysis revealed bias between collection techniques. UPE prediction model using the independent variables of eGFR, sPi, albumin and spot uPi provided R (2) = 0.443. No direct correlation was noted between spot uPiCr and 24-h UPE. Normalization of uPi to uCr on spot urine samples may be inappropriate when evaluating urinary phosphate excretion in adults and thus, a spot uPiCr is not a suitable surrogate for measuring UPE. A UPE prediction model utilising spot urine biochemistry cannot be advocated at present.

Validation study of the Tanaka and Kawasaki equations to estimate the daily sodium excretion by a spot urine sample.

PubMed

Mill, José Geraldo; Rodrigues, Sérgio Lamêgo; Baldo, Marcelo Perim; Malta, Deborah Carvalho; Szwarcwald, Celia Landmann

2015-12-01

To validate Tanaka and Kawasaki's formulas to calculate the salt intake by the sodium/creatinine ratio in spot of urine. Two hundred and seventy two adults (20 - 69 years old; 52.6% women) with 24 h urine collection and two urinary spots collected on the same day (while fasting - spot 1 - or not fasting - spot 2). Anthropometry, blood pressure and fasting blood were measured on the same day. The analysis of agreement between salt consumption measured in the 24 h urine test and urinary spots were determined by the Pearson's correlation (r) and the Bland & Altman method. The mean salt consumption measured by the 24 h sodium excretion was 10.4 ± 5.3 g/day. The correlation between the measured 24 h sodium excretion and the estimation based on spots 1 and 2, respectively, was only moderated according to Tanaka (r = 0.51 and r = 0.55; p < 0.001) and to Kawasaki (r = 0.52 and r = 0.54; p < 0.001). We observed an increasing underestimation of salt consumption by Tanaka to increasing salt consumption and conversely, an overestimation of consumption by the Kawasaki formula. The estimation of salt consumption (difference between measured and calculated salt consumption lower than 1 g/day) was adequate only when the consumption was between 9 - 12 g/day (Tanaka) and 12 - 18 g/day (Kawasaki). Spot urine sampling is adequate to estimate salt consumption only among individuals with an actual consumption near the population mean.

Development and validation of dried matrix spot sampling for the quantitative determination of amyloid β peptides in cerebrospinal fluid.

PubMed

Delaby, Constance; Gabelle, Audrey; Meynier, Philippe; Loubiere, Vincent; Vialaret, Jérôme; Tiers, Laurent; Ducos, Jacques; Hirtz, Christophe; Lehmann, Sylvain

2014-05-01

The use of dried blood spots on filter paper is well documented as an affordable and practical alternative to classical venous sampling for various clinical needs. This technique has indeed many advantages in terms of collection, biological safety, storage, and shipment. Amyloid β (Aβ) peptides are useful cerebrospinal fluid (CSF) biomarkers for Alzheimer disease diagnosis. However, Aβ determination is hindered by preanalytical difficulties in terms of sample collection and stability in tubes. We compared the quantification of Aβ peptides (1-40, 1-42, and 1-38) by simplex and multiplex ELISA, following either a standard operator method (liquid direct quantification) or after spotting CSF onto dried matrix paper card. The use of dried matrix spot (DMS) overcame preanalytical problems and allowed the determination of Aβ concentrations that were highly commutable (Bland-Altman) with those obtained using CSF in classical tubes. Moreover, we found a positive and significant correlation (r2=0.83, Pearson coefficient p=0.0329) between the two approaches. This new DMS method for CSF represents an interesting alternative that increases the quality and efficiency in preanalytics. This should enable the better exploitation of Aβ analytes for Alzheimer's diagnosis.

Comparison between dried blood spot and plasma sampling for therapeutic drug monitoring of antiepileptic drugs in children with epilepsy: A step towards home sampling.

PubMed

Linder, Camilla; Wide, Katarina; Walander, Malin; Beck, Olof; Gustafsson, Lars L; Pohanka, Anton

2017-05-01

To investigate if dried blood spots could be used for therapeutic drug monitoring of the antiepileptic drugs, carbamazepine, lamotrigine and valproic acid in children with epilepsy. Fingerprick blood samples from 46 children at a neuropediatric outpatient clinic was collected on filterpaper at the same time as capillary plasma sampling. A validated dried blood spot liquid chromatography tandem mass spectrometry method for carbamazepine, lamotrigine and valproic acid was compared with the routine plasma laboratory methods. Method agreement was evaluated and plasma concentrations were estimated by different conversion approaches. Strong correlation was shown between dried blood spot and plasma concentrations for all three drugs, with R2 values>0.89. Regression analysis showed a proportional bias with 35% lower dried blood spot concentrations for valproic acid (n=33) and concentrations were 18% higher for carbamazepine (n=17). A ratio approach was used to make a conversion from dried blood spots to estimated plasma for these two drugs. Dried blood spot concentrations were directly comparable with plasma for lamotrigine (n=20). This study supports that dried blood spot concentrations can be used as an alternative to plasma in a children population for three commonly used antiepileptic drugs with the possibility to expand by adding other antiepileptic drugs. Clinical decisions can be made based on converted (carbamazepine, valproic acid) or unconverted (lamotrigine) dried blood spot concentrations. Dried blood spot sampling, in the future taken at home, will simplify an effective therapeutic drug monitoring for this group of patients who often have concomitant disorders and also reduce costs for society. Copyright © 2016 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Hematocrit-Independent Quantitation of Stimulants in Dried Blood Spots: Pipet versus Microfluidic-Based Volumetric Sampling Coupled with Automated Flow-Through Desorption and Online Solid Phase Extraction-LC-MS/MS Bioanalysis.

PubMed

Verplaetse, Ruth; Henion, Jack

2016-07-05

A workflow overcoming microsample collection issues and hematocrit (HCT)-related bias would facilitate more widespread use of dried blood spots (DBS). This report describes comparative results between the use of a pipet and a microfluidic-based sampling device for the creation of volumetric DBS. Both approaches were successfully coupled to HCT-independent, fully automated sample preparation and online liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) analysis allowing detection of five stimulants in finger prick blood. Reproducible, selective, accurate, and precise responses meeting generally accepted regulated bioanalysis guidelines were observed over the range of 5-1000 ng/mL whole blood. The applied heated flow-through solvent desorption of the entire spot and online solid phase extraction (SPE) procedure were unaffected by the blood's HCT value within the tested range of 28.0-61.5% HCT. Enhanced stability for mephedrone on DBS compared to liquid whole blood was observed. Finger prick blood samples were collected using both volumetric sampling approaches over a time course of 25 h after intake of a single oral dose of phentermine. A pharmacokinetic curve for the incurred phentermine was successfully produced using the described validated method. These results suggest that either volumetric sample collection method may be amenable to field-use followed by fully automated, HCT-independent DBS-SPE-LC-MS/MS bioanalysis for the quantitation of these representative controlled substances. Analytical data from DBS prepared with a pipet and microfluidic-based sampling devices were comparable, but the latter is easier to operate, making this approach more suitable for sample collection by unskilled persons.

Wilson Corners SWMU 001 2014 Annual Long Term Monitoring Report Kennedy Space Center, Florida

NASA Technical Reports Server (NTRS)

Langenbach, James

2015-01-01

This document presents the findings of the 2014 Long Term Monitoring (LTM) that was completed at the Wilson Corners site, located at the National Aeronautics and Space Administration (NASA) John F. Kennedy Space Center (KSC), Florida. The goals of the 2014 annual LTM event were to evaluate the groundwater flow direction and gradient and to monitor the vertical and downgradient horizontal extent of the volatile organic compounds (VOCs) in groundwater at the site. The LTM activities consisted of an annual groundwater sampling event in December 2014, which included the collection of water levels from the LTM wells. During the annual groundwater sampling event, depth to groundwater was measured and VOC samples were collected using passive diffusion bags (PDBs) from 30 monitoring wells. In addition to the LTM sampling, additional assessment sampling was performed at the site using low-flow techniques based on previous LTM results and assessment activities. Assessment of monitoring well MW0052DD was performed by collecting VOC samples using low-flow techniques before and after purging 100 gallons from the well. Monitoring well MW0064 was sampled to supplement shallow VOC data north of Hot Spot 2 and east of Hot Spot 4. Monitoring well MW0089 was sampled due to its proximity to MW0090. MW0090 is screened in a deeper interval and had an unexpected detection of trichloroethene (TCE) during the 2013 LTM, which was corroborated during the March 2014 verification sampling. Monitoring well MW0130 was sampled to provide additional VOC data beneath the semi-confining clay layer in the Hot Spot 2 area.

Spot sputum samples are at least as good as early morning samples for identifying Mycobacterium tuberculosis.

PubMed

Murphy, Michael E; Phillips, Patrick P J; Mendel, Carl M; Bongard, Emily; Bateson, Anna L C; Hunt, Robert; Murthy, Saraswathi; Singh, Kasha P; Brown, Michael; Crook, Angela M; Nunn, Andrew J; Meredith, Sarah K; Lipman, Marc; McHugh, Timothy D; Gillespie, Stephen H

2017-10-27

The use of early morning sputum samples (EMS) to diagnose tuberculosis (TB) can result in treatment delay given the need for the patient to return to the clinic with the EMS, increasing the chance of patients being lost during their diagnostic workup. However, there is little evidence to support the superiority of EMS over spot sputum samples. In this new analysis of the REMoxTB study, we compare the diagnostic accuracy of EMS with spot samples for identifying Mycobacterium tuberculosis pre- and post-treatment. Patients who were smear positive at screening were enrolled into the study. Paired sputum samples (one EMS and one spot) were collected at each trial visit pre- and post-treatment. Microscopy and culture on solid LJ and liquid MGIT media were performed on all samples; those missing corresponding paired results were excluded from the analyses. Data from 1115 pre- and 2995 post-treatment paired samples from 1931 patients enrolled in the REMoxTB study were analysed. Patients were recruited from South Africa (47%), East Africa (21%), India (20%), Asia (11%), and North America (1%); 70% were male, median age 31 years (IQR 24-41), 139 (7%) co-infected with HIV with a median CD4 cell count of 399 cells/μL (IQR 318-535). Pre-treatment spot samples had a higher yield of positive Ziehl-Neelsen smears (98% vs. 97%, P = 0.02) and LJ cultures (87% vs. 82%, P = 0.006) than EMS, but there was no difference for positivity by MGIT (93% vs. 95%, P = 0.18). Contaminated and false-positive MGIT were found more often with EMS rather than spot samples. Surprisingly, pre-treatment EMS had a higher smear grading and shorter time-to-positivity, by 1 day, than spot samples in MGIT culture (4.5 vs. 5.5 days, P < 0.001). There were no differences in time to positivity in pre-treatment LJ culture, or in post-treatment MGIT or LJ cultures. Comparing EMS and spot samples in those with unfavourable outcomes, there were no differences in smear or culture results, and positive results were not detected earlier in Kaplan-Meier analyses in either EMS or spot samples. Our data do not support the hypothesis that EMS samples are superior to spot sputum samples in a clinical trial of patients with smear positive pulmonary TB. Observed small differences in mycobacterial burden are of uncertain significance and EMS samples do not detect post-treatment positives any sooner than spot samples.

Metals, Parasites, and Environmental Conditions Affecting Breeding Populations of Spotted Salamanders (Ambystoma maculatum) in Northern Arkansas, USA.

PubMed

DeMali, Heather M; Trauth, Stanley E; Bouldin, Jennifer L

2016-06-01

The spotted salamander (Ambystoma maculatum) is indigenous to northern Arkansas, and several breeding sites are known to exist in the region. Spotted salamanders (n = 17) were collected and examined for parasites and only three females harbored nematodes (Physaloptera spp.). Chronic aquatic bioassays were conducted using water collected from eight breeding ponds during different hydroperiod events. No lethal or sublethal effects were measured in Ceriodaphnia dubia; however, decreased growth and survival were seen in Pimephales promelas. Aqueous, sediment, and salamander hepatic samples were analyzed for As, Cd, Cu, Pb, and Ni. Metal analysis revealed possible increased metal exposure following precipitation, with greatest metal concentrations measured in sediment samples. Hepatic metal concentrations were similar in parasitized and non-parasitized individuals, and greatest Pb concentrations were measured following normal precipitation events. Determining environmental stressors of amphibians, especially during their breeding and subsequent larval life stage, is imperative to improve species conservation.

Laser dissection sampling modes for direct mass spectral analysis [using a hybrid optical microscopy/laser ablation liquid vortex capture/electrospray ionization system

DOE PAGES

Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

2016-02-01

Here, laser microdissection coupled directly with mass spectrometry provides the capability of on-line analysis of substrates with high spatial resolution, high collection efficiency, and freedom on shape and size of the sampling area. Establishing the merits and capabilities of the different sampling modes that the system provides is necessary in order to select the best sampling mode for characterizing analytically challenging samples. The capabilities of laser ablation spot sampling, laser ablation raster sampling, and laser 'cut and drop' sampling modes of a hybrid optical microscopy/laser ablation liquid vortex capture electrospray ionization mass spectrometry system were compared for the analysis ofmore » single cells and tissue. Single Chlamydomonas reinhardtii cells were monitored for their monogalactosyldiacylglycerol (MGDG) and diacylglyceryltrimethylhomo-Ser (DGTS) lipid content using the laser spot sampling mode, which was capable of ablating individual cells (4-15 m) even when agglomerated together. Turbid Allium Cepa cells (150 m) having unique shapes difficult to precisely measure using the other sampling modes could be ablated in their entirety using laser raster sampling. Intact microdissections of specific regions of a cocaine-dosed mouse brain tissue were compared using laser 'cut and drop' sampling. Since in laser 'cut and drop' sampling whole and otherwise unmodified sections are captured into the probe, 100% collection efficiencies were achieved. Laser ablation spot sampling has the highest spatial resolution of any sampling mode, while laser ablation raster sampling has the highest sampling area adaptability of the sampling modes. In conclusion, laser ablation spot sampling has the highest spatial resolution of any sampling mode, useful in this case for the analysis of single cells. Laser ablation raster sampling was best for sampling regions with unique shapes that are difficult to measure using other sampling modes. Laser 'cut and drop' sampling can be used for cases where the highest sensitivity is needed, for example, monitoring drugs present in trace amounts in tissue.« less

Laser dissection sampling modes for direct mass spectral analysis [using a hybrid optical microscopy/laser ablation liquid vortex capture/electrospray ionization system

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

Cahill, John F.; Kertesz, Vilmos; Van Berkel, Gary J.

Here, laser microdissection coupled directly with mass spectrometry provides the capability of on-line analysis of substrates with high spatial resolution, high collection efficiency, and freedom on shape and size of the sampling area. Establishing the merits and capabilities of the different sampling modes that the system provides is necessary in order to select the best sampling mode for characterizing analytically challenging samples. The capabilities of laser ablation spot sampling, laser ablation raster sampling, and laser 'cut and drop' sampling modes of a hybrid optical microscopy/laser ablation liquid vortex capture electrospray ionization mass spectrometry system were compared for the analysis ofmore » single cells and tissue. Single Chlamydomonas reinhardtii cells were monitored for their monogalactosyldiacylglycerol (MGDG) and diacylglyceryltrimethylhomo-Ser (DGTS) lipid content using the laser spot sampling mode, which was capable of ablating individual cells (4-15 m) even when agglomerated together. Turbid Allium Cepa cells (150 m) having unique shapes difficult to precisely measure using the other sampling modes could be ablated in their entirety using laser raster sampling. Intact microdissections of specific regions of a cocaine-dosed mouse brain tissue were compared using laser 'cut and drop' sampling. Since in laser 'cut and drop' sampling whole and otherwise unmodified sections are captured into the probe, 100% collection efficiencies were achieved. Laser ablation spot sampling has the highest spatial resolution of any sampling mode, while laser ablation raster sampling has the highest sampling area adaptability of the sampling modes. In conclusion, laser ablation spot sampling has the highest spatial resolution of any sampling mode, useful in this case for the analysis of single cells. Laser ablation raster sampling was best for sampling regions with unique shapes that are difficult to measure using other sampling modes. Laser 'cut and drop' sampling can be used for cases where the highest sensitivity is needed, for example, monitoring drugs present in trace amounts in tissue.« less

Stability of Phosphatidylethanol in Dry Blood Spot Cards.

PubMed

Bakhireva, Ludmila N; Shrestha, Shikhar; Gutierrez, Hilda L; Berry, Mike; Schmitt, Cheryl; Sarangarm, Dusadee

2016-05-01

The analysis of phosphatidylethanol, a promising direct ethanol metabolite, in dry blood spots (PEth-DBS) is advantageous due to ease of storage, transportation and minimal invasiveness of capillary blood collection. One potential application of PEth-DBS is to confirm prenatal alcohol exposure in newborns suspected of FASD; however, stability of PEth-DBS is largely unknown. Phlebotomized samples from 31 adults with a history of alcoholism, admitted to the University of New Mexico Emergency Department, were analyzed for blood alcohol content and pipetted onto DBS cards (13 spots per patient). The first spot was analyzed within 2 weeks of collection for a baseline PEth; the remaining 12 spots were allocated into three temperature conditions (room temperature, 4°C, -80°C) for the repeated measures analysis. In addition, 5 newborn DBS samples with a baseline PEth>LOD were obtained from a prospective cohort at UNM and re-analyzed at 4 months after storage at -80°C. A mixed linear model was fitted to examine the effects of temperature, time and temperature-time interaction on PEth degradation over the first 9 months. The baseline PEth levels were 592.8 ± 86.7 ng/ml and 18.3 ± 4.8 ng/ml in adult and newborn samples, respectively. All DBS samples remained positive in successive samples in all temperature conditions. Results of mixed linear model demonstrated a significant effect of temperature (P < 0.001) on PEth degradation over 9 months. PEth-DBS appears to be relatively stable, especially when stored at lower temperatures. These initial results are encouraging and highlight the PEth-DBS potential in retrospective assessment of alcohol exposure. © The Author 2015. Medical Council on Alcohol and Oxford University Press. All rights reserved.

Stability of Phosphatidylethanol in Dry Blood Spot Cards

PubMed Central

Bakhireva, Ludmila N.; Shrestha, Shikhar; Gutierrez, Hilda L.; Berry, Mike; Schmitt, Cheryl; Sarangarm, Dusadee

2016-01-01

Background The analysis of phosphatidylethanol, a promising direct ethanol metabolite, in dry blood spots (PEth-DBS) is advantageous due to ease of storage, transportation and minimal invasiveness of capillary blood collection. One potential application of PEth-DBS is to confirm prenatal alcohol exposure in newborns suspected of FASD; however, stability of PEth-DBS is largely unknown. Methods Phlebotomized samples from 31 adults with a history of alcoholism, admitted to the University of New Mexico Emergency Department, were analyzed for blood alcohol content and pipetted onto DBS cards (13 spots per patient). The first spot was analyzed within 2 weeks of collection for a baseline PEth; the remaining 12 spots were allocated into three temperature conditions (room temperature, 4°C, −80°C) for the repeated measures analysis. In addition, 5 newborn DBS samples with a baseline PEth>LOD were obtained from a prospective cohort at UNM and re-analyzed at 4 months after storage at −80°C. A mixed linear model was fitted to examine the effects of temperature, time and temperature–time interaction on PEth degradation over the first 9 months. Results The baseline PEth levels were 592.8 ± 86.7 ng/ml and 18.3 ± 4.8 ng/ml in adult and newborn samples, respectively. All DBS samples remained positive in successive samples in all temperature conditions. Results of mixed linear model demonstrated a significant effect of temperature (P < 0.001) on PEth degradation over 9 months. Conclusions PEth-DBS appears to be relatively stable, especially when stored at lower temperatures. These initial results are encouraging and highlight the PEth-DBS potential in retrospective assessment of alcohol exposure. PMID:26519350

Dried Blood Spot RNA Transcriptomes Correlate with Transcriptomes Derived from Whole Blood RNA.

PubMed

Reust, Mary J; Lee, Myung Hee; Xiang, Jenny; Zhang, Wei; Xu, Dong; Batson, Tatiana; Zhang, Tuo; Downs, Jennifer A; Dupnik, Kathryn M

2018-05-01

Obtaining RNA from clinical samples collected in resource-limited settings can be costly and challenging. The goals of this study were to 1) optimize messenger RNA extraction from dried blood spots (DBS) and 2) determine how transcriptomes generated from DBS RNA compared with RNA isolated from blood collected in Tempus tubes. We studied paired samples collected from eight adults in rural Tanzania. Venous blood was collected on Whatman 903 Protein Saver cards and in tubes with RNA preservation solution. Our optimal DBS RNA extraction used 8 × 3-mm DBS punches as the starting material, bead beater disruption at maximum speed for 60 seconds, extraction with Illustra RNAspin Mini RNA Isolation kit, and purification with Zymo RNA Concentrator kit. Spearman correlations of normalized gene counts in DBS versus whole blood ranged from 0.887 to 0.941. Bland-Altman plots did not show a trend toward over- or under-counting at any gene size. We report a method to obtain sufficient RNA from DBS to generate a transcriptome. The DBS transcriptome gene counts correlated well with whole blood transcriptome gene counts. Dried blood spots for transcriptome studies could be an option when field conditions preclude appropriate collection, storage, or transport of whole blood for RNA studies.

Food habits of the Northern Spotted Owl (Strix occidentalis caurina) at six nest sites in Washington?s east Cascades

Treesearch

Kenneth R. Bevis; Jo Ellen Richards; Gina M. King; Eric E. Hanson

1997-01-01

This paper reports on 245 pellet samples containing 479 identified prey items collected at six Northern Spotted Owl (Strix occidentalis caurina) breeding sites in the eastern portion of its range. The majority of prey (biomass) came from four species; northern flying squirrels (Glaucomys sabrinus), bushy-tailed woodrats (...

Simultaneous LC-MS/MS quantitation of acetaminophen and its glucuronide and sulfate metabolites in human dried blood spot samples collected by subjects in a pilot clinical study.

PubMed

Li, Wenkui; Doherty, John P; Kulmatycki, Kenneth; Smith, Harold T; Tse, Francis Ls

2012-06-01

In support of a pilot clinical trial using acetaminophen as the model compound to assess dried blood spot (DBS) sampling as the method for clinical pharmacokinetic sample collection, a novel sensitive LC-MS/MS method was developed and validated for the simultaneous determination of acetaminophen and its major metabolites, acetaminophen glucuronide and sulfate, in human DBS samples collected by subjects via fingerprick. The validated assay dynamic range was from 50.0 to 5000 ng/ml for each compound using a 1/8´´ (3-mm) disc punched from a DBS sample. Baseline separation of the three analytes was achieved to eliminate the possible impact of insource fragmentation of the conjugated metabolites on the analysis of the parent. The overall extraction efficiency was from 61.3 to 78.8% for the three analytes by direct extraction using methanol. The validated method was successfully implemented in the pilot clinical study with the obtained pharmacokinetic parameters in agreement with the values reported in literature.

Evaluation of Off-season Potential Breeding Sources for Spotted Wing Drosophila (Drosophila suzukii Matsumura) in Michigan.

PubMed

Bal, Harit K; Adams, Christopher; Grieshop, Matthew

2017-12-05

It has been suggested that fruit wastes including dropped and unharvested fruits, and fruit byproducts (i.e., pomace) found in fruit plantings and cideries or wine-making facilities could serve as potential off-season breeding sites for spotted wing Drosophila (Drosophila suzukii Matsumura (Diptera: Drosophilidae)). This idea, however, has yet to be widely tested. The goal of our study was to determine the potential of dropped fruit and fruit wastes as Fall spotted wing Drosophila breeding resources in Michigan, USA. Fruit waste samples were collected from 15 farms across the lower peninsula of Michigan and were evaluated for spotted wing Drosophila and other drosophilid emergence and used in host suitability bioassays. All of the dropped apples, pears, grapes, and raspberries and 40% of apple and 100% of grape fruit pomace evaluated were found to contain spotted wing Drosophila with the highest numbers collected from dropped grapes and pears. Greater spotted wing Drosophila recovery was found in fruit wastes at sites attached with cideries and wine-making facilities and with multiple cultivated fruit crops than sites with no cideries and only one crop. Females oviposited in raspberry, pear, apple, grape, apple pomace and grape pomace samples with the highest rates of reproduction in raspberries. Our results demonstrate that fruit wastes including dropped berry, pomme and stone fruits, as well as fruit compost may be important late season reproductive resources for spotted wing Drosophila. © The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Estimation of Daily Sodium and Potassium Excretion Using Spot Urine and 24-Hour Urine Samples in a Black Population (Benin).

PubMed

Mizéhoun-Adissoda, Carmelle; Houehanou, Corine; Chianéa, Thierry; Dalmay, François; Bigot, André; Preux, Pierre-Marie; Bovet, Pascal; Houinato, Dismand; Desport, Jean-Claude

2016-07-01

The 24-hour urine collection method is considered the gold standard for the estimation of ingested potassium and sodium. Because of the impracticalities of collecting all urine over a 24-hour period, spot urine is often used for epidemiological investigations. This study aims to assess the agreement between spot urine and 24-hour urine measurements to determine sodium and potassium intake. A total of 402 participants aged 25 to 64 years were randomly selected in South Benin. Spot urine was taken during the second urination of the day. Twenty-four-hour urine was also collected. Samples (2-mL) were taken and then stored at -20°C. The analysis was carried out using potentiometric dosage. The agreement between spot urine and 24-hour urine measurements was established using Bland-Altman plots. A total of 354 results were analyzed. Daily sodium chloride and potassium chloride urinary excretion means were 10.2±4.9 g/24 h and 2.9±1.4 g/24 h, respectively. Estimated daily sodium chloride and potassium chloride means from the spot urine were 10.7±7.0 g/24 h and 3.9±2.1 g/24 h, respectively. Concordance coefficients were 0.61 at d=-0.5 g, (d±2SD=-11 g and 10.1 g) for sodium chloride and 0.61 at d=-1 g, (d±2SD=-3.8 g and 1.8 g) for potassium chloride. Spot urine method is acceptable for estimating 24-hour urinary sodium and potassium excretion to assess sodium and potassium intake in a black population. However, the confidence interval for the mean difference, which is too large, makes the sodium chloride results inadmissible at a clinical level. © 2015 Wiley Periodicals, Inc.

Correlation of Serum and Dried Blood Spot Results for Quantitation of Schistosoma Circulating Anodic Antigen: a Proof of Principle

PubMed Central

Downs, Jennifer A.; Corstjens, Paul L.A.M.; Mngara, Julius; Lutonja, Peter; Isingo, Raphael; Urassa, Mark; Kornelis, Dieuwke; van Dam, Govert J.

2015-01-01

Circulating Anodic Antigen (CAA) testing is a powerful, increasingly-used tool for diagnosis of active schistosome infection. We sought to determine the feasibility and reliability of measuring CAA in blood spots collected on Whatman 903 Protein Saver cards, which are the predominant filter papers used worldwide for dried blood spot (DBS) research and clinical care. CAA was eluted from blood spots collected from 19 individuals onto Whatman 903 cards in Mwanza, Tanzania, and the assay was optimized to achieve CAA ratios comparable to those obtained from the spots’ corresponding serum samples. The optimized assay was then used to determine the correlation of serum samples (n=16) with DBS from cards that had been stored for 8 years at ambient temperature.Using a DBS volume equivalent to approximately four times the quantity of serum, CAA testing in DBS had a sensitivity of 76% and a specificity of 79% compared to CAA testing in serum. CAA testing was reliable in samples eluted from Whatman 903 cards that had been stored for 8 years at ambient temperature. The overall kappa coefficient was 0.53 (standard error 0.17, p<0.001). We conclude that CAA can be reliably and accurately measured in DBS collected onto the filter paper that is most commonly used for clinical care and research, and that can be stored for prolonged periods of time. This finding opens new avenues for future work among more than 700 million individuals living in areas worldwide in which schistosomes are endemic. PMID:26149541

Diets and foraging behavior of northern spotted owls in Oregon.

Treesearch

Eric D. Forsman; Robert G. Anthony; E. Charles Meslow; Cynthia J. Zabel

2004-01-01

We describe local, regional, and annual variation in diets of northern Spotted Owls (Strix occidaatalis caurina) in Oregon based on 24497 prey collected at 1118 owl territories in 1970-2003. The sample included 91.5% mammals, 4.3% birds, 4.1% insects, and 0.1% other prey. The diet included 2131 species, including 49 mammals, 41 birds, 3 reptiles, 1...

Application of Bayesian methods to habitat selection modeling of the northern spotted owl in California: new statistical methods for wildlife research

Treesearch

Howard B. Stauffer; Cynthia J. Zabel; Jeffrey R. Dunk

2005-01-01

We compared a set of competing logistic regression habitat selection models for Northern Spotted Owls (Strix occidentalis caurina) in California. The habitat selection models were estimated, compared, evaluated, and tested using multiple sample datasets collected on federal forestlands in northern California. We used Bayesian methods in interpreting...

Prevalence of white spot syndrome virus (WSSV) in wild shrimp Penaeus monodon in the Philippines.

PubMed

de la Peña, Leobert D; Lavilla-Pitogo, Celia R; Villar, Corina Belle R; Paner, Milagros G; Sombito, Christopher D; Capulos, Geimbo C

2007-10-15

Prevalence of white spot syndrome virus (WSSV) was determined using polymerase chain reaction (PCR) methodology on DNA extracted from the gills of wild black tiger shrimp Penaeus monodon collected from 7 sampling sites in the Philippines. These 7 sampling sites are the primary sources of spawners and broodstock for hatchery use. During the dry season, WSSV was detected in shrimp from all sites except Bohol, but during the wet season it was not detected in any site except Palawan. None of the WSSV-PCR positive shrimp showed signs of white spots in the cuticle. Prevalence of WSSV showed seasonal variations, i.e. prevalence in dry season (April to May) was higher than in the wet season (August to October). These results suggest that WSSV has already become established in the local marine environment and in wild populations of P. monodon. Thus, broodstock collected during the dry season could serve as the main source of WSSV contamination in shrimp farms due to vertical transmission of the virus in hatcheries.

Validation and Assessment of Three Methods to Estimate 24-h Urinary Sodium Excretion from Spot Urine Samples in High-Risk Elder Patients of Stroke from the Rural Areas of Shaanxi Province

PubMed Central

Ma, Wenxia; Yin, Xuejun; Zhang, Ruijuan; Liu, Furong; Yang, Danrong; Fan, Yameng; Rong, Jie; Tian, Maoyi; Yu, Yan

2017-01-01

Background: 24-h urine collection is regarded as the “gold standard” for monitoring sodium intake at the population level, but ensuring high quality urine samples is difficult to achieve. The Kawasaki, International Study of Sodium, Potassium, and Blood Pressure (INTERSALT) and Tanaka methods have been used to estimate 24-h urinary sodium excretion from spot urine samples in some countries, but few studies have been performed to compare and validate these methods in the Chinese population. Objective: To compare and validate the Kawasaki, INTERSALT and Tanaka formulas in predicting 24-h urinary sodium excretion using spot urine samples in 365 high-risk elder patients of strokefrom the rural areas of Shaanxi province. Methods: Data were collected from a sub-sample of theSalt Substitute and Stroke Study. 365 high-risk elder patients of stroke from the rural areas of Shaanxi province participated and their spot and 24-h urine specimens were collected. The concentrations of sodium, potassium and creatinine in spot and 24-h urine samples wereanalysed. Estimated 24-h sodium excretion was predicted from spot urine concentration using the Kawasaki, INTERSALT, and Tanaka formulas. Pearson correlation coefficients and agreement by Bland-Altman method were computed for estimated and measured 24-h urinary sodium excretion. Results: The average 24-h urinary sodium excretion was 162.0 mmol/day, which representing a salt intake of 9.5 g/day. Three predictive equations had low correlation with the measured 24-h sodium excretion (r = 0.38, p < 0.01; ICC = 0.38, p < 0.01 for the Kawasaki; r = 0.35, p < 0.01; ICC = 0.31, p < 0.01 for the INTERSALT; r = 0.37, p < 0.01; ICC = 0.34, p < 0.01 for the Tanaka). Significant biases between estimated and measured 24-h sodium excretion were observed (all p < 0.01 for three methods). Among the three methods, the Kawasaki method was the least biased compared with the other two methods (mean bias: 31.90, 95% Cl: 23.84, 39.97). Overestimation occurred when the Kawasaki and Tanaka methods were used while the INTERSALT method underestimated 24-h sodium excretion. Conclusion: The Kawasaki, INTERSALT and Tanaka methods for estimation of 24-h urinary sodium excretion from spot urine specimens were inadequate for the assessment of sodium intake at the population level in high-risk elder patients of stroke from the rural areas of Shaanxi province, although the Kawasaki method was the least biased compared with the other two methods. PMID:29019912

Validation and Assessment of Three Methods to Estimate 24-h Urinary Sodium Excretion from Spot Urine Samples in High-Risk Elder Patients of Stroke from the Rural Areas of Shaanxi Province.

PubMed

Ma, Wenxia; Yin, Xuejun; Zhang, Ruijuan; Liu, Furong; Yang, Danrong; Fan, Yameng; Rong, Jie; Tian, Maoyi; Yu, Yan

2017-10-11

Background : 24-h urine collection is regarded as the "gold standard" for monitoring sodium intake at the population level, but ensuring high quality urine samples is difficult to achieve. The Kawasaki, International Study of Sodium, Potassium, and Blood Pressure (INTERSALT) and Tanaka methods have been used to estimate 24-h urinary sodium excretion from spot urine samples in some countries, but few studies have been performed to compare and validate these methods in the Chinese population. Objective : To compare and validate the Kawasaki, INTERSALT and Tanaka formulas in predicting 24-h urinary sodium excretion using spot urine samples in 365 high-risk elder patients of strokefrom the rural areas of Shaanxi province. Methods : Data were collected from a sub-sample of theSalt Substitute and Stroke Study. 365 high-risk elder patients of stroke from the rural areas of Shaanxi province participated and their spot and 24-h urine specimens were collected. The concentrations of sodium, potassium and creatinine in spot and 24-h urine samples wereanalysed. Estimated 24-h sodium excretion was predicted from spot urine concentration using the Kawasaki, INTERSALT, and Tanaka formulas. Pearson correlation coefficients and agreement by Bland-Altman method were computed for estimated and measured 24-h urinary sodium excretion. Results : The average 24-h urinary sodium excretion was 162.0 mmol/day, which representing a salt intake of 9.5 g/day. Three predictive equations had low correlation with the measured 24-h sodium excretion (r = 0.38, p < 0.01; ICC = 0.38, p < 0.01 for the Kawasaki; r = 0.35, p < 0.01; ICC = 0.31, p < 0.01 for the INTERSALT; r = 0.37, p < 0.01; ICC = 0.34, p < 0.01 for the Tanaka). Significant biases between estimated and measured 24-h sodium excretion were observed (all p < 0.01 for three methods). Among the three methods, the Kawasaki method was the least biased compared with the other two methods (mean bias: 31.90, 95% Cl: 23.84, 39.97). Overestimation occurred when the Kawasaki and Tanaka methods were used while the INTERSALT method underestimated 24-h sodium excretion. Conclusion : The Kawasaki, INTERSALT and Tanaka methods for estimation of 24-h urinary sodium excretion from spot urine specimens were inadequate for the assessment of sodium intake at the population level in high-risk elder patients of stroke from the rural areas of Shaanxi province, although the Kawasaki method was the least biased compared with the other two methods.

Effects of reservoir hydrology on reproduction by largemouth bass and spotted bass in Normandy Reservoir, Tennessee

USGS Publications Warehouse

Sammons, S.M.; Dorsey, L.G.; Bettoli, P.W.; Fiss, F.C.

1999-01-01

Age-O largemouth bass Micropterus salmoides and spotted bass M. punctulatus were collected from Normandy Reservoir, Tennessee, 1992-1996, to evaluate effects of reservoir hydrology and hatching of shad Dorosoma spp. on hatching and first-year growth and survival of these two species. Fish were collected in cove rotenone samples in early August and electrofishing samples biweekly throughout the summer; hatch dates and age-specific growth for both species were determined from cove samples with sagittal otoliths. Hatching of both species ranged from early April to early June. Initiation of largemouth bass spawning, but not spotted bass spawning, was positively related to the first day water levels achieved full pool. Mean hatch dates of both species were positively related to the first day of full pool. Timing of spawning for both species was not related to water temperature, Largemouth bass exhibited bimodal length-frequency distributions by midsummer in two wet years and length frequencies were unimodal in dry years; spotted bass always formed unimodal length-frequency distributions. When largemouth bass exhibited bimodal length distributions, earlier hatched fish grew faster than later hatched fish. Spotted bass grew at similar rates, regardless of hatch date, every year except during 1992 when later hatched fish grew faster than earlier hatched fish. Weekly survival of largemouth bass in their first summer was positively related to reservoir water level. First-year growth of both species was not directly affected by the timing of threadfin shad D. petenense or gizzard shad D. cepedianum hatching.

Reliable Quantification of the Potential for Equations Based on Spot Urine Samples to Estimate Population Salt Intake: Protocol for a Systematic Review and Meta-Analysis.

PubMed

Huang, Liping; Crino, Michelle; Wu, Jason Hy; Woodward, Mark; Land, Mary-Anne; McLean, Rachael; Webster, Jacqui; Enkhtungalag, Batsaikhan; Nowson, Caryl A; Elliott, Paul; Cogswell, Mary; Toft, Ulla; Mill, Jose G; Furlanetto, Tania W; Ilich, Jasminka Z; Hong, Yet Hoi; Cohall, Damian; Luzardo, Leonella; Noboa, Oscar; Holm, Ellen; Gerbes, Alexander L; Senousy, Bahaa; Pinar Kara, Sonat; Brewster, Lizzy M; Ueshima, Hirotsugu; Subramanian, Srinivas; Teo, Boon Wee; Allen, Norrina; Choudhury, Sohel Reza; Polonia, Jorge; Yasuda, Yoshinari; Campbell, Norm Rc; Neal, Bruce; Petersen, Kristina S

2016-09-21

Methods based on spot urine samples (a single sample at one time-point) have been identified as a possible alternative approach to 24-hour urine samples for determining mean population salt intake. The aim of this study is to identify a reliable method for estimating mean population salt intake from spot urine samples. This will be done by comparing the performance of existing equations against one other and against estimates derived from 24-hour urine samples. The effects of factors such as ethnicity, sex, age, body mass index, antihypertensive drug use, health status, and timing of spot urine collection will be explored. The capacity of spot urine samples to measure change in salt intake over time will also be determined. Finally, we aim to develop a novel equation (or equations) that performs better than existing equations to estimate mean population salt intake. A systematic review and meta-analysis of individual participant data will be conducted. A search has been conducted to identify human studies that report salt (or sodium) excretion based upon 24-hour urine samples and spot urine samples. There were no restrictions on language, study sample size, or characteristics of the study population. MEDLINE via OvidSP (1946-present), Premedline via OvidSP, EMBASE, Global Health via OvidSP (1910-present), and the Cochrane Library were searched, and two reviewers identified eligible studies. The authors of these studies will be invited to contribute data according to a standard format. Individual participant records will be compiled and a series of analyses will be completed to: (1) compare existing equations for estimating 24-hour salt intake from spot urine samples with 24-hour urine samples, and assess the degree of bias according to key demographic and clinical characteristics; (2) assess the reliability of using spot urine samples to measure population changes in salt intake overtime; and (3) develop a novel equation that performs better than existing equations to estimate mean population salt intake. The search strategy identified 538 records; 100 records were obtained for review in full text and 73 have been confirmed as eligible. In addition, 68 abstracts were identified, some of which may contain data eligible for inclusion. Individual participant data will be requested from the authors of eligible studies. Many equations for estimating salt intake from spot urine samples have been developed and validated, although most have been studied in very specific settings. This meta-analysis of individual participant data will enable a much broader understanding of the capacity for spot urine samples to estimate population salt intake.

Determination of Efavirenz in Human Dried Blood Spots by Reversed-Phase High Performance Liquid Chromatography with UV Detection

PubMed Central

Hoffman, Justin T; Rossi, Steven S; Espina-Quinto, Rowena; Letendre, Scott; Capparelli, Edmund V

2013-01-01

Background Previously published methods for determination of efavirenz (EFV) in human dried blood spots (DBS) employ costly and complex liquid chromatography/mass spectrometry. We describe the validation and evaluation of a simple and inexpensive high-performance liquid chromatography (HPLC) method for EFV quantification in human DBS and dried plasma spots (DPS), using ultraviolet (UV) detection appropriate for resource-limited settings. Methods 100μl of heparinized whole blood or plasma were spotted onto blood collection cards, dried, punched, and eluted. Eluates are injected onto a C-18 reversed phase HPLC column. EFV is separated isocratically using a potassium phosphate and ACN mobile phase. UV detection is at 245nm. Quantitation is by use of external calibration standards. Following validation, the method was evaluated using whole blood and plasma from HIV-positive patients undergoing EFV therapy. Results Mean recovery of drug from dried blood spots is 91.5%. The method is linear over the validated concentration range of 0.3125 – 20.0μg/mL. A good correlation (Spearman r=0.96) between paired plasma and DBS EFV concentrations from the clinical samples was observed, and hematocrit level was not found to be a significant determinant of the EFV DBS level. The mean observed CDBS/Cplasma ratio was 0.68. A good correlation (Spearman r=0.96) between paired plasma and DPS EFV concentrations from the clinical samples was observed. The mean percent deviation of DPS samples from plasma samples is 1.68%. Conclusions Dried whole blood spot or dried plasma spot sampling is well suited for monitoring EFV therapy in resource limited settings, particularly when high sensitivity is not essential. PMID:23503446

Dipstick Spot urine pH does not accurately represent 24 hour urine PH measured by an electrode.

PubMed

Omar, Mohamed; Sarkissian, Carl; Jianbo, Li; Calle, Juan; Monga, Manoj

2016-01-01

To determine whether spot urine pH measured by dipstick is an accurate representation of 24 hours urine pH measured by an electrode. We retrospectively reviewed urine pH results of patients who presented to the urology stone clinic. For each patient we recorded the most recente pH result measured by dipstick from a spot urine sample that preceded the result of a 24-hour urine pH measured by the use of a pH electrode. Patients were excluded if there was a change in medications or dietary recommendations or if the two samples were more than 4 months apart. A difference of more than 0.5 pH was considered na inaccurate result. A total 600 patients were retrospectively reviewed for the pH results. The mean difference in pH between spot urine value and the 24 hours collection values was 0.52±0.45 pH. Higher pH was associated with lower accuracy (p<0.001). The accuracy of spot urine samples to predict 24-hour pH values of <5.5 was 68.9%, 68.2% for 5.5 to 6.5 and 35% for >6.5. Samples taken more than 75 days apart had only 49% the accuracy of more recent samples (p<0.002). The overall accuracy is lower than 80% (p<0.001). Influence of diurnal variation was not significant (p=0.588). Spot urine pH by dipstick is not an accurate method for evaluation of the patients with urolithiasis. Patients with alkaline urine are more prone to error with reliance on spot urine pH.

Outbreak of hepatitis E virus infection in Darfur, Sudan: effectiveness of real-time reverse transcription-PCR analysis of dried blood spots.

PubMed

Mérens, Audrey; Guérin, Philippe Jean; Guthmann, Jean-Paul; Nicand, Elisabeth

2009-06-01

Biological samples collected in refugee camps during an outbreak of hepatitis E were used to compare the accuracy of hepatitis E virus RNA amplification by real-time reverse transcription-PCR (RT-PCR) for sera and dried blood spots (concordance of 90.6%). Biological profiles (RT-PCR and serology) of asymptomatic individuals were also analyzed.

Comparison of population iodine estimates from 24-hour urine and timed-spot urine samples.

PubMed

Perrine, Cria G; Cogswell, Mary E; Swanson, Christine A; Sullivan, Kevin M; Chen, Te-Ching; Carriquiry, Alicia L; Dodd, Kevin W; Caldwell, Kathleen L; Wang, Chia-Yih

2014-04-01

Median urine iodine concentration (UIC; μg/L) in spot urine samples is recommended for monitoring population iodine status. Other common measures are iodine:creatinine ratio (I/Cr; μg/g) and estimated 24-hour urine iodine excretion (UIE; I/Cr × predicted 24-hour Cr; μg/day). Despite different units, these measures are often used interchangeably, and it is unclear how they compare with the reference standard 24-hour UIE. Volunteers aged 18-39 years collected all their urine samples for 24 hours (n=400). Voids from morning, afternoon, evening, overnight, and a composite 24-hour sample were analyzed for iodine. We calculated median observed 24-hour UIE and 24-hour UIC, and spot UIC, I/Cr, and two measures of estimated UIE calculated using predicted 24-hour Cr from published estimates by Kesteloot and Joosens (varies by age and sex) and published equations by Mage et al. (varies by age, sex, race, and anthropometric measures). We examined mean differences and relative difference across iodine excretion levels using Bland-Altman plots. Median 24-hour UIE was 173.6 μg/day and 24-hour UIC was 144.8 μg/L. From timed-spot urine samples, estimates were: UIC 147.3-156.2 μg/L; I/Cr 103.6-114.3 μg/g, estimated 24-hour UIE (Kesteloot and Joosens) 145.7-163.3 μg/day; and estimated 24-hour UIE (Mage) 176.5-187.7 μg/day. Iodine measures did not vary consistently by timing of spot urine collection. Compared with observed 24-hour UIE, on average, estimated (Mage) 24-hour UIE was not significantly different, while estimated 24-hour UIE (Kesteloot and Joosens) was significantly different for some ethnicity/sex groups. Compared with 24-hour UIC, on average, spot UIC did not differ. Estimates of UIC, I/Cr, and estimated 24-hour UIE (I/Cr × predicted 24-hour Cr) from spot urine samples should not be used interchangeably. Estimated 24-hour UIE, where predicted 24-hour Cr varies by age, sex, ethnicity, and anthropometric measures and was calculated with prediction equations using data from the sample, was more comparable to observed 24-hour UIE than when predicted 24-hour Cr was from published estimates from a different population. However, currently no cutoffs exist to interpret population estimated 24-hour UIE values.

Cost-effective antigen testing for delimitation, monitoring and evaluation in bancroftian filariasis.

PubMed

Das, L K; Pani, S P; Vanamail, P; Vijayalakshmi, G; Debritto, L J

2012-01-31

This study was focussed on identifying a cost-effective method for delimitation, monitoring and evaluation in bancroftian filariasis. Finger prick blood samples were collected between 20.00 and 23.00 hours for the detection of microfilariae (mf) from the available population in a village which was endemic for lymphatic filariasis. Simultaneously, from each individual, four spots of 25-μl blood samples were collected on Whatman number 3 filter paper and air dried. Dried filter paper spots were pooled in quantities of 1, 5, 10, 15, 20 and 25 on unknown and simulated mf and antigen prevalence. Pooled samples were assayed for circulating filarial antigen (CFA) using TropBIO Og4C3 ELISA kits. The community mf and CFA rates were 3.4% and 25.9%, respectively. The pool sizes of 20 and 25 showed CFA positivity in all the above categories tested. The results of the pooled blood spot samples suggest that, in areas with mf and CFA prevalence rates between 1 and 10%, pools of 20 or 25 could be considered as the ideal pool size for the detection of filarial infection in the community. CFA prevalence at the level of 5-6% following desirable rounds of mass drug administration (MDA) indicates that the community mf prevalence is likely to be at the 1% level.

Importing, Working With, and Sharing Microstructural Data in the StraboSpot Digital Data System, Including an Example Dataset from the Pilbara Craton, Western Australia.

NASA Astrophysics Data System (ADS)

Roberts, N.; Cunningham, H.; Snell, A.; Newman, J.; Tikoff, B.; Chatzaras, V.; Walker, J. D.; Williams, R. T.

2017-12-01

There is currently no repository where a geologist can survey microstructural datasets that have been collected from a specific field area or deformation experiment. New development of the StraboSpot digital data system provides a such a repository as well as visualization and analysis tools. StraboSpot is a graph database that allows field geologists to share primary data and develop new types of scientific questions. The database can be accessed through: 1) a field-based mobile application that runs on iOS and Android mobile devices; and 2) a desktop system. We are expanding StraboSpot to include the handling of a variety of microstructural data types. Presented here is the detailed vocabulary and logic used for the input of microstructural data, and how this system operates with the anticipated workflow of users. Microstructural data include observations and interpretations from photomicrographs, scanning electron microscope images, electron backscatter diffraction, and transmission electron microscopy data. The workflow for importing microstructural data into StraboSpot is organized into the following tabs: Images, Mineralogy & Composition; Sedimentary; Igneous; Metamorphic; Fault Rocks; Grain size & configuration; Crystallographic Preferred Orientation; Reactions; Geochronology; Relationships; and Interpretations. Both the sample and the thin sections are also spots. For the sample spot, the user can specify whether a sample is experimental or natural; natural samples are inherently linked to their field context. For the thin section (sub-sample) spot, the user can select between different options for sample preparation, geometry, and methods. A universal framework for thin section orientation is given, which allows users to overlay different microscope images of the same area and keeps georeferenced orientation. We provide an example dataset of field and microstructural data from the Mt Edgar dome, a granitic complex in the Paleoarchean East Pilbara craton, Australia. StraboSpot provides a single place for georeferenced geologic data at every spatial scale, in which data are interconnected. Incorporating microstructural data into an open-access platform will give field and experimental geologists a library of microstructural data across a range of tectonic and experimental contexts.

Filter Paper Blood Spot Enzyme Linked Immunoassay for Adiponectin and Application in the Evaluation of Determinants of Child Insulin Sensitivity

PubMed Central

Martin, Richard M.; Patel, Rita; Oken, Emily; Thompson, Jennifer; Zinovik, Alexander; Kramer, Michael S.; Vilchuck, Konstantin; Bogdanovich, Natalia; Sergeichick, Natalia; Foo, Ying; Gusina, Nina

2013-01-01

Background Adiponectin is an adipocyte-derived hormone that acts as a marker of insulin sensitivity. Bloodspot sampling by fingerstick onto filter paper may increase the feasibility of large-scale studies of the determinants of insulin sensitivity. We first describe the validation of an enzyme-linked immunoassay (ELISA) for quantifying adiponectin from dried blood spots and then demonstrate its application in a large trial (PROBIT). Methods We quantified adiponectin from 3-mm diameter discs (≈3 µL of blood) punched from dried blood spots obtained from: i) whole blood standards (validation); and ii) PROBIT trial samples (application) in which paediatricians collected blood spots from 13,879 children aged 11.5 years from 31 sites across Belarus. We examined the distribution of bloodspot adiponectin by demographic and anthropometric factors, fasting insulin and glucose. Results In the validation study, mean intra-assay coefficients of variation (n = 162) were 15%, 13% and 10% for ‘low’ (6.78 µg/ml), ‘medium’ (18.18 µg/ml) and 'high’ (33.13 µg/ml) internal quality control (IQC) samples, respectively; the respective inter-assay values (n = 40) were 23%, 21% and 14%. The correlation coefficient between 50 paired whole bloodspot versus plasma samples, collected simultaneously, was 0.87 (95% CI: 0.78 to 0.93). Recovery of known quantities of adiponectin (between 4.5 to 36 µg/ml) was 100.3–133%. Bloodspot adiponectin was stable for at least 30 months at −80°C. In PROBIT, we successfully quantified fasting adiponectin from dried blood spots in 13,329 of 13,879 (96%) children. Mean adiponectin (standard deviation) concentrations were 17.34 µg/ml (7.54) in boys and 18.41 µg/ml (7.92) in girls and were inversely associated with body mass index, fat mass, triceps and subscapular skin-fold thickness, waist circumference, height and fasting glucose. Conclusions Bloodspot ELISA is suitable for measuring adiponectin in very small volumes of blood collected on filter paper and can be applied to large-scale studies. PMID:23936498

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

PubMed Central

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

2014-01-01

The quantification of antituberculosis drug concentrations in multinational trials currently requires the collection of modest blood volumes, centrifugation, aliquoting of plasma, freezing, and keeping samples frozen during shipping. We prospectively enrolled healthy individuals into the Tuberculosis Trials Consortium Study 29B, a phase I dose escalation study of rifapentine, a rifamycin under evaluation in tuberculosis treatment trials. We developed a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantifying rifapentine in whole blood on dried blood spots (DBS) to facilitate pharmacokinetic/pharmacodynamic analyses in clinical trials. Paired plasma and whole-blood samples were collected by venipuncture, and whole blood was spotted on Whatman protein saver 903 cards. The methods were optimized for plasma and then validated for DBS. The analytical measuring range for quantification of rifapentine and its metabolite was 50 to 80,000 ng/ml in whole-blood DBS. The analyte was stable on the cards for 11 weeks with a desiccant at room temperature and protected from light. The method concordance for paired plasma and whole-blood DBS samples was determined after correcting for participant hematocrit or population-based estimates of bias from Bland-Altman plots. The application of either correction factor resulted in acceptable correlation between plasma and whole-blood DBS (Passing-Bablok regression corrected for hematocrit; y = 0.98x + 356). Concentrations of rifapentine may be determined from whole-blood DBS collected via venipuncture after normalization in order to account for the dilutional effects of red blood cells. Additional studies are focused on the application of this methodology to capillary blood collected by finger stick. The simplicity of processing, storage, shipping, and low blood volume makes whole-blood DBS attractive for rifapentine pharmacokinetic evaluations, especially in international and pediatric trials. PMID:25182637

Stability of Proteins in Dried Blood Spot Biobanks*

PubMed Central

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

2017-01-01

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

Effectiveness of a Communication for Behavioral Impact (COMBI) Intervention to Reduce Salt Intake in a Vietnamese Province Based on Estimations From Spot Urine Samples.

PubMed

Do, Ha Thi Phuong; Santos, Joseph Alvin; Trieu, Kathy; Petersen, Kristina; Le, Mai Bach; Lai, Duc Truong; Bauman, Adrian; Webster, Jacqui

2016-11-01

This study evaluated the effectiveness of the Communication for Behavioral Impact (COMBI)-Eat Less Salt intervention conducted in Viet Tri, Vietnam. The behavior change intervention was implemented in four wards and four communes for one year, which included mass media communication, school interventions, community programs, and focus on high-risk groups. Mean sodium excretion was estimated from spot urine samples using different equations. A subsample provided 24-hour urine to validate estimates from spot urine. Information about salt-related knowledge and behaviors was also collected. There were 513 participants at both baseline and follow-up. Mean sodium excretion estimated from spot urines fell significantly from 8.48 g/d at baseline to 8.05 g/d at follow-up (P=.001). All spot equations demonstrated a significant reduction in sodium levels; however, the change was smaller than the measured 24-hour urine. Participants showed improved knowledge and behaviors following the intervention. The COMBI intervention was effective in lowering average population salt intake and improving knowledge and behaviors. ©2016 Wiley Periodicals, Inc.

Use of Dried Capillary Blood Sampling for Islet Autoantibody Screening in Relatives: A Feasibility Study.

PubMed

Bingley, Polly J; Rafkin, Lisa E; Matheson, Della; Steck, Andrea K; Yu, Liping; Henderson, Courtney; Beam, Craig A; Boulware, David C

2015-12-01

Islet autoantibody testing provides the basis for assessment of risk of progression to type 1 diabetes. We set out to determine the feasibility and acceptability of dried capillary blood spot-based screening to identify islet autoantibody-positive relatives potentially eligible for inclusion in prevention trials. Dried blood spot (DBS) and venous samples were collected from 229 relatives participating in the TrialNet Pathway to Prevention Study. Both samples were tested for glutamic acid decarboxylase, islet antigen 2, and zinc transporter 8 autoantibodies, and venous samples were additionally tested for insulin autoantibodies and islet cell antibodies. We defined multiple autoantibody positive as two or more autoantibodies in venous serum and DBS screen positive if one or more autoantibodies were detected. Participant questionnaires compared the sample collection methods. Of 44 relatives who were multiple autoantibody positive in venous samples, 42 (95.5%) were DBS screen positive, and DBS accurately detected 145 of 147 autoantibody-negative relatives (98.6%). Capillary blood sampling was perceived as more painful than venous blood draw, but 60% of participants would prefer initial screening using home fingerstick with clinic visits only required if autoantibodies were found. Capillary blood sampling could facilitate screening for type 1 diabetes prevention studies.

Determination of efavirenz in human dried blood spots by reversed-phase high-performance liquid chromatography with UV detection.

PubMed

Hoffman, Justin T; Rossi, Steven S; Espina-Quinto, Rowena; Letendre, Scott; Capparelli, Edmund V

2013-04-01

Previously published methods for determination of efavirenz (EFV) in human dried blood spots (DBS) use costly and complex liquid chromatography/mass spectrometry. We describe the validation and evaluation of a simple and inexpensive high-performance liquid chromatography method for EFV quantification in human DBS and dried plasma spots (DPS), using ultraviolet detection appropriate for resource-limited settings. One hundred microliters of heparinized whole blood or plasma were spotted onto blood collection cards, dried, punched, and eluted. Eluates are injected onto a C-18 reversed phase high-performance liquid chromatography column. EFV is separated isocratically using a potassium phosphate and acetonitrile mobile phase. Ultraviolet detection is at 245 nm. Quantitation is by use of external calibration standards. Following validation, the method was evaluated using whole blood and plasma from HIV-positive patients undergoing EFV therapy. Mean recovery of drug from DBS is 91.5%. The method is linear over the validated concentration range of 0.3125-20.0 μg/mL. A good correlation (Spearman r = 0.96) between paired plasma and DBS EFV concentrations from the clinical samples was observed, and hematocrit level was not found to be a significant determinant of the EFV DBS level. The mean observed C DBS/C plasma ratio was 0.68. A good correlation (Spearman r = 0.96) between paired plasma and DPS EFV concentrations from the clinical samples was observed. The mean percent deviation of DPS samples from plasma samples is 1.68%. Dried whole blood spot or dried plasma spot sampling is well suited for monitoring EFV therapy in resource-limited settings, particularly when high sensitivity is not essential.

Filter Paper Blood Spot Enzyme Linked Immunoassay for Insulin and Application in the Evaluation of Determinants of Child Insulin Resistance

PubMed Central

Martin, Richard M.; Patel, Rita; Zinovik, Alexander; Kramer, Michael S.; Oken, Emily; Vilchuck, Konstantin; Bogdanovich, Natalia; Sergeichick, Natalia; Gunnarsson, Robert; Grufman, Lisa; Foo, Ying; Gusina, Nina

2012-01-01

Background In large-scale epidemiology, bloodspot sampling by fingerstick onto filter paper has many advantages, including ease and low costs of collection, processing and transport. We describe the development of an enzyme-linked immunoassay (ELISA) for quantifying insulin from dried blood spots and demonstrate its application in a large trial. Methods We adapted an existing commercial kit (Mercodia Human Insulin ELISA, 10-1113-01) to quantify insulin from two 3-mm diameter discs (≈6 µL of blood) punched from whole blood standards and from trial samples. Paediatricians collected dried blood spots in a follow-up of 13,879 fasted children aged 11.5 years (interquartile range 11.3–11.8 years) from 31 trial sites across Belarus. We quantified bloodspot insulin levels and examined their distribution by demography and anthropometry. Results Mean intra-assay (n = 157) coefficients of variation were 15% and 6% for ‘low’ (6.7 mU/L) and ‘high’ (23.1 mU/L) values, respectively; the respective inter-assay values (n = 33) were 23% and 11%. The intraclass correlation coefficient between 50 paired whole bloodspot versus serum samples, collected simultaneously, was 0.90 (95% confidence interval 0.85 to 0.95). Bloodspot insulin was stable for at least 31 months at −80°C, for one week at +30°C and following four freeze-thaw cycles. Paediatricians collected a median of 8 blood spots from 13,487 (97%) children. The geometric mean insulin (log standard deviation) concentrations amongst 12,812 children were 3.0 mU/L (1.1) in boys and 4.0 mU/L (1.0) in girls and were positively associated with pubertal stage, measures of central and peripheral adiposity, height and fasting glucose. Conclusions Our simple and convenient bloodspot assay is suitable for the measurement of insulin in very small volumes of blood collected on filter paper cards and can be applied to large-scale epidemiology studies of the early-life determinants of circulating insulin. PMID:23056434

Analysis of the stability of urea in dried blood spots collected and stored on filter paper.

PubMed

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

2013-05-01

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

Prevalence and distribution of White Spot Syndrome Virus in cultured shrimp.

PubMed

Hossain, A; Nandi, S P; Siddique, M A; Sanyal, S K; Sultana, M; Hossain, M A

2015-02-01

White Spot Syndrome Virus (WSSV) is a dsDNA virus causing White Spot Syndrome Disease (WSSD) in shrimp with almost 100% morality rate within 3-10 days. In Bangladesh, WSSD is one of the major impediments of shrimp farming. This study first investigated the prevalence and distribution of WSSV in cultured shrimps of the coastal regions in Bangladesh. A total of 60 shrimp samples, collected from the 25 shrimp farms of different coastal regions (Satkhira, Khulna, Bagerhat and Cox's Bazar), were analysed during 2013-2014 by conventional PCR using VP28 and VP664 gene-specific primers; 39 of 60 samples were found WSSV positive. SYBR green real-time PCR using 71-bp amplicon for VP664 gene correlated well with conventional PCR data. The prevalence rates of WSSV among the collected 60 samples were Satkhira 79%, Khulna 50%, Bagerhat 38% and Cox's Bazar 25%. Sequencing of WSSV-positive PCR amplicons of VP28 showed 99% similarity with WSSV NCBI Ref/Seq Sequences. Molecular analysis of the VP28 gene sequences of WSSV revealed that Bangladeshi strains phylogenetically affiliated to the strains belong to India. This work concluded that WSSV infections are widely distributed in the coastal regions cultured shrimp in Bangladesh. © 2014 The Society for Applied Microbiology.

Self-clearance from BVDV infections--a frequent finding in dairy herds in an endemically infected region in Peru.

PubMed

Ståhl, K; Lindberg, A; Rivera, H; Ortiz, C; Moreno-López, J

2008-03-17

In this cross-sectional study, a stratified two-stage random sampling procedure was employed to select 221 dairy herds for bulk tank milk (BTM) sampling, and a subset of 55 dairy herds for individual blood sampling of a number of young animals (spot test), to predict presence or absence of current BVDV infection, and for data collection. The prediction was based on the high probability of seropositivity in groups of animals where PI animals are present because of the efficient spread of virus from PI animals to the surrounding group. BTM samples were collected in August 2003 (n=192) and February 2004 (n=195), and the 55 herds selected for spot testing and data collection were visited in December 2003. All samples were tested for presence of BVDV specific antibodies using a commercial indirect ELISA (SVANOVA Biotech AB, Uppsala, Sweden). The results demonstrated a very high level of exposure to BVDV in the region, and the proportion of herds with high antibody levels in the BTM was above 95% on both occasions. Despite this, almost two thirds of the herds had spot test results indicating absence of current infection, suggesting a high probability of self-clearance. A logistic regression model with the results from the spot tests as dependent variable was used to investigate possible herd and management factors associated with self-clearance, and suggested that this may occur regardless of herd size. Even though it is well established that the process of identification and elimination of PI animals is required within a systematic BVDV eradication programme, the present study strongly suggests that many herds may be cleared without intervention even in regions with high cattle density and high BVDV prevalence. Consequently, in any BVDV infected population (regardless of the herd-level BVDV seroprevalence), and at any given point of time, a large proportion of the herds will be free from infection due to self-clearance. Self-clearance is therefore a process that works in favour of any effort to control BVDV, which should be taken into account when planning and assessing the cost-effectiveness of a systematic control programme.

Optimised laser microdissection of the human ocular surface epithelial regions for microarray studies

PubMed Central

2013-01-01

Background The most important challenge of performing insitu transcriptional profiling of the human ocular surface epithelial regions is obtaining samples in sufficient amounts, without contamination from adjacent tissue, as the region of interest is microscopic and closely apposed to other tissues regions. We have effectively collected ocular surface (OS) epithelial tissue samples from the Limbal Epithelial Crypt (LEC), limbus, cornea and conjunctiva of post-mortem cadaver eyes with laser microdissection (LMD) technique for gene expression studies with spotted oligonucleotide microarrays and Gene 1.0 ST arrays. Methods Human donor eyes (4 pairs for spotted oligonucleotide microarrays, 3 pairs for Gene 1.0 ST arrays) consented for research were included in this study with due ethical approval of the Nottingham Research Ethics Committee. Eye retrieval was performed within 36 hours of post-mortem period. The dissected corneoscleral buttons were immersed in OCT media and frozen in liquid nitrogen and stored at −80°C till further use. Microscopic tissue sections of interest were taken on PALM slides and stained with Toluidine Blue for laser microdissection with PALM microbeam systems. Optimisation of the laser microdissection technique was crucial for efficient and cost effective sample collection. Results The starting concentration of RNA as stipulated by the protocol of microarray platforms was taken as the cut-off concentration of RNA samples in our studies. The area of LMD tissue processed for spotted oligonucleotide microarray study ranged from 86,253 μm2 in LEC to 392,887 μm2 in LEC stroma. The RNA concentration of the LMD samples ranged from 22 to 92 pg/μl. The recommended starting concentration of the RNA samples used for Gene 1.0 ST arrays was 6 ng/5 μl. To achieve the desired RNA concentration the area of ocular surface epithelial tissue sample processed for the Gene 1.0 ST array experiments was approximately 100,0000 μm2 to 130,0000 μm2. RNA concentration of these samples ranged from 10.88 ng/12 μl to 25.8 ng/12 μl, with the RNA integrity numbers (RIN) for these samples from 3.3 to 7.9. RNA samples with RIN values below 2, that had failed to amplify satisfactorily were discarded. Conclusions The optimised protocol for sample collection and laser microdissection improved the RNA yield of the insitu ocular surface epithelial regions for effective microarray studies on spotted oligonucleotide and affymetrix platforms. PMID:24160452

Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking.

PubMed

Choi, Eun-Hye; Lee, Sang Kwang; Ihm, Chunhwa; Sohn, Young-Hak

2014-12-01

Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper. The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used. DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening. Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

White spot syndrome virus isolates of tiger shrimp Penaeus monodon (Fabricious) in India are similar to exotic isolates as revealed by polymerase chain reaction and electron microscopy.

PubMed

Mishra, S S; Shekhar, M S

2005-07-01

Microbiological analysis of samples collected from cases of white spot disease outbreaks in cultured shrimp in different farms located in three regions along East Coast of India viz. Chidambram (Tamil Nadu), Nellore (Andhra Pradesh) and Balasore (Orissa), revealed presence of Vibrio alginolyticus, Vibrio parahaemolyticus, and Aeromonas spp. but experimental infection trials in Penaeus monodon with these isolates did not induce any acute mortality or formation of white spots on carapace. Infection trials using filtered tissue extracts by oral and injection method induced mortality in healthy P. monodon with all samples and 100% mortality was noted by the end of 7 day post-inoculation. Histopathological analysis demonstrated degenerated cells characterized by hypertrophied nuclei in gills, hepatopancreas and lymphoid organ with presence of intranuclear basophilic or eosino-basophilic bodies in tubular cells and intercellular spaces. Analysis of samples using 3 different primer sets as used by other for detection of white spot syndrome virus (WSSV) generated 643, 1447 and 520bp amplified DNA products in all samples except in one instance. Variable size virions with mean size in the range of 110 x 320 +/- 20 nm were observed under electron microscope. It could be concluded that the viral isolates in India involved with white spot syndrome in cultured shrimp are similar to RV-PJ and SEMBV in Japan, WSBV in Taiwan and WSSV in Malaysia, Indonesia, Thailand, China and Japan.

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

PubMed

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

2013-03-01

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

Analysis of the variability of human normal urine by 2D-GE reveals a "public" and a "private" proteome.

PubMed

Molina, Laurence; Salvetat, Nicolas; Ameur, Randa Ben; Peres, Sabine; Sommerer, Nicolas; Jarraya, Fayçal; Ayadi, Hammadi; Molina, Franck; Granier, Claude

2011-12-10

The characterization of the normal urinary proteome is steadily progressing and represents a major interest in the assessment of clinical urinary biomarkers. To estimate quantitatively the variability of the normal urinary proteome, urines of 20 healthy people were collected. We first evaluated the impact of the sample conservation temperature on urine proteome integrity. Keeping the urine sample at RT or at +4°C until storage at -80°C seems the best way for long-term storage of samples for 2D-GE analysis. The quantitative variability of the normal urinary proteome was estimated on the 20 urines mapped by 2D-GE. The occurrence of the 910 identified spots was analysed throughout the gels and represented in a virtual 2D gel. Sixteen percent of the spots were found to occur in all samples and 23% occurred in at least 90% of urines. About 13% of the protein spots were present only in 10% or less of the samples, thus representing the most variable part of the normal urinary proteome. Twenty proteins corresponding to a fraction of the fully conserved spots were identified by mass spectrometry. In conclusion, a "public" urinary proteome, common to healthy individuals, seems to coexist with a "private" urinary proteome, which is more specific to each individual. Copyright © 2011 Elsevier B.V. All rights reserved.

Rickettsial infections of fleas collected from small mammals on four islands in Indonesia.

PubMed

Barbara, Kathryn A; Farzeli, Arik; Ibrahim, Ima N; Antonjaya, Ungke; Yunianto, Andre; Winoto, Imelda; Ester; Perwitasari, Dian; Widjaya, Susana; Richards, Allen L; Williams, Maya; Blair, Patrick J

2010-11-01

Ectoparasites were sampled from small mammals collected in West Java, West Sumatra, North Sulawesi, and East Kalimantan, Indonesia, in 2007-2008 and were screened for evidence of infection from bacteria in the Rickettsaceae family. During eight trap nights at eight sites, 208 fleas were collected from 96 of 507 small mammals trapped from four orders (379 Rodentia; 123 Soricomorpha; two Carnivora; three Scandentia). Two species of fleas were collected: Xenopsylla cheopis (n = 204) and Nosopsyllus spp. (n = 4). Among the 208 fleas collected, 171 X. cheopis were removed from rats (Rattus spp.) and 33 X. cheopis from shrews (Suncus murinus). X. cheopis were pooled and tested for DNA from rickettsial agents Rickettsia typhi, Rickettsia felis, and spotted fever group rickettsiae. R. typhi, the agent of murine typhus, was detected in X. cheopis collected from small mammals in West Java and East Kalimantan. R. felis was detected in X. cheopis collected from small mammals in Manado, North Sulawesi. R. felis and spotted fever group rickettsiae were detected in a pool of X. cheopis collected from an animal in East Kalimantan. Sixteen percent of the X. cheopis pools were found positive for Rickettsia spp.; four (10.8%) R. typhi, one (2.7%) R. felis, and one (2.7%) codetection of R. felis and a spotted fever group rickettsia. These data suggest that rickettsial infections remain a threat to human health across Indonesia.

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

PubMed Central

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

2013-01-01

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

Active learning approach for detection of hard exudates, cotton wool spots, and drusen in retinal images

NASA Astrophysics Data System (ADS)

Sánchez, Clara I.; Niemeijer, Meindert; Kockelkorn, Thessa; Abràmoff, Michael D.; van Ginneken, Bram

2009-02-01

Computer-aided Diagnosis (CAD) systems for the automatic identification of abnormalities in retinal images are gaining importance in diabetic retinopathy screening programs. A huge amount of retinal images are collected during these programs and they provide a starting point for the design of machine learning algorithms. However, manual annotations of retinal images are scarce and expensive to obtain. This paper proposes a dynamic CAD system based on active learning for the automatic identification of hard exudates, cotton wool spots and drusen in retinal images. An uncertainty sampling method is applied to select samples that need to be labeled by an expert from an unlabeled set of 4000 retinal images. It reduces the number of training samples needed to obtain an optimum accuracy by dynamically selecting the most informative samples. Results show that the proposed method increases the classification accuracy compared to alternative techniques, achieving an area under the ROC curve of 0.87, 0.82 and 0.78 for the detection of hard exudates, cotton wool spots and drusen, respectively.

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

NASA Astrophysics Data System (ADS)

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

2012-11-01

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

Using dried blood spot sampling to improve data quality and reduce animal use in mouse pharmacokinetic studies.

PubMed

Wickremsinhe, Enaksha R; Perkins, Everett J

2015-03-01

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

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

PubMed Central

Wickremsinhe, Enaksha R; Perkins, Everett J

2015-01-01

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

Comparing identified and statistically significant lipids and polar metabolites in 15-year old serum and dried blood spot samples for longitudinal studies: Comparing lipids and metabolites in serum and DBS samples

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

Kyle, Jennifer E.; Casey, Cameron P.; Stratton, Kelly G.

The use of dried blood spots (DBS) has many advantages over traditional plasma and serum samples such as smaller blood volume required, storage at room temperature, and ability for sampling in remote locations. However, understanding the robustness of different analytes in DBS samples is essential, especially in older samples collected for longitudinal studies. Here we analyzed DBS samples collected in 2000-2001 and stored at room temperature and compared them to matched serum samples stored at -80°C to determine if they could be effectively used as specific time points in a longitudinal study following metabolic disease. Four hundred small molecules weremore » identified in both the serum and DBS samples using gas chromatograph-mass spectrometry (GC-MS), liquid chromatography-MS (LC-MS) and LC-ion mobility spectrometry-MS (LC-IMS-MS). The identified polar metabolites overlapped well between the sample types, though only one statistically significant polar metabolite in a case-control study was conserved, indicating degradation occurs in the DBS samples affecting quantitation. Differences in the lipid identifications indicated that some oxidation occurs in the DBS samples. However, thirty-six statistically significant lipids correlated in both sample types indicating that lipid quantitation was more stable across the sample types.« less

Trichinella nelsoni in carnivores from the Serengeti ecosystem, Tanzania.

PubMed

Pozio, E; De Meneghi, D; Roelke-Parker, M E; La Rosa, G

1997-12-01

A survey of trichinellosis among sylvatic carnivore mammals from the Serengeti ecosystem (Tanzania) demonstrated the presence of Trichinella nelsoni in 5 of 9 species examined. Muscle samples were collected from carcasses of 56 carnivores from 1993 to 1995 and frozen before transport and examination. Following artificial digestion of the samples, collected larvae were analyzed by the random amplified polymorphic DNA technique. Trichinella nelsoni was identified in 1 bat-eared fox (Otocyon megalotis), 1 cheetah (Acinonyx jubatus), 1 leopard (Panthera pardus), 3 lions (Panthera leo), and 3 spotted hyenas (Crocuta crocuta). The numbers of bat-eared foxes (6), cheetahs (5), and leopards (3) examined were too small to reveal the roles of these carnivore species in the ecology of T. nelsoni. The numbers of lions and spotted hyenas examined, with a prevalence of 12% and 23%, respectively, suggest that these species may be reservoirs of T. nelsoni in the area under study.

Aethalometer™ Instrument Handbook

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

Sedlacek, Arthur J.

2016-04-01

The Aethalometer is an instrument that provides a real-time readout of the concentration of “Black” or “Elemental” carbon aerosol particles (BC or E) in an air stream (see Figure 1 and Figure 2). It is a self-contained instrument that measures the rate of change of optical transmission through a spot on a filter where aerosol is being continuously collected and uses the information to calculate the concentration of optically absorbing material in the sampled air stream. The instrument measures the transmitted light intensities through the “sensing” portion of the filter, on which the aerosol spot is being collected, and amore » “reference” portion of the filter as a check on the stability of the optical source. A mass flowmeter monitors the sample air flow rate. The data from these three measurements is used to determine the mean BC content of the air stream.« less

Measurement of Crystalline Silica Aerosol Using Quantum Cascade Laser-Based Infrared Spectroscopy.

PubMed

Wei, Shijun; Kulkarni, Pramod; Ashley, Kevin; Zheng, Lina

2017-10-24

Inhalation exposure to airborne respirable crystalline silica (RCS) poses major health risks in many industrial environments. There is a need for new sensitive instruments and methods for in-field or near real-time measurement of crystalline silica aerosol. The objective of this study was to develop an approach, using quantum cascade laser (QCL)-based infrared spectroscopy (IR), to quantify airborne concentrations of RCS. Three sampling methods were investigated for their potential for effective coupling with QCL-based transmittance measurements: (i) conventional aerosol filter collection, (ii) focused spot sample collection directly from the aerosol phase, and (iii) dried spot obtained from deposition of liquid suspensions. Spectral analysis methods were developed to obtain IR spectra from the collected particulate samples in the range 750-1030 cm -1 . The new instrument was calibrated and the results were compared with standardized methods based on Fourier transform infrared (FTIR) spectrometry. Results show that significantly lower detection limits for RCS (≈330 ng), compared to conventional infrared methods, could be achieved with effective microconcentration and careful coupling of the particulate sample with the QCL beam. These results offer promise for further development of sensitive filter-based laboratory methods and portable sensors for near real-time measurement of crystalline silica aerosol.

Evaluation of the Sequential Spot Sampler (S3) for time-resolved measurement of PM2.5 sulfate and nitrate through lab and field measurements

NASA Astrophysics Data System (ADS)

Hecobian, A.; Evanoski-Cole, A.; Eiguren-Fernandez, A.; Sullivan, A. P.; Lewis, G. S.; Hering, S. V.; Collett, J. L., Jr.

2016-02-01

The Sequential Spot Sampler (S3), a newly developed instrument to collect aerosols for time-resolved chemical composition measurements, was evaluated in the laboratory and field for the measurement of particulate sulfate and nitrate. The S3 uses a multi-temperature condensation growth tube to grow individual aerosols to droplets which are then deposited as a ˜ 1 mm diameter dry spot at the end of the growth tube in a 100 µL well of a multi-well plate. The well plate advances automatically to provide a sequence of time-resolved samples. The collected aerosols are subsequently analyzed in the laboratory. The sample is concentrated during the collection process, and the laboratory extraction and analysis steps can be automated. The well plate, as received from the field, is placed onto a needle-based autosampler that adds liquid for sample extraction and injects sample extract from each well onto an ion chromatograph for analysis. Laboratory evaluation for sulfate and nitrate ions showed that poly ether ether ketone (PEEK) used as well plate material does not contribute any artifacts; a 60 min extraction procedure leads to the recovery of sulfate and nitrate from the dry spots at above 95 % extraction efficiency; and samples stored frozen and analyzed up to 23 months later show less than a 10 % change in sulfate and nitrate concentrations. The limit of detection was 0.5 µg m-3 for sulfate and 0.2 µg m-3 for nitrate for a 1 h sampling period. In a month-long field study conducted in southern California, two S3s were deployed alongside a URG denuder-filter-pack and a Particle-Into-Liquid Sampler combined with an Ion Chromatograph (PILS-IC). Collocated S3 sampler concentrations compared by linear regression show good agreement, with r2 = 0.99 and slope = 0.99 (±0.004) µg m-3 for sulfate and r2 = 0.99 and slope = 1.0 (±0.006) µg m-3 for nitrate. When compared to the URG denuder-filter-pack and the PILS-IC, the S3 sulfate and nitrate concentrations yielded correlations above 0.84 for the square of the correlation coefficient and regression slopes close to 1.

Temporal variability of urinary cadmium in spot urine samples and first morning voids.

PubMed

Vacchi-Suzzi, Caterina; Porucznik, Christina A; Cox, Kyley J; Zhao, Yuan; Ahn, Hongshik; Harrington, James M; Levine, Keith E; Demple, Bruce; Marsit, Carmen J; Gonzalez, Adam; Luft, Benjamin; Meliker, Jaymie R

2017-05-01

Cadmium is a carcinogenic heavy metal. Urinary levels of cadmium are considered to be an indicator of long-term body burden, as cadmium accumulates in the kidneys and has a half-life of at least 10 years. However, the temporal stability of the biomarker in urine samples from a non-occupationally exposed population has not been rigorously established. We used repeated measurements of urinary cadmium (U-Cd) in spot urine samples and first morning voids from two separate cohorts, to assess the temporal stability of the samples. Urine samples from two cohorts including individuals of both sexes were measured for cadmium and creatinine. The first cohort (Home Observation of Perinatal Exposure (HOPE)) consisted of 21 never-smokers, who provided four first morning urine samples 2-5 days apart, and one additional sample roughly 1 month later. The second cohort (World Trade Center-Health Program (WTC-HP)) consisted of 78 individuals, including 52 never-smokers, 22 former smokers and 4 current smokers, who provided 2 spot urine samples 6 months apart, on average. Intra-class correlation was computed for groups of replicates from each individual to assess temporal variability. The median creatinine-adjusted U-Cd level (0.19 and 0.21 μg/g in the HOPE and WTC-HP, respectively) was similar to levels recorded in the United States by the National Health and Nutrition Examination Survey. The intra-class correlation (ICC) was high (0.76 and 0.78 for HOPE and WTC-HP, respectively) and similar between cohorts, irrespective of whether samples were collected days or months apart. Both single spot or first morning urine cadmium samples show good to excellent reproducibility in low-exposure populations.

Real-time PCR detection of Plasmodium directly from whole blood and filter paper samples

PubMed Central

2011-01-01

Background Real-time PCR is a sensitive and specific method for the analysis of Plasmodium DNA. However, prior purification of genomic DNA from blood is necessary since PCR inhibitors and quenching of fluorophores from blood prevent efficient amplification and detection of PCR products. Methods Reagents designed to specifically overcome PCR inhibition and quenching of fluorescence were evaluated for real-time PCR amplification of Plasmodium DNA directly from blood. Whole blood from clinical samples and dried blood spots collected in the field in Colombia were tested. Results Amplification and fluorescence detection by real-time PCR were optimal with 40× SYBR® Green dye and 5% blood volume in the PCR reaction. Plasmodium DNA was detected directly from both whole blood and dried blood spots from clinical samples. The sensitivity and specificity ranged from 93-100% compared with PCR performed on purified Plasmodium DNA. Conclusions The methodology described facilitates high-throughput testing of blood samples collected in the field by fluorescence-based real-time PCR. This method can be applied to a broad range of clinical studies with the advantages of immediate sample testing, lower experimental costs and time-savings. PMID:21851640

Multiplexed Quantitation of Endogenous Proteins in Dried Blood Spots by Multiple Reaction Monitoring - Mass Spectrometry

PubMed Central

Chambers, Andrew G.; Percy, Andrew J.; Yang, Juncong; Camenzind, Alexander G.; Borchers, Christoph H.

2013-01-01

Dried blood spot (DBS) sampling, coupled with multiple reaction monitoring mass spectrometry (MRM-MS), is a well-established approach for quantifying a wide range of small molecule biomarkers and drugs. This sampling procedure is simpler and less-invasive than those required for traditional plasma or serum samples enabling collection by minimally trained personnel. Many analytes are stable in the DBS format without refrigeration, which reduces the cost and logistical challenges of sample collection in remote locations. These advantages make DBS sample collection desirable for advancing personalized medicine through population-wide biomarker screening. Here we expand this technology by demonstrating the first multiplexed method for the quantitation of endogenous proteins in DBS samples. A panel of 60 abundant proteins in human blood was targeted by monitoring proteotypic tryptic peptides and their stable isotope-labeled analogs by MRM. Linear calibration curves were obtained for 40 of the 65 peptide targets demonstrating multiple proteins can be quantitatively extracted from DBS collection cards. The method was also highly reproducible with a coefficient of variation of <15% for all 40 peptides. Overall, this assay quantified 37 proteins spanning a range of more than four orders of magnitude in concentration within a single 25 min LC/MRM-MS analysis. The protein abundances of the 33 proteins quantified in matching DBS and whole blood samples showed an excellent correlation, with a slope of 0.96 and an R2 value of 0.97. Furthermore, the measured concentrations for 80% of the proteins were stable for at least 10 days when stored at −20 °C, 4 °C and 37 °C. This work represents an important first step in evaluating the integration of DBS sampling with highly-multiplexed MRM for quantitation of endogenous proteins. PMID:23221968

Spot protein-creatinine ratio and spot albumin-creatinine ratio in the assessment of pre-eclampsia: a diagnostic accuracy study with decision-analytic model-based economic evaluation and acceptability analysis.

PubMed

Waugh, Jason; Hooper, Richard; Lamb, Edmund; Robson, Stephen; Shennan, Andrew; Milne, Fiona; Price, Christopher; Thangaratinam, Shakila; Berdunov, Vladislav; Bingham, Jenn

2017-10-01

The National Institute for Health and Care Excellence (NICE) guidelines highlighted the need for 'large, high-quality prospective studies comparing the various methods of measuring proteinuria in women with new-onset hypertensive disorders during pregnancy'. The primary objective was to evaluate quantitative assessments of spot protein-creatinine ratio (SPCR) and spot albumin-creatinine ratio (SACR) in predicting severe pre-eclampsia (PE) compared with 24-hour urine protein measurement. The secondary objectives were to investigate interlaboratory assay variation, to evaluate SPCR and SACR thresholds in predicting adverse maternal and fetal outcomes and to assess the cost-effectiveness of these models. This was a prospective diagnostic accuracy cohort study, with decision-analytic modelling and a cost-effectiveness analysis. The setting was 36 obstetric units in England, UK. Pregnant women (aged ≥ 16 years), who were at > 20 weeks' gestation with confirmed gestational hypertension and trace or more proteinuria on an automated dipstick urinalysis. Women provided a spot urine sample for protein analysis (the recruitment sample) and were asked to collect a 24-hour urine sample, which was stored for secondary analysis. A further spot sample of urine was taken immediately before delivery. Outcome data were collected from hospital records. There were four index tests on a spot sample of urine: (1) SPCR test (conducted at the local laboratory); (2) SPCR test [conducted at the central laboratory using the benzethonium chloride (BZC) assay]; (3) SPCR test [conducted at the central laboratory using the pyrogallol red (PGR) assay]; and (4) SACR test (conducted at the central laboratory using an automated chemistry analyser). The comparator tests on 24-hour urine collection were a central test using the BZC assay and a central test using the PGR assay. The primary reference standard was the NICE definition of severe PE. Secondary reference standards were a clinician diagnosis of severe PE, which is defined as treatment with magnesium sulphate or with severe PE protocol; adverse perinatal outcome; one or more of perinatal or infant mortality, bronchopulmonary dysplasia, necrotising enterocolitis or grade III/IV intraventricular haemorrhage; and economic cost and outcomes. Health service data on service use and costs followed published economic models. In total, 959 women were available for primary analysis and 417 of them had severe PE. The diagnostic accuracy of the four assays on spot urine samples against the reference standards was similar. The three SPCR tests had sensitivities in excess of 90% at prespecified thresholds, with poor specificities and negative likelihood ratios of ≥ 0.1. The SACR test had a significantly higher sensitivity of 99% (confidence interval 98% to 100%) and lower specificity. Receiver operating characteristic (ROC) curves were similar (area under ROC curve between 0.87 and 0.89); the area under the central laboratory's SACR curve was significantly higher ( p  = 0.004). The central laboratory's SACR test was the most cost-effective option, generating an additional 0.03 quality-adjusted life-years at an additional cost of £45.07 compared with the local laboratory's SPCR test. The probabilistic analysis showed it to have a 100% probability of being cost-effective at the standard willingness-to-pay threshold recommended by NICE. Implementation of NICE guidelines has led to an increased intervention rate in the study population that affected recruitment rates and led to revised sample size calculations. Evidence from this clinical study does not support the recommendation of 24-hour urine sample collection in hypertensive pregnant women. The SACR test had better diagnostic performance when predicting severe pre-eclampsia. All four tests could potentially be used as rule-out tests for the NICE definition of severe PE. Testing SACR at a threshold of 8 mg/mmol should be studied as a 'rule-out' test of proteinuria. Current Controlled Trials ISRCTN82607486. This project was funded by the National Institute Health Research (NIHR) Health Technology Assessment programme and will be published in full in Health Technology Assessment ; Vol. 21, No. 61. See the NIHR Journals Library website for further project information.

Pooled HIV-1 viral load testing using dried blood spots to reduce the cost of monitoring antiretroviral treatment in a resource-limited setting.

PubMed

Pannus, Pieter; Fajardo, Emmanuel; Metcalf, Carol; Coulborn, Rebecca M; Durán, Laura T; Bygrave, Helen; Ellman, Tom; Garone, Daniela; Murowa, Michael; Mwenda, Reuben; Reid, Tony; Preiser, Wolfgang

2013-10-01

Rollout of routine HIV-1 viral load monitoring is hampered by high costs and logistical difficulties associated with sample collection and transport. New strategies are needed to overcome these constraints. Dried blood spots from finger pricks have been shown to be more practical than the use of plasma specimens, and pooling strategies using plasma specimens have been demonstrated to be an efficient method to reduce costs. This study found that combination of finger-prick dried blood spots and a pooling strategy is a feasible and efficient option to reduce costs, while maintaining accuracy in the context of a district hospital in Malawi.

Risk factors associated with high potential for serious crashes.

DOT National Transportation Integrated Search

2015-09-01

Crashes are random events and low traffic volumes therefore dont always make crash hot-spot : identification possible. This project has used extensive data collection and analysis for a large sample : of Oregons low volume roads to develop a ri...

Evaluation of Dried Urine Spot Method to Screen Cotinine among Tobacco Dependents: An Exploratory Study.

PubMed

Jain, Raka; Quraishi, Rizwana; Verma, Arpita

2017-01-01

Assessment of cotinine, a metabolite of nicotine in body fluids, is an important approach for validating the self-report among tobacco users. Adaptation of assays on dried urine spots (DUSs) has advantages of ease of collection, transportation, minimal invasiveness, and requirement of small volume. The aim of the present study was to develop an efficient method for testing cotinine in DUSs and evaluating its clinical applicability. This involved optimization of conditions for detection, recovery, and stability of cotinine from dried urine, spotted on filter paper. Enzyme-linked immunosorbent assay was used for screening, whereas confirmation was done by gas chromatography. For clinical applicability, urine samples of tobacco users were tested. Water was found to be a suitable extracting solvent as compared to carbonate-bicarbonate buffer (pH 9.2) and saline. Screening was achieved by two punches taken from a 20 μl (diameter 1.3 cm) spotted urine samples, and confirmation was achieved by five complete circles each of 20 μl sample volume. The recovery was found to be 97% in water. Limit of detection for the method was found to be 100 ng/ml. No signs of significant degradation were found under all storage conditions. All the urine samples of tobacco users were found to be positive by a conventional method as well as DUSs, and the method proved to be efficient. DUS samples are a useful alternative for biological monitoring of recent nicotine use, especially in developing countries where sample logistics could be an important concern.

Noninvasive measures of reproductive function and disturbance in the barred owl, great horned owl, and northern spotted owl.

PubMed

Wasser, Samuel K; Hunt, Kathleen E

2005-06-01

There is an urgent need for noninvasive methods to study reproduction and environmental stress in at-risk species such as the northern spotted owl (Strix occidentalis caurina). Two related owl species (barred owl and great horned owl) were used as surrogates to validate hormone assays for fecal metabolites of progesterone, 17beta-estradiol, testosterone, and corticosterone. Infusions of radiolabeled hormones showed that the owls excreted most hormone within 6 h. Feces and urine contained roughly equal amounts of hormone, and most fecal hormone metabolites were quite polar. The testosterone and corticosterone assays in this study bound to the major excreted metabolites of these hormones, but two progesterone assays did not appreciably bind to the major progesterone metabolites. All assays showed excellent parallelism with hydrolyzed and unhydrolyzed samples and with previously dried or undried fecal samples. Thus, samples do not require hydrolysis or prior drying. Samples from a female barred owl had significantly higher fecal estrogen, lower fecal testosterone, and higher fecal estrogen/testosterone ratio than samples from two male barred owls. The fecal estrogen/testosterone ratio was the most accurate predictor of owl gender, particularly if two or more samples are available from the same individual. Fecal corticosterone metabolites also demonstrated considerable utility for wild northern spotted owls. Fecal glucocorticoid levels varied by gender and breeding stage, being highest in male northern spotted owls early in the breeding season and highest in females when nestlings were fledging. Collectively, these studies show that noninvasive fecal hormone measurements show great promise for noninvasive assessment of reproduction and stress in wild owls.

Biomarker evidence of tobacco smoke exposure in children participating in lead screening.

PubMed

Joseph, Anne; Spector, Logan; Wickham, Katherine; Janis, Gregory; Winickoff, Jonathan; Lindgren, Bruce; Murphy, Sharon

2013-12-01

We assessed tobacco smoke exposure (TSE), defined according to detection of cotinine, in dried blood spots collected from children for lead screening. Dried blood spots collected from a national sample of 1541 Black and White children and submitted to a commercial laboratory for lead analysis were analyzed for cotinine. We used an anonymous administrative data set including information on children's characteristics to conduct univariate and multivariate analyses. Cotinine was detected in 61% of dried blood spots; 17% of samples had cotinine levels above 3 nanograms per gram. Median cotinine levels were significantly higher among Black than White children (0.66 ng/g vs 0.30 ng/g) and among Medicaid recipients (0.94 ng/g vs < 0.3 ng/g). In multivariate analyses, significant increases in cotinine levels were associated with Black (vs White) race, older age, Medicaid coverage, higher state smoking rate, and higher average winter temperature. Detectable cotinine levels were significantly associated with higher lead levels. TSE is highly prevalent among children undergoing lead screening, and exposure levels are greater among Black children and children on Medicaid. TSE may contribute to lead exposure. Concurrent lead screening and biological screening for TSE may be a feasible approach to increasing childhood TSE detection.

Measurement of daily sodium excretion in patients with chronic kidney disease; special reference to the difference between the amount measured from 24 h collected urine sample and the estimated amount from a spot urine.

PubMed

Amano, Hoichi; Kobayashi, Seiji; Terawaki, Hiroyuki; Ogura, Makoto; Kawaguchi, Yoshindo; Yokoo, Takashi

2018-11-01

It is important to grasp a patient's daily sodium intake in the management of chronic kidney disease, as sodium intake is widely recommended at 6 g/day or less. There are multiple equations widely known for estimating the daily sodium excretion from a spot urine sample, but these are aimed at healthy people. There are few reports that validate equations in patients with chronic kidney disease. The purpose of this study is to evaluate whether the amount of measured daily sodium excretion from a sample collected for 24-h urine (24HU) is equal to that of using an equation from a spot urine sample (SU) in patients with chronic kidney disease. One hundred sixty-two patients with chronic kidney disease from Kanagawa Prefecture Shiomidai Hospital, Japan and the Jikei University Kashiwa Hospital, Japan participated in the study. Daily sodium excretion was measured from 24HU and compared with it from SU by using the formula according to Tanaka et al. Sodium excretion by 24HU was 2744 mg/day and estimating daily sodium excretion from SU was 3315 mg/day. The coefficient of determination was 0.17 (p < .001) in multivariate regression analysis. The coefficient of determination was extremely low. Thus, there is a considerable difference between the amount of sodium excretion calculated from a 24HU and that from a SU in patients with chronic kidney disease.

Front-Loading Sputum Microscopy Services: An Opportunity to Optimise Smear-Based Case Detection of Tuberculosis in High Prevalence Countries

PubMed Central

Ramsay, Andy; Yassin, Mohammed Ahmed; Cambanis, Alexis; Hirao, Susumu; Almotawa, Ahmad; Gammo, Mohamed; Lawson, Lovett; Arbide, Izabel; Al-Aghbari, Nasher; Al-Sonboli, Najla; Sherchand, Jeevan Bahadur; Gauchan, Punita; Cuevas, Luis Eduardo

2009-01-01

Setting. Ethiopia, Nepal, Nigeria, and Yemen. Objective. To reduce the time to complete sputum microscopy. Design. Cross-sectional surveys enrolling 923 patients with chronic cough in the 4 countries and using similar protocols. Spot-morning-spot sputum specimens were collected. An additional sputum specimen (Xspot) was collected one hour after the first, and the yields of the first two or the three specimens collected as spot-morning-spot or spot-Xspot-morning were compared. Results. 216 patients had ≥ one positive smear. 210 (97%) were identified by the spot-morning-spot, and 210 (97%) were identified by the spot-Xspot-morning specimens, with 203 and 200 identified by the first 2 specimens of each approach, respectively. Neither difference was significant. Conclusions. The time to complete smear microscopy could be reduced. PMID:20309419

Glyphosate in Irish adults - A pilot study in 2017.

PubMed

Connolly, Alison; Leahy, Michelle; Jones, Kate; Kenny, Laura; Coggins, Marie A

2018-08-01

Glyphosate is the highest volume herbicide used globally and has recently been classified as a 2 A 'probably carcinogenic to humans' by the International Agency for Research on Cancer (IARC). There is limited data to evaluate the public health impacts from glyphosate exposure. The objective of this study is to conduct an exploratory glyphosate exposure assessment study among Irish adults, who were non-occupational users of glyphosate. A convenient sampling method was used, collecting one first morning void spot urine sample from each participant. A biomonitoring survey involving the collection and analysis of 20 ml spot urine samples from 50 Irish adults was conducted in June 2017. Participants completed a short questionnaire to collect information on demographics, dietary habits and lifestyle. Glyphosate was extracted using solid phase extraction (SPE) and analysed by liquid chromatography tandem mass spectrometry (LC-MC/MS). Of the 50 urine samples analysed, 10 (20%) contained detectable levels of glyphosate (0.80-1.35 µg L -1 ). Exposure concentrations are higher than those reported in comparable studies of European and American adults. Glyphosate was detectable in 20% of the samples collected from Irish adults. The low proportion of detectable glyphosate levels could be due to lower localised use of pesticides, having a small sample size or the higher analytical detection limit used in this study (0.5 µg L -1 ), which could underestimate the true exposure and warrants further investigation. Given the widespread use of glyphosate, further information on population exposure is required to advance our understanding of the relationship between chronic low dose exposure to glyphosate and human health risk. Copyright © 2018 Elsevier Inc. All rights reserved.

Urinalysis of MMX-mesalazine as a tool to monitor 5-ASA adherence in daily IBD practice.

PubMed

Römkens, Tessa E H; Te Morsche, Rene; Peters, Wilbert; Burger, David M; Hoentjen, Frank; Drenth, Joost P H

2018-03-01

Adherence is pivotal but challenging in ulcerative colitis (UC) treatment. Many methods to assess adherence are subjective or have limitations. (Nac-)5-aminosalicylic acid (5-ASA) urinalysis by high-performance liquid chromatography (HPLC) seems feasible and reproducible in healthy volunteers. We performed a prospective study in adult quiescent UC patients to evaluate the feasibility of spot (Nac-)5-ASA urinalysis by HPLC to assess adherence in daily inflammatory bowel disease (IBD) care. Twenty-nine patients (51.7% male, mean age 52 ± 11 years) were included (median FU 9 months) and weekly spot urine samples were collected. We found large variation in spot (Nac-)5-ASA urinary excretion that was unrelated to brand, dosing schedule or dosage of 5-ASA. In conclusion, spot (Nac-)5-ASA urinalysis is not applicable to assess 5-ASA adherence in daily IBD care. © 2017 The British Pharmacological Society.

Analysis of Hemoglobin A1c from Dried Blood Spot Samples with the Tina-quant® II Immunoturbidimetric Method

PubMed Central

Jones, Trevor G.; Warber, Kimbrough D.; Roberts, Billy D.

2010-01-01

Background Hemoglobin A1c (HbA1c) has been endorsed as a tool for the diagnosis of diabetes. This test requires instrumentation that may not be available in underdeveloped areas. Dried blood spot (DBS) samples collected by finger stick procedures offer a mechanism to transport samples to laboratories that do measure HbA1c. Methods Whole blood (ethylenediaminetetraacetic acid) was applied to Ahlstrom 226 filter paper. These DBS samples were compared to whole blood samples using the Roche Tina-quant® II immunoturbidometric assay. Hemoglobin A1c stability on DBS was assessed at three temperatures—4, 25, and 40°C—for up to 9 days. A 44-day study was also done for DBS at 20–25°C. Results The Tina-quant® II DBS method showed excellent agreement with whole blood HbA1c results (r2 = 0.99) with a slight positive mean bias of 0.08 ± 0.04% HbA1c (95% confidence interval). The variation in HbA1c on DBS samples subjected to different temperatures and times did not exceed 5.6%. Conclusions Dried blood spot samples represent an alternative to whole blood for HbA1c by measurement when transporting whole blood is not feasible. PMID:20307383

Detection of ethyl glucuronide in blood spotted on different surfaces.

PubMed

Winkler, M; Kaufmann, E; Thoma, D; Thierauf, A; Weinmann, W; Skopp, G; Alt, A

2011-07-15

This study aims to show that sensitive detection of ethyl glucuronide in dried blood spotted onto various surfaces after a period of 24h is feasible. At present, there is insufficient information how tightly ethyl glucuronide (EtG) binds to various materials and how easily it can be eluted. 4ml aliquots of blood samples obtained from seven volunteers after consumption of alcoholic beverages were applied to six different surfaces. After drying and a 24h-storage at 20±2°C the samples were re-dissolved in water, and EtG was subsequently analyzed by a LC-MS Paul-type ion trap. A comparison was made between dried and corresponding fluid samples. EtG was detectable in all subjects' samples following consumption of alcohol. EtG was also detectable after a storage time of four weeks at 4°C in whole blood that had been preserved with EDTA. EtG was detectable in all samples dried on different surfaces and its concentration remained relatively constant irrespective of the particular condition of the material. Detection of EtG in blood spots from the scene may indicate recent alcohol consumption in cases where collection of blood remained undone or could not be performed. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

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

PubMed

Watanabe, Kyoko; Varesio, Emmanuel; Hopfgartner, Gérard

2014-08-15

An assay was developed and validated for the quantification of eight protease inhibitors (indinavir (IDV), ritonavir (RTV), lopinavir (LPV), saquinavir (SQV), amprenavir (APV), nelfinavir (NFV), atazanavir (AZV) and darunavir (DRV)) in dried plasma spots using parallel ultra-high performance liquid chromatography and mass spectrometry detection in the multiple reaction monitoring mode. For each analyte an isotopically labeled internal standard was used and the assay based on liquid-solid extraction the area response ratio (analyte/IS) was found to be linear; from 0.025 μg/ml to 20 μg/ml for IDV, SQV, DRV, AZV, LPV, from 0.025 μg/ml to 10 μg/ml for NFV, APV and from 0.025 μg/ml to 5 μg/ml for RTV using 15 μl of plasma spotted on filter paper placed in a sample tube. The total analysis time was of 4 min and inter-assay accuracies and precisions were in the range of 87.7-109% and 2.5-11.8%, respectively. On dried plasma spots all analytes were found to be stable for at least 7 days. Practicability of the assay to blood was also demonstrated. The sample drying process could be reduced to 5 min using a commercial microwave system without any analyte degradation. Together with quantification, confirmatory analysis was performed on representative clinical samples. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparative value of blood and skin samples for diagnosis of spotted fever group rickettsial infection in model animals.

PubMed

Levin, Michael L; Snellgrove, Alyssa N; Zemtsova, Galina E

2016-07-01

The definitive diagnosis of spotted fever group (SFG) rickettsioses in humans is challenging due to the retrospective nature and cross reactivity of the serological methods and the absence of reliable and consistent samples for molecular diagnostics. Existing data indicate the transient character of bacteremia in experimentally infected animals. The ability of arthropod vectors to acquire rickettsial infection from the laboratory animals in the absence of systemic infection and known tropism of rickettsial agents to endothelial cells of peripheral blood vessels underline the importance of local infection and consequently the diagnostic potential of skin samples. In order to evaluate the diagnostic sensitivity of rickettsial DNA detection in blood and skin samples, we compared results of PCR testing in parallel samples collected from model laboratory animals infected with Rickettsia rickettsii, Rickettsia parkeri and Rickettsia slovaca-like agent at different time points after infection. Skin samples were collected from ears - away from the site of tick placement and without eschars. Overall, testing of skin samples resulted in a higher proportion of positive results than testing of blood samples. Presented data from model animals demonstrates that testing of skin samples from sites of rickettsial proliferation can provide definitive molecular diagnosis of up to 60-70% of tick-borne SFG rickettsial infections during the acute stage of illness. Detection of pathogen DNA in cutaneous samples is a valuable alternative to blood-PCR at least in model animals. Published by Elsevier GmbH.

LED Fluorescence Microscopy for the Diagnosis of Pulmonary Tuberculosis: A Multi-Country Cross-Sectional Evaluation

PubMed Central

Cuevas, Luis Eduardo; Al-Sonboli, Najla; Lawson, Lovett; Yassin, Mohammed Ahmed; Arbide, Isabel; Al-Aghbari, Nasher; Bahadur Sherchand, Jeevan; Al-Absi, Amin; Emenyonu, Emmanuel Nnamdi; Merid, Yared; Okobi, Mosis Ifenyi; Onuoha, Juliana Olubunmi; Aschalew, Melkamsew; Aseffa, Abraham; Harper, Greg; Anderson de Cuevas, Rachel Mary; Theobald, Sally Jane; Nathanson, Carl-Michael; Joly, Jean; Faragher, Brian; Squire, Stephen Bertel; Ramsay, Andrew

2011-01-01

Background The diagnosis of tuberculosis (TB) in resource-limited settings relies on Ziehl-Neelsen (ZN) smear microscopy. LED fluorescence microscopy (LED-FM) has many potential advantages over ZN smear microscopy, but requires evaluation in the field. The aim of this study was to assess the sensitivity/specificity of LED-FM for the diagnosis of pulmonary TB and whether its performance varies with the timing of specimen collection. Methods and Findings Adults with cough ≥2 wk were enrolled consecutively in Ethiopia, Nepal, Nigeria, and Yemen. Sputum specimens were examined by ZN smear microscopy and LED-FM and compared with culture as the reference standard. Specimens were collected using a spot-morning-spot (SMS) or spot-spot-morning (SSM) scheme to explore whether the collection of the first two smears at the health care facility (i.e., “on the spot”) the first day of consultation followed by a morning sample the next day (SSM) would identify similar numbers of smear-positive patients as smears collected via the SMS scheme (i.e., one on-the-spot-smear the first day, followed by a morning specimen collected at home and a second on-the-spot sample the second day). In total, 529 (21.6%) culture-positive and 1,826 (74.6%) culture-negative patients were enrolled, of which 1,156 (49%) submitted SSM specimens and 1,199 (51%) submitted SMS specimens. Single LED-FM smears had higher sensitivity but lower specificity than single ZN smears. Using two LED-FM or two ZN smears per patient was 72.8% (385/529, 95% CI 68.8%–76.5%) and 65.8% (348/529, 95% CI 61.6%–69.8%) sensitive (p<0.001) and 90.9% (1,660/1,826, 95% CI 89.5%–92.2%) and 98% (1,790/1,826, 95% CI 97.3%–98.6%) specific (p<0.001). Using three LED-FM or three ZN smears per patient was 77% (408/529, 95% CI 73.3%–80.6%) and 70.5% (373/529, 95% CI 66.4%–74.4%, p<0.001) sensitive and 88.1% (95% CI 86.5%–89.6%) and 96.5% (95% CI 96.8%–98.2%, p<0.001) specific. The sensitivity/specificity of ZN smear microscopy and LED-FM did not vary between SMS and SSM. Conclusions LED-FM had higher sensitivity but, in this study, lower specificity than ZN smear microscopy for diagnosis of pulmonary TB. Performance was independent of the scheme used for collecting specimens. The introduction of LED-FM needs to be accompanied by appropriate training, quality management, and monitoring of performance in the field. Trial Registration Current Controlled Trials ISRCTN53339491 Please see later in the article for the Editors' Summary PMID:21765809

COMPARISON OF A GENUS-SPECIFIC CONVENTIONAL PCR AND A SPECIES-SPECIFIC NESTED-PCR FOR MALARIA DIAGNOSIS USING FTA COLLECTED SAMPLES FROM KINGDOM OF SAUDI ARABIA.

PubMed

Al-Harthi, Saeed A

2015-12-01

Molecular tools are increasingly accepted as the most sensitive and reliable techniques for malaria diagnosis and epidemiological surveys. Also, collection of finger prick blood spots onto filter papers is the most simple and affordable method for samples preservation and posterior molecular analysis, especially in rural endemic regions where malaria remains a major health problem. Two malaria molecular diagnostic tests, a Plasmodium genus-specific conventional PCR and a Plasmodium species-specific Nested PCR, were evaluated using DNA templates prepared from Whatman-FTA cards' dry blood spots using both, Methanol-fixation/Heat-extraction and FTA commercial purification kit. A total of 121 blood samples were collected from six Saudi south-western endemic districts both, as thick and thin films for routine microscopic screening and onto FTA cards for molecular studies. Out of the 121 samples, 75 were P. falciparum positive by at least one technique. No other species of Plasmodium were detected. P. falciparum parasites were identified in 69/75 (92%) samples by microscopic screening in health care centers. P. genus-specific PCR was able to amplify P. falciparum DNA in 41/75 (55%) and 59/75 (79%) samples using Methanol-fixation/Heat-extraction and FTA purification kit, respectively. P. species-specific Nested PCR revealed 68/75 (91%) and 75/75 (100%) positive samples using DNA templates were isolated by Methanol-fixation/Heat- extraction and FTA purification methods, respectively. The species-specific Nested PCR applied to Whatman-FTA preserved and processed blood samples represents the best alternative to classical microscopy for malaria diagnosis, particularly in epidemiological screening.

Measure of viral load by using the Abbott Real-Time HIV-1 assay on dried blood and plasma spot specimens collected in 2 rural dispensaries in Cameroon.

PubMed

Mbida, André Dieudonné; Sosso, Samuel; Flori, Pierre; Saoudin, Henia; Lawrence, Philip; Monny-Lobé, Marcel; Oyono, Yves; Ndzi, Edward; Cappelli, Giulia; Lucht, Frédéric; Pozzetto, Bruno; Oukem-Boyer, Odile Ouwe Missi; Bourlet, Thomas

2009-09-01

This study aimed to evaluate the use of dried blood spots (DBSs) and dried plasma spots (DPSs) locally collected in 2 rural dispensaries in Cameroon for the quantification of HIV-1 RNA. Forty-one subjects were sampled and spots of whole blood and plasma were deposited onto Whatman 903 cards and dried at ambient temperature under local conditions. Two sets of DBS and DPS cards were done per patient. The rest of the liquid plasma (LP) was frozen until use. LPs were tested at the "Chantal Biya" International Reference Centre (Yaoundé, Cameroon) by the Abbott Real-Time HIV-1 assay (Abbott Molecular Diagnostics, Wiesbaden, Germany). One series of DBS and DPS was transported and tested between 2 and 6 weeks later at the Virology Laboratory of Saint-Etienne (France). The second series was routed by mail and tested after up to 3 months of storage at ambient temperature. From the first series, the correlation rate between viral loads obtained from LP and DBS, and from LP and DPS, was 0.98 and 0.99, respectively; specificity of DBS and DPS results was 100%. The results obtained from the second series indicate a great stability of DBS after long-term storage. This study demonstrates that DBSs collected under local conditions in resource-limited settings are suitable for the differed quantification of HIV-1 RNA.

Validation and Assessment of Three Methods to Estimate 24-h Urinary Sodium Excretion from Spot Urine Samples in Chinese Adults

PubMed Central

Peng, Yaguang; Li, Wei; Wang, Yang; Chen, Hui; Bo, Jian; Wang, Xingyu; Liu, Lisheng

2016-01-01

24-h urinary sodium excretion is the gold standard for evaluating dietary sodium intake, but it is often not feasible in large epidemiological studies due to high participant burden and cost. Three methods—Kawasaki, INTERSALT, and Tanaka—have been proposed to estimate 24-h urinary sodium excretion from a spot urine sample, but these methods have not been validated in the general Chinese population. This aim of this study was to assess the validity of three methods for estimating 24-h urinary sodium excretion using spot urine samples against measured 24-h urinary sodium excretion in a Chinese sample population. Data are from a substudy of the Prospective Urban Rural Epidemiology (PURE) study that enrolled 120 participants aged 35 to 70 years and collected their morning fasting urine and 24-h urine specimens. Bias calculations (estimated values minus measured values) and Bland-Altman plots were used to assess the validity of the three estimation methods. 116 participants were included in the final analysis. Mean bias for the Kawasaki method was -740 mg/day (95% CI: -1219, 262 mg/day), and was the lowest among the three methods. Mean bias for the Tanaka method was -2305 mg/day (95% CI: -2735, 1875 mg/day). Mean bias for the INTERSALT method was -2797 mg/day (95% CI: -3245, 2349 mg/day), and was the highest of the three methods. Bland-Altman plots indicated that all three methods underestimated 24-h urinary sodium excretion. The Kawasaki, INTERSALT and Tanaka methods for estimation of 24-h urinary sodium excretion using spot urines all underestimated true 24-h urinary sodium excretion in this sample of Chinese adults. Among the three methods, the Kawasaki method was least biased, but was still relatively inaccurate. A more accurate method is needed to estimate the 24-h urinary sodium excretion from spot urine for assessment of dietary sodium intake in China. PMID:26895296

Nonlinear vibrational microscopy

DOEpatents

Holtom, Gary R.; Xie, Xiaoliang Sunney; Zumbusch, Andreas

2000-01-01

The present invention is a method and apparatus for microscopic vibrational imaging using coherent Anti-Stokes Raman Scattering or Sum Frequency Generation. Microscopic imaging with a vibrational spectroscopic contrast is achieved by generating signals in a nonlinear optical process and spatially resolved detection of the signals. The spatial resolution is attained by minimizing the spot size of the optical interrogation beams on the sample. Minimizing the spot size relies upon a. directing at least two substantially co-axial laser beams (interrogation beams) through a microscope objective providing a focal spot on the sample; b. collecting a signal beam together with a residual beam from the at least two co-axial laser beams after passing through the sample; c. removing the residual beam; and d. detecting the signal beam thereby creating said pixel. The method has significantly higher spatial resolution then IR microscopy and higher sensitivity than spontaneous Raman microscopy with much lower average excitation powers. CARS and SFG microscopy does not rely on the presence of fluorophores, but retains the resolution and three-dimensional sectioning capability of confocal and two-photon fluorescence microscopy. Complementary to these techniques, CARS and SFG microscopy provides a contrast mechanism based on vibrational spectroscopy. This vibrational contrast mechanism, combined with an unprecedented high sensitivity at a tolerable laser power level, provides a new approach for microscopic investigations of chemical and biological samples.

Performances of fourth generation HIV antigen/antibody assays on filter paper for detection of early HIV infections.

PubMed

Kania, Dramane; Truong, Tam Nguyen; Montoya, Ana; Nagot, Nicolas; Van de Perre, Philippe; Tuaillon, Edouard

2015-01-01

Point-of-care testing and diagnosis of HIV acute infections play important roles in preventing transmission, but HIV rapid diagnosis tests have poor capacity to detect early infections. Filter paper can be used for capillary blood collection and HIV testing using 4th generation immunoassays. Antigen/antibody combined immunoassays were evaluated for their capacity to identify early HIV infections using filter paper in comparison with rapid test. Thirty nine serum samples collected from HIV seroconverters were spotted onto filter paper and tested by the Roche Elecsys(®) HIV Combi PT test and the DiaSorin Liaison XL Murex HIV Ab/Ag assay. Fourth generation immunoassays identified 34 out of 39 HIV early infections using dried serum spot, whereas the Determine™ HIV-1/2 rapid test detected 24 out of 39 HIV positive serum (87.2% vs 61.5% respectively, p = 0.009). p24 antigen was detected by the Liaison XL in 19 dried serum samples (48.7%). In the group characterized by a negative western blot, 7 out of 8 (87.5%) and 6 out of 8 (75.0%) samples were found positive for HIV using the Elecsys and the Liaison XL, respectively. None of these eight samples classified in this group of early acute infections were found positive by the rapid test. Fourth generation Ag/Ab immunoassays performed on dried serum spot had good performance for HIV testing during the early phases of HIV infection. This method may be useful to detect HIV early infections in hard-to-reach populations and individuals living in remote areas before rapid tests become positive. Copyright © 2014 Elsevier B.V. All rights reserved.

Exposures to volatile organic compounds (VOCs) and associated health risks of socio-economically disadvantaged population in a "hot spot" in Camden, New Jersey

NASA Astrophysics Data System (ADS)

Wu, Xiangmei (May); Fan, Zhihua (Tina); Zhu, Xianlei; Jung, Kyung Hwa; Ohman-Strickland, Pamela; Weisel, Clifford P.; Lioy, Paul J.

2012-09-01

To address disparities in health risks associated with ambient air pollution for racial/ethnic minority groups, this study characterized personal and ambient concentrations of volatile organic compounds (VOCs) in a suspected hot spot of air pollution - the Village of Waterfront South (WFS), and an urban reference community - the Copewood/Davis Streets (CDS) neighborhood in Camden, New Jersey. Both are minority-dominant, impoverished communities. We collected 24-h integrated personal air samples from 54 WFS residents and 53 CDS residents, with one sample on a weekday and one on a weekend day during the summer and winter seasons of 2004-2006. Ambient air samples from the center of each community were also collected simultaneously during personal air sampling. Toluene, ethylbenzene, and xylenes (TEX) presented higher (p < 0.05) ambient levels in WFS than in CDS, particularly during weekdays. A stronger association between personal and ambient concentrations of MTBE and TEX was found in WFS than in CDS. Fourteen to forty-two percent of the variation in personal MTBE, hexane, benzene, and TEX was explained by local outdoor air pollution. These observations indicated that local sources impacted the community air pollution and personal exposure in WFS. The estimated cancer risks resulting from two locally emitted VOCs, benzene and ethylbenzene, and non-cancer neurological and respiratory effects resulting from hexane, benzene, toluene, and xylenes exceeded the US EPA risk benchmarks in both communities. These findings emphasized the need to address disparity in health risks associated with ambient air pollution for the socio-economically disadvantaged groups. This study also demonstrated that air pollution hot spots similar to WFS can provide robust setting to investigate health effects of ambient air pollution.

Variability of Urinary Phthalate Metabolite and Bisphenol A Concentrations before and during Pregnancy

PubMed Central

Braun, Joe M.; Smith, Kristen W.; Williams, Paige L.; Calafat, Antonia M.; Berry, Katharine; Ehrlich, Shelley

2012-01-01

Background: Gestational phthalate and bisphenol A (BPA) exposure may increase the risk of adverse maternal/child health outcomes, but there are few data on the variability of urinary biomarkers before and during pregnancy. Objective: We characterized the variability of urinary phthalate metabolite and BPA concentrations before and during pregnancy and the ability of a single spot urine sample to classify average gestational exposure. Methods: We collected 1,001 urine samples before and during pregnancy from 137 women who were partners in couples attending a Boston fertility clinic and who had a live birth. Women provided spot urine samples before (n ≥ 2) and during (n ≥ 2) pregnancy. We measured urinary concentrations of monoethyl phthalate (MEP), mono-n-butyl phthalate (MBP), mono-iso-butyl phthalate, monobenzyl phthalate (MBzP), four metabolites of di-(2-ethylhexyl) phthalate (DEHP), and BPA. After adjusting for specific gravity, we characterized biomarker variability using intraclass correlation coefficients (ICCs) and conducted several surrogate category analyses to determine whether a single spot urine sample could adequately classify average gestational exposure. Results: Absolute concentrations of phthalate metabolites and BPA were similar before and during pregnancy. Variability was higher during pregnancy than before pregnancy for BPA and MBzP, but similar during and before pregnancy for MBP, MEP, and ΣDEHP. During pregnancy, MEP (ICC = 0.50) and MBP (ICC = 0.45) were less variable than BPA (ICC = 0.12), MBzP (ICC = 0.25), and ΣDEHP metabolites (ICC = 0.08). Surrogate analyses suggested that a single spot urine sample may reasonably classify MEP and MBP concentrations during pregnancy, but more than one sample may be necessary for MBzP, DEHP, and BPA. Conclusions: Urinary phthalate metabolites and BPA concentrations were variable before and during pregnancy, but the magnitude of variability was biomarker specific. A single spot urine sample adequately classified MBP and MEP concentrations during pregnancy. The present results may be related to unique features of the women studied, and replication in other pregnancy cohorts is recommended. PMID:22262702

Quantitation of pregabalin in dried blood spots and dried plasma spots by validated LC-MS/MS methods.

PubMed

Kostić, Nađa; Dotsikas, Yannis; Jović, Nebojša; Stevanović, Galina; Malenović, Anđelija; Medenica, Mirjana

2015-05-10

In this paper, novel LC-MS/MS methods for the determination of antiepileptic drug pregabalin in dried matrix spots (DMS) are presented. This attractive technique of sample collection in micro amount was utilized in the form of dried blood spots (DBS) and dried plasma spots (DPS). Following a pre-column derivatization procedure, using n-propyl chloroformate in the presence of n-propanol, and consecutive liquid-liquid extraction, derivatized pregabalin and its internal standard, 4-aminocyclohexanecarboxylic acid, were detected in positive ion mode by applying two SRM transitions per analyte. A YMC-Pack Octyl column (50mm×4.0mm, 3μm particle size) maintained at 30°C, was utilized with running mobile phase composed of acetonitrile: 0.15% formic acid (85:15, v/v). Flow rate was 550μL/min and total run time 2min. Established methods were fully validated over the concentration range of 0.200-20.0μg/mL for DBS and 0.400-40.0μg/mL for DPS, respectively, while specificity, accuracy, precision, recovery, matrix-effect, stability, dilution integrity and spot homogeneity were found within acceptance criteria. Validated methods were applied for the determination of pregabalin levels in dried blood and plasma samples obtained from patients with epilepsy, after per os administration of commercial capsules. Comparison of drug level in blood and plasma, as well as correction steps undertaken in order to overcome hematocrit issue, when analyzing DBS, are also given. Copyright © 2015 Elsevier B.V. All rights reserved.

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

PubMed

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

2016-12-01

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

Comparing bacterial community composition of healthy and dark spot-affected Siderastrea siderea in Florida and the Caribbean

USGS Publications Warehouse

Kellogg, Christina A.; Piceno, Yvette M.; Tom, Lauren M.; DeSantis, Todd Z.; Gray, Michael A.; Andersen, Gary L.

2014-01-01

Coral disease is one of the major causes of reef degradation. Dark Spot Syndrome (DSS) was described in the early 1990's as brown or purple amorphous areas of tissue on a coral and has since become one of the most prevalent diseases reported on Caribbean reefs. It has been identified in a number of coral species, but there is debate as to whether it is in fact the same disease in different corals. Further, it is questioned whether these macroscopic signs are in fact diagnostic of an infectious disease at all. The most commonly affected species in the Caribbean is the massive starlet coral Siderastrea siderea. We sampled this species in two locations, Dry Tortugas National Park and Virgin Islands National Park. Tissue biopsies were collected from both healthy colonies and those with dark spot lesions. Microbial-community DNA was extracted from coral samples (mucus, tissue, and skeleton), amplified using bacterial-specific primers, and applied to PhyloChip G3 microarrays to examine the bacterial diversity associated with this coral. Samples were also screened for the presence of a fungal ribotype that has recently been implicated as a causative agent of DSS in another coral species, but the amplifications were unsuccessful. S. siderea samples did not cluster consistently based on health state (i.e., normal versus dark spot). Various bacteria, including Cyanobacteria and Vibrios, were observed to have increased relative abundance in the discolored tissue, but the patterns were not consistent across all DSS samples. Overall, our findings do not support the hypothesis that DSS in S. siderea is linked to a bacterial pathogen or pathogens. This dataset provides the most comprehensive overview to date of the bacterial community associated with the scleractinian coral S. siderea.

Comparing bacterial community composition of healthy and dark spot-affected Siderastrea siderea in Florida and the Caribbean

DOE PAGES

Kellogg, Christina A.; Piceno, Yvette M.; Tom, Lauren M.; ...

2014-10-07

Coral disease is one of the major causes of reef degradation. Dark Spot Syndrome (DSS) was described in the early 1990's as brown or purple amorphous areas of tissue on a coral and has since become one of the most prevalent diseases reported on Caribbean reefs. It has been identified in a number of coral species, but there is debate as to whether it is in fact the same disease in different corals. Further, it is questioned whether these macroscopic signs are in fact diagnostic of an infectious disease at all. The most commonly affected species in the Caribbean ismore » the massive starlet coral Siderastrea siderea. We sampled this species in two locations, Dry Tortugas National Park and Virgin Islands National Park. Tissue biopsies were collected from both healthy colonies and those with dark spot lesions. Microbial-community DNA was extracted from coral samples (mucus, tissue, and skeleton), amplified using bacterial-specific primers, and applied to PhyloChip G3 microarrays to examine the bacterial diversity associated with this coral. Samples were also screened for the presence of a fungal ribotype that has recently been implicated as a causative agent of DSS in another coral species, but the amplifications were unsuccessful. S. siderea samples did not cluster consistently based on health state (i.e., normal versus dark spot). Various bacteria, including Cyanobacteria and Vibrios, were observed to have increased relative abundance in the discolored tissue, but the patterns were not consistent across all DSS samples. Overall, our findings do not support the hypothesis that DSS in S. siderea is linked to a bacterial pathogen or pathogens. This dataset provides the most comprehensive overview to date of the bacterial community associated with the scleractinian coral S. siderea.« less

Comparing bacterial community composition of healthy and dark spot-affected Siderastrea siderea in Florida and the Caribbean

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

Kellogg, Christina A.; Piceno, Yvette M.; Tom, Lauren M.

Coral disease is one of the major causes of reef degradation. Dark Spot Syndrome (DSS) was described in the early 1990's as brown or purple amorphous areas of tissue on a coral and has since become one of the most prevalent diseases reported on Caribbean reefs. It has been identified in a number of coral species, but there is debate as to whether it is in fact the same disease in different corals. Further, it is questioned whether these macroscopic signs are in fact diagnostic of an infectious disease at all. The most commonly affected species in the Caribbean ismore » the massive starlet coral Siderastrea siderea. We sampled this species in two locations, Dry Tortugas National Park and Virgin Islands National Park. Tissue biopsies were collected from both healthy colonies and those with dark spot lesions. Microbial-community DNA was extracted from coral samples (mucus, tissue, and skeleton), amplified using bacterial-specific primers, and applied to PhyloChip G3 microarrays to examine the bacterial diversity associated with this coral. Samples were also screened for the presence of a fungal ribotype that has recently been implicated as a causative agent of DSS in another coral species, but the amplifications were unsuccessful. S. siderea samples did not cluster consistently based on health state (i.e., normal versus dark spot). Various bacteria, including Cyanobacteria and Vibrios, were observed to have increased relative abundance in the discolored tissue, but the patterns were not consistent across all DSS samples. Overall, our findings do not support the hypothesis that DSS in S. siderea is linked to a bacterial pathogen or pathogens. This dataset provides the most comprehensive overview to date of the bacterial community associated with the scleractinian coral S. siderea.« less

Comparing Bacterial Community Composition of Healthy and Dark Spot-Affected Siderastrea siderea in Florida and the Caribbean

PubMed Central

Kellogg, Christina A.; Piceno, Yvette M.; Tom, Lauren M.; DeSantis, Todd Z.; Gray, Michael A.; Andersen, Gary L.

2014-01-01

Coral disease is one of the major causes of reef degradation. Dark Spot Syndrome (DSS) was described in the early 1990's as brown or purple amorphous areas of tissue on a coral and has since become one of the most prevalent diseases reported on Caribbean reefs. It has been identified in a number of coral species, but there is debate as to whether it is in fact the same disease in different corals. Further, it is questioned whether these macroscopic signs are in fact diagnostic of an infectious disease at all. The most commonly affected species in the Caribbean is the massive starlet coral Siderastrea siderea. We sampled this species in two locations, Dry Tortugas National Park and Virgin Islands National Park. Tissue biopsies were collected from both healthy colonies and those with dark spot lesions. Microbial-community DNA was extracted from coral samples (mucus, tissue, and skeleton), amplified using bacterial-specific primers, and applied to PhyloChip G3 microarrays to examine the bacterial diversity associated with this coral. Samples were also screened for the presence of a fungal ribotype that has recently been implicated as a causative agent of DSS in another coral species, but the amplifications were unsuccessful. S. siderea samples did not cluster consistently based on health state (i.e., normal versus dark spot). Various bacteria, including Cyanobacteria and Vibrios, were observed to have increased relative abundance in the discolored tissue, but the patterns were not consistent across all DSS samples. Overall, our findings do not support the hypothesis that DSS in S. siderea is linked to a bacterial pathogen or pathogens. This dataset provides the most comprehensive overview to date of the bacterial community associated with the scleractinian coral S. siderea. PMID:25289937

Spatial Variation of Soil Lead in an Urban Community Garden: Implications for Risk-Based Sampling.

PubMed

Bugdalski, Lauren; Lemke, Lawrence D; McElmurry, Shawn P

2014-01-01

Soil lead pollution is a recalcitrant problem in urban areas resulting from a combination of historical residential, industrial, and transportation practices. The emergence of urban gardening movements in postindustrial cities necessitates accurate assessment of soil lead levels to ensure safe gardening. In this study, we examined small-scale spatial variability of soil lead within a 15 × 30 m urban garden plot established on two adjacent residential lots located in Detroit, Michigan, USA. Eighty samples collected using a variably spaced sampling grid were analyzed for total, fine fraction (less than 250 μm), and bioaccessible soil lead. Measured concentrations varied at sampling scales of 1-10 m and a hot spot exceeding 400 ppm total soil lead was identified in the northwest portion of the site. An interpolated map of total lead was treated as an exhaustive data set, and random sampling was simulated to generate Monte Carlo distributions and evaluate alternative sampling strategies intended to estimate the average soil lead concentration or detect hot spots. Increasing the number of individual samples decreases the probability of overlooking the hot spot (type II error). However, the practice of compositing and averaging samples decreased the probability of overestimating the mean concentration (type I error) at the expense of increasing the chance for type II error. The results reported here suggest a need to reconsider U.S. Environmental Protection Agency sampling objectives and consequent guidelines for reclaimed city lots where soil lead distributions are expected to be nonuniform. © 2013 Society for Risk Analysis.

Characterization study of cesium concentrated particles in the soils near the Fukushima Daiichi nuclear power plant

NASA Astrophysics Data System (ADS)

Satou, Yukihiko; Sueki, Keisuke; Sasa, Kimikazu; Adachi, Kouji; Igarashi, Yasuhito

2015-04-01

Radionuclides from the Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident contaminated a vast area. Two types of contamination, spread and spot types, were observed in soils with autoradiography using an imaging plate. Other samples such as dust filters, vegetation, X-ray films, and so on, also indicate the spot type contamination in the early stage of the FDNPP accident. The source of spot type contamination is well known as hot particles at the Chernobyl Nuclear Power Plant (ChNPP) accident in 1986. Hot particles were divided into two groups, fuel hot particles and fission product particles, and they were emitted directly from reactor core with phreatic explosion and fire. In contrast, the official reports of the FDNPP accident did not conforme core explosion. In addition, the emitted total amount of Uranium was very few (Yamamoto et al., 2014). Thus, the spot type contaminations were not identified as the same of hot particles yet. Therefore, the present study aimed to pick up and identify the spot contaminations in soils. Surface soil samples were collected at 20 km northwest from the FDNPP in June 2013. Soils were spread in plastic bags for autoradiography with imaging plate analysis. Then, the soil particles were collected on a sticky carbon tape and analyzed by SEM-EDS to detect radioactive particles. Finally, particles were confirmed to contain photo peaks in the γ-spectrum by a germanium semiconductor detector. Four radioactive particles were isolated from the soil samples in the present study. Detected γ-ray emission radionuclides were only Cs-134 and Cs-137. The X-ray spectra on the SEM-EDS of all particles showed a Cs peak as well as O, Fe, Zn, and Rb peaks, and these elements were distributed uniformly within the particles. In addition, uniform distribution of Si was also shown. Moreover, U was detected from one of the particles, but U concentration was very low and existed locally in the particle. These characters are very similar to previous studies reported particles isolated from air dust filters collected in Tsukuba, Ibaraki, 170 km south from the FDNPP (Adachi et al., 2013, Abe et al., 2014) but are different from former reported hot particles in the ChNPP accident. References Yamamoto, M. et al., J. Environ. Radioact. 2014, 132, 31-46. Adachi, K. et al., Sci. Rep. 2013, 3, 2554. Abe, Y. et al., Anal. Chem. 2014, 86, 8521-8525.

Effects of bacterial infestation caused by human wastes on the skin structures of Mugil platanus Günther, 1880 (Mugilidae).

PubMed

Langer, S L; Vargas, V M F; Flores-Lopes, F; Malabarba, L R

2009-05-01

Manifestation of infectious pathologies in fishes usually increases in environments where organic wastes are disposed. Specimens of Mugil platanus Günther, 1880 and water samples collected at three points of the Tramandaí river were analyzed during a one year period. The macroscopic observation revealed ulcerations in the caudal peduncle area covered with a mass of amorphous and whitened tissues. Histopathologic analysis showed the presence of negative gram bacteria, probably responsible for alterations of the normal structure of the epidermic tissues. Non-parametric statistical analysis for ammonia concentration showed a significant variation among the three collected spots as well as in the multiple comparison between two spots. In this study, we describe cutaneous lesions observed in Mugil platanus specimens and tested their correlation with environmental ammonia concentration.

[Validity of the assessment of urinary protein excretion by spot urine in patients with chronic kidney disease].

PubMed

Hayashi, Ami; Okada, Tomonari; Matsumoto, Hiroshi; Nagaoka, Yume; Wada, Toshikazu; Gondo, Asako; Nango, Tomoka; Miyaoka, Yoshitaka; Watanabe, Kanna; Iwata, Azusa; Nakao, Toshiyuki

2013-01-01

We investigate the validity of the assessment of urinary protein excretion by spot urine samples collected by different methods in outpatients with chronic kidney disease (CKD). SUBJECTS AND METHODS We obtained 24-hour urine and two spot urine samples, including the first morning urine and daytime urine in 159 CKD patients. Urinary protein excretion was assessed by the protein/creatinine ratio from spot urine samples (morning: m-UP (g/gCr), daytime: d-UP (g/gCr) ]. We examined the correlations and the differences among m-UP, d-UP and the actual urinary protein excretion obtained by 24-hour urine (a-UP(g/day) . Significant correlations were found between m-UP and a-UP, and between d-UP and a-UP (r = 0.88, 0.85; p < 0.001). Correlations between m-UP and a-UP were greater relative to those between d-UP and a-UP in patients with less than 3.5 g/day of a-UP and in patients with CKD stages 1 to approximately 3. The percent difference between m-UP and a-UP was--16.0 +/- 40.5%, and that between d-UP and a-UP was 27.1 +/- 72.9%. The absolute value of the percent difference between d-UP and a-UP tended to be greater than that between m-UP and a-UP (34.9 +/- 25.9% vs. 49.9 +/- 59.9%, p = 0.06). Urinary protein/creatinie ratio of the first morning urine is better approximate the urinary protein excretion obtained by 24-hour urine compared with that of spot urine in the daytime.

Identification of Biomarkers of Mercury Contamination in Brachyplatystoma filamentosum of the Madeira River, Brazil, Using Metalloproteomic Strategies.

PubMed

de Queiroz, João Vitor; Vieira, José Cavalcante Souza; de Oliveira, Grasieli; Braga, Camila Pereira; da Cunha Bataglioli, Izabela; da Silva, Janaína Macedo; de Paula Araújo, Wellington Luiz; de Magalhães Padilha, Pedro

2018-05-08

Predator fish can accumulate high levels of mercury, which qualifies them as potential indicators of this toxic metal. The predatory species Brachyplatystoma filamentosum, popularly known as filhote, is among the most consumed species in the Brazilian Amazon. Continuing the metalloproteomic studies of mercury in Amazonian fishes that have been developed in the last 5 years, the present paper provides the data of protein characterization associated with mercury in muscle and liver samples of filhote (Brachyplatystoma filamentosum) collected in the Madeira River, Brazilian Amazon. The mercury concentration in the muscle and liver samples was determined by graphite furnace atomic absorption spectrometry (GFAAS). The protein fraction was extracted in an aqueous medium, and later, a fractional precipitation procedure was performed to obtain the protein pellets. Then, the proteome of the tissue samples of this fish species was separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and a mercury mapping of the protein spots was carried out by GFAAS after acid digestion. Protein spots that had mercury were characterized by mass spectrometry with electrospray ionization in sequence (ESI-MS/MS) after tryptic digestion. It was possible to characterize 11 mercury-associated protein spots that presented biomarker characteristics and could be used to monitor mercury in fish species of the Amazon region. Thus, the metalloproteomic strategies used in the present study allowed us to characterize 11 mercury-associated protein spots. It should be noted that the protein spots identified as GFRP, TMEM186, TMEM57B, and BHMT, which have coordination sites for elements with characteristics of soft acids, such as mercury, can be used as biomarkers of mercury contamination in monitoring studies of this toxic metal in fish species from the Amazon region.

Thin-layer chromatography and colorimetric analysis of multi-component explosive mixtures

DOEpatents

Pagoria, Philip F.; Mitchell, Alexander R.; Whipple, Richard E.; Carman, M. Leslie

2014-08-26

A thin-layer chromatography method for detection and identification of common military and peroxide explosives in samples includes the steps of provide a reverse-phase thin-layer chromatography plate; prepare the plate by marking spots on which to deposit the samples by touching the plate with a marker; spot one micro liter of a first standard onto one of the spots, spot one micro liter of a second standard onto another of the spots, and spot samples onto other of spots producing a spotted plate; add eluent to a developing chamber; add the spotted plate to the developing chamber; remove the spotted plate from the developing chamber producing a developed plate; place the developed plate in an ultraviolet light box; add a visualization agent to a dip tank; dip the developed plate in the dip tank and remove the developed plate quickly; and detect explosives by viewing said developed plate.

A Struggle for Dominance: Relational Communication Messages in Television Programming.

ERIC Educational Resources Information Center

Barbatsis, Gretchen S.; And Others

Television's messages about sex role behavior were analyzed by collecting and coding spot samples of the ten top ranked programs in prime viewing time and proportionate numbers of daytime soap operas and Saturday morning children's programs. The content analysis was based on a relational coding system developed to assess interpersonal…

Distribution of spotted fever group rickettsiae in select tissues of experimentally infected and field-collected Gulf Coast ticks.

PubMed

Edwards, Kristine T; Goddard, Jerome; Varela-Stokes, Andrea

2011-05-01

Salivary glands, midgut, Malpighian tubules, and ovaries were dissected from infected, colony-derived Amblyomma maculatum (Gulf Coast ticks) injected as nymphs with either Rickettsia parkeri (a spotted fever group rickettsia [SFGR]; treatment) or phosphate-buffered saline (negative control). For comparison, similar tissues were dissected from hemolymph-positive, field-collected ticks. Tissues were analyzed by indirect fluorescent antibody (IFA) tests. All phosphate-buffered saline-injected ticks were IFA negative, whereas SFGR were detected by IFA in 100% of the salivary glands and ovaries and 78 and 75% of midgut and Malpighian tubule samples, respectively, of R. parkeri-injected ticks. Nearly 22% (10/46) of the field-collected ticks were hemolymph positive. Of those, SFGR were detected by IFA in 80% of the salivary glands, 67% of the ovaries, and 60% in the midgut and Malpighian tubules. This is the first study to assess the distribution of SFGR in select tissues of A. maculatum ticks.

Effect of storage conditions on the weight and appearance of dried blood spot samples on various cellulose-based substrates.

PubMed

Denniff, Philip; Spooner, Neil

2010-11-01

Before shipping and storage, dried blood spot (DBS) samples must be dried in order to protect the integrity of the spots. In this article, we examine the time required to dry blood spot samples and the effects of different environmental conditions on their integrity. Under ambient laboratory conditions, DBS samples on Whatman 903(®), FTA(®) and FTA(®) Elute substrates are dry within 90 min of spotting. An additional 5% of moisture is lost during subsequent storage with desiccant. When exposed to elevated conditions of temperature and relative humidity, the DBS samples absorb moisture. DBS samples on FTA lose this moisture on being returned to ambient conditions. DBS samples on 903 show no visible signs of deterioration when stored at elevated conditions. However, these conditions cause the DBS to diffuse through the FTA Elute substrate. Blood spots are dry within 90 min of spotting. However, the substrates examined behave differently when exposed to conditions of high relative humidity and temperature, in some cases resulting in the integrity of the substrate and DBS sample being compromised. It is recommended that these factors be investigated as part of method development and validation.

Comparison of the manual, semiautomatic, and automatic selection and leveling of hot spots in whole slide images for Ki-67 quantification in meningiomas.

PubMed

Swiderska, Zaneta; Korzynska, Anna; Markiewicz, Tomasz; Lorent, Malgorzata; Zak, Jakub; Wesolowska, Anna; Roszkowiak, Lukasz; Slodkowska, Janina; Grala, Bartlomiej

2015-01-01

Background. This paper presents the study concerning hot-spot selection in the assessment of whole slide images of tissue sections collected from meningioma patients. The samples were immunohistochemically stained to determine the Ki-67/MIB-1 proliferation index used for prognosis and treatment planning. Objective. The observer performance was examined by comparing results of the proposed method of automatic hot-spot selection in whole slide images, results of traditional scoring under a microscope, and results of a pathologist's manual hot-spot selection. Methods. The results of scoring the Ki-67 index using optical scoring under a microscope, software for Ki-67 index quantification based on hot spots selected by two pathologists (resp., once and three times), and the same software but on hot spots selected by proposed automatic methods were compared using Kendall's tau-b statistics. Results. Results show intra- and interobserver agreement. The agreement between Ki-67 scoring with manual and automatic hot-spot selection is high, while agreement between Ki-67 index scoring results in whole slide images and traditional microscopic examination is lower. Conclusions. The agreement observed for the three scoring methods shows that automation of area selection is an effective tool in supporting physicians and in increasing the reliability of Ki-67 scoring in meningioma.

Comparison of the Manual, Semiautomatic, and Automatic Selection and Leveling of Hot Spots in Whole Slide Images for Ki-67 Quantification in Meningiomas

PubMed Central

Swiderska, Zaneta; Korzynska, Anna; Markiewicz, Tomasz; Lorent, Malgorzata; Zak, Jakub; Wesolowska, Anna; Roszkowiak, Lukasz; Slodkowska, Janina; Grala, Bartlomiej

2015-01-01

Background. This paper presents the study concerning hot-spot selection in the assessment of whole slide images of tissue sections collected from meningioma patients. The samples were immunohistochemically stained to determine the Ki-67/MIB-1 proliferation index used for prognosis and treatment planning. Objective. The observer performance was examined by comparing results of the proposed method of automatic hot-spot selection in whole slide images, results of traditional scoring under a microscope, and results of a pathologist's manual hot-spot selection. Methods. The results of scoring the Ki-67 index using optical scoring under a microscope, software for Ki-67 index quantification based on hot spots selected by two pathologists (resp., once and three times), and the same software but on hot spots selected by proposed automatic methods were compared using Kendall's tau-b statistics. Results. Results show intra- and interobserver agreement. The agreement between Ki-67 scoring with manual and automatic hot-spot selection is high, while agreement between Ki-67 index scoring results in whole slide images and traditional microscopic examination is lower. Conclusions. The agreement observed for the three scoring methods shows that automation of area selection is an effective tool in supporting physicians and in increasing the reliability of Ki-67 scoring in meningioma. PMID:26240787

Urinary spot albumin:creatinine ratio for documenting proteinuria in women with preeclampsia.

PubMed

Huang, Qitao; Gao, Yunfei; Yu, Yanhong; Wang, Wei; Wang, Shuoshi; Zhong, Mei

2012-01-01

To assess whether a single urinary spot urinary albumin:creatinine ratio (ACR) can be used to estimate 24-hour urinary protein excretion in women with preeclampsia. ACR and 24-hour urinary protein excretion were measured in 50 consecutive patients with preeclampsia. ACR was determined in a spot midstream urine sample and the amount of protein excretion was quantified in a 24-hour urine collection performed the following day. The correlation between the spot ACR and 24-hour urine protein excretion was assessed, and the diagnostic value of ACR was expressed in terms of specificity and sensitivity. Receiver operating characteristic curve analysis was used to determine the best cutoff values of the spot ACR for mild preeclampsia (proteinuria ≥ 0.3 g/24 h) and severe preeclampsia (defined in China as proteinuria ≥ 2 g/24 h). A strong correlation was evident between the spot ACR and 24-hour urinary protein excretion (r = .938; P < .001). The optimal spot ACR cutoff point was 22.8 mg/mmol for 0.3 g/24 h of protein excretion (mild preeclampsia) with a sensitivity and specificity of 82.4% and 99.4%, respectively, and 155.6 mg/mmol for 2 g/24 h of protein excretion (severe preeclampsia) with a sensitivity and specificity of 90.6% and 99.6%, respectively. Compared with 24-hour urinary protein excretion, the spot urinary ACR may be a simple, convenient, and accurate indicator of significant proteinuria in women with preeclampsia.

Postnatal gestational age estimation using newborn screening blood spots: a proposed validation protocol

PubMed Central

Murphy, Malia S Q; Hawken, Steven; Atkinson, Katherine M; Milburn, Jennifer; Pervin, Jesmin; Gravett, Courtney; Stringer, Jeffrey S A; Rahman, Anisur; Lackritz, Eve; Chakraborty, Pranesh; Wilson, Kumanan

2017-01-01

Background Knowledge of gestational age (GA) is critical for guiding neonatal care and quantifying regional burdens of preterm birth. In settings where access to ultrasound dating is limited, postnatal estimates are frequently used despite the issues of accuracy associated with postnatal approaches. Newborn metabolic profiles are known to vary by severity of preterm birth. Recent work by our group and others has highlighted the accuracy of postnatal GA estimation algorithms derived from routinely collected newborn screening profiles. This protocol outlines the validation of a GA model originally developed in a North American cohort among international newborn cohorts. Methods Our primary objective is to use blood spot samples collected from infants born in Zambia and Bangladesh to evaluate our algorithm’s capacity to correctly classify GA within 1, 2, 3 and 4 weeks. Secondary objectives are to 1) determine the algorithm's accuracy in small-for-gestational-age and large-for-gestational-age infants, 2) determine its ability to correctly discriminate GA of newborns across dichotomous thresholds of preterm birth (≤34 weeks, <37 weeks GA) and 3) compare the relative performance of algorithms derived from newborn screening panels including all available analytes and those restricted to analyte subsets. The study population will consist of infants born to mothers already enrolled in one of two preterm birth cohorts in Lusaka, Zambia, and Matlab, Bangladesh. Dried blood spot samples will be collected and sent for analysis in Ontario, Canada, for model validation. Discussion This study will determine the validity of a GA estimation algorithm across ethnically diverse infant populations and assess population specific variations in newborn metabolic profiles. PMID:29104765

Effect of ambient humidity on the rate at which blood spots dry and the size of the spot produced.

PubMed

Denniff, Philip; Woodford, Lynsey; Spooner, Neil

2013-08-01

For shipping and storage, dried blood spot (DBS) samples must be sufficiently dry to protect the integrity of the sample. When the blood is spotted the humidity has the potential to affect the size of the spot created and the speed at which it dries. The area of DBS produced on three types of substrates were not affected by the humidity under which they were generated. DBS samples reached a steady moisture content 150 min after spotting and 90 min for humidities less than 60% relative humidity. All packaging materials examined provided some degree of protection from external extreme conditions. However, none of the packaging examined provided a total moisture barrier to extreme environmental conditions. Humidity was shown not to affect the spot area and DBS samples were ready for shipping and storage 2 h after spotting. The packing solutions examined all provided good protection from external high humidity conditions.

Hyperspectral imaging and multivariate analysis in the dried blood spots investigations

NASA Astrophysics Data System (ADS)

Majda, Alicja; Wietecha-Posłuszny, Renata; Mendys, Agata; Wójtowicz, Anna; Łydżba-Kopczyńska, Barbara

2018-04-01

The aim of this study was to apply a new methodology using the combination of the hyperspectral imaging and the dry blood spot (DBS) collecting. Application of the hyperspectral imaging is fast and non-destructive. DBS method offers the advantage also on the micro-invasive blood collecting and low volume of required sample. During experimental step, the reflected light was recorded by two hyperspectral systems. The collection of 776 spectral bands in the VIS-NIR range (400-1000 nm) and 256 spectral bands in the SWIR range (970-2500 nm) was applied. Pixel has the size of 8 × 8 and 30 × 30 µm for VIS-NIR and SWIR camera, respectively. The obtained data in the form of hyperspectral cubes were treated with chemometric methods, i.e., minimum noise fraction and principal component analysis. It has been shown that the application of these methods on this type of data, by analyzing the scatter plots, allows a rapid analysis of the homogeneity of DBS, and the selection of representative areas for further analysis. It also gives the possibility of tracking the dynamics of changes occurring in biological traces applied on the surface. For the analyzed 28 blood samples, described method allowed to distinguish those blood stains because of time of apply.

Epidemiology of Rickettsia sp. strain Atlantic rainforest in a spotted fever-endemic area of southern Brazil.

PubMed

Barbieri, Amalia R M; Filho, Jonas M; Nieri-Bastos, Fernanda A; Souza, Julio C; Szabó, Matias P J; Labruna, Marcelo B

2014-10-01

The present study was performed in Vila Itoupava, an area of the state of Santa Catarina, southern Brazil, in which a tick-borne spotted fever illness has been endemic since 2003. Notably, both the etiological agent and the vector of these spotted fever cases remain unknown. During January 2011, humans, domestic dogs, and their ticks were sampled in households that are typically surrounded by highly preserved Atlantic rainforest fragments. Ticks collected from dogs were Amblyomma ovale (34% prevalence), Amblyomma aureolatum (18.9%), and Rhipicephalus sanguineus (3.8%). A total of 7.8% (6/77) A. ovale and 9.3% (4/43) A. aureolatum were infected by Rickettsia sp. strain Atlantic rainforest, a Rickettsia parkeri-like agent recently shown to cause spotted fever illness in southeastern Brazil. Overall, 67.3% (35/52) of the dogs were seroreactive to spotted fever group rickettsiae, mostly with highest endpoint titers to R. parkeri. Among humans, 46.7% (7/15) reacted serologically to rickettsiae at low to moderate endpoint titers. Because canine seroreactivity to R. parkeri was strongly associated with frequent contact with forests (the preferred habitat for A. ovale and A. aureolatum), it is concluded that sampled dogs have been infected by strain Atlantic rainforest through the parasitism of these tick species. The present study provides epidemiological evidence that the spotted fever in the study area has been caused by Rickettsia sp. strain Atlantic rainforest, transmitted to humans by either A. ovale or A. aureolatum. Further studies encompassing direct diagnostic methods on clinical specimens from patients are needed to confirm the above epidemiological evidence. Copyright © 2014 Elsevier GmbH. All rights reserved.

Quantification of multiple elements in dried blood spot samples.

PubMed

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

2017-08-01

Dried blood spots (DBS) is a unique matrix that offers advantages compared to conventional blood collection making it increasingly popular in large population studies. We here describe development and validation of a method to determine multiple elements in DBS. Elements were extracted from punches and analyzed using inductively coupled plasma-mass spectrometry (ICP-MS). The method was evaluated with quality controls with defined element concentration and blood spiked with elements to assess accuracy and imprecision. DBS element concentrations were compared with concentrations in venous blood. Samples with different hematocrit were spotted onto filter paper to assess hematocrit effect. The established method was precise and accurate for measurement of most elements in DBS. There was a significant but relatively weak correlation between measurement of the elements Mg, K, Fe, Cu, Zn, As and Se in DBS and venous whole blood. Hematocrit influenced the DBS element measurement, especially for K, Fe and Zn. Trace elements can be measured with high accuracy and low imprecision in DBS, but contribution of signal from the filter paper influences measurement of some elements present at low concentrations. Simultaneous measurement of K and Fe in DBS extracts may be used to estimate sample hematocrit. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Reliable and fast quantitative analysis of active ingredient in pharmaceutical suspension using Raman spectroscopy.

PubMed

Park, Seok Chan; Kim, Minjung; Noh, Jaegeun; Chung, Hoeil; Woo, Youngah; Lee, Jonghwa; Kemper, Mark S

2007-06-12

The concentration of acetaminophen in a turbid pharmaceutical suspension has been measured successfully using Raman spectroscopy. The spectrometer was equipped with a large spot probe which enabled the coverage of a representative area during sampling. This wide area illumination (WAI) scheme (coverage area 28.3 mm2) for Raman data collection proved to be more reliable for the compositional determination of these pharmaceutical suspensions, especially when the samples were turbid. The reproducibility of measurement using the WAI scheme was compared to that of using a conventional small-spot scheme which employed a much smaller illumination area (about 100 microm spot size). A layer of isobutyric anhydride was placed in front of the sample vials to correct the variation in the Raman intensity due to the fluctuation of laser power. Corrections were accomplished using the isolated carbonyl band of isobutyric anhydride. The acetaminophen concentrations of prediction samples were accurately estimated using a partial least squares (PLS) calibration model. The prediction accuracy was maintained even with changes in laser power. It was noted that the prediction performance was somewhat degraded for turbid suspensions with high acetaminophen contents. When comparing the results of reproducibility obtained with the WAI scheme and those obtained using the conventional scheme, it was concluded that the quantitative determination of the active pharmaceutical ingredient (API) in turbid suspensions is much improved when employing a larger laser coverage area. This is presumably due to the improvement in representative sampling.

High-resolution droplet-based fractionation of nano-LC separations onto microarrays for MALDI-MS analysis.

PubMed

Küster, Simon K; Pabst, Martin; Jefimovs, Konstantins; Zenobi, Renato; Dittrich, Petra S

2014-05-20

We present a robust droplet-based device, which enables the fractionation of ultralow flow rate nanoflow liquid chromatography (nano-LC) eluate streams at high frequencies and high peak resolution. This is achieved by directly interfacing the separation column to a micro T-junction, where the eluate stream is compartmentalized into picoliter droplets. This immediate compartmentalization prevents peak dispersion during eluate transport and conserves the chromatographic performance. Subsequently, nanoliter eluate fractions are collected at a rate of one fraction per second on a high-density microarray to retain the separation with high temporal resolution. Chromatographic separations of up to 45 min runtime can thus be archived on a single microarray possessing 2700 sample spots. The performance of this device is demonstrated by fractionating the separation of a tryptic digest of a known protein mixture onto the microarray chip and subsequently analyzing the sample archive using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Resulting peak widths are found to be significantly reduced compared to standard continuous flow spotting technologies as well as in comparison to a conventional nano-LC-electrospray ionization-mass spectrometry interface. Moreover, we demonstrate the advantage of our high-definition nanofractionation device by applying two different MALDI matrices to all collected fractions in an alternating fashion. Since the information that is obtained from a MALDI-MS measurement depends on the choice of MALDI matrix, we can extract complementary information from neighboring spots containing almost identical composition but different matrices.

Urinary iodine concentration identifies pregnant women as iodine deficient yet school-aged children as iodine sufficient in rural Niger.

PubMed

Hess, Sonja Y; Ouédraogo, Césaire T; Young, Rebecca R; Bamba, Ibrahim F; Stinca, Sara; Zimmermann, Michael B; Wessells, K Ryan

2017-05-01

To assess iodine status among pregnant women in rural Zinder, Niger and to compare their status with the iodine status of school-aged children from the same households. Seventy-three villages in the catchment area of sixteen health centres were randomly selected to participate in the cross-sectional survey. Salt iodization is mandatory in Niger, requiring 20-60 ppm iodine at the retail level. A spot urine sample was collected from randomly selected pregnant women (n 662) and one school-aged child from the same household (n 373). Urinary iodine concentration (UIC) was assessed as an indicator of iodine status in both groups. Dried blood spots (DBS) were collected from venous blood samples of pregnant women and thyroglobulin (Tg), thyroid-stimulating hormone and total thyroxine were measured. Iodine content of household salt samples (n 108) was assessed by titration. Median iodine content of salt samples was 5·5 ppm (range 0-41 ppm), 98 % had an iodine content 40 µg/l. In this region of Niger, most salt is inadequately iodized. UIC in pregnant women indicated iodine deficiency, whereas UIC of school-aged children indicated marginally adequate iodine status. Thus, estimating population iodine status based solely on monitoring of UIC among school-aged children may underestimate the risk of iodine deficiency in pregnant women.

Validation of an LC-MS/MS assay to simultaneously monitor the intracellular active metabolites of tenofovir, emtricitabine, and lamivudine in dried blood spots.

PubMed

Schauer, Amanda P; Sykes, Craig; Cottrell, Mackenzie L; Prince, Heather; Kashuba, Angela D M

2018-02-05

The ability to monitor adherence to antiretroviral therapy is critical for the interpretation of outcomes from clinical studies of HIV, and for optimizing patient care. The antiretrovirals tenofovir (TFV), emtricitabine (FTC), and lamivudine (3TC) are commonly included in drug regimens for HIV prevention and treatment. The active form of the drugs tenofovir diphosphate (TFVdp), emtricitabine triphosphate (FTCtp), and lamivudine triphosphate (3TCtp) are found intracellularly in erythrocytes and peripheral blood mononuclear cells (PBMCs). The ability to collect and analyze dried blood spot (DBS) samples is an attractive alternative to PBMC sampling in many resource limited settings. We developed and validated an assay to quantify all three intracellular metabolites over the range of 100-25000 fmol/sample. This assay utilizes a simple protein precipitation/liquid-liquid extraction of a single 3-mm DBS punch (from a Whatman 903 Protein Saver card) with isotopically labeled 13 C 5 -TFVdp included as the internal standard. Following extraction, samples are analyzed by anion exchange chromatography on a Thermo Biobasic AX 5μm column with detection by electrospray ionization in the positive mode on a AB Sciex API-5000 triple quadrupole mass spectrometer with a total run time of 8min. The assay was linear over the entire range (R 2 >0.996). The assay was accurate (inter-assay%bias within ±3.0%) and precise (inter-assay % CV≤9.8%). The assay was also reproducible from multiple punches within a spot as well as punches from separate blood spots. Stability was established at room temperature for 3days, and at -80°C for up to 63days. Clinical samples were analyzed from subjects on Truvada ® , Stribild ® , Descovy ® , and Triumeq ® regimens and intracellular metabolites were detected in all samples as expected, indicating the assay performs well for all current formulations of TFV, FTC, and 3TC. Copyright © 2017 Elsevier B.V. All rights reserved.

Telemetry-based mortality estimates of juvenile spot in two North Carolina estuarine creeks

USGS Publications Warehouse

Friedl, Sarah E.; Buckel, Jeffery A.; Hightower, Joseph E.; Scharf, Frederick S.; Pollock, Kenneth H.

2013-01-01

We estimated natural mortality rates (M) of age-1 Spot Leiostomus xanthurus by using a sonic telemetry approach. Sonic transmitters were surgically implanted into a total of 123 age-1 Spot in two North Carolina estuarine creeks during spring 2009 and 2010, and the fish were monitored by using a stationary acoustic receiver array and manual tracking. Fates of telemetered Spot were inferred based on telemetry information from estimated locations and swimming speeds. Potential competitors of age-1 Spot were assessed through simultaneous otter trawl sampling, while potential predators of Spot were collected using gill nets and trammel nets. The number of inferred natural mortalities was zero in 2009 (based on 29 telemetered Spot at risk) and four in 2010 (based on 52 fish at risk), with fish being at risk for up to about 70 d each year. Catches of potential competitors or predators did not differ between years, and age-1 Spot were not found in analyzed stomach contents of potential predators. Our estimated 30-d M of 0.03 (95% credible interval = 0.01–0.07) was lower than that predicted from weight-based (M = 0.07) and life-history-based (M = 0.06–0.36) estimates. Our field-based estimate of M for age-1 Spot in this estuarine system can assist in the assessment and management of Spot by allowing a direct comparison with M-values predicted from fish size or life history characteristics. The field telemetry and statistical analysis techniques developed here provide guidance for future telemetry studies of relatively small fish in open, dynamic habitat systems, as they highlight strengths and weaknesses of using a telemetry approach to estimate M.

Three Novel Herpesviruses of Endangered Clemmys and Glyptemys Turtles

PubMed Central

Ossiboff, Robert J.; Raphael, Bonnie L.; Ammazzalorso, Alyssa D.; Seimon, Tracie A.; Newton, Alisa L.; Chang, Tylis Y.; Zarate, Brian; Whitlock, Alison L.; McAloose, Denise

2015-01-01

The rich diversity of the world’s reptiles is at risk due to significant population declines of broad taxonomic and geographic scope. Significant factors attributed to these declines include habitat loss, pollution, unsustainable collection and infectious disease. To investigate the presence and significance of a potential pathogen on populations of critically endangered bog turtles (Glyptemys muhlenbergii) as well sympatric endangered wood (G. insculpta) and endangered spotted (Clemmys guttata) turtles in the northeastern United States, choanal and cloacal swabs collected from 230 turtles from 19 sites in 5 states were screened for herpesvirus by polymerase chain reaction. We found a high incidence of herpesvirus infection in bog turtles (51.5%; 105/204) and smaller numbers of positive wood (5) and spotted (1) turtles. Sequence and phylogenetic analysis revealed three previously uncharacterized alphaherpesviruses. Glyptemys herpesvirus 1 was the predominant herpesvirus detected and was found exclusively in bog turtles in all states sampled. Glyptemys herpesvirus 2 was found only in wood turtles. Emydid herpesvirus 2 was found in a small number of bog turtles and a single spotted turtle from one state. Based on these findings, Glyptemys herpesvirus 1 appears to be a common infection in the study population, whereas Glyptemys herpesvirus 2 and Emydid herpesvirus 2 were not as frequently detected. Emydid herpesvirus 2 was the only virus detected in more than one species. Herpesviruses are most often associated with subclinical or mild infections in their natural hosts, and no sampled turtles showed overt signs of disease at sampling. However, infection of host-adapted viruses in closely related species can result in significant disease. The pathogenic potential of these viruses, particularly Emydid herpesvirus 2, in sympatric chelonians warrants additional study in order to better understand the relationship of these viruses with their endangered hosts. PMID:25875510

Three novel herpesviruses of endangered Clemmys and Glyptemys turtles.

PubMed

Ossiboff, Robert J; Raphael, Bonnie L; Ammazzalorso, Alyssa D; Seimon, Tracie A; Newton, Alisa L; Chang, Tylis Y; Zarate, Brian; Whitlock, Alison L; McAloose, Denise

2015-01-01

The rich diversity of the world's reptiles is at risk due to significant population declines of broad taxonomic and geographic scope. Significant factors attributed to these declines include habitat loss, pollution, unsustainable collection and infectious disease. To investigate the presence and significance of a potential pathogen on populations of critically endangered bog turtles (Glyptemys muhlenbergii) as well sympatric endangered wood (G. insculpta) and endangered spotted (Clemmys guttata) turtles in the northeastern United States, choanal and cloacal swabs collected from 230 turtles from 19 sites in 5 states were screened for herpesvirus by polymerase chain reaction. We found a high incidence of herpesvirus infection in bog turtles (51.5%; 105/204) and smaller numbers of positive wood (5) and spotted (1) turtles. Sequence and phylogenetic analysis revealed three previously uncharacterized alphaherpesviruses. Glyptemys herpesvirus 1 was the predominant herpesvirus detected and was found exclusively in bog turtles in all states sampled. Glyptemys herpesvirus 2 was found only in wood turtles. Emydid herpesvirus 2 was found in a small number of bog turtles and a single spotted turtle from one state. Based on these findings, Glyptemys herpesvirus 1 appears to be a common infection in the study population, whereas Glyptemys herpesvirus 2 and Emydid herpesvirus 2 were not as frequently detected. Emydid herpesvirus 2 was the only virus detected in more than one species. Herpesviruses are most often associated with subclinical or mild infections in their natural hosts, and no sampled turtles showed overt signs of disease at sampling. However, infection of host-adapted viruses in closely related species can result in significant disease. The pathogenic potential of these viruses, particularly Emydid herpesvirus 2, in sympatric chelonians warrants additional study in order to better understand the relationship of these viruses with their endangered hosts.

Day-to-day variability in spot urine protein-creatinine ratio measurements.

PubMed

Naresh, Chetana N; Hayen, Andrew; Craig, Jonathan C; Chadban, Steven J

2012-10-01

Accurate measurement of proteinuria is important in the diagnosis and management of chronic kidney disease (CKD). The reference standard test, 24-hour urinary protein excretion, is inconvenient and vulnerable to collection errors. Spot urine protein-creatinine ratio (PCR) is a convenient alternative and is in widespread use. However, day-to-day variability in PCR measurements has not been evaluated. Prospective cohort study of day-to-day variability in spot urine PCR measurement. Clinically stable outpatients with CKD (n = 145) attending a university hospital CKD clinic in Australia between July 2007 and April 2010. Spot urine PCR. Spot PCR variability was assessed and repeatability limits were determined using fractional polynomials. Spot PCRs were measured from urine samples collected at 9:00 am on consecutive days and 24-hour urinary protein excretion was collected concurrently. Paired results were analyzed from 145 patients: median age, 56 years; 59% men; and median 24-hour urinary protein excretion, 0.7 (range, 0.06-35.7) g/d. Day-to-day variability was substantial and increased in absolute terms, but decreased in relative terms with increasing baseline PCR. For patients with a low baseline PCR (20 mg/mmol [177 mg/g]), a change greater than ±160% (repeatability limits, 0-52 mg/mmol [0-460 mg/g]) is required to indicate a real change in proteinuria status with 95% certainty, whereas for those with a high baseline PCR (200 mg/mmol [1,768 mg/g]), a change of ±50% (decrease to <100 mg/mmol [<884 mg/g] or increase to >300 mg/mmol [>2,652 mg/g]) represents significant change. These study results need to be replicated in other ethnic groups. Changes in PCR observed in patients with CKD, ranging from complete resolution to doubling of PCR values, could be due to inherent biological variation and may not indicate a change in disease status. This should be borne in mind when using PCR in the diagnosis and management of CKD. Copyright © 2012 National Kidney Foundation, Inc. All rights reserved.

Temporal variability in urinary levels of drinking water disinfection byproducts dichloroacetic acid and trichloroacetic acid among men

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

Wang, Yi-Xin; Zeng, Qiang; Wang, Le

Urinary haloacetic acids (HAAs), such as dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA), have been suggested as potential biomarkers of exposure to drinking water disinfection byproducts (DBPs). However, variable exposure to and the short elimination half-lives of these biomarkers can result in considerable variability in urinary measurements, leading to exposure misclassification. Here we examined the variability of DCAA and TCAA levels in the urine among eleven men who provided urine samples on 8 days over 3 months. The urinary concentrations of DCAA and TCAA were measured by gas chromatography coupled with electron capture detection. We calculated the intraclass correlation coefficientsmore » (ICCs) to characterize the within-person and between-person variances and computed the sensitivity and specificity to assess how well single or multiple urine collections accurately determined personal 3-month average DCAA and TCAA levels. The within-person variance was much higher than the between-person variance for all three sample types (spot, first morning, and 24-h urine samples) for DCAA (ICC=0.08–0.37) and TCAA (ICC=0.09–0.23), regardless of the sampling interval. A single-spot urinary sample predicted high (top 33%) 3-month average DCAA and TCAA levels with high specificity (0.79 and 0.78, respectively) but relatively low sensitivity (0.47 and 0.50, respectively). Collecting two or three urine samples from each participant improved the classification. The poor reproducibility of the measured urinary DCAA and TCAA concentrations indicate that a single measurement may not accurately reflect individual long-term exposure. Collection of multiple urine samples from one person is an option for reducing exposure classification errors in studies exploring the effects of DBP exposure on reproductive health. - Highlights: • We evaluated the variability of DCAA and TCAA levels in the urine among men. • Urinary DCAA and TCAA levels varied greatly over a 3-month period. • Single measurement may not accurately reflect personal long-term exposure levels. • Collecting multiple samples from one person improved the exposure classification.« less

First evidence of dengue infection in domestic dogs living in different ecological settings in Thailand.

PubMed

Thongyuan, Suporn; Kittayapong, Pattamaporn

2017-01-01

Dengue is a vector-borne disease transmitted by Aedes mosquitoes. It is considered an important public health problem in many countries worldwide. However, only a few studies have been conducted on primates and domestic animals that could potentially be a reservoir of dengue viruses. Since domestic dogs share both habitats and vectors with humans, this study aimed to investigate whether domestic dogs living in different ecological settings in dengue endemic areas in Thailand could be naturally infected with dengue viruses. Serum samples were collected from domestic dogs in three different ecological settings of Thailand: urban dengue endemic areas of Nakhon Sawan Province; rubber plantation areas of Rayong Province; and Koh Chang, an island tourist spot of Trat Province. These samples were screened for dengue viral genome by using semi-nested RT-PCR. Positive samples were then inoculated in mosquito and dog cell lines for virus isolation. Supernatant collected from cell culture was tested for the presence of dengue viral genome by semi-nested RT-PCR, then double-strand DNA products were double-pass custom-sequenced. Partial nucleotide sequences were aligned with the sequences already recorded in GenBank, and a phylogenetic tree was constructed. In the urban setting, 632 domestic dog serum samples were screened for dengue virus genome by RT-PCR, and six samples (0.95%) tested positive for dengue virus. Four out of six dengue viruses from positive samples were successfully isolated. Dengue virus serotype 2 and serotype 3 were found to have circulated in domestic dog populations. One of 153 samples (0.65%) collected from the rubber plantation area showed a PCR-positive result, and dengue serotype 3 was successfully isolated. Partial gene phylogeny revealed that the isolated dengue viruses were closely related to those strains circulating in human populations. None of the 71 samples collected from the island tourist spot showed a positive result. We concluded that domestic dogs can be infected with dengue virus strains circulating in dengue endemic areas. The role of domestic dogs in dengue transmission needs to be further investigated, i.e., whether they are potential reservoirs or incidental hosts of dengue viruses.

Brazilian spotted fever: real-time PCR for diagnosis of fatal cases.

PubMed

dos Santos, Fabiana Cristina Pereira; do Nascimento, Elvira Maria Mendes; Katz, Gizelda; Angerami, Rodrigo Nogueira; Colombo, Silvia; de Souza, Eliana Rodrigues; Labruna, Marcelo Bahia; da Silva, Marcos Vinicius

2012-12-01

Suspicion of Brazilian spotted fever (BSF) should occur in endemic regions upon surveillance of the acute febrile icteric hemorrhagic syndrome (AFIHS). However, limitations associated with currently available laboratory tests pose a challenge to early diagnosis, especially in fatal cases. Two real-time PCR (qPCR) protocols were evaluated to diagnose BSF in 110 fatal AFIHS cases, collected in BSF-endemic regions in 2009-2010. Of these, 24 were positive and 86 negative by indirect immunofluorescence (IFA) assay (cut-off IgG and/or IgM ≥ 128). DNA from these samples was used in the qPCR protocols: one to detect Rickettsia spp. (citrate synthase gene) and another to determine spotted fever group (SFG) Rickettsia species (OmpA gene). Of the 24 IFA-positive samples, 5 (21%) were positive for OmpA and 9 (38%) for citrate synthase. In the IFA-negative group (n=86), OmpA and citrate synthase were positive in 23 (27%) and 27 (31%), respectively. These results showed that the 2 qPCR protocols were about twice as sensitive as the IFA test alone (93% concordance). In conclusion, qPCR is a sensitive method for the diagnosis of fatal BSF cases and should be considered for routine surveillance of AFIHS in places like Brazil, where spotted fever-related lethality is high and other endemic diseases like dengue and leptospirosis can mislead diagnosis. Copyright © 2012 Elsevier GmbH. All rights reserved.

Sampling probe for microarray read out using electrospray mass spectrometry

DOEpatents

Van Berkel, Gary J.

2004-10-12

An automated electrospray based sampling system and method for analysis obtains samples from surface array spots having analytes. The system includes at least one probe, the probe including an inlet for flowing at least one eluting solvent to respective ones of a plurality of spots and an outlet for directing the analyte away from the spots. An automatic positioning system is provided for translating the probe relative to the spots to permit sampling of any spot. An electrospray ion source having an input fluidicly connected to the probe receives the analyte and generates ions from the analyte. The ion source provides the generated ions to a structure for analysis to identify the analyte, preferably being a mass spectrometer. The probe can be a surface contact probe, where the probe forms an enclosing seal along the periphery of the array spot surface.

Dried blood spot measurement: application in tacrolimus monitoring using limited sampling strategy and abbreviated AUC estimation.

PubMed

Cheung, Chi Yuen; van der Heijden, Jaques; Hoogtanders, Karin; Christiaans, Maarten; Liu, Yan Lun; Chan, Yiu Han; Choi, Koon Shing; van de Plas, Afke; Shek, Chi Chung; Chau, Ka Foon; Li, Chun Sang; van Hooff, Johannes; Stolk, Leo

2008-02-01

Dried blood spot (DBS) sampling and high-performance liquid chromatography tandem-mass spectrometry have been developed in monitoring tacrolimus levels. Our center favors the use of limited sampling strategy and abbreviated formula to estimate the area under concentration-time curve (AUC(0-12)). However, it is inconvenient for patients because they have to wait in the center for blood sampling. We investigated the application of DBS method in tacrolimus level monitoring using limited sampling strategy and abbreviated AUC estimation approach. Duplicate venous samples were obtained at each time point (C(0), C(2), and C(4)). To determine the stability of blood samples, one venous sample was sent to our laboratory immediately. The other duplicate venous samples, together with simultaneous fingerprick blood samples, were sent to the University of Maastricht in the Netherlands. Thirty six patients were recruited and 108 sets of blood samples were collected. There was a highly significant relationship between AUC(0-12), estimated from venous blood samples, and fingerprick blood samples (r(2) = 0.96, P < 0.0001). Moreover, there was an excellent correlation between whole blood venous tacrolimus levels in the two centers (r(2) = 0.97; P < 0.0001). The blood samples were stable after long-distance transport. DBS sampling can be used in centers using limited sampling and abbreviated AUC(0-12) strategy as drug monitoring.

Spectral induced polarization as a tool to map subsurface biogeochemical hot spots: a first laboratory evaluation in the Fe-S system

NASA Astrophysics Data System (ADS)

Nordsiek, Sven; Gilfedder, Ben; Frei, Sven

2017-04-01

Zones of intense biogeochemical reactivity (hot spots) arise in the saturated subsurface at the interface between regions with oxidizing and reducing conditions. Hot spots are both sinks and sources of different chemical compounds, thus they are of particular importance for element cycling in the subsurface. However, the investigation of hot spot structures is difficult, because they are not directly identifiable from the surface and can only be investigated by invasive methods in the subsurface. Additionally, they often form in sensitive wetland ecosystems where only non-destructive measurements are applicable to avoid significant degradation of these sensitive environments. Under these circumstances, geophysical methods may provide useful tools to identify biogeochemically active regions. One of the most important biogeochemical reactions in wetlands is the reduction of sulphate and formation and accumulation of FexSy minerals (where x and y delineate mineral stoichiometry). These reactions only occur in specific hot spots where specific chemical and microbial conditions are met. Within a research project concerning biogeochemical transformations and turnover in wetlands, we investigate the applicability of the geoelectrical method of spectral induced polarization (SIP) to locate and monitor regions containing polarizing FexSy particles as indicator for biogeochemical hot spots. After developing and testing a sample holder and a set of non-polarizing electrodes for laboratory SIP measurements, we performed experiments on natural soil samples taken from the hyporheic zone of a local river channel. The collected material originates from a location known for biogeochemical activity. The sample contains a high percentage of dark grayish/black sediment interpreted as FexSy, and possibly pyrite (FeS2). The material was homogenized and split into four samples. The FexSy concentration was adjusted to three different levels by oxidation using H2O2. For all samples we recorded the SIP spectra in the frequency range between 0.26 Hz and 25 Hz repeatedly. The sensitivity of SIP measurements to the FexSy content is promising. For a sample of the sediment saturated with its natural fluid (conductivity > 600 μS/cm), we observed phase shifts of more than 7 mrad at frequencies between 1 and 10 Hz. We expect a lower fluid conductivity and thus a larger maximum phase shift for measurements in wetlands. The application of the SIP method in field surveys at places with biogeochemical activity will be the next step in our study.

Estimating the occupancy of spotted owl habitat areas by sampling and adjusting for bias

Treesearch

David L. Azuma; James A. Baldwin; Barry R. Noon

1990-01-01

A basic sampling scheme is proposed to estimate the proportion of sampled units (Spotted Owl Habitat Areas (SOHAs) or randomly sampled 1000-acre polygon areas (RSAs)) occupied by spotted owl pairs. A bias adjustment for the possibility of missing a pair given its presence on a SOHA or RSA is suggested. The sampling scheme is based on a fixed number of visits to a...

Front-loaded smear microscopy for the diagnosis of pulmonary TB in Tripoli, Libya.

PubMed

Gammo, Mohamed; Lamaric, Widad; Hadida, Mukhtar; Abuazza, Aida; Askar, Nabil A; Yassin, Mohammed A; Cuevas, Luis E

2013-02-01

Diagnosis of pulmonary TB by conventional smear microscopy requires patient attendance on 2 consecutive days. We investigated whether collecting sputum on-the-spot for smear microscopy on the day the patient presented was as sensitive and specific for diagnosis as the conventional spot-morning-spot scheme. We enrolled 412 adults who presented between January 2009 and October 2010 at the National Centre for TB Control in Tripoli, Libya, with cough of >2 weeks' duration, into a cross-sectional survey collecting four sputum specimens: on-the-spot and Xspot on Day 1; morning and on-the-spot on Day 2. 97 (24%) of 401 culture results were culture positive. Spot-Xspot and spot-morning smear microscopy had, respectively, 65% and 66% sensitivity and 97% and 96% specificity (p > 0.5). Spot-Xspot-morning and Spot-morning-spot smear-microscopy had, respectively, 67% and 66% sensitivity and 96% and 96% specificity (p > 0.5). For the diagnosis of pulmonary TB, the sensitivity and specificity of front-loaded (same-day) smear microscopy is similar to that of the standard smear microscopy scheme.

Standardized Methods for Detection of Poliovirus Antibodies.

PubMed

Weldon, William C; Oberste, M Steven; Pallansch, Mark A

2016-01-01

Testing for neutralizing antibodies against polioviruses has been an established gold standard for assessing individual protection from disease, population immunity, vaccine efficacy studies, and other vaccine clinical trials. Detecting poliovirus specific IgM and IgA in sera and mucosal specimens has been proposed for evaluating the status of population mucosal immunity. More recently, there has been a renewed interest in using dried blood spot cards as a medium for sample collection to enhance surveillance of poliovirus immunity. Here, we describe the modified poliovirus microneutralization assay, poliovirus capture IgM and IgA ELISA assays, and dried blood spot polio serology procedures for the detection of antibodies against poliovirus serotypes 1, 2, and 3.

First report of Lasmenia sp. causing rachis necrosis, flower abortion, fruit rot and leaf spots on rambutan in Puerto Rico

USDA-ARS?s Scientific Manuscript database

Rambutan is an exotic tropical fruit that has increased in commercial importance for growers in Puerto Rico. In 2008 and 2009, fruit rot and lesions on both leaves and inflorescences were observed. A total of 276 diseased samples from these plant parts were collected at commercial orchards, Agricult...

Use of a commercial ELISA for the detection of measles-specific immunoglobulin G (IgG) in dried blood spots collected from children living in low-resource settings.

PubMed

Colson, K Ellicott; Potter, Alan; Conde-Glez, Carlos; Hernandez, Bernardo; Ríos-Zertuche, Diego; Zúñiga-Brenes, Paola; Iriarte, Emma; Mokdad, Ali H

2015-09-01

Seroepidemiological monitoring of population immunity to vaccine-preventable diseases is critical to prevent future outbreaks. Dried blood spots (DBS), drops of capillary blood dried on filter paper, are an affordable, minimally invasive alternative to venipuncture for collecting blood in field settings. However, few proven methods exist to analyze DBS for the presence of protective antibodies. This study validates a novel technique for measuring measles-specific immunoglobulin G (IgG) in capillary DBS using a commercial ELISA. The predictive performance of a new method for analyzing DBS was tested by comparing matched serum and DBS samples from 50 children. The accuracy, precision, and reliability of the procedure were evaluated, and the optimal cut points to classify positive and negative samples were determined. The method was then applied to 1,588 DBS collected during a large survey of children in Mexico and Nicaragua. Measles-specific IgG in serum samples were 62% negative, 10% equivocal, and 28% positive. In comparisons with matched serum, DBS results were 100% sensitive and 96 · 8% specific, and agreed in 46 of 50 (92%) cases. The inter-assay and intra-assay coefficients of variation from kit-provided controls were greater than desired (24.8% and 8.4%, respectively). However, in predictive simulations the average misclassification was only 3.9%. Procedures were found to be acceptable to surveyors and participants. Analyzing DBS collected in low-resources settings is a feasible and accurate means of measuring population immunity to measles and should be used to generate objective measures of health status and health system performance. © 2015 Wiley Periodicals, Inc.

Isolation of human genomic DNA for genetic analysis from premature neonates: a comparison between newborn dried blood spots, whole blood and umbilical cord tissue

PubMed Central

2013-01-01

Background Genotyping requires biological sample collection that must be reliable, convenient and acceptable for patients and clinicians. Finding the most optimal procedure of sample collection for premature neonates who have a very limited blood volume is a particular challenge. The aim of the current study was to evaluate the use of umbilical cord (UC) tissue and newborn dried blood spot (DBS)-extracted genomic DNA (gDNA) as an alternative to venous blood-derived gDNA from premature neonates for molecular genetic analysis. All samples were obtained from premature newborn infants between 24-32 weeks of gestation. Paired blood and UC samples were collected from 31 study participants. gDNA was extracted from ethylenediaminetetraacetic acid (EDTA) anticoagulant-treated blood samples (~500 μl) and newborn DBSs (n = 723) using QIAamp DNA Micro kit (Qiagen Ltd., Crawley, UK); and from UC using Qiagen DNAeasy Blood and Tissue kit (Qiagen Ltd., Crawley, UK). gDNA was quantified and purity confirmed by measuring the A260:A280 ratio. PCR amplification and pyrosequencing was carried out to determine suitability of the gDNA for molecular genetic analysis. Minor allele frequency of two unrelated single nucleotide polymorphisms (SNPs) was calculated using the entire cohort. Results Both whole blood samples and UC tissue provided good quality and yield of gDNA, which was considerably less from newborn DBS. The gDNA purity was also reduced after 3 years of storage of the newborn DBS. PCR amplification of three unrelated genes resulted in clear products in all whole blood and UC samples and 86%-100% of newborn DBS. Genotyping using pyrosequencing showed 100% concordance in the paired UC and whole blood samples. Minor allele frequencies of the two SNPs indicated that no maternal gDNA contamination occurred in the genotyping of the UC samples. Conclusions gDNAs from all three sources are suitable for standard PCR and pyrosequencing assays. Given that UC provide good quality and quantity gDNA with 100% concordance in the genetic analysis with whole blood, it can replace blood sampling from premature infants. This is likely to reduce the stress and potential side effects associated with invasive sample collection and thus, greatly facilitate participant recruitment for genetic studies. PMID:24168095

Seasonal change in nutrient composition of spotted knapweed and preference by sheep

Treesearch

Amy C. Ganguli; Michael B. Hale; Karen L. Launchbaugh

2010-01-01

Spotted knapweed (Centaurea stoebe ssp. micranthos) is one of the most troublesome rangeland weeds in western North America. As part of a larger investigation evaluating the use of targeted sheep grazing to control spotted knapweed, we examined sheep preference and forage quality of dried spotted knapweed in different phenological stages. We collected spotted knapweed...

Continuous Flow Liquid Microjunction Surface Sampling Probe Connected On-line with HPLC/MS for Spatially Resolved Analysis of Small Molecules and Proteins

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

Van Berkel, Gary J; Kertesz, Vilmos

RATIONALE: A continuous flow liquid microjunction surface sampling probe extracts soluble material from surfaces for direct ionization and detection by MS. Demonstrated here is the on-line coupling of such a probe with HPLC/MS enabling extraction, separation and detection of small molecules and proteins from surfaces in a spatially resolved (~0.5 mm diameter spots) manner. Methods: A continuous flow liquid microjunction surface sampling probe was connected to a 6-port, 2-position valve for extract collection and injection to an HPLC column. A QTRAP 5500 hybrid triple quadrupole linear ion trap equipped with a Turbo V ion source operated in positive ESI modemore » was used for all experiments. System operation was tested with extraction, separation and detection of propranolol and associated metabolites from drug dosed tissues and proteins from dried sheep blood spots on paper. Results: Confirmed in the tissue were the parent drug and two different hydroxypropranolol glucuronides. The mass spectrometric response for these compounds from different locations in the liver showed an increase with increasing extraction time (5, 20 and 40 s extractions). For on-line separation and detection/identification of extracted proteins from dried sheep blood spots, two major protein peaks dominated the chromatogram and could be correlated with the expected masses for the hemoglobin and chains. Conclusions: Spatially resolved sampling, separation, and detection of small molecules and proteins from surfaces can be accomplished using a continuous flow liquid microjunction surface sampling probe coupled on-line with HPLC/MS detection.« less

Pre-cut Filter Paper for Detecting Anti-Japanese Encephalitis Virus IgM from Dried Cerebrospinal Fluid Spots

PubMed Central

Bharucha, Tehmina; Chanthongthip, Anisone; Phuangpanom, Soumphou; Phonemixay, Ooyanong; Sengvilaipaseuth, Onanong; Vongsouvath, Manivanh; Lee, Sue; Newton, Paul N.; Dubot-Pérès, Audrey

2016-01-01

Background The use of filter paper as a simple, inexpensive tool for storage and transportation of blood, ‘Dried Blood Spots’ or Guthrie cards, for diagnostic assays is well-established. In contrast, there are a paucity of diagnostic evaluations of dried cerebrospinal fluid (CSF) spots. These have potential applications in low-resource settings, such as Laos, where laboratory facilities for central nervous system (CNS) diagnostics are only available in Vientiane. In Laos, a major cause of CNS infection is Japanese encephalitis virus (JEV). We aimed to develop a dried CSF spot protocol and to evaluate its diagnostic performance using the World Health Organisation recommended anti-JEV IgM antibody capture enzyme-linked immunosorbent assay (JEV MAC-ELISA). Methodology and Principal Findings Sample volumes, spotting techniques and filter paper type were evaluated using a CSF-substitute of anti-JEV IgM positive serum diluted in Phosphate Buffer Solution (PBS) to end-limits of detection by JEV MAC-ELISA. A conventional protocol, involving eluting one paper punch in 200μl PBS, did not detect the end-dilution, nor did multiple punches utilising diverse spotting techniques. However, pre-cut filter paper enabled saturation with five times the volume of CSF-substitute, sufficiently improving sensitivity to detect the end-dilution. The diagnostic accuracy of this optimised protocol was compared with routine, neat CSF in a pilot, retrospective study of JEV MAC-ELISA on consecutive CSF samples, collected 2009–15, from three Lao hospitals. In comparison to neat CSF, 132 CSF samples stored as dried CSF spots for one month at 25–30°C showed 81.6% (65.7–92.3 95%CI) positive agreement, 96.8% (91.0–99.3 95%CI) negative agreement, with a kappa coefficient of 0.81 (0.70–0.92 95%CI). Conclusions/Significance The novel design of pre-cut filter paper saturated with CSF could provide a useful tool for JEV diagnostics in settings with limited laboratory access. It has the potential to improve national JEV surveillance and inform vaccination policies. The saturation of filter paper has potential use in the wider context of pathogen detection, including dried spots for detecting other analytes in CSF, and other body fluids. PMID:26986061

Early infant HIV diagnosis and entry to HIV care cascade in Thailand: an observational study.

PubMed

Sirirungsi, Wasna; Khamduang, Woottichai; Collins, Intira Jeannie; Pusamang, Artit; Leechanachai, Pranee; Chaivooth, Suchada; Ngo-Giang-Huong, Nicole; Samleerat, Tanawan

2016-06-01

Early infant diagnosis of HIV is crucial for timely initiation of antiretroviral therapy (ART) in infected children who are at high risk of mortality. Early infant diagnosis with dried blood spot testing was provided by the National AIDS Programme in Thailand from 2007. We report ART initiation and vital status in children with HIV after 7 years of rollout in Thailand. Dried blood spot samples were collected from HIV-exposed children in hospitals in Thailand and mailed to the Faculty of Associated Medical Sciences, Chiang Mai University, where HIV DNA was assessed with real-time PCR to establish HIV infection. We linked data from children with an HIV infection to the National AIDS Programme database to ascertain ART and vital status. Between April 5, 2007, and Oct 1, 2014, 16 046 dried blood spot samples were sent from 8859 children in 364 hospitals in Thailand. Median age at first dried blood spot test was 2·1 (IQR 1·8-2·5) months. Of 7174 (81%) children with two or more samples, 223 (3%) were HIV positive (including five unconfirmed). Of 1685 (19%) children with one sample, 70 (4%) were unconfirmed positive. Of 293 (3%) children who were HIV positive, 220 (75%) registered for HIV care and 170 (58%) initiated ART. Median age at ART initiation decreased from 14·2 months (IQR 10·2-25·6) in 2007 to 6·1 months (4·2-9·2) in 2013, and the number of children initiating ART aged younger than 1 year increased from five (33%) of 15 children initiating ART in 2007 to ten (83%) of 12 initiating ART in 2013. 15 (9%) of 170 children who initiated ART died and 16 (32%) of 50 who had no ART record died. Early infant diagnosis with dried blood spot testing had high uptake in primary care settings. Further improvement of linkage to HIV care is needed to ensure timely treatment of all children with an HIV infection. None. Copyright © 2016 Elsevier Ltd. All rights reserved.

7 CFR 27.97 - Ascertaining the accuracy of price quotations.

Code of Federal Regulations, 2010 CFR

2010-01-01

... cotton in each spot market shall be responsible for providing accurate and timely price, quality, and... ascertaining the accuracy of the price quotations in each designated spot market. The Cotton Division will... will collect and analyze pertinent information on the prices and values of spot cotton from each spot...

Geodynamic implications for zonal and meridional isotopic patterns across the northern Lau and North Fiji Basins

NASA Astrophysics Data System (ADS)

Price, Allison A.; Jackson, Matthew G.; Blichert-Toft, Janne; Kurz, Mark D.; Gill, Jim; Blusztajn, Jerzy; Jenner, Frances; Brens, Raul; Arculus, Richard

2017-03-01

We present new Sr-Nd-Pb-Hf-He isotopic data for 65 volcanic samples from the northern Lau and North Fiji Basins. This includes 47 lavas obtained from 40 dredge sites spanning an east-west transect across the Lau and North Fiji basins, 10 ocean island basalt (OIB)-type lavas collected from seven Fijian islands, and eight OIB lavas sampled on Rotuma. For the first time, we are able to map clear north-south and east-west geochemical gradients in 87Sr/86Sr across the northern Lau and North Fiji Basins: lavas with the most geochemically enriched radiogenic isotopic signatures are located in the northeast Lau Basin, while signatures of geochemical enrichment are diminished to the south and west away from the Samoan hot spot. Based on these geochemical patterns and plate reconstructions of the region, these observations are best explained by the addition of Samoa, Rurutu, and Rarotonga hot spot material over the past 4 Ma. We suggest that underplated Samoan material has been advected into the Lau Basin over the past ˜4 Ma. As the slab migrated west (and toward the Samoan plume) via rollback over time, younger and hotter (and therefore less viscous) underplated Samoan plume material was entrained. Thus, entrainment efficiency of underplated plume material was enhanced, and Samoan plume signatures in the Lau Basin became stronger as the trench approached the Samoan hot spot. The addition of subducted volcanoes from the Cook-Austral Volcanic Lineament first from the Rarotonga hot spot, then followed by the Rurutu hot spot, contributes to the extreme geochemical signatures observed in the northeast Lau Basin.

Determination of PCDD/Fs in breast milk of women living in the vicinities of Da Nang Agent Orange hot spot (Vietnam) and estimation of the infant's daily intake.

PubMed

Hue, N T M; Nam, V D; Thuong, N V; Huyen, N T; Phuong, N T H; Hung, N X; Tuan, N H; Son, L K; Minh, N H

2014-09-01

Seventeen toxic congeners of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were determined in breast milks using the high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS) method. Twenty seven breast milk samples were collected from primiparae who have lived over 5 years in wards namely Chinh Gian, An Khe, Khue Trung, and Hoa Thuan Tay which are located near the Da Nang Agent Orange hot spot (the AO/Dioxin hot spot). The samples were then analyzed for PCDD/F residues in order to assess the human exposure to dioxins from the AO/Dioxin hot spot, especially health risk to the breast-fed infants. The average TEQ levels in the four studied cohorts ranged from 8.1 to 26 pg/g lipid, with the highest level up to 51 pg TEQ/g lipid found in the An Khe ward. The TEQ level was correlated with geographical position and ranking in the order of Khue Trung, Hoa Thuan Tay, Chinh Gian and An Khe. The mean estimated PCDD/Fs infant's daily intake in the cohort of Khue Trung, Hoa Thuan Tay, Chinh Gian and An Khe was about 41, 122, 124, and 134 pg TEQ/kg bw/day, respectively, which are much higher than the tolerable daily intake proposed by the World Health Organization (4 pg TEQ/kg bw/day). Copyright © 2014 Elsevier B.V. All rights reserved.

Fipronil washoff to municipal wastewater from dogs treated with spot-on products.

PubMed

Teerlink, Jennifer; Hernandez, Jorge; Budd, Robert

2017-12-01

Fipronil and fipronil degradates have been reported in treated wastewater effluent at concentrations that exceed USEPA Aquatic Life Benchmarks, posing a potential risk to the surface waters to which they discharge. Fipronil is a common insecticide found in spot-on flea and tick treatment products that have the potential for down-the-drain transport and direct washoff into surface water. Volunteers currently treating their dogs with a fipronil-containing spot-on product were recruited. Dogs were washed either 2, 7, or 28days after product application, and rinsate from 34 discrete bathing events were analyzed by LC-MS/MS for fipronil and fipronil degradates (collectively known as fiproles). Total fipronil application dosage ranged from 67.1-410.0mg per dog following manufacturers' recommendation based on dog body weight. Total mass of fiproles measured in rinsate ranged from 3.6-230.6mg per dog (0.2 ̶ 86.0% of mass applied). Average percentage of fiproles detected in rinsate generally decreased with increasing time from initial application: 21±22, 16±13, and 4±5% respectively for 2, 7, and 28days post application. Fipronil was the dominant fiprole, >63% of total fiproles for all samples and >92% of total fiproles in 2 and 7day samples. Results confirm a direct pathway of pesticides to municipal wastewater through the use of spot-on products on dogs and subsequent bathing by either professional groomers or by pet owners in the home. Comparisons of mass loading calculated using California sales data and recent wastewater monitoring results suggest fipronil-containing spot-on products are a potentially important source of fipronil to wastewater treatment systems in California. This study highlights the potential for other active ingredients (i.e., bifenthrin, permethrin, etofenprox, imidacloprid) contained in spot-on and other pet products (i.e., shampoos, sprays) to enter wastewater catchments through bathing activities, posing a potential risk to the aquatic organisms downstream of wastewater discharge. Copyright © 2017 Elsevier B.V. All rights reserved.

Impact of various factors on radioactivity distribution in different DBS papers.

PubMed

Ren, Xiao; Paehler, Tobias; Zimmer, Manfred; Guo, Zuyu; Zane, Patricia; Emmons, Gary T

2010-08-01

Dried blood spot (DBS) sampling could potentially become the preferred blood collection technique in toxicological and clinical studies. Autoradiography was performed to study compound distribution within a dbs under different conditions using five papers, 31ETF, Grade 226, 903(®), FTA(®) and FTA(®) Elute. The results showed an uneven distribution in all papers with common distribution patterns regardless of compounds: decreased concentrations along the edge, the volcano effect in the middle and the speckle pattern in the center. Treated papers were more readily influenced by environmental factors. Autoradiography enables visualization of a compound's distribution and can guide bioanalytical assay development by allowing convenient evaluation of factors, such as choice of paper, spotting volume, punch size, punch location, temperature and humidity.

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

NASA Astrophysics Data System (ADS)

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

2013-08-01

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

Late-onset Pompe disease: what is the prevalence of limb-girdle muscular weakness presentation?

PubMed

Lorenzoni, Paulo José; Kay, Cláudia Suemi Kamoi; Higashi, Nádia Sugano; D'Almeida, Vânia; Werneck, Lineu Cesar; Scola, Rosana Herminia

2018-04-01

Pompe disease is an inherited disease caused by acid alpha-glucosidase (GAA) deficiency. A single center observational study aimed at assessing the prevalence of late-onset Pompe disease in a high-risk Brazilian population, using the dried blood spot test to detect GAA deficiency as a main screening tool. Dried blood spots were collected for GAA activity assay from 24 patients with "unexplained" limb-girdle muscular weakness without vacuolar myopathy in their muscle biopsy. Samples with reduced enzyme activity were also investigated for GAA gene mutations. Of the 24 patients with dried blood spots, one patient (4.2%) showed low GAA enzyme activity (NaG/AaGIA: 40.42; %INH: 87.22%). In this patient, genetic analysis confirmed two heterozygous mutations in the GAA gene (c.-32-13T>G/p.Arg854Ter). Our data confirm that clinicians should look for late-onset Pompe disease in patients whose clinical manifestation is an "unexplained" limb-girdle weakness even without vacuolar myopathy in muscle biopsy.

The basel cocktail for simultaneous phenotyping of human cytochrome P450 isoforms in plasma, saliva and dried blood spots.

PubMed

Donzelli, Massimiliano; Derungs, Adrian; Serratore, Maria-Giovanna; Noppen, Christoph; Nezic, Lana; Krähenbühl, Stephan; Haschke, Manuel

2014-03-01

Phenotyping cocktails use a combination of cytochrome P450 (CYP)-specific probe drugs to simultaneously assess the activity of different CYP isoforms. To improve the clinical applicability of CYP phenotyping, the main objectives of this study were to develop a new cocktail based on probe drugs that are widely used in clinical practice and to test whether alternative sampling methods such as collection of dried blood spots (DBS) or saliva could be used to simplify the sampling process. In a randomized crossover study, a new combination of commercially available probe drugs (the Basel cocktail) was tested for simultaneous phenotyping of CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6 and CYP3A4. Sixteen subjects received low doses of caffeine, efavirenz, losartan, omeprazole, metoprolol and midazolam in different combinations. All subjects were genotyped, and full pharmacokinetic profiles of the probe drugs and their main metabolites were determined in plasma, dried blood spots and saliva samples. The Basel cocktail was well tolerated, and bioequivalence tests showed no evidence of mutual interactions between the probe drugs. In plasma, single timepoint metabolic ratios at 2 h (for CYP2C19 and CYP3A4) or at 8 h (for the other isoforms) after dosing showed high correlations with corresponding area under the concentration-time curve (AUC) ratios (AUC0-24h parent/AUC0-24h metabolite) and are proposed as simple phenotyping metrics. Metabolic ratios in dried blood spots (for CYP1A2 and CYP2C19) or in saliva samples (for CYP1A2) were comparable to plasma ratios and offer the option of minimally invasive or non-invasive phenotyping of these isoforms. This new combination of phenotyping probe drugs can be used without mutual interactions. The proposed sampling timepoints have the potential to facilitate clinical application of phenotyping but require further validation in conditions of altered CYP activity. The use of DBS or saliva samples seems feasible for phenotyping of the selected CYP isoforms.

A simple non-enzymatic method for the preparation of white spot syndrome virus (WSSV) DNA from the haemolymph of Marsupenaeus japonicus using FTA matrix cards.

PubMed

Sudhakaran, R; Mekata, T; Kono, T; Supamattaya, K; Linh, N T H; Suzuki, Y; Sakai, M; Itami, T

2009-07-01

White spot syndrome virus (WSSV) is an important shrimp pathogen responsible for large economic losses for the shrimp culture industry worldwide. The nucleic acids of the virus must be adequately preserved and transported from the field to the laboratory before molecular diagnostic analysis is performed. Here, we developed a new method to isolate WSSV-DNA using Flinders Technology Associates filter paper (FTA matrix card; Whatman) without centrifugation or hazardous steps involved. FTA technology is a new method allowing the simple collection, shipment and archiving of nucleic acids from haemolymph samples providing DNA protection against nucleases, oxidation, UV damage, microbial and fungal attack. DNA samples prepared from 10-fold dilutions of moribund shrimp haemolymph using FTA matrix cards were analysed using semi-quantitative and quantitative polymerase chain reaction (PCR) and were compared with two commercially available DNA isolation methods, the blood GenomicPrep Mini Spin Kit (GE Healthcare) and the DNAzol (Invitrogen). Sequence analysis was performed for the DNA samples prepared using the various isolation procedures and no differences in the sequence among these methods were identified. Results based on the initial copy number of DNA prepared from the GenomicPrep Mini Spin Kit are a little more sensitive than the DNA prepared from FTA matrix cards, whereas the DNAzol method is not suitable for blood samples. Our data shows the efficiency of retention capacity of WSSV-DNA samples from impregnated FTA matrix cards. Matrix cards were easy to store and ship for long periods of time. They provide ease of handling and are a reliable alternative for sample collection and for molecular detection and characterization of WSSV isolates.

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

PubMed Central

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

2016-01-01

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

Dried Blood Spots Combined With Ultra-High-Performance Liquid Chromatography-Mass Spectrometry for the Quantification of the Antipsychotics Risperidone, Aripiprazole, Pipamperone, and Their Major Metabolites.

PubMed

Tron, Camille; Kloosterboer, Sanne M; van der Nagel, Bart C H; Wijma, Rixt A; Dierckx, Bram; Dieleman, Gwen C; van Gelder, Teun; Koch, Birgit C P

2017-08-01

Risperidone, aripiprazole, and pipamperone are antipsychotic drugs frequently prescribed for the treatment of comorbid behavioral problems in children with autism spectrum disorders. Therapeutic drug monitoring (TDM) could be useful to decrease side effects and to improve patient outcome. Dried blood spot (DBS) sample collection seems to be an attractive technique to develop TDM of these drugs in a pediatric population. The aim of this work was to develop and validate a DBS assay suitable for TDM and home sampling. Risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone were extracted from DBS and analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry using a C18 reversed-phase column with a mobile phase consisting of ammonium acetate/formic acid in water or methanol. The suitability of DBS for TDM was assessed by studying the influence of specific parameters: extraction solution, EDTA carryover, hematocrit, punching location, spot volume, and hemolysis. The assay was validated with respect to conventional guidelines for bioanalytical methods. The method was linear, specific without any critical matrix effect, and with a mean recovery around 90%. Accuracy and imprecision were within the acceptance criteria in samples with hematocrit values from 30% to 45%. EDTA or hemolysis did not skew the results, and no punching carryover was observed. No significant influence of the spot volume or the punch location was observed. The antipsychotics were all stable in DBS stored 10 days at room temperature and 1 month at 4 or -80°C. The method was successfully applied to quantify the 3 antipsychotics and their metabolites in patient samples. A UHPLC-MS/MS method has been successfully validated for the simultaneous quantification of risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone in DBS. The assay provided good analytical performances for TDM and clinical research applications.

Transcriptome Profiling of Buffalograss Challenged with the Leaf Spot Pathogen Curvularia inaequalis.

PubMed

Amaradasa, Bimal S; Amundsen, Keenan

2016-01-01

Buffalograss (Bouteloua dactyloides) is a low maintenance U. S. native turfgrass species with exceptional drought, heat, and cold tolerance. Leaf spot caused by Curvularia inaequalis negatively impacts buffalograss visual quality. Two leaf spot susceptible and two resistant buffalograss lines were challenged with C. inaequalis. Samples were collected from treated and untreated leaves when susceptible lines showed symptoms. Transcriptome sequencing was done and differentially expressed genes were identified. Approximately 27 million raw sequencing reads were produced per sample. More than 86% of the sequencing reads mapped to an existing buffalograss reference transcriptome. De novo assembly of unmapped reads was merged with the existing reference to produce a more complete transcriptome. There were 461 differentially expressed transcripts between the resistant and susceptible lines when challenged with the pathogen and 1552 in its absence. Previously characterized defense-related genes were identified among the differentially expressed transcripts. Twenty one resistant line transcripts were similar to genes regulating pattern triggered immunity and 20 transcripts were similar to genes regulating effector triggered immunity. There were also nine up-regulated transcripts in resistance lines which showed potential to initiate systemic acquired resistance (SAR) and three transcripts encoding pathogenesis-related proteins which are downstream products of SAR. This is the first study characterizing changes in the buffalograss transcriptome when challenged with C. inaequalis.

Proteomic features of delayed ocular symptoms caused by exposure to sulfur mustard: As studied by protein profiling of corneal epithelium.

PubMed

Pajoohesh, Maryam; Naderi, Mostafa; Naderi-Manesh, Hossein

2017-11-01

Exposure to mustard gas can lead to variations in the proteome of corneal epithelium cells and after a latency period produces delayed symptoms in the eyes of chemical victims. Hence, a comparative proteome analysis was conducted between the corneal epithelial cells of chemical victims from Iran-Iraq war (1980-1988) and healthy donors. To this end, corneal epithelium samples from victims and healthy individuals were collected, and the proteome of these samples were prepared for two-dimensional electrophoresis and the analysis of spots by statistical software. Selected spots were further analyzed by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Twenty four proteins were identified of which eighteen proteins showed downregulation while six proteins were upregulated in the victims in comparison to the normal individuals. Also, six protein spots were confirmed by western-blot analysis. In conclusion, all the twenty-four identified proteins are involved in pathways which their up- or down-regulation leads to the accumulation of undesired substrates, cell death and apoptosis. Bioinformatics' tools indicated that these identified proteins were involved in various metabolic processes, DNA damage response, immune response and etc. The present study provides a suitable platform for further clinical studies. Copyright © 2017 Elsevier B.V. All rights reserved.

Tick-borne diseases in North Carolina: is "Rickettsia amblyommii" a possible cause of rickettsiosis reported as Rocky Mountain spotted fever?

PubMed

Apperson, Charles S; Engber, Barry; Nicholson, William L; Mead, Daniel G; Engel, Jeffrey; Yabsley, Michael J; Dail, Kathy; Johnson, Joey; Watson, D Wesley

2008-10-01

Cases of Rocky Mountain spotted fever (RMSF) in North Carolina have escalated markedly since 2000. In 2005, we identified a county in the Piedmont region with high case numbers of RMSF. We collected ticks and examined them for bacterial pathogens using molecular methods to determine if a novel tick vector or spotted fever group rickettsiae (SFGR) might be emerging. Amblyomma americanum, the lone star tick, comprised 99.6% of 6,502 specimens collected in suburban landscapes. In contrast, Dermacentor variabilis, the American dog tick, a principal vector of Rickettsia rickettsii, comprised < 1% of the ticks collected. Eleven of 25 lone star tick pools tested were infected with "Rickettsia amblyommii," an informally named SFGR. Sera from patients from the same county who were presumptively diagnosed by local physicians with a tick-borne illness were tested by an indirect immunofluorescence antibody (IFA) assay to confirm clinical diagnoses. Three of six patients classified as probable RMSF cases demonstrated a fourfold or greater rise in IgG class antibody titers between paired acute and convalescent sera to "R. amblyommii" antigens, but not to R. rickettsii antigens. White-tailed deer, Odocoileus virginianus, are preferred hosts of lone star ticks. Blood samples collected from hunter-killed deer from the same county were tested by IFA test for antibodies to Ehrlichia chaffeensis and "R. amblyommii." Twenty-eight (87%) of 32 deer were positive for antibodies to E. chaffeensis, but only 1 (3%) of the deer exhibited antibodies to "R. amblyommii," suggesting that deer are not the source of "R. amblyommii" infection for lone star ticks. We propose that some cases of rickettsiosis reported as RMSF may have been caused by "R. amblyommii" transmitted through the bite of A. americanum.

Paper-based SERS swab for rapid trace detection on real-world surfaces.

PubMed

Lee, Chang H; Tian, Limei; Singamaneni, Srikanth

2010-12-01

One of the important but often overlooked considerations in the design of surface-enhanced Raman scattering (SERS) substrates for trace detection is the efficiency of sample collection. Conventional designs based on rigid substrates such as silicon, alumina, and glass resist conformal contact with the surface under investigation, making the sample collection inefficient. We demonstrate a novel SERS substrate based on common filter paper adsorbed with gold nanorods, which allows conformal contact with real-world surfaces, thus dramatically enhancing the sample collection efficiency compared to conventional rigid substrates. We demonstrate the detection of trace amounts of analyte (140 pg spread over 4 cm2) by simply swabbing the surface under investigation with the novel SERS substrate. The hierarchical fibrous structure of paper serves as a 3D vasculature for easy uptake and transport of the analytes to the electromagnetic hot spots in the paper. Simple yet highly efficient and cost-effective SERS substrate demonstrated here brings SERS-based trace detection closer to real-world applications.

Continuous-flow liquid microjunction surface sampling probe connected on-line with high-performance liquid chromatography/mass spectrometry for spatially resolved analysis of small molecules and proteins.

PubMed

Van Berkel, Gary J; Kertesz, Vilmos

2013-06-30

A continuous-flow liquid microjunction surface sampling probe extracts soluble material from surfaces for direct ionization and detection by mass spectrometry. Demonstrated here is the on-line coupling of such a probe with high-performance liquid chromatography/mass spectrometry (HPLC/MS) enabling extraction, separation and detection of small molecules and proteins from surfaces in a spatially resolved (~0.5 mm diameter spots) manner. A continuous-flow liquid microjunction surface sampling probe was connected to a six-port, two-position valve for extract collection and injection to an HPLC column. A QTRAP® 5500 hybrid triple quadrupole linear ion trap equipped with a Turbo V™ ion source operated in positive electrospray ionization (ESI) mode was used for all experiments. The system operation was tested with the extraction, separation and detection of propranolol and associated metabolites from drug dosed tissues, caffeine from a coffee bean, cocaine from paper currency, and proteins from dried sheep blood spots on paper. Confirmed in the tissue were the parent drug and two different hydroxypropranolol glucuronides. The mass spectrometric response for these compounds from different locations in the liver showed an increase with increasing extraction time (5, 20 and 40 s). For on-line separation and detection/identification of extracted proteins from dried sheep blood spots, two major protein peaks dominated the chromatogram and could be correlated with the expected masses for the hemoglobin α and β chains. Spatially resolved sampling, separation, and detection of small molecules and proteins from surfaces can be accomplished using a continuous-flow liquid microjunction surface sampling probe coupled on-line with HPLC/MS detection. Published in 2013. This article is a U.S. Government work and is in the public domain in the USA.

Iridium emissions from Hawaiian volcanoes

NASA Technical Reports Server (NTRS)

Finnegan, D. L.; Zoller, W. H.; Miller, T. M.

1988-01-01

Particle and gas samples were collected at Mauna Loa volcano during and after its eruption in March and April, 1984 and at Kilauea volcano in 1983, 1984, and 1985 during various phases of its ongoing activity. In the last two Kilauea sampling missions, samples were collected during eruptive activity. The samples were collected using a filterpack system consisting of a Teflon particle filter followed by a series of 4 base-treated Whatman filters. The samples were analyzed by INAA for over 40 elements. As previously reported in the literature, Ir was first detected on particle filters at the Mauna Loa Observatory and later from non-erupting high temperature vents at Kilauea. Since that time Ir was found in samples collected at Kilauea and Mauna Loa during fountaining activity as well as after eruptive activity. Enrichment factors for Ir in the volcanic fumes range from 10,000 to 100,000 relative to BHVO. Charcoal impregnated filters following a particle filter were collected to see if a significant amount of the Ir was in the gas phase during sample collection. Iridium was found on charcoal filters collected close to the vent, no Ir was found on the charcoal filters. This indicates that all of the Ir is in particulate form very soon after its release. Ratios of Ir to F and Cl were calculated for the samples from Mauna Loa and Kilauea collected during fountaining activity. The implications for the KT Ir anomaly are still unclear though as Ir was not found at volcanoes other than those at Hawaii. Further investigations are needed at other volcanoes to ascertain if basaltic volcanoes other than hot spots have Ir enrichments in their fumes.

A validated method for the quantification of fosfomycin on dried plasma spots by HPLC-MS/MS: application to a pilot pharmacokinetic study in humans.

PubMed

Parker, Suzanne L; Lipman, Jeffrey; Dimopoulos, George; Roberts, Jason A; Wallis, Steven C

2015-11-10

Quantification of fosfomycin in the plasma samples of patients is the basis of clinical pharmacokinetic studies from which evidence based dosing regimens can be devised to maximise antibiotic effectiveness against a pathogen. We have developed and validated a LC-MS/MS method to quantify fosfomycin using dried plasma spot sampling. Following HILIC chromatography, fosfomycin and ethylphosphonic acid, used as internal standard, were measured using negative-ion multiple reaction monitoring. The method was linear over the calibration range of 5-2000mg/L of fosfomycin. Intra-day assay results for dried plasma spot quality control samples at 15.6, 79.9 and 1581mg/L of fosfomycin had precision of ±4.2, 8.2, and 2.0%, respectively, and accuracy of +3.9, -0.1, and -1.2%, respectively. Recovery of fosfomycin from dried plasma spots was calculated as 83.6% and the dried plasma spot samples were found to be stable stored at room temperature for three months and when stored for four hours at 50°C. A Bland-Altman plot comparing DPS to plasma sampling found a negative bias of 16.6%, with all but one sample within the mean limits of agreement (-2.6 to 30.6%). Dried plasma spot sampling provides a useful tool for pharmacokinetic research of fosfomycin. Copyright © 2015 Elsevier B.V. All rights reserved.

Statistical Analysis of Variation in the Human Plasma Proteome

DOE PAGES

Corzett, Todd H.; Fodor, Imola K.; Choi, Megan W.; ...

2010-01-01

Quantifying the variation in the human plasma proteome is an essential prerequisite for disease-specific biomarker detection. We report here on the longitudinal and individual variation in human plasma characterized by two-dimensional difference gel electrophoresis (2-D DIGE) using plasma samples from eleven healthy subjects collected three times over a two week period. Fixed-effects modeling was used to remove dye and gel variability. Mixed-effects modeling was then used to quantitate the sources of proteomic variation. The subject-to-subject variation represented the largest variance component, while the time-within-subject variation was comparable to the experimental variation found in a previous technical variability study where onemore » human plasma sample was processed eight times in parallel and each was then analyzed by 2-D DIGE in triplicate. Here, 21 protein spots had larger than 50% CV, suggesting that these proteins may not be appropriate as biomarkers and should be carefully scrutinized in future studies. Seventy-eight protein spots showing differential protein levels between different individuals or individual collections were identified by mass spectrometry and further characterized using hierarchical clustering. The results present a first step toward understanding the complexity of longitudinal and individual variation in the human plasma proteome, and provide a baseline for improved biomarker discovery.« less

Statistical analysis of variation in the human plasma proteome.

PubMed

Corzett, Todd H; Fodor, Imola K; Choi, Megan W; Walsworth, Vicki L; Turteltaub, Kenneth W; McCutchen-Maloney, Sandra L; Chromy, Brett A

2010-01-01

Quantifying the variation in the human plasma proteome is an essential prerequisite for disease-specific biomarker detection. We report here on the longitudinal and individual variation in human plasma characterized by two-dimensional difference gel electrophoresis (2-D DIGE) using plasma samples from eleven healthy subjects collected three times over a two week period. Fixed-effects modeling was used to remove dye and gel variability. Mixed-effects modeling was then used to quantitate the sources of proteomic variation. The subject-to-subject variation represented the largest variance component, while the time-within-subject variation was comparable to the experimental variation found in a previous technical variability study where one human plasma sample was processed eight times in parallel and each was then analyzed by 2-D DIGE in triplicate. Here, 21 protein spots had larger than 50% CV, suggesting that these proteins may not be appropriate as biomarkers and should be carefully scrutinized in future studies. Seventy-eight protein spots showing differential protein levels between different individuals or individual collections were identified by mass spectrometry and further characterized using hierarchical clustering. The results present a first step toward understanding the complexity of longitudinal and individual variation in the human plasma proteome, and provide a baseline for improved biomarker discovery.

Thermal regimes of Mexican spotted owl nest stands

Treesearch

Joseph L. Ganey

2004-01-01

To evaluate the hypothesis that spotted owls (Strix occidentalis) select habitats with cool microclimates to avoid high daytime temperatures, I sampled thermal regimes in nest areas used by Mexican spotted owls (S. o. lucida) in northern Arizona. I sampled air temperature at 30-min intervals in 30 pairs of nest and random sites...

Detection of spotted fever group (SFG) rickettsiae in Dermacentor reticulatus (Acari: Ixodidae) in Poland.

PubMed

Stańczak, Joanna

2006-05-01

Dermacentor reticulatus ticks from Poland were investigated by molecular methods for the presence of rickettsiae. During 2003/2004, a total of 285 adult ticks was assayed using primers RpCS.877 and RpCS.1258 derived from the citrate synthase (gltA) gene, and 116 samples (40.7%) were positive for rickettsial DNA. Ten out of these positive samples were further assayed using SLO1F and SLO1R primers derived form the rOmpA-encoding gene to confirm that detected rickettsiae belong to the spotted fever group (SFG). The obtained sequence of a fragment of the gltA gene of Rickettsia sp. isolated from Polish D. reticulatus demonstrated 96-98% similarities to Rickettsia slovaca, Rickettsia sibirica, Rickettsia honei, and other SFG rickettsiae. The nucleotide sequences of the amplified fragments of the ompA gene were 98% homologous to RpA4 Rickettsia sp. reported from ticks collected in territories of the former Soviet Union.

Time Resolved Near Field Optical Microscopy

NASA Astrophysics Data System (ADS)

Stark, J. B.

1996-03-01

We use broadband pulses to image the carrier dynamics of semiconductor microstructures on a 150 nm spatial scale, with a time resolution of 60 femtoseconds. Etched disks of GaAs/AlGaAs multiple quantum well material, 10 microns in diameter, are excited with a 30 fs pump from a Ti:Sapphire laser, and probed using a near-field optical microscope. The nonlinear transmission of the microdisks is measured using a double-modulation technique, sensitive to transmission changes of 0.0005 within a 150 nm diameter spot on the sample. This spot is scanned to produce an image of the sample. The nonlinear response is produced by the occupation of phase space by the excited distribution. Images of this evolving distribution are collected at time intervals following excitation, measuring the relaxation of carriers at each point in the microdisk. The resulting data can be viewed as a movie of the carrier dynamics of nonequilibrium distributions in excited semiconductor structures. Work done in collaboration with U. Mohideen and R. E. Slusher.

Use of Dried Blood Spots to Elucidate Full-Length Transmitted/Founder HIV-1 Genomes

PubMed Central

Salazar-Gonzalez, Jesus F.; Salazar, Maria G.; Tully, Damien C.; Ogilvie, Colin B.; Learn, Gerald H.; Allen, Todd M.; Heath, Sonya L.; Goepfert, Paul; Bar, Katharine J.

2016-01-01

Background Identification of HIV-1 genomes responsible for establishing clinical infection in newly infected individuals is fundamental to prevention and pathogenesis research. Processing, storage, and transportation of the clinical samples required to perform these virologic assays in resource-limited settings requires challenging venipuncture and cold chain logistics. Here, we validate the use of dried-blood spots (DBS) as a simple and convenient alternative to collecting and storing frozen plasma. Methods We performed parallel nucleic acid extraction, single genome amplification (SGA), next generation sequencing (NGS), and phylogenetic analyses on plasma and DBS. Results We demonstrated the capacity to extract viral RNA from DBS and perform SGA to infer the complete nucleotide sequence of the transmitted/founder (TF) HIV-1 envelope gene and full-length genome in two acutely infected individuals. Using both SGA and NGS methodologies, we showed that sequences generated from DBS and plasma display comparable phylogenetic patterns in both acute and chronic infection. SGA was successful on samples with a range of plasma viremia, including samples as low as 1,700 copies/ml and an estimated ∼50 viral copies per blood spot. Further, we demonstrated reproducible efficiency in gp160 env sequencing in DBS stored at ambient temperature for up to three weeks or at -20°C for up to five months. Conclusions These findings support the use of DBS as a practical and cost-effective alternative to frozen plasma for clinical trials and translational research conducted in resource-limited settings. PMID:27819061

Contamination of dried blood spots - an underestimated risk in newborn screening.

PubMed

Winter, Theresa; Lange, Anja; Hannemann, Anke; Nauck, Matthias; Müller, Cornelia

2018-01-26

Newborn screening (NBS) is an established screening procedure in many countries worldwide, aiming at the early detection of inborn errors of metabolism. For decades, dried blood spots have been the standard specimen for NBS. The procedure of blood collection is well described and standardized and includes many critical pre-analytical steps. We examined the impact of contamination of some anticipated common substances on NBS results obtained from dry spot samples. This possible pre-analytical source of uncertainty has been poorly examined in the past. Capillary blood was obtained from 15 adult volunteers and applied to 10 screening filter papers per volunteer. Nine filter papers were contaminated without visible trace. The contaminants were baby diaper rash cream, baby wet wipes, disinfectant, liquid infant formula, liquid infant formula hypoallergenic (HA), ultrasonic gel, breast milk, feces, and urine. The differences between control and contaminated samples were evaluated for 45 NBS quantities. We estimated if the contaminations might lead to false-positive NBS results. Eight of nine investigated contaminants significantly altered NBS analyte concentrations and potentially caused false-positive screening outcomes. A contamination with feces was most influential, affecting 24 of 45 tested analytes followed by liquid infant formula (HA) and urine, affecting 19 and 13 of 45 analytes, respectively. A contamination of filter paper samples can have a substantial effect on the NBS results. Our results underline the importance of good pre-analytical training to make the staff aware of the threat and ensure reliable screening results.

Determination of total mercury in fillets of sport fishes collected from Folsom Reservoir, California, 2006

USGS Publications Warehouse

May, Thomas W.; Brumbaugh, William G.

2007-01-01

This report presents the results of a study by the U.S. Geological Survey, done in cooperation with the U.S. Bureau of Reclamation, to determine mercury concentrations in selected sport fishes from Folsom Reservoir in California. Fillets were collected from each fish sample, and after homogenization and lyophilization of fish fillets, mercury concentrations were determined with a direct mercury analyzer utilizing the process of thermal combustion-gold amalgamation atomic absorption spectroscopy. Mercury concentrations in fillets ranged from 0.031 to 0.20 micrograms per gram wet weight in rainbow trout (Oncorhynchus mykiss) samples and 0.071 to 0.16 micrograms per gram wet weight in bluegill (Lepomis macrochirus) samples. Mercury concentration was 0.98 microgram per gram wet weight in a single spotted bass (Micropterus punctulatus) sample, which was the only one in the sample set which exceeded the U.S. Environmental Protection Agency's fish consumption advisory of 0.30 microgram per gram wet weight.

Leveraging Digital Health Technologies and Outpatient Sampling in Clinical Drug Development: A Phase 1 Exploratory Study.

PubMed

Dockendorf, Marissa F; Murthy, Gowri; Bateman, Kevin P; Kothare, Prajakti A; Anderson, Melanie; Xie, Iris; Sachs, Jeff R; Burlage, Rubi; Goldman, Andra; Moyer, Matthew; Shah, Jyoti; Ruba, Rachel; Shipley, Lisa; Harrelson, Jane

2018-06-09

Merck & Co., Inc. (Kenilworth, New Jersey) is investing in approaches to enrich clinical trial data and augment decision-making through utilization of digital health technologies, outpatient sampling, and real-time data access. As part of this strategy, a Phase 1 study was conducted to explore a few technologies of interest. In this fixed-sequence, two-period trial, 16 healthy subjects were administered 50-mg once-daily sitagliptin packaged in a bottle that electronically captured the date and time study medication was dispensed (Period 1) and in a traditional pharmacy bottle (Period 2). Dried blood spot (DBS) samples were collected for sitagliptin concentration analysis on select study days, both in-clinic and at-home, with collection time recorded using an electronic diary in Period 1 and by clinic staff in Period 2. Study results demonstrated the feasibility and subject acceptance of collecting digital adherence data and outpatient DBS samples in clinical trials and highlighted areas for future improvements. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

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

PubMed

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

2015-05-01

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

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

PubMed Central

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

2015-01-01

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

Phthalate metabolites in Norwegian mothers and children: Levels, diurnal variation and use of personal care products.

PubMed

Sakhi, Amrit Kaur; Sabaredzovic, Azemira; Cequier, Enrique; Thomsen, Cathrine

2017-12-01

Exposure to phthalates has been associated with reproductive and developmental toxicity. Data on levels of these compounds in the Norwegian population is limited. In this study, urine samples were collected from 48 mothers and their children in two counties in Norway. Eleven different phthalate metabolites originating from six commonly used phthalates in consumer products were determined. Concentrations of phthalate metabolites were significantly higher in children compared to mothers except for mono-ethyl phthalate (MEP). The mothers provided several urine samples during 24hours (h) and diurnal variation showed that the concentrations in the morning urine samples (24-8h) were significantly higher than at other time-periods for most of the phthalate metabolites. Intraclass correlation coefficients (ICCs) for 24-hour time-period were in the range of 0.49-0.81. These moderate to high ICCs indicate that one spot urine sample can be used to estimate the exposure to phthalates. Since a significant effect of time of day was observed, it is still advisable to standardize the collection time point to reduce the variation. For the mothers, the use of personal care products (PCPs) were less associated with morning urine samples than early day (8-12h) and evening (16-24h) urine samples. The use of perfume and hair products were positively associated with the urinary concentrations of low molecular weight phthalates. Use of shower soap and shampoo were positively associated with urinary concentration of di(2-ethylhexyl) phthalate (DEHP) metabolites. For children, face cream use was positively associated with phthalate metabolites in the morning samples, and hand soap use was negatively associated with concentration of urinary DEHP metabolites in afternoon/evening samples. Since different PCPs were associated with the urinary phthalate metabolites in different time-periods during a day, more than one spot urine sample might be required to study associations between urinary phthalate metabolites and the use of PCPs. Copyright © 2017 Elsevier B.V. All rights reserved.

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

PubMed

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

2015-10-01

Microbes play an important role in the degradation of fish products, thus better knowledge of the microbiological conditions throughout the fish production chain may help to optimise product quality and resource utilisation. This paper presents the results of a ten-year spot sampling programme (2005-2014) of the commercially most important pelagic fish species harvested in Norway. Fish-, surface-, and storage water samples were collected from fishing vessels and processing factories. Totally 1,181 samples were assessed with respect to microbiological quality, hygiene and food safety. We introduce a quality and safety assessment scheme for fresh pelagic fish recommending limits for heterotrophic plate counts (HPC), thermos tolerant coliforms, enterococci and Listeria monocytogenes. According to the scheme, in 25 of 41 samplings, sub-optimal conditions were found with respect to quality, whereas in 21 and 9 samplings, samples were not in compliance concerning hygiene and food safety, respectively. The present study has revealed that the quality of pelagic fish can be optimised by improving the hygiene conditions at some critical points at an early phase of the production chain. Thus, the proposed assessment scheme may provide a useful tool for the industry to optimise quality and maintain consumer safety of pelagic fishery products. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Using Spot Biomarker Data to Inform Chronic Exposure and Health Risk

EPA Science Inventory

The U.S. NHANES and other health surveys frequently collect and analyze spot biological specimens to inform chemical exposures. These spot measurements can be compared to biomarker-based reference levels, such as “Biomonitoring Equivalents” (BEs), to evaluate health r...

Spot urine sodium excretion as prognostic marker in acutely decompensated heart failure: the spironolactone effect.

PubMed

Ferreira, João Pedro; Girerd, Nicolas; Medeiros, Pedro Bettencourt; Santos, Mário; Carvalho, Henrique Cyrne; Bettencourt, Paulo; Kénizou, David; Butler, Javed; Zannad, Faiez; Rossignol, Patrick

2016-06-01

Loop diuretic resistance characterized by inefficient sodium excretion complicates many patients with acutely decompensated heart failure (ADHF). Mineralocorticoid receptor antagonists (MRAs) in natriuretic doses may improve spot urine sodium excretion and outcomes. Our primary aim was to assess the association of high-dose spironolactone with short-term spot urine sodium excretion, and our secondary aim was to determine if this higher short-term spot urine sodium excretion is associated with reduction in the composite clinical outcome (of cardiovascular mortality and/or ADHF hospitalization) event rate at 180 days. Single-centre, non-randomized, open-label study enrolling 100 patients with ADHF. Patients were treated with standard ADHF therapy alone (n = 50) or oral spironolactone 100 mg/day plus standard ADHF therapy (n = 50). Spot urine samples were collected at day 1 and day 3 of hospitalization. Spironolactone group had significantly higher spot urine sodium levels compared to standard care group at day 3 (84.13 ± 28.71 mmol/L vs 70.74 ± 34.43 mmol/L, p = 0.04). The proportion of patients with spot urinary sodium <60 mmol/L was lower in spironolactone group at day 3 (18.8 vs 45.7, p = 0.01). In multivariate analysis, spironolactone was independently associated with increased spot urinary sodium and urinary sodium/potassium ratio of >2 at day 3 (both, p < 0.05). Higher spot urine sodium levels were associated with a lower event rate [HR for urinary sodium >100 mmol/L = 0.16 (0.06-0.42), p < 0.01, compared to <60], and provided a significant prognostic gain measured by net reclassification indexes. Spot urinary sodium levels >60 mmol/L and urinary sodium/potassium ratio >2 measured at day 3 of hospitalization for ADHF are associated with improved mid-term outcomes. Spironolactone is associated with increased spot urinary sodium and sodium/potassium ratio >2.

Crowdsourcing the identification of organisms: A case-study of iSpot.

PubMed

Silvertown, Jonathan; Harvey, Martin; Greenwood, Richard; Dodd, Mike; Rosewell, Jon; Rebelo, Tony; Ansine, Janice; McConway, Kevin

2015-01-01

Accurate species identification is fundamental to biodiversity science, but the natural history skills required for this are neglected in formal education at all levels. In this paper we describe how the web application ispotnature.org and its sister site ispot.org.za (collectively, "iSpot") are helping to solve this problem by combining learning technology with crowdsourcing to connect beginners with experts. Over 94% of observations submitted to iSpot receive a determination. External checking of a sample of 3,287 iSpot records verified > 92% of them. To mid 2014, iSpot crowdsourced the identification of 30,000 taxa (>80% at species level) in > 390,000 observations with a global community numbering > 42,000 registered participants. More than half the observations on ispotnature.org were named within an hour of submission. iSpot uses a unique, 9-dimensional reputation system to motivate and reward participants and to verify determinations. Taxon-specific reputation points are earned when a participant proposes an identification that achieves agreement from other participants, weighted by the agreers' own reputation scores for the taxon. This system is able to discriminate effectively between competing determinations when two or more are proposed for the same observation. In 57% of such cases the reputation system improved the accuracy of the determination, while in the remainder it either improved precision (e.g. by adding a species name to a genus) or revealed false precision, for example where a determination to species level was not supported by the available evidence. We propose that the success of iSpot arises from the structure of its social network that efficiently connects beginners and experts, overcoming the social as well as geographic barriers that normally separate the two.

Technical Stability and Biological Variability in MicroRNAs from Dried Blood Spots: A Lung Cancer Therapy-Monitoring Showcase.

PubMed

Kahraman, Mustafa; Laufer, Thomas; Backes, Christina; Schrörs, Hannah; Fehlmann, Tobias; Ludwig, Nicole; Kohlhaas, Jochen; Meese, Eckart; Wehler, Thomas; Bals, Robert; Keller, Andreas

2017-09-01

Different work flows have been proposed to use miRNAs as blood-borne biomarkers. In particular, the method used for collecting blood from patients can considerably influence the diagnostic results. We explored whether dried blood spots (DBSs) facilitate stable miRNA measurements and compared its technical stability with biological variability. First, we tested the stability of DBS samples by generating from 1 person 18 whole-genome-wide miRNA profiles of DBS samples that were exposed to different temperature and humidity conditions. Second, we investigated technical reproducibility by performing 7 replicates of DBS again from 1 person. Third, we investigated DBS samples from 53 patients with lung cancer undergoing different therapies. Across these 3 stages, 108 genome-wide miRNA profiles from DBS were generated and evaluated biostatistically. In the stability analysis, we observed that temperature and humidity had an overall limited influence on the miRNomes (average correlation between the different conditions of 0.993). Usage of a silica gel slightly diminished DBS' technical reproducibility. The 7 technical replicates had an average correlation of 0.996. The correlation with whole-blood PAXGene miRNomes of the same individual was remarkable (correlation of 0.88). Finally, evaluation of the samples from the 53 patients with lung cancer exposed to different therapies showed that the biological variations exceeded the technical variability significantly ( P < 0.0001), yielding 51 dysregulated miRNAs. We present a stable work flow for profiling of whole miRNomes on the basis of samples collected from DBS. Biological variations exceeded technical variations significantly. DBS-based miRNA profiles will potentially further the translational character of miRNA biomarker studies. © 2017 American Association for Clinical Chemistry.

7 CFR 28.423 - Middling Spotted Color.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 2 2014-01-01 2014-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

7 CFR 28.423 - Middling Spotted Color.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

7 CFR 28.423 - Middling Spotted Color.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 2 2013-01-01 2013-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

7 CFR 28.423 - Middling Spotted Color.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 2 2012-01-01 2012-01-01 false Middling Spotted Color. 28.423 Section 28.423... REGULATIONS COTTON CLASSING, TESTING, AND STANDARDS Standards Spotted Cotton § 28.423 Middling Spotted Color. Middling Spotted Color is color which is within the range represented by a set of samples in the custody of...

7 CFR 28.423 - Middling Spotted Color.

Code of Federal Regulations, 2010 CFR

2010-01-01

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

Dermoscopy of black-spot poison ivy.

PubMed

Rader, Ryan K; Mu, Ruipu; Shi, Honglan; Stoecker, William V; Hinton, Kristen A

2012-10-15

Black-spot poison ivy is an uncommon presentation of poison ivy (Toxicodendron) allergic contact dermatitis. A 78-year-old sought evaluation of a black spot present on her right hand amid pruritic vesicles. The presentation of a black spot on the skin in a clinical context suggesting poison ivy is indicative of black-spot poison ivy. Dermoscopy revealed a jagged, centrally homogeneous, dark brown lesion with a red rim. A skin sample was obtained and compared against a poison ivy standard using ultra-fast liquid chromatography-tandem mass spectrometry (UFLC-MS/MS). This finding confirmed the presence of multiple urushiol congeners in the skin sample. Black-spot poison ivy may be added to the list of diagnoses that show a specific dermoscopic pattern.

The measurement of mangrove characteristics in southwest Florida using SPOT multispectral data

NASA Technical Reports Server (NTRS)

Jensen, John R.; Lin, Hongyue; Yang, Xinghe; Ramsey, Elijah, III; Davis, Bruce A.; Thoemke, Chris W.

1991-01-01

An intensive in situ sampling program near Marco Island, Florida during 19-23 October 1988 collected information on mangrove type, maximum canopy height, and percent canopy closure. These data were correlated with selected vegetation index information derived from analysis of SPOT multispectral (XS) data obtained on 21 October 1988. The Normalized Difference (ND) vegetation index information was the most highly correlated index with percent canopy closure (r = 0.91). Percent canopy closure information can be used as a surrogate for mangrove density which is of great value when predicting which parts of the mangrove ecosystem are at greatest risk after an oil spill occurs. Such information is very valuable when constructing oil spill Environmental Sensitivity Index (ESI) Maps for tropical regions of the world.

Industrial-hygiene report, perchloroethylene, Carman's Classic Cleaners, Richmond, California, July 28, 1978

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

Ludwig, H.R.

1978-03-01

The survey was part of an industrywide mortality and industrial hygiene study of dry cleaning workers exposure to perchlorethylene. To gather data for this larger study, an on site visit was made to the Carman's Classic Cleaners located in Richmond, California. Carman's Classic Cleaners has been using PCE since the present owner had the building constructed in 1954. About 621 kilograms of clothing were dry cleaned each week, with about 129 liters of PCE consumed monthly. The workforce of six includes two pressers, a full time employee who works at the counter and also sews, a part time maintenance man,more » the owner, and his wife. The owner did all the dry cleaning and spotting while his wife marked clothing and worked the counter. Air samples were collected using battery operated pumps and charcoal tubes. The time weighted average exposure to PCE during the 5 hours of operation was 25 parts per million (ppm) for the dry cleaner and 2ppm for the presser. Samples collected in the vicinity of the front counter, washer, and spotting table indicated PCE levels of 3, 29, and 21ppm, respectively. Five minute peak samples during clothing transfer indicated PCE levels of 22, 38, and 17ppm. These exposures were well within the current standards. However, as PCE is a suspected carcinogen, it is necessary that levels be lowered as much as possible. The author recommends measures to help bring this about.« less

78 FR 18563 - Proposed Collection; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-27

... Tracker (SPOT) System; OMB Control Number 0704-0460. Needs and Uses: In accordance with section 861 of... data into the Synchronized Predeployment and Operational Tracker (SPOT) System before deployment... designated the SPOT as the joint Web- based database to assist the Combatant Commander (CCDR) in maintaining...

Proteomic analysis of heat treated bitter gourd (Momordica charantia L. var. Hong Kong Green) using 2D-DIGE.

PubMed

Ng, Zhi Xiang; Chua, Kek Heng; Kuppusamy, Umah Rani

2014-04-01

This study aimed to investigate the changes in the proteome of bitter gourd prior to and after subjecting to boiling and microwaving. A comparative analysis of the proteome profiles of raw and thermally treated bitter gourds was performed using 2D-DIGE. The protein content and number of protein spots in raw sample was higher when compared to the cooked samples. Qualitative analysis revealed that 103 (boiled sample) and 110 (microwaved sample) protein spots were up regulated whereas 120 (boiled sample) and 107 (microwaved sample) protein spots were down regulated. Ten protein spots with the highest significant fold change in the cooked samples were involved in carbohydrate/energy metabolisms and stress responses. Small heat shock proteins, superoxide dismutase, quinone oxidoreductase, UDP-glucose pyrophosphorylase and phosphoglycerate kinase play a role in heat-stress-mediated protection of bitter gourd. This study suggests that appropriate heat treatment (cooking methods) can lead to induction of selected proteins in bitter gourd. Copyright © 2013 Elsevier Ltd. All rights reserved.

5C.07: A METHOD TO ESTIMATE 24-HOUR SODIUM EXCRETION THROUGH SPOT URINE SAMPLES AND ITS APPLICATION VALUE FOR TARGET-ORGAN DAMAGE ASSESSMENT.

PubMed

Wang, H; Zhao, L; Xi, Y; Sun, N

2015-06-01

24-h urine sodium excretion is considered the most reliable method to evaluate the salt intakes. However, this method is cumbersome. So we want to develop formulas to estimate 24-h urinary sodium excretion using spot urinary samples in Chinese hypertensive population and explore the application value of this method in salt intake assessment and target organ damage. 1. We enrolled 510 cases of hospitalized patients with hypertension, 2/3 of them were arranged randomly to formula group to develop a new formula and the remainings were used to test the performance of the formula. All participants were instructed to collect a 24-h urine sample, a second morning voiding urine sample (SMU), and a post-meridiem urine sample in the late afternoon or early evening, prior to the evening meal (PMU). All samples were sent to measure sodium and creatinine concentration.2. We compared the differences of office blood pressure, 24-hour ambulatory blood pressure and left ventricular hypertrophy, vascular stiffness and urine protein among groups of different sodium intake. 24hour sodium excretion formulas was obtained using SMU and PMU respectively, which have good cosistency. The difference between the estimated and measured values in sodium excretion is 12.66mmol/day (SMU) and 9.41mmol/day (PM), to be equal to 0.7 g (SMU) and 0.6 g (PM) salt intake. Comparing with Kawasaki and Tanaka method, the new formula shows the lower degree of deviation, and higher accuracy and precision. Blood pressure of high urinary sodium group is higher than that in low urinary sodium group (P < 0.05). Left ventricular hypertrophy and urinary albumin/creatinine aggravated with the salt intake increase, this has eliminated the influence of other factors. All of morphologies of the relationship between ambulatory arterial stiffness index, pulse wave velocity and carotid intima-media thickness with quartiles of sodium intake resembled a J-shaped curve. In Chinese hypertensive population, the formulas to estimate 24-h urinary sodium using spot urinary samples spot urine are considered useful for estimating the mean level of population salt intake, and have a role in evaluating target organ damage.

Comparability of HbA1c and lipids measured with dried blood spot versus venous samples: a systematic review and meta-analysis

PubMed Central

2014-01-01

Background Levels of haemoglobin A1c (HbA1c) and blood lipids are important determinants of risk in patients with diabetes. Standard analysis methods based upon venous blood samples can be logistically challenging in resource-poor settings where much of the diabetes epidemic is occurring. Dried blood spots (DBS) provide a simple alternative method for sample collection but the comparability of data from analyses based on DBS is not well established. Methods We conducted a systematic review and meta-analysis to define the association of findings for HbA1c and blood lipids for analyses based upon standard methods compared to DBS. The Cochrane, Embase and Medline databases were searched for relevant reports and summary regression lines were estimated. Results 705 abstracts were found by the initial electronic search with 6 further reports identified by manual review of the full papers. 16 studies provided data for one or more outcomes of interest. There was a close agreement between the results for HbA1c assays based on venous and DBS samples (DBS = 0.9858venous + 0.3809), except for assays based upon affinity chromatography. Significant adjustment was required for assays of total cholesterol (DBS = 0.6807venous + 1.151) but results for triglycerides (DBS = 0.9557venous + 0.1427) were directly comparable. Conclusions For HbA1c and selected blood lipids, assays based on DBS samples are clearly associated with assays based on standard venous samples. There are, however, significant uncertainties about the nature of these associations and there is a need for standardisation of the sample collection, transportation, storage and analysis methods before the technique can be considered mainstream. This should be a research priority because better elucidation of metabolic risks in resource poor settings, where venous sampling is infeasible, will be key to addressing the global epidemic of cardiovascular diseases. PMID:25045323

Rickettsia parkeri rickettsiosis and its clinical distinction from Rocky Mountain spotted fever.

PubMed

Paddock, Christopher D; Finley, Richard W; Wright, Cynthia S; Robinson, Howard N; Schrodt, Barbara J; Lane, Carole C; Ekenna, Okechukwu; Blass, Mitchell A; Tamminga, Cynthia L; Ohl, Christopher A; McLellan, Susan L F; Goddard, Jerome; Holman, Robert C; Openshaw, John J; Sumner, John W; Zaki, Sherif R; Eremeeva, Marina E

2008-11-01

Rickettsia parkeri rickettsiosis, a recently identified spotted fever transmitted by the Gulf Coast tick (Amblyomma maculatum), was first described in 2004. We summarize the clinical and epidemiological features of 12 patients in the United States with confirmed or probable disease attributable to R. parkeri and comment on distinctions between R. parkeri rickettsiosis and other United States rickettsioses. Clinical specimens from patients in the United States who reside within the range of A. maculatum for whom an eschar or vesicular rash was described were evaluated by > or =1 laboratory assays at the Centers for Disease Control and Prevention (Atlanta, GA) to identify probable or confirmed infection with R. parkeri. During 1998-2007, clinical samples from 12 patients with illnesses epidemiologically and clinically compatible with R. parkeri rickettsiosis were submitted for diagnostic evaluation. Using indirect immunofluorescence antibody assays, immunohistochemistry, polymerase chain reaction assays, and cell culture isolation, we identified 6 confirmed and 6 probable cases of infection with R. parkeri. The aggregate clinical characteristics of these patients revealed a disease similar to but less severe than classically described Rocky Mountain spotted fever. Closer attention to the distinct clinical features of the various spotted fever syndromes that exist in the United States and other countries of the Western hemisphere, coupled with more frequent use of specific confirmatory assays, may unveil several unique diseases that have been identified collectively as Rocky Mountain spotted fever during the past century. Accurate assessments of these distinct infections will ultimately provide a more valid description of the currently recognized distribution, incidence, and case-fatality rate of Rocky Mountain spotted fever.

Impact of cryopreservation on bull () semen proteome.

PubMed

Westfalewicz, B; Dietrich, M A; Ciereszko, A

2015-11-01

Cryopreservation of bull spermatozoa is a well-established technique, allowing artificial insemination of cattle on a commercial scale. However, the extent of proteome changes in seminal plasma and spermatozoa during cryopreservation are not yet fully known. The objective of this study was to compare the proteomes of fresh, equilibrated, and cryopreserved bull semen (spermatozoa and seminal plasma) to establish the changes in semen proteins during the cryopreservation process. Semen was collected from 6 mature Holstein Friesian bulls. After sample processing, comparative analysis and identification of proteins was performed using 2-dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization mass spectrometry. Analysis of spermatozoa extracts revealed that 25 identified protein spots, representing 16 proteins, underwent significant ( < 0.05) changes in abundance due to equilibration and cryopreservation. Eighteen protein spots decreased in abundance, 5 protein spots increased in abundance, and 2 protein spots showed different, specific patterns of abundance changes. Analysis of seminal fluid containing seminal plasma showed that 6 identified protein spots, representing 4 proteins, underwent significant ( < 0.05) changes in abundance due to equilibration and cryopreservation. Two protein spots increased in abundance and 4 decreased in abundance. Semen extending and equilibration seems to be responsible for a significant portion of the proteome changes related to cryopreservation technology. Most sperm proteins affected by equilibration and cryopreservation are membrane bound, and loss of those proteins may reduce natural spermatozoa coating. Further research is needed to unravel the mechanisms of the particular protein changes described in this study and establish the relationship between those changes and sperm quality.

Hyperspectral analysis of columbia spotted frog habitat

USGS Publications Warehouse

Shive, J.P.; Pilliod, D.S.; Peterson, C.R.

2010-01-01

Wildlife managers increasingly are using remotely sensed imagery to improve habitat delineations and sampling strategies. Advances in remote sensing technology, such as hyperspectral imagery, provide more information than previously was available with multispectral sensors. We evaluated accuracy of high-resolution hyperspectral image classifications to identify wetlands and wetland habitat features important for Columbia spotted frogs (Rana luteiventris) and compared the results to multispectral image classification and United States Geological Survey topographic maps. The study area spanned 3 lake basins in the Salmon River Mountains, Idaho, USA. Hyperspectral data were collected with an airborne sensor on 30 June 2002 and on 8 July 2006. A 12-year comprehensive ground survey of the study area for Columbia spotted frog reproduction served as validation for image classifications. Hyperspectral image classification accuracy of wetlands was high, with a producer's accuracy of 96 (44 wetlands) correctly classified with the 2002 data and 89 (41 wetlands) correctly classified with the 2006 data. We applied habitat-based rules to delineate breeding habitat from other wetlands, and successfully predicted 74 (14 wetlands) of known breeding wetlands for the Columbia spotted frog. Emergent sedge microhabitat classification showed promise for directly predicting Columbia spotted frog egg mass locations within a wetland by correctly identifying 72 (23 of 32) of known locations. Our study indicates hyperspectral imagery can be an effective tool for mapping spotted frog breeding habitat in the selected mountain basins. We conclude that this technique has potential for improving site selection for inventory and monitoring programs conducted across similar wetland habitat and can be a useful tool for delineating wildlife habitats. ?? 2010 The Wildlife Society.

Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS

PubMed Central

Shokati, Touraj; Bodenberger, Nicholas; Gadpaille, Holly; Schniedewind, Björn; Vinks, Alexander A.; Jiang, Wenlei; Alloway, Rita R.; Christians, Uwe

2015-01-01

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United States. Tacrolimus is a narrow therapeutic index drug and as such requires therapeutic drug monitoring and dose adjustment based on its whole blood trough concentrations. To facilitate home therapeutic drug and adherence monitoring, the collection of dried blood spots is an attractive concept. After a finger stick, the patient collects a blood drop on filter paper at home. After the blood is dried, it is mailed to the analytical laboratory where tacrolimus is quantified using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in combination with a simple manual protein precipitation step and online column extraction. For tacrolimus analysis, a 6-mm disc is punched from the saturated center of the blood spot. The blood spot is homogenized using a bullet blender and then proteins are precipitated with methanol/0.2 M ZnSO4 containing the internal standard D2,13C-tacrolimus. After vortexing and centrifugation, 100 µl of supernatant is injected into an online extraction column and washed with 5 ml/min of 0.1 formic acid/acetonitrile (7:3, v:v) for 1 min. Hereafter, the switching valve is activated and the analytes are back-flushed onto the analytical column (and separated using a 0.1% formic acid/acetonitrile gradient). Tacrolimus is quantified in the positive multi reaction mode (MRM) using a tandem mass spectrometer. The assay is linear from 1 to 50 ng/ml. Inter-assay variability (3.6%-6.1%) and accuracy (91.7%-101.6%) as assessed over 20 days meet acceptance criteria. Average extraction recovery is 95.5%. There are no relevant carry-over, matrix interferences and matrix effects. Tacrolimus is stable in dried blood spots at RT and at +4 °C for 1 week. Extracted samples in the autosampler are stable at +4 °C for at least 72 hr. PMID:26575262

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

PubMed

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

2015-02-01

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

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

PubMed

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

2016-11-01

In many agricultural catchments of Europe and North America, pesticides occur at generally low concentrations with significant temporal variation. This poses several challenges for both monitoring and understanding ecological risks/impacts of these chemicals. This study aimed to compare the performance of passive and spot sampling strategies given the constraints of typical regulatory monitoring. Nine pesticides were investigated in a river currently undergoing regulatory monitoring (River Ugie, Scotland). Within this regulatory framework, spot and passive sampling were undertaken to understand spatiotemporal occurrence, mass loads and ecological risks. All the target pesticides were detected in water by both sampling strategies. Chlorotoluron was observed to be the dominant pesticide by both spot (maximum: 111.8ng/l, mean: 9.35ng/l) and passive sampling (maximum: 39.24ng/l, mean: 4.76ng/l). The annual pesticide loads were estimated to be 2735g and 1837g based on the spot and passive sampling data, respectively. The spatiotemporal trend suggested that agricultural activities were the primary source of the compounds with variability in loads explained in large by timing of pesticide applications and rainfall. The risk assessment showed chlorotoluron and chlorpyrifos posed the highest ecological risks with 23% of the chlorotoluron spot samples and 36% of the chlorpyrifos passive samples resulting in a Risk Quotient greater than 0.1. This suggests that mitigation measures might need to be taken to reduce the input of pesticides into the river. The overall comparison of the two sampling strategies supported the hypothesis that passive sampling tends to integrate the contaminants over a period of exposure and allows quantification of contamination at low concentration. The results suggested that within a regulatory monitoring context passive sampling was more suitable for flux estimation and risk assessment of trace contaminants which cannot be diagnosed by spot sampling and for determining if long-term average concentrations comply with specified standards. Copyright © 2016 Elsevier B.V. All rights reserved.

Analysis of the Cross-Reactivity of Various 56 kDa Recombinant Protein Antigens with Serum Samples Collected after Orientia tsutsugamushi Infection by ELISA

DTIC Science & Technology

2011-01-01

Rickettsia typhi and TT118 spotted fever group rickettsiae among Malaysian blood donors and febrile patients in the urban areas . Southeast Asian J...Jackson EB , 1948 . Studies on scrub typhus (tsutsugamushi disease). Part III: Heterogeneity of strains of Rickettsia tsutsugamushi as...specific and crossreactive lymphocyte proliferative responses during chronic immunizing infections with Rickettsia tsutsugamushi . Infect Immun 40

Blood Substrate Collection and Handling Procedures under Pseudo-Field Conditions: Evaluation of Suitability for Inflammatory Biomarker Measurement.

PubMed

Sugden, Karen; Danese, Andrea; Shalev, Idan; Williams, Benjamin S; Caspi, Avshalom

2015-01-01

Routine incorporation of blood-based biomarker measurements in population studies has been hampered by challenges in obtaining samples suitable for biomarker assessment outside of laboratory settings. Here, we assessed the suitability of venous blood left unprocessed for 4, 24, or 48 hours post-collection at either room temperature or 4°C for quantification of two biomarkers, Interleukin-6 (IL-6) and C-reactive protein (CRP). Blood samples were collected in both K2EDTA tubes and a dedicated plasma-preservation tube, P100. Dried blood spot (DBS) samples from the same subjects were also collected in order to compare delayed-processing plasma performance against a popular alternative collection method. We found that K2EDTA mean plasma concentrations of both IL-6 and CRP were not significantly different from concentrations in plasma processed immediately; this was observed for tubes stored up to 48 hours pre-processing at either temperature. Concentrations of IL-6 measured in P100 tubes showed significant time-dependent increases when stored at room temperature; otherwise, levels of IL-6 and CRP were similar to those found in samples processed immediately. Levels of CRP in DBS were correlated with plasma CRP levels, even when pre-processed blood was stored for up to 48 hours. These data indicate that plasma is suitable for IL-6 and CRP estimation under data collection conditions that involve processing delays.

Rapid LC-MS/MS quantification of the major benzodiazepines and their metabolites on dried blood spots using a simple and cost-effective sample pretreatment.

PubMed

Déglon, Julien; Versace, François; Lauer, Estelle; Widmer, Christèle; Mangin, Patrice; Thomas, Aurélien; Staub, Christian

2012-06-01

Dried blood spots (DBS) sampling has gained popularity in the bioanalytical community as an alternative to conventional plasma sampling, as it provides numerous benefits in terms of sample collection and logistics. The aim of this work was to show that these advantages can be coupled with a simple and cost-effective sample pretreatment, with subsequent rapid LC-MS/MS analysis for quantitation of 15 benzodiazepines, six metabolites and three Z-drugs. For this purpose, a simplified offline procedure was developed that consisted of letting a 5-µl DBS infuse directly into 100 µl of MeOH, in a conventional LC vial. The parameters related to the DBS pretreatment, such as extraction time or internal standard addition, were investigated and optimized, demonstrating that passive infusion in a regular LC vial was sufficient to quantitatively extract the analytes of interest. The method was validated according to international criteria in the therapeutic concentration ranges of the selected compounds. The presented strategy proved to be efficient for the rapid analysis of the selected drugs. Indeed, the offline sample preparation was reduced to a minimum, using a small amount of organic solvent and consumables, without affecting the accuracy of the method. Thus, this approach enables simple and rapid DBS analysis, even when using a non-DBS-dedicated autosampler, while lowering the costs and environmental impact.

Development of dried serum spot sampling techniques for the assessment of trace elements in serum samples by LA-ICP-MS.

PubMed

Chantada-Vázquez, María Pilar; Moreda-Piñeiro, Jorge; Cantarero-Roldán, Alicia; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

2018-08-15

A novel approach for serum analysis by dried matrix spot (DMS) technique is proposed. The methodology consists of sampling filter paper discs (2.7 mm in diameter) containing the large amount of serum retained after a single spotting. Several oxidizers (sodium chlorate, sodium azide, acetic acid, formic acid, 1-butyl-3-methylimidazoliumm chloride/bromide) were tested (oxidizers premixed with the sample before spotting, and papers previously soaked in concentrated additive/oxidizer solutions). Direct multi-element determination (Al, Be, Ca, Cu, Fe, K, Li, Mg, Mn, Mo, Na, P, Rb, Se, V, and Zn) in dried serum spots at very low levels was therefore assessed by laser ablation (LA) coupled with inductively coupled plasma - mass spectrometry (ICP-MS). Laser ablation was performed using a focused Nd: YAG laser beam in lineal scan mode (wavelength 213 nm, laser fluency 2.2 J cm -2 , repetition rate 20 Hz, laser spot diameter 90 µm, depth 0 µm, scanning speed 12 µm s -1 ). Matrix-matched calibration mode and 13 C as internal standard (for signal intensities normalization) was used throughout the work. Limits of quantification were found to be from 21 µg L -1 to 221 mg L -1 . Repeatability (seven ablations of the same dried serum spot) and reproducibility (two ablations of seven dried serum spot from the same material) offered RSDs below 12% for all analytes, which seems satisfactory for clinical purposes. The method was validated by analyzing several certified reference materials (Seronorm™ level I and II trace elements in serum), and it was applied to several DMS from serum samples from healthy adults. Copyright © 2018 Elsevier B.V. All rights reserved.

Prevalence of glucose-6-phosphate dehydrogenase deficiency in neonates in Egypt.

PubMed

Elella, Soheir Abo; Tawfik, Mahaa; Barseem, Naglaa; Moustafa, Wafaa

2017-01-01

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked disorder which causes neonatal jaundice in most cases, and under certain conditions, can cause a spectrum of hemolytic manifestations. To determine the local prevalence of G6PD deficiency in newborns. Cross-sectional. University hospital. Infants born during 2015 were prospectively screened for G6PD deficiency. Dried blood spot samples on filter paper were collected in collaboration with the central laboratories of the Ministry of Health. Quantitative measurement of G6PD enzyme activity was measured from the blood samples using fluorometric analysis. A value.

A cathodoluminescence study on zircons with a complex thermal history traces back Permian crustal events in the Ivrea Zone (South Alpine, Northern Italy)

NASA Astrophysics Data System (ADS)

Peressini, G.; Poller, U.

2003-04-01

In the context of a U-Pb SHRIMP data-set, a cathodoluminescence (CL) study has been performed on zircons from the Ivrea-Verbano Zone; this is a tectonically bounded section of intermediate to deep crust, tilted and obducted in Alpine time, in which a large deep-crustal intrusion of mantle origin, the Mafic Complex (MC), was emplaced in the Early Permian. Zircons from 16 samples from the different units of the MC have been studied using CL. Three samples collected from the Paragneiss Bearing Belt (PBB) yield some partially reset spot-ages, reflecting the field observation that country rock slabs are frequent in that area. On the other side, unlike in zircons from granites, the cores are invisible under CL-imaging, and this makes the U-Pb spot-age results unpredictable, with a continuous range of ages in the same sample, spanning for an interval of over 35 Ma, followed by some much older peaks, up to 600 Ma. A fourth sample, collected 2 km far, but still within the PBB, defines instead a unique, well-defined age at 287±3 Ma, with no older peaks, its zircons showing a CL pattern typical for metamorphic grains. A different case is displayed by a sample collected from the deeper Amphibole Gabbro unit: each single grain records a complex story of magmatic growth with variable diffusivity conditions. All of them show a second major overprint, that lead to both (re)crystallization and resorption, always corresponding to much lower U and Th contents, with no sensible modification of the Th-U ratio. The age of the second event, though, is not distinguishable from that of first crystallization of the grains, and has not been at such a temperature as to obliterate the fine zoning pattern of the primary grain. The CL patterns of each single grain, composed of different domains, allow considerations on the environmental conditions of growth and (re)crystallization. CL is a very powerful tool itself, revealing crystal-chemical processes. The integration of the CL-study with the SHRIMP spot-age determination relates them to the sequence of geologic events. The study, performed on samples coming from all over the Complex, has allowed a better understanding of processes in the Carboniferous-Permian lower crust of the Ivrea Zone: the knowledge of the Complex, its structure and the field relationships has become a key for interpreting the results. Where the CL displays a composite pattern, mechanisms are needed to explain geochemical fingerprints: therefore, trace and REE analyses by ion probe are in progress to integrate the SHRIMP data.

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

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

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

Rapid DNA extraction protocol for detection of alpha-1 antitrypsin deficiency from dried blood spots by real-time PCR.

PubMed

Struniawski, R; Szpechcinski, A; Poplawska, B; Skronski, M; Chorostowska-Wynimko, J

2013-01-01

The dried blood spot (DBS) specimens have been successfully employed for the large-scale diagnostics of α1-antitrypsin (AAT) deficiency as an easy to collect and transport alternative to plasma/serum. In the present study we propose a fast, efficient, and cost effective protocol of DNA extraction from dried blood spot (DBS) samples that provides sufficient quantity and quality of DNA and effectively eliminates any natural PCR inhibitors, allowing for successful AAT genotyping by real-time PCR and direct sequencing. DNA extracted from 84 DBS samples from chronic obstructive pulmonary disease patients was genotyped for AAT deficiency variants by real-time PCR. The results of DBS AAT genotyping were validated by serum IEF phenotyping and AAT concentration measurement. The proposed protocol allowed successful DNA extraction from all analyzed DBS samples. Both quantity and quality of DNA were sufficient for further real-time PCR and, if necessary, for genetic sequence analysis. A 100% concordance between AAT DBS genotypes and serum phenotypes in positive detection of two major deficiency S- and Z- alleles was achieved. Both assays, DBS AAT genotyping by real-time PCR and serum AAT phenotyping by IEF, positively identified PI*S and PI*Z allele in 8 out of the 84 (9.5%) and 16 out of 84 (19.0%) patients, respectively. In conclusion, the proposed protocol noticeably reduces the costs and the hand-on-time of DBS samples preparation providing genomic DNA of sufficient quantity and quality for further real-time PCR or genetic sequence analysis. Consequently, it is ideally suited for large-scale AAT deficiency screening programs and should be method of choice.

Simple and rapid analytical method for detection of amino acids in blood using blood spot on filter paper, fast-GC/MS and isotope dilution technique.

PubMed

Kawana, Shuichi; Nakagawa, Katsuhiro; Hasegawa, Yuki; Yamaguchi, Seiji

2010-11-15

A simple and rapid method for quantitative analysis of amino acids, including valine (Val), leucine (Leu), isoleucine (Ile), methionine (Met) and phenylalanine (Phe), in whole blood has been developed using GC/MS. In this method, whole blood was collected using a filter paper technique, and a 1/8 in. blood spot punch was used for sample preparation. Amino acids were extracted from the sample, and the extracts were purified using cation-exchange resins. The isotope dilution method using ²H₈-Val, ²H₃-Leu, ²H₃-Met and ²H₅-Phe as internal standards was applied. Following propyl chloroformate derivatization, the derivatives were analyzed using fast-GC/MS. The extraction recoveries using these techniques ranged from 69.8% to 87.9%, and analysis time for each sample was approximately 26 min. Calibration curves at concentrations from 0.0 to 1666.7 μmol/l for Val, Leu, Ile and Phe and from 0.0 to 333.3 μmol/l for Met showed good linearity with regression coefficients=1. The method detection limits for Val, Leu, Ile, Met and Phe were 24.2, 16.7, 8.7, 1.5 and 12.9 μmol/l, respectively. This method was applied to blood spot samples obtained from patients with phenylketonuria (PKU), maple syrup urine disease (MSUD), hypermethionine and neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD), and the analysis results showed that the concentrations of amino acids that characterize these diseases were increased. These results indicate that this method provides a simple and rapid procedure for precise determination of amino acids in whole blood. Copyright © 2010 Elsevier B.V. All rights reserved.

A liquid chromatography-tandem mass spectrometry method for the determination of cocaine and metabolites in blood and in dried blood spots collected from postmortem samples and evaluation of the stability over a 3-month period.

PubMed

Moretti, Matteo; Visonà, Silvia Damiana; Freni, Francesca; Tomaciello, Ilaria; Vignali, Claudia; Groppi, Angelo; Tajana, Luca; Osculati, Antonio Marco Maria; Morini, Luca

2018-05-04

The aims of this study were (1) to identify and quantify cocaine (COC), benzoylecgonine (BE), ecgonine methyl ester (EME), and cocaethylene (CE) in DBS; (2) to compare dried blood spots (DBSs) analytical results with the routine blood analyses; (3) to monitor analytes stability on DBS within a 3-month period. Eighty-five μL of blood from postmortem cases were put on a card for DBS analysis and kept in the dark, at room temperature. Samples were extracted through solid-phase extraction (SPE) cartridges and injected in the liquid chromatography-tandem mass spectrometry (LC-MS/MS) system. The analytical procedure is simple, sensitive, and specific. Limits of detection (LODs) and quantification (LOQs) were calculated at 1.0 and 5.0 ng/mL(g) for COC and CE, and at 0.5 and 2 ng/mL for EME and BE, respectively. Validation parameters fulfilled all the acceptance criteria. Fifty-five postmortem cases were evaluated. Eighteen cases were positive for COC (44-2456 ng/mL) and BE (228-4700 ng/mL), 12 for EME (92-1500 ng/mL), and 11 cases for CE (11-273 ng/mL). Stability was evaluated on 8 cases collected in the period January 2017-January 2018. For each case, 5 DBSs were collected at T0. Four DBSs were analyzed within the 4 following weeks and 1 sample was analyzed after 3 months. The concentrations on DBSs, stored at room temperature, always matched the ones obtained on blood samples kept at -20°C (<20% variation, both at T0 and after 3 months). BE and COC concentrations remained stable after a 3-month storage, EME concentrations slightly increased after 3 weeks in the 2 analyzed samples, while CE provided a less homogeneous stability depending on the sample. Copyright © 2018 John Wiley & Sons, Ltd.

A universal reference sample derived from clone vector for improved detection of differential gene expression

PubMed Central

Khan, Rishi L; Gonye, Gregory E; Gao, Guang; Schwaber, James S

2006-01-01

Background Using microarrays by co-hybridizing two samples labeled with different dyes enables differential gene expression measurements and comparisons across slides while controlling for within-slide variability. Typically one dye produces weaker signal intensities than the other often causing signals to be undetectable. In addition, undetectable spots represent a large problem for two-color microarray designs and most arrays contain at least 40% undetectable spots even when labeled with reference samples such as Stratagene's Universal Reference RNAs™. Results We introduce a novel universal reference sample that produces strong signal for all spots on the array, increasing the average fraction of detectable spots to 97%. Maximizing detectable spots on the reference image channel also decreases the variability of microarray data allowing for reliable detection of smaller differential gene expression changes. The reference sample is derived from sequence contained in the parental EST clone vector pT7T3D-Pac and is called vector RNA (vRNA). We show that vRNA can also be used for quality control of microarray printing and PCR product quality, detection of hybridization anomalies, and simplification of spot finding and segmentation tasks. This reference sample can be made inexpensively in large quantities as a renewable resource that is consistent across experiments. Conclusion Results of this study show that vRNA provides a useful universal reference that yields high signal for almost all spots on a microarray, reduces variation and allows for comparisons between experiments and laboratories. Further, it can be used for quality control of microarray printing and PCR product quality, detection of hybridization anomalies, and simplification of spot finding and segmentation tasks. This type of reference allows for detection of small changes in differential expression while reference designs in general allow for large-scale multivariate experimental designs. vRNA in combination with reference designs enable systems biology microarray experiments of small physiologically relevant changes. PMID:16677381

Characterization of Plasmodium Lactate Dehydrogenase and Histidine-Rich Protein 2 Clearance Patterns via Rapid On-Bead Detection from a Single Dried Blood Spot

PubMed Central

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

2018-01-01

Abstract. A rapid, on-bead enzyme-linked immunosorbent assay for Plasmodium lactate dehydrogenase (pLDH) and Plasmodium falciparum histidine-rich protein 2 (HRP2) was adapted for use with dried blood spot (DBS) samples. This assay detected both biomarkers from a single DBS sample with only 45 minutes of total incubation time and detection limits of 600 ± 500 pM (pLDH) and 69 ± 30 pM (HRP2), corresponding to 150 and 24 parasites/μL, respectively. This sensitive and reproducible on-bead detection method was used to quantify pLDH and HRP2 in patient DBS samples from rural Zambia collected at multiple time points after treatment. Biomarker clearance patterns relative to parasite clearance were determined; pLDH clearance followed closely with parasite clearance, whereas most patients maintained detectable levels of HRP2 for 35–52 days after treatment. Furthermore, weak-to-moderate correlations between biomarker concentration and parasite densities were found for both biomarkers. This work demonstrates the utility of the developed assay for epidemiological study and surveillance of malaria. PMID:29557342

DBS-platform for biomonitoring and toxicokinetics of toxicants: proof of concept using LC-MS/MS analysis of fipronil and its metabolites in blood

NASA Astrophysics Data System (ADS)

Raju, Kanumuri Siva Rama; Taneja, Isha; Rashid, Mamunur; Sonkar, Ashish Kumar; Wahajuddin, Muhammad; Singh, Sheelendra Pratap

2016-03-01

A simple, sensitive and high throughput LC-MS/MS method was developed and validated for quantification of fipronil, fipronil sulfone and fipronil desulfinyl in rat and human dried blood spots (DBS). DBS samples were prepared by spiking 10 μl blood on DMPK-C cards followed by drying at room temperature. The whole blood spots were then punched from the card and extracted using acetonitrile. The total chromatographic run time of the method was only 2 min. The lower limit of quantification of the method was 0.1 ng/ml for all the analytes. The method was successfully applied to determine fipronil desulfinyl in DBS samples obtained from its toxicokinetic study in rats following intravenous dose (1 mg/kg). In conclusion, the proposed DBS methodology has significant potential in toxicokinetics and biomonitoring studies of environmental toxicants. This microvolume DBS technique will be an ideal tool for biomonitoring studies, particularly in paediatric population. Small volume requirements, minimally invasive blood sampling method, easier storage and shipping procedure make DBS a suitable technique for such studies. Further, DBS technique contributes towards the principles of 3Rs resulting in significant reduction in the number of rodents used and refinement in sample collection for toxicokinetic studies.

Occurrence of pathogenic fungi to Amblyomma cajennense in a rural area of Central Brazil and their activities against vectors of Rocky Mountain spotted fever.

PubMed

D'Alessandro, Walmirton B; Humber, Richard A; Luz, Christian

2012-08-13

Two isolates of Beauveria bassiana and one of Purpureocillium lilacinum (=Paecilomyces lilacinus) were found infecting Amblyomma cajennense engorged females collected on horses (0.15% infection rate from a total of 1982 specimens) and another two isolates of P. lilacinum and one Metarhizium anisopliae detected in soils (2.1% from 144 samples) collected in typical pasture habitats of this tick in Central Brazil from October 2009 to March 2011. Fungi were isolated from soils with Rhipicephalus sanguineus as surrogate baits. No fungi were found in ticks or soils during the driest months (May to August). Testing pathogenicity of fungi all R. sanguineus females were killed regardless of the isolate and fungi sporulated abundantly on the cadavers. A. cajennense was less susceptible to infection with P. lilacinum within 20 days than R. sanguineus. All three fungal species probably act as natural antagonists of A. cajennense particularly in the rainy season and have interest for integrate control of vectors of Rocky Mountain spotted fever. Copyright © 2012 Elsevier B.V. All rights reserved.

Rickettsial Infections of Fleas Collected From Small Mammals on Four Islands in Indonesia

DTIC Science & Technology

2010-01-01

cheopis from shrews ( Suncus murinus). X. cheopis were pooled and tested for DNA from rickettsial agents Rickettsia typhi, Rickettsia felis, and spotted...fever group rickettsiae . R. typhi, the agent of murine typhus, was detected in X. cheopis collected from small mammals in West Java and East...Kalimantan. R.felis was detected in X. cheopis collected from small mammals in Manado, North Sulawesi. R. felis and spotted fever group rickettsiae were

Three-dimensional rotation electron diffraction: software RED for automated data collection and data processing

PubMed Central

Wan, Wei; Sun, Junliang; Su, Jie; Hovmöller, Sven; Zou, Xiaodong

2013-01-01

Implementation of a computer program package for automated collection and processing of rotation electron diffraction (RED) data is described. The software package contains two computer programs: RED data collection and RED data processing. The RED data collection program controls the transmission electron microscope and the camera. Electron beam tilts at a fine step (0.05–0.20°) are combined with goniometer tilts at a coarse step (2.0–3.0°) around a common tilt axis, which allows a fine relative tilt to be achieved between the electron beam and the crystal in a large tilt range. An electron diffraction (ED) frame is collected at each combination of beam tilt and goniometer tilt. The RED data processing program processes three-dimensional ED data generated by the RED data collection program or by other approaches. It includes shift correction of the ED frames, peak hunting for diffraction spots in individual ED frames and identification of these diffraction spots as reflections in three dimensions. Unit-cell parameters are determined from the positions of reflections in three-dimensional reciprocal space. All reflections are indexed, and finally a list with hkl indices and intensities is output. The data processing program also includes a visualizer to view and analyse three-dimensional reciprocal lattices reconstructed from the ED frames. Details of the implementation are described. Data collection and data processing with the software RED are demonstrated using a calcined zeolite sample, silicalite-1. The structure of the calcined silicalite-1, with 72 unique atoms, could be solved from the RED data by routine direct methods. PMID:24282334

Three-dimensional rotation electron diffraction: software RED for automated data collection and data processing.

PubMed

Wan, Wei; Sun, Junliang; Su, Jie; Hovmöller, Sven; Zou, Xiaodong

2013-12-01

Implementation of a computer program package for automated collection and processing of rotation electron diffraction (RED) data is described. The software package contains two computer programs: RED data collection and RED data processing. The RED data collection program controls the transmission electron microscope and the camera. Electron beam tilts at a fine step (0.05-0.20°) are combined with goniometer tilts at a coarse step (2.0-3.0°) around a common tilt axis, which allows a fine relative tilt to be achieved between the electron beam and the crystal in a large tilt range. An electron diffraction (ED) frame is collected at each combination of beam tilt and goniometer tilt. The RED data processing program processes three-dimensional ED data generated by the RED data collection program or by other approaches. It includes shift correction of the ED frames, peak hunting for diffraction spots in individual ED frames and identification of these diffraction spots as reflections in three dimensions. Unit-cell parameters are determined from the positions of reflections in three-dimensional reciprocal space. All reflections are indexed, and finally a list with hkl indices and intensities is output. The data processing program also includes a visualizer to view and analyse three-dimensional reciprocal lattices reconstructed from the ED frames. Details of the implementation are described. Data collection and data processing with the software RED are demonstrated using a calcined zeolite sample, silicalite-1. The structure of the calcined silicalite-1, with 72 unique atoms, could be solved from the RED data by routine direct methods.

International Study to Predict Optimized Treatment for Depression (iSPOT-D), a randomized clinical trial: rationale and protocol.

PubMed

Williams, Leanne M; Rush, A John; Koslow, Stephen H; Wisniewski, Stephen R; Cooper, Nicholas J; Nemeroff, Charles B; Schatzberg, Alan F; Gordon, Evian

2011-01-05

Clinically useful treatment moderators of Major Depressive Disorder (MDD) have not yet been identified, though some baseline predictors of treatment outcome have been proposed. The aim of iSPOT-D is to identify pretreatment measures that predict or moderate MDD treatment response or remission to escitalopram, sertraline or venlafaxine; and develop a model that incorporates multiple predictors and moderators. The International Study to Predict Optimized Treatment - in Depression (iSPOT-D) is a multi-centre, international, randomized, prospective, open-label trial. It is enrolling 2016 MDD outpatients (ages 18-65) from primary or specialty care practices (672 per treatment arm; 672 age-, sex- and education-matched healthy controls). Study-eligible patients are antidepressant medication (ADM) naïve or willing to undergo a one-week wash-out of any non-protocol ADM, and cannot have had an inadequate response to protocol ADM. Baseline assessments include symptoms; distress; daily function; cognitive performance; electroencephalogram and event-related potentials; heart rate and genetic measures. A subset of these baseline assessments are repeated after eight weeks of treatment. Outcomes include the 17-item Hamilton Rating Scale for Depression (primary) and self-reported depressive symptoms, social functioning, quality of life, emotional regulation, and side-effect burden (secondary). Participants may then enter a naturalistic telephone follow-up at weeks 12, 16, 24 and 52. The first half of the sample will be used to identify potential predictors and moderators, and the second half to replicate and confirm. First enrolment was in December 2008, and is ongoing. iSPOT-D evaluates clinical and biological predictors of treatment response in the largest known sample of MDD collected worldwide. International Study to Predict Optimised Treatment - in Depression (iSPOT-D) ClinicalTrials.gov Identifier: NCT00693849. URL: http://clinicaltrials.gov/ct2/show/NCT00693849?term=International+Study+to+Predict+Optimized+Treatment+for+Depression&rank=1

Sexing young snowy owls

USGS Publications Warehouse

Seidensticker, M.T.; Holt, D.W.; Detienne, J.; Talbot, S.; Gray, K.

2011-01-01

We predicted sex of 140 Snowy Owl (Bubo scandiacus) nestlings out of 34 nests at our Barrow, Alaska, study area to develop a technique for sexing these owls in the field. We primarily sexed young, flightless owls (3844 d old) by quantifying plumage markings on the remiges and tail, predicting sex, and collecting blood samples to test our field predictions using molecular sexing techniques. We categorized and quantified three different plumage markings: two types of bars (defined as markings that touch the rachis) and spots (defined as markings that do not touch the rachis). We predicted sex in the field assuming that males had more spots than bars and females more bars than spots on the remiges and rectrices. Molecular data indicated that we correctly sexed 100% of the nestlings. We modeled the data using random forests and classification trees. Both models indicated that the number and type of markings on the secondary feathers were the most important in classifying nestling sex. The statistical models verified our initial qualitative prediction that males have more spots than bars and females more bars than spots on flight feathers P6P10 for both wings and tail feathers T1 and T2. This study provides researchers with an easily replicable and highly accurate method for sexing young Snowy Owls in the field, which should aid further studies of sex-ratios and sex-related variation in behavior and growth of this circumpolar owl species. ?? 2011 The Raptor Research Foundation, Inc.

Combining near infrared spectra of feces and geostatistics to generate forage nutritional quality maps across landscapes.

PubMed

Pierre-Olivier, Jean; Bradley, Robert L; Tremblay, Jean-Pierre; Côté, Steeve D

2015-09-01

An important asset for the management of wild ungulates is recognizing the spatial distribution of forage quality across heterogeneous landscapes. To do so typically requires knowledge of which plant species are eaten, in what abundance they are eaten, and what their nutritional quality might be. Acquiring such data, however, may be difficult and time consuming. Here, we are proposing a rapid and cost-effective forage quality monitoring tool that combines near infrared (NIR) spectra of fecal samples and easily obtained data on plant community composition. Our approach rests on the premise that NIR spectra of fecal samples collected within low population density exclosures reflect the optimal forage quality of a given landscape. Forage quality can thus be based on the Mahalanobis distance of fecal spectral scans across the landscape relative to fecal spectral scans inside exclosures (referred to as DISTEX). The Gi* spatial autocorrelation statistic can then be applied among neighboring DISTEX values to detect and map "hot spots" and "cold spots" of nutritional quality over the landscape. We tested our approach in a heterogeneous boreal landscape on Anticosti Island (Québec, Canada), where white-tailed deer (Odocoileus virginianus) populations over the landscape have ranged from 20 to 50 individuals/km2 for at least 80 years, resulting in a loss of most palatable and nutritious plant species. Our results suggest that hot spots of forage quality occur when old-growth balsam fir stands comprise >39.8% of 300 ha neighborhoods, whereas cold spots occur in laggs (i.e., transition zones from forest to peatland). In terms of ground-level indicator plant species, the presence of Canada bunchberry (Cornus canadensis) was highly correlated with hot spots, whereas tamarack (Larix laricina) was highly correlated with cold spots. Mean DISTEX values were positively and significantly correlated with the neutral detergent fiber and acid detergent lignin contents of feces. While our approach would need more independent field trials before it is fully validated, its low cost and ease of execution should make it a valuable tool for advancing both the basic and applied ecology of large herbivores.

Population structure and the rapid increase of QoI fungicide resistance in frogeye leaf spot (Cercospora sojina) from Tennessee

USDA-ARS?s Scientific Manuscript database

Frogeye leaf spot (FLS), caused by Cercospora sojina, causes significant damage to soybean in the US. In this study, C. sojina isolates collected from Jackson and Milan, TN in 2015 and historical isolates collected before 2015 were included. Fifty novel single nucleotide polymorphism (SNP) markers, ...

BOREAS Level-3s SPOT Imagery: Scaled At-sensor Radiance in LGSOWG Format

NASA Technical Reports Server (NTRS)

Strub, Richard; Nickeson, Jaime; Newcomer, Jeffrey A.; Hall, Forrest G. (Editor); Cihlar, Josef

2000-01-01

For BOReal Ecosystem-Atmosphere Study (BOREAS), the level-3s Satellite Pour l'Observation de la Terre (SPOT) data, along with the other remotely sensed images, were collected in order to provide spatially extensive information over the primary study areas. This information includes radiant energy, detailed land cover, and biophysical parameter maps such as Fraction of Photosynthetically Active Radiation (FPAR) and Leaf Area Index (LAI). The SPOT images acquired for the BOREAS project were selected primarily to fill temporal gaps in the Landsat Thematic Mapper (TM) image data collection. CCRS collected and supplied the level-3s images to BOREAS Information System (BORIS) for use in the remote sensing research activities. Spatially, the level-3s images cover 60- by 60-km portions of the BOREAS Northern Study Area (NSA) and Southern Study Area (SSA). Temporally, the images cover the period of 17-Apr-1994 to 30-Aug-1996. The images are available in binary image format files. Due to copyright issues, the SPOT images may not be publicly available.

Writing News Spots about Science: A Way to Promote Scientific Literacy

ERIC Educational Resources Information Center

Marks, Ralf; Otten, Juliane; Eilks, Ingo

2010-01-01

Use of the "Journalist Method" in science education is described, in which pupils are asked to create short news presentations (we call them news spots) for a fictitious television newscast about science-related issues. The writing of such news spots by cooperative teams of pupils is supported by collections of background information…

Molecular and serological evidence of flea-associated typhus group and spotted fever group rickettsial infections in Madagascar.

PubMed

Rakotonanahary, Rado J L; Harrison, Alan; Maina, Alice N; Jiang, Ju; Richards, Allen L; Rajerison, Minoarisoa; Telfer, Sandra

2017-03-04

Rickettsiae are obligate intracellular bacteria responsible for many febrile syndromes around the world, including in sub-Saharan Africa. Vectors of these pathogens include ticks, lice, mites and fleas. In order to assess exposure to flea-associated Rickettsia species in Madagascar, human and small mammal samples from an urban and a rural area, and their associated fleas were tested. Anti-typhus group (TGR)- and anti-spotted fever group rickettsiae (SFGR)-specific IgG were detected in 24 (39%) and 21 (34%) of 62 human serum samples, respectively, using indirect ELISAs, with six individuals seropositive for both. Only two (2%) Rattus rattus out of 86 small mammals presented antibodies against TGR. Out of 117 fleas collected from small mammals, Rickettsia typhi, a TGR, was detected in 26 Xenopsylla cheopis (24%) collected from rodents of an urban area (n = 107), while two of these urban X. cheopis (2%) were positive for Rickettsia felis, a SFGR. R. felis DNA was also detected in eight (31%) out of 26 Pulex irritans fleas. The general population in Madagascar are exposed to rickettsiae, and two flea-associated Rickettsia pathogens, R. typhi and R. felis, are present near or in homes. Although our results are from a single district, they demonstrate that rickettsiae should be considered as potential agents of undifferentiated fever in Madagascar.

Use of finger-prick dried blood spots (fpDBS) and capillary electrophoresis for carbohydrate deficient transferrin (CDT) screening in forensic toxicology.

PubMed

Bertaso, Anna; Sorio, Daniela; Vandoros, Anthula; De Palo, Elio F; Bortolotti, Federica; Tagliaro, Franco

2016-10-01

Continued progress in chronic alcohol abuse investigation requires the development of less invasive procedures for screening purposes. The application of finger-prick and related dried blood spots (fpDBS) for carbohydrate deficient transferrin (CDT) detection appears suitable for this aim. Therefore, the goal of this project was to develop a screening method for CDT using fpDBS with CZE analysis. Blood samples prepared by finger-prick were placed on DBS cards and left to air dry; each dried fpDBS disc was shredded into small pieces and suspended in acid solution (60 μL of HCl 120 mmol/L). After centrifugation (10 min at 1500 × g), the collected sample was adjusted to pH 3.5. After an overnight incubation, the pH was neutralised and an iron rich solution was added. After 1 h, CZE analysis was carried out. A group of 47 individuals was studied. Parallel serum samples were collected from each investigated subject and the %CDT for each sample was measured using HPLC and CZE techniques. The fpDBS transferrin sialo isoform electropherograms were similar to those obtained with serum. Moreover, fpDBS CZE CDT percentage levels demonstrated significant statistical correlation with those obtained from serum for both HPLC and CZE %CDT (p < 0.01; r 2 = 0.8913 and 0.8976, respectively), with %CDT from 0.8 to 13.7% for fpDBS and from 0.7 to 12.7% for serum. The newly developed fpDBS procedure for CDT analysis provides a simple and inexpensive tool for use in population screening. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Chlamydia psittaci and C. avium in feral pigeon (Columba livia domestica) droppings in two cities in the Netherlands.

PubMed

Burt, Sara A; Röring, Romy E; Heijne, Marloes

2018-06-05

Feral pigeons (Columba livia domestica) live and breed in many city centres and contact with their droppings can be a hazard for human health if the birds carry Chlamydia psittaci. The aim of this study was to establish whether pigeon droppings in two Dutch cities (Utrecht and Haarlem) contain C. psittaci and/or C. avium, which could be a potential hazard for transmission to humans. In May 2017 seven feral pigeon 'hot spots' with between 5 and 40+ pigeons present were identified in two cities by visual observations over two days. During the following ten days fresh droppings were collected at these hot spots and the samples were pooled per three droppings to achieve 40-41 samples per city. Samples were analysed for Chlamydia DNA with a broad range 23S Chlamydiaceae Real-Time PCR and positive samples were tested with a specific C. psittaci and C. avium Real-Time PCR. Positive C. psittaci samples were genotyped. C. psittaci and C. avium were detected in both cities. For C. psittaci the prevalences in Utrecht and Haarlem were 2.4% and 7.5%, respectively; for C. avium 36.6% and 20.0%, respectively. One sample contained both species. All C. psittaci samples belonged to genotype B. C. psittaci and C. avium are present in feral pigeon droppings in Utrecht and Haarlem. Human contact with droppings from infected pigeons or inhalation of dust from dried droppings represent a potential hazard to public health.

Paper based Flexible and Conformal SERS Substrate for Rapid Trace Detection on Real-world Surfaces

NASA Astrophysics Data System (ADS)

Singamaneni, Srikanth; Lee, Chang; Tian, Limei

2011-03-01

One of the important but often overlooked considerations in the design of surface enhanced Raman scattering (SERS) substrates for trace detection is the efficiency of sample collection. Conventional designs based on rigid substrates such as silicon, alumina, and glass resist conformal contact with the surface under investigation, making the sample collection inefficient. We demonstrate a novel SERS substrate based on common filter paper adsorbed with gold nanorods, which allows conformal contact with real-world surfaces, thus dramatically enhancing the sample collection efficiency compared to conventional rigid substrates. We demonstrate the detection of trace amounts of analyte (140 pg spread over 4 cm2) by simply swabbing the surface under investigation with the novel SERS substrate. The hierarchical fibrous structure of paper serves as a 3D vasculature for easy uptake and transport of the analytes to the electromagnetic hot spots in the paper. Simple yet highly efficient and cost effective SERS substrate demonstrated here brings SERS based trace detection closer to real-world applications. We acknowledge the financial support from Center for Materials Innovation at Washington University.

Dried blood spot analysis for therapeutic drug monitoring of pazopanib.

PubMed

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

2015-12-01

Dried blood spot (DBS) sampling is potentially a more patient-friendly and flexible alternative to venous sampling of pazopanib. This study determined the agreement between pazopanib DBS and plasma concentrations to facilitate implementation of pazopanib DBS sampling into clinical practice. Paired DBS and plasma samples were collected in 12 patients. Pazopanib plasma concentrations were calculated from DBS concentrations using the formula: plasma concentration = DBSconcentration /(1 - hematocrit). Passing-Bablok and Bland-Altman analyses were used to determine the agreement between calculated and measured plasma concentrations. We predefined a clinical acceptance limit of 25% for the Bland-Altman analysis. Passing-Bablok analysis showed a small constant (intercept estimate, -8.53 [95%CI, -12.22 to -4.41]) and slightly proportional (slope estimate, 1.15 [95%CI, 1.04-1.24]) bias between calculated and measured concentrations. This bias was clinically nonrelevant, as shown by Bland-Altman analysis; the mean ratio of calculated to measured concentrations was 0.94 (95%CI, 0.65-1.23). The clinical acceptance limits were well within these 95% limits of agreement. More specifically, 92.6% of the data points were within the predefined acceptance limits. Pazopanib plasma concentrations can be accurately calculated from DBS concentrations. Although validation of DBS cards prepared by patients themselves is required, these results show that DBS sampling can be used to monitor pazopanib therapy in clinical practice. © 2015, The American College of Clinical Pharmacology.

Malaria PCR Detection in Cambodian Low-Transmission Settings: Dried Blood Spots versus Venous Blood Samples

PubMed Central

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

2015-01-01

In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas. PMID:25561570

75 FR 5996 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2010-02-05

... and OMB Number: Synchronized Predeployment and Operational Tracker (SPOT) System; OMB Control Number... Operational Tracker (SPOT) System before deployment outside the United States. Data collection on contractors...

Understanding biological mechanisms underlying adverse birth outcomes in developing countries: protocol for a prospective cohort (AMANHI bio-banking) study.

PubMed

Baqui, Abdullah H; Khanam, Rasheda; Rahman, Mohammad Sayedur; Ahmed, Aziz; Rahman, Hasna Hena; Moin, Mamun Ibne; Ahmed, Salahuddin; Jehan, Fyezah; Nisar, Imran; Hussain, Atiya; Ilyas, Muhammad; Hotwani, Aneeta; Sajid, Muhammad; Qureshi, Shahida; Zaidi, Anita; Sazawal, Sunil; Ali, Said M; Deb, Saikat; Juma, Mohammed Hamad; Dhingra, Usha; Dutta, Arup; Ame, Shaali Makame; Hayward, Caroline; Rudan, Igor; Zangenberg, Mike; Russell, Donna; Yoshida, Sachiyo; Polašek, Ozren; Manu, Alexander; Bahl, Rajiv

2017-12-01

The AMANHI study aims to seek for biomarkers as predictors of important pregnancy-related outcomes, and establish a biobank in developing countries for future research as new methods and technologies become available. AMANHI is using harmonised protocols to enrol 3000 women in early pregnancies (8-19 weeks of gestation) for population-based follow-up in pregnancy up to 42 days postpartum in Bangladesh, Pakistan and Tanzania, with collection taking place between August 2014 and June 2016. Urine pregnancy tests will be used to confirm reported or suspected pregnancies for screening ultrasound by trained sonographers to accurately date the pregnancy. Trained study field workers will collect very detailed phenotypic and epidemiological data from the pregnant woman and her family at scheduled home visits during pregnancy (enrolment, 24-28 weeks, 32-36 weeks & 38+ weeks) and postpartum (days 0-6 or 42-60). Trained phlebotomists will collect maternal and umbilical blood samples, centrifuge and obtain aliquots of serum, plasma and the buffy coat for storage. They will also measure HbA1C and collect a dried spot sample of whole blood. Maternal urine samples will also be collected and stored, alongside placenta, umbilical cord tissue and membrane samples, which will both be frozen and prepared for histology examination. Maternal and newborn stool (for microbiota) as well as paternal and newborn saliva samples (for DNA extraction) will also be collected. All samples will be stored at -80°C in the biobank in each of the three sites. These samples will be linked to numerous epidemiological and phenotypic data with unique study identification numbers. AMANHI biobank proves that biobanking is feasible to implement in LMICs, but recognises that biobank creation is only the first step in addressing current global challenges.

Crowdsourcing the identification of organisms: A case-study of iSpot

PubMed Central

Silvertown, Jonathan; Harvey, Martin; Greenwood, Richard; Dodd, Mike; Rosewell, Jon; Rebelo, Tony; Ansine, Janice; McConway, Kevin

2015-01-01

Abstract Accurate species identification is fundamental to biodiversity science, but the natural history skills required for this are neglected in formal education at all levels. In this paper we describe how the web application ispotnature.org and its sister site ispot.org.za (collectively, “iSpot”) are helping to solve this problem by combining learning technology with crowdsourcing to connect beginners with experts. Over 94% of observations submitted to iSpot receive a determination. External checking of a sample of 3,287 iSpot records verified > 92% of them. To mid 2014, iSpot crowdsourced the identification of 30,000 taxa (>80% at species level) in > 390,000 observations with a global community numbering > 42,000 registered participants. More than half the observations on ispotnature.org were named within an hour of submission. iSpot uses a unique, 9-dimensional reputation system to motivate and reward participants and to verify determinations. Taxon-specific reputation points are earned when a participant proposes an identification that achieves agreement from other participants, weighted by the agreers’ own reputation scores for the taxon. This system is able to discriminate effectively between competing determinations when two or more are proposed for the same observation. In 57% of such cases the reputation system improved the accuracy of the determination, while in the remainder it either improved precision (e.g. by adding a species name to a genus) or revealed false precision, for example where a determination to species level was not supported by the available evidence. We propose that the success of iSpot arises from the structure of its social network that efficiently connects beginners and experts, overcoming the social as well as geographic barriers that normally separate the two. PMID:25685027

Sensitivity and specificity of QuantiFERON-TB Gold Plus compared with QuantiFERON-TB Gold In-Tube and T-SPOT.TB on active tuberculosis in Japan.

PubMed

Takasaki, Jin; Manabe, Toshie; Morino, Eriko; Muto, Yoshikazu; Hashimoto, Masao; Iikura, Motoyasu; Izumi, Shinyu; Sugiyama, Haruhito; Kudo, Koichiro

2018-03-01

The QuantiFERON-TB Gold Plus (QFT-Plus) was introduced in 2015 as a new generation of interferon-gamma release assays (IGRAs) designed to detect Mycobacterium tuberculosis infection (TB). Examination of its diagnostic accuracy is crucial before it is launched in Japan. We examined 99 patients with laboratory-confirmed active TB (patients) and 117 healthy volunteers with no risk of TB infection (controls) at a medical center in Tokyo, Japan. Blood samples were collected from both the patients and controls and tested using three types of IGRAs: the QFT-Plus, the QuantiFERON-TB Gold In-Tube (QFT-GIT), and the T-SPOT.TB (T-SPOT). The sensitivity and specificity of each IGRA were examined and compared. The sensitivity of the QFT-Plus was 98.9% (95% confidence interval [CI], 0.934-0.998) and similar to that of the QFT-GIT (97.9%; 95% CI, 0.929-0.998) and T-SPOT (96.9%; 95% CI, 0.914-0.994). The specificity of the QFT-Plus was the same as that of the QFT-GIT and T-SPOT (98.1%; 95% CI, 0.934-0.998). One patient with uncontrolled diabetes mellitus showed negative results on all three IGRAs. The QFT-Plus showed a high degree of agreement with the QFT-GIT and T-SPOT, with high sensitivity and specificity. Severe diabetes mellitus may influence the results of IGRAs. Larger studies are needed to validate the accuracy of the GFT-Plus and determine whether it can contribute as adjunctive method for the early diagnosis of active TB in Japan. Copyright © 2017 Japanese Society of Chemotherapy and The Japanese Association for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

78 FR 38695 - Submission for OMB Review; Comment Request

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-27

..., Associated Form and OMB Number: Synchronized Predeployment and Operational Tracker (SPOT) System; OMB Control... Operational Tracker (SPOT) System before deployment outside of the United States. Data collection on...

Development of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of ticks and fleas for spotted fever group rickettsia

PubMed Central

Martin, Jaclyn; Carrillo, Yisel; Talley, Justin L.; Ochoa-Corona, Francisco M.

2018-01-01

Background The importance of tick and flea-borne rickettsia infections is increasingly recognized worldwide. While increased focus has shifted in recent years to the development of point-of-care diagnostics for various vector-borne diseases in humans and animals, little research effort has been devoted to their integration into vector surveillance and control programs, particularly in resource-challenged countries. One technology which may be helpful for large scale vector surveillance initiatives is loop-mediated isothermal amplification (LAMP). The aim of this study was to develop a LAMP assay to detect spotted fever group (SFG) rickettsia DNA from field-collected ticks and fleas and compare with published end-point PCR results. Methodology/Principal findings A Spotted Fever Group rickettsia-specific loop-mediated isothermal amplification (SFGR-LAMP) assay was developed using primers based on a region of the R. rickettsii 17kDa protein gene. The sensitivity, specificity, and reproducibility of the assay were evaluated. The assay was then compared with the results of end-point PCR assays for pooled tick and flea samples obtained from field-based surveillance studies. The sensitivity of the SFGR-LAMP assay was 0.00001 ng/μl (25μl volume) which was 10 times more sensitive than the 17kDa protein gene end-point PCR used as the reference method. The assay only recognized gDNA from SFG and transitional group (TRG) rickettsia species tested but did not detect gDNA from typhus group (TG) rickettsia species or closely or distantly related bacterial species. The SFGR-LAMP assay detected the same positives from a set of pooled tick and flea samples detected by end-point PCR in addition to two pooled flea samples not detected by end-point PCR. Conclusions/significance To our knowledge, this is the first study to develop a functional LAMP assay to initially screen for SFG and TRG rickettsia pathogens in field-collected ticks and fleas. With a high sensitivity and specificity, the results indicate the potential use as a field-based surveillance tool for tick and flea-borne rickettsial pathogens in resource-challenged countries. PMID:29390021

Development of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of ticks and fleas for spotted fever group rickettsia.

PubMed

Noden, Bruce H; Martin, Jaclyn; Carrillo, Yisel; Talley, Justin L; Ochoa-Corona, Francisco M

2018-01-01

The importance of tick and flea-borne rickettsia infections is increasingly recognized worldwide. While increased focus has shifted in recent years to the development of point-of-care diagnostics for various vector-borne diseases in humans and animals, little research effort has been devoted to their integration into vector surveillance and control programs, particularly in resource-challenged countries. One technology which may be helpful for large scale vector surveillance initiatives is loop-mediated isothermal amplification (LAMP). The aim of this study was to develop a LAMP assay to detect spotted fever group (SFG) rickettsia DNA from field-collected ticks and fleas and compare with published end-point PCR results. A Spotted Fever Group rickettsia-specific loop-mediated isothermal amplification (SFGR-LAMP) assay was developed using primers based on a region of the R. rickettsii 17kDa protein gene. The sensitivity, specificity, and reproducibility of the assay were evaluated. The assay was then compared with the results of end-point PCR assays for pooled tick and flea samples obtained from field-based surveillance studies. The sensitivity of the SFGR-LAMP assay was 0.00001 ng/μl (25μl volume) which was 10 times more sensitive than the 17kDa protein gene end-point PCR used as the reference method. The assay only recognized gDNA from SFG and transitional group (TRG) rickettsia species tested but did not detect gDNA from typhus group (TG) rickettsia species or closely or distantly related bacterial species. The SFGR-LAMP assay detected the same positives from a set of pooled tick and flea samples detected by end-point PCR in addition to two pooled flea samples not detected by end-point PCR. To our knowledge, this is the first study to develop a functional LAMP assay to initially screen for SFG and TRG rickettsia pathogens in field-collected ticks and fleas. With a high sensitivity and specificity, the results indicate the potential use as a field-based surveillance tool for tick and flea-borne rickettsial pathogens in resource-challenged countries.

Use of newborn screening program blood spots for exposure assessment: declining levels of perluorinated compounds in New York State infants.

PubMed

Spliethoff, Henry M; Tao, Lin; Shaver, Shannon M; Aldous, Kenneth M; Pass, Kenneth A; Kannan, Kurunthachalam; Eadon, George A

2008-07-15

Temporal biomonitoring studies can assess changes in population exposures to contaminants, but collection of biological specimens with adequate representation and sufficient temporal resolution can be resource-intensive. Newborn Screening Programs (NSPs) collect blood as dried spots on filter paper from nearly all infants born in the United States (U.S.). In this study, we investigated the use of NSP blood spots for temporal biomonitoring by analyzing perfluorooctane sulfonate (PFOS), perfluorooctane sulfonamide (PFOSA), perfluorohexane sulfonate (PFHxS), perfluorooctanoic acid (PFOA), and perfluorononanoic acid (PFNA) in 110 New York State (NYS) NSP blood spot composite specimens collected between 1997 and 2007, representing a total of 2640 infants. All analytes were detected in > or =90% of the specimens. Concentrations of PFOS, PFOSA, PFHxS, and PFOA exhibited significant exponential declines after the year 2000, coinciding with the phase-out in PFOS production in the U.S. Calculated disappearance half-lives for PFOS, PFHxS, and PFOA (4.4, 8.2, and 4.1 years, respectively) were similar to biological half-lives reported for retired fluorochemical workers. Our results suggest sharp decreases in perinatal exposure of NYS infants to PFOS, PFOSA, PFHxS, and PFOA and demonstrate, for the first time, the utility of NSP blood spots for assessment of temporal trends in exposure.

Assessing Statistically Significant Heavy-Metal Concentrations in Abandoned Mine Areas via Hot Spot Analysis of Portable XRF Data

PubMed Central

Kim, Sung-Min; Choi, Yosoon

2017-01-01

To develop appropriate measures to prevent soil contamination in abandoned mining areas, an understanding of the spatial variation of the potentially toxic trace elements (PTEs) in the soil is necessary. For the purpose of effective soil sampling, this study uses hot spot analysis, which calculates a z-score based on the Getis-Ord Gi* statistic to identify a statistically significant hot spot sample. To constitute a statistically significant hot spot, a feature with a high value should also be surrounded by other features with high values. Using relatively cost- and time-effective portable X-ray fluorescence (PXRF) analysis, sufficient input data are acquired from the Busan abandoned mine and used for hot spot analysis. To calibrate the PXRF data, which have a relatively low accuracy, the PXRF analysis data are transformed using the inductively coupled plasma atomic emission spectrometry (ICP-AES) data. The transformed PXRF data of the Busan abandoned mine are classified into four groups according to their normalized content and z-scores: high content with a high z-score (HH), high content with a low z-score (HL), low content with a high z-score (LH), and low content with a low z-score (LL). The HL and LH cases may be due to measurement errors. Additional or complementary surveys are required for the areas surrounding these suspect samples or for significant hot spot areas. The soil sampling is conducted according to a four-phase procedure in which the hot spot analysis and proposed group classification method are employed to support the development of a sampling plan for the following phase. Overall, 30, 50, 80, and 100 samples are investigated and analyzed in phases 1–4, respectively. The method implemented in this case study may be utilized in the field for the assessment of statistically significant soil contamination and the identification of areas for which an additional survey is required. PMID:28629168

Assessing Statistically Significant Heavy-Metal Concentrations in Abandoned Mine Areas via Hot Spot Analysis of Portable XRF Data.

PubMed

Kim, Sung-Min; Choi, Yosoon

2017-06-18

To develop appropriate measures to prevent soil contamination in abandoned mining areas, an understanding of the spatial variation of the potentially toxic trace elements (PTEs) in the soil is necessary. For the purpose of effective soil sampling, this study uses hot spot analysis, which calculates a z -score based on the Getis-Ord Gi* statistic to identify a statistically significant hot spot sample. To constitute a statistically significant hot spot, a feature with a high value should also be surrounded by other features with high values. Using relatively cost- and time-effective portable X-ray fluorescence (PXRF) analysis, sufficient input data are acquired from the Busan abandoned mine and used for hot spot analysis. To calibrate the PXRF data, which have a relatively low accuracy, the PXRF analysis data are transformed using the inductively coupled plasma atomic emission spectrometry (ICP-AES) data. The transformed PXRF data of the Busan abandoned mine are classified into four groups according to their normalized content and z -scores: high content with a high z -score (HH), high content with a low z -score (HL), low content with a high z -score (LH), and low content with a low z -score (LL). The HL and LH cases may be due to measurement errors. Additional or complementary surveys are required for the areas surrounding these suspect samples or for significant hot spot areas. The soil sampling is conducted according to a four-phase procedure in which the hot spot analysis and proposed group classification method are employed to support the development of a sampling plan for the following phase. Overall, 30, 50, 80, and 100 samples are investigated and analyzed in phases 1-4, respectively. The method implemented in this case study may be utilized in the field for the assessment of statistically significant soil contamination and the identification of areas for which an additional survey is required.

Blood Substrate Collection and Handling Procedures under Pseudo-Field Conditions: Evaluation of Suitability for Inflammatory Biomarker Measurement

PubMed Central

Danese, Andrea; Shalev, Idan; Williams, Benjamin S.; Caspi, Avshalom

2015-01-01

Routine incorporation of blood-based biomarker measurements in population studies has been hampered by challenges in obtaining samples suitable for biomarker assessment outside of laboratory settings. Here, we assessed the suitability of venous blood left unprocessed for four, 24 or 48 hours post-collection at either room temperature or 4°C for quantification of two biomarkers, Interleukin-6 (IL-6) and C-Reactive Protein (CRP). Blood samples were collected in both K2EDTA tubes and a dedicated plasma-preservation tube, P100. Dried Blood Spot (DBS) samples from the same subjects were also collected in order to compare delayed-processing plasma performance against a popular alternative collection method. K2EDTA mean plasma concentrations of both IL-6 and CRP were not significantly different from concentrations in plasma processed immediately; this was observed for tubes stored up to 48 hours pre-processing at either temperature. Concentrations of IL-6 measured in P100 tubes showed significant time-dependent increases when stored at room temperature; otherwise, levels of IL-6 and CRP were similar to those processed immediately. Levels of CRP in DBS were correlated with plasma CRP levels, even when pre-processed blood was stored for up to 48 hours. These data indicate that plasma is suitable for IL-6 and CRP estimation under data-collection conditions that involve processing delays. PMID:26652682

78 FR 72099 - Agency Information Collection Activities: Submitted for Office of Management and Budget Review...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-12-02

... from leases in California or Alaska. Value is the average of the daily mean ANS spot prices published.... . . . (1) To calculate the daily mean spot price . . . (2) Use only the days . . . (3) You must adjust the... to your refinery and the NYMEX 330 2 660 price or ANS spot price is an unreasonable value. (1...

78 FR 14824 - Agency Information Collection Activities: Submitted for Office of Management and Budget Review...

Federal Register 2010, 2011, 2012, 2013, 2014

2013-03-07

... Alaska. Value is the average of the daily mean ANS spot prices published in any ONRR-approved publication during the trading month most concurrent with the production month. (1) To calculate the daily mean spot... ANS spot price is an unreasonable value. (1) . . . you may apply to the ONRR Director to establish a...

The factors that influence job satisfaction among royal Malaysian customs department employee

NASA Astrophysics Data System (ADS)

Ammar Shafi, Muhammad; Saifullah Rusiman, Mohd; Nor, Maria Elena; Khamis, Azme; Nabilah Syuhada Abdullah, Siti; Syafiq Azmi, Mohd; Sakinah Zainal Abidin, Munirah; Ali, Maselan

2018-04-01

This research aims to spot the factors that influence job satisfaction among Royal Malaysian Customs Department employees. Primary data was used in this research and it was collected from the employees who work in five different departments at Royal Malaysian Customs Department Tower Johor. Those departments were customs department, Internal Taxes, Technical Services, Management and Prevention. The research used stratified random sampling to collect the sample and Structural Equation Modelling (SEM) to measure the relationship between variables using AMOS software. About 127 employees are selected as the respondents from five departments to represent the sample. The result showed that ‘Organizational Commitment’ (p-value = 0.001) has significant and direct effect toward job satisfaction compared to the ‘Stress Condition’ (p-value = 0.819) and ‘Motivation’ factor (p-value = 0.978). It was also concluded that ‘Organizational Commitment’ was the most influential factor toward job satisfaction among Royal Malaysian Customs Department employees at Tower Custom Johor, Johor Bahru.

Water-quality and amphibian population data for Maryland, Washington, D.C., and Virginia, 2001-2004

USGS Publications Warehouse

Rice, K.C.; Jung, R.E.

2004-01-01

Data on the chemical composition of water and on amphibian populations were collected at least annually from vernal pool and stream sites in Maryland, Washington, D.C., and Virginia, from 2001 through 2004. The data were collected as part of long-term monitoring projects of the Northeast Region of the Amphibian Research and Monitoring Initiative (ARMI) of the U.S. Geological Survey. Water samples were analyzed for temperature, specific conductance, pH, dissolved-oxygen concentration, acid-neutralizing capacity, and concentrations of total Kjeldahl nitrogen and total phosphorus; in 2004, samples also were analyzed for nitrite plus nitrate concentrations and total nitrogen concentrations. Field and laboratory analytical results of water samples and quality-assurance information are presented. Amphibian population data include the presence of amphibian species and the maximum number of egg masses of wood frogs and spotted salamanders at vernal pools, and counts of amphibians made during stream transect and stream quadrat surveys.

Metal nanoparticles in DBS card materials modification

NASA Astrophysics Data System (ADS)

Metelkin, A.; Frolov, G.; Kuznetsov, D.; Kolesnikov, E.; Chuprunov, K.; Kondakov, S.; Osipov, A.; Samsonova, J.

2015-11-01

In the recent years the method of collecting and storing Dried Blood Spots (DBS) on special cellulose membrane (paper) has gained wide popularity. But possible damage of biosamples caused by microorganisms in case of their incomplete drying is a disadvantage of the method. It can be overcome by treating sample-collection membranes with colloidal solutions of metal nanoparticles, having antibacterial effect. The team studied antibacterial properties of nonwoven material samples with various coatings (alcohol sols of copper, aluminium, iron, titanium, silver and vanadium nanoparticles). Colloidal solutions of nanoparticles were obtained by means of electroerosion method with further low-temperature plasma condensation. Antibacterial activity of fiberglass and cellulose membrane samples with nanoparticle coatings was studied using B. cereus and plaque bacteria cultures. It was revealed that nanostructured coatings can suppress bacterial activity; in addition they can diffuse from the membrane surface into medium which leads to widening the areas of inhibiting testing cultures’ growth. Thus, membrane materials treatment with alcohol-sols of metal nanoparticles can be seen as promising for conferring antibacterial properties to DBS carriers.

Estimating the Potential Impacts of Large Mesopredators on Benthic Resources: Integrative Assessment of Spotted Eagle Ray Foraging Ecology in Bermuda

PubMed Central

Ajemian, Matthew J.; Powers, Sean P.; Murdoch, Thaddeus J. T.

2012-01-01

Declines of large sharks and subsequent release of elasmobranch mesopredators (smaller sharks and rays) may pose problems for marine fisheries management as some mesopredators consume exploitable shellfish species. The spotted eagle ray (Aetobatus narinari) is the most abundant inshore elasmobranch in subtropical Bermuda, but its predatory role remains unexamined despite suspected abundance increases and its hypothesized specialization for mollusks. We utilized a combination of acoustic telemetry, benthic invertebrate sampling, gut content analysis and manipulative experiments to assess the impact of spotted eagle rays on Bermudian shellfish resources. Residency and distribution of adult spotted eagle rays was monitored over two consecutive summers in Harrington Sound (HS), an enclosed inshore lagoon that has historically supported multiple recreational and commercial shellfish species. Telemetered rays exhibited variable fidelity (depending on sex) to HS, though generally selected regions that supported relatively high densities of potential mollusk prey. Gut content analysis from rays collected in HS revealed a diet of mainly bivalves and a few gastropods, with calico clam (Macrocallista maculata) representing the most important prey item. Manipulative field and mesocosm experiments with calico clams suggested that rays selected prey patches based on density, though there was no evidence of rays depleting clam patches to extirpation. Overall, spotted eagle rays had modest impacts on local shellfish populations at current population levels, suggesting a reduced role in transmitting cascading effects from apex predator loss. However, due to the strong degree of coupling between rays and multiple protected mollusks in HS, ecosystem-based management that accounts for ray predation should be adopted. PMID:22802956

Estimating the potential impacts of large mesopredators on benthic resources: integrative assessment of spotted eagle ray foraging ecology in Bermuda.

PubMed

Ajemian, Matthew J; Powers, Sean P; Murdoch, Thaddeus J T

2012-01-01

Declines of large sharks and subsequent release of elasmobranch mesopredators (smaller sharks and rays) may pose problems for marine fisheries management as some mesopredators consume exploitable shellfish species. The spotted eagle ray (Aetobatus narinari) is the most abundant inshore elasmobranch in subtropical Bermuda, but its predatory role remains unexamined despite suspected abundance increases and its hypothesized specialization for mollusks. We utilized a combination of acoustic telemetry, benthic invertebrate sampling, gut content analysis and manipulative experiments to assess the impact of spotted eagle rays on Bermudian shellfish resources. Residency and distribution of adult spotted eagle rays was monitored over two consecutive summers in Harrington Sound (HS), an enclosed inshore lagoon that has historically supported multiple recreational and commercial shellfish species. Telemetered rays exhibited variable fidelity (depending on sex) to HS, though generally selected regions that supported relatively high densities of potential mollusk prey. Gut content analysis from rays collected in HS revealed a diet of mainly bivalves and a few gastropods, with calico clam (Macrocallista maculata) representing the most important prey item. Manipulative field and mesocosm experiments with calico clams suggested that rays selected prey patches based on density, though there was no evidence of rays depleting clam patches to extirpation. Overall, spotted eagle rays had modest impacts on local shellfish populations at current population levels, suggesting a reduced role in transmitting cascading effects from apex predator loss. However, due to the strong degree of coupling between rays and multiple protected mollusks in HS, ecosystem-based management that accounts for ray predation should be adopted.

Spotting stellar activity cycles in Gaia astrometry

NASA Astrophysics Data System (ADS)

Morris, Brett M.; Agol, Eric; Davenport, James R. A.; Hawley, Suzanne L.

2018-06-01

Astrometry from Gaia will measure the positions of stellar photometric centroids to unprecedented precision. We show that the precision of Gaia astrometry is sufficient to detect starspot-induced centroid jitter for nearby stars in the Tycho-Gaia Astrometric Solution (TGAS) sample with magnetic activity similar to the young G-star KIC 7174505 or the active M4 dwarf GJ 1243, but is insufficient to measure centroid jitter for stars with Sun-like spot distributions. We simulate Gaia observations of stars with 10 year activity cycles to search for evidence of activity cycles, and find that Gaia astrometry alone likely cannot detect activity cycles for stars in the TGAS sample, even if they have spot distributions like KIC 7174505. We review the activity of the nearby low-mass stars in the TGAS sample for which we anticipate significant detections of spot-induced jitter.

MA_MISS: Mars Multispectral Imager for Subsurface Studies

NASA Astrophysics Data System (ADS)

De Sanctis, M. C.; Coradini, A.; Ammannito, E.; Boccaccini, A.; Di Iorio, T.; Battistelli, E.; Capanni, A.

2012-04-01

A Drilling system, coupled with an in situ analysis package, is installed on the ExoMars Pasteur Rover to perform in situ investigations up to 2m in the Mars soil. Ma_Miss (Mars Multispectral Imager for Subsurface Studies) is a spectrometer devoted to observe the lateral wall of the borehole generated by the Drilling system. The instrument is fully integrated with the Drill and shares its structure and electronics. For the first time in Mars exploration experiments the water/geochemical environment will be investigated as function of depth in the shallow subsurface. Samples from the subsurface of Martian soil are unaltered by weathering process, oxidation and erosion. Subsurface access can be the key to look for signs of present and past environmental conditions, associated to the possibility for life (water, volatiles and weathering process). The analysis of uncontaminated samples by means of instrumented Drill and in situ observations is the solution for unambiguous interpretation of the original environment that leading to the formation of rocks. Ma_Miss experiment is perfectly suited to perform multispectral imaging of the drilled layers. Ma_Miss is a miniaturized near-infrared imaging spectrometer in the range 0.4-2.2 µm with 20nm spectral sampling. The task of illuminating the borehole wall and collecting the diffused light from the illuminated spot on the target requires a transparent window on the Drill tool, which shall prevent the dust contamination of the optical and mechanical elements inside. Hardness of sapphire is the closest to diamond one, thus avoiding the risk of scratches on its surface. The Sapphire window is cylindrical, and bounded such as to realize a continuous auger profile. Ma_Miss Optical Head performs the double task of illuminating the borehole wall with a spot around 1 mm diameter and of collecting the scattered light coming from a 0.1 mm diameter spot of the target. The signal from the Optical Head to the spectrometer is transferred through the different elements of the Drill by means of fiber optics and an optical rotary joint implemented in the roto-translation group of the Drill. Ma_Miss Optical Head has been tested in the breadboard to capture the diffused light from the observed target and transfer the signal to a laboratory spectrometer for analysis. The Optical Head of Ma_Miss has been tested after integration in ExoMars Drill. The drilling experiment has been carried out in realistic media (tuff, red brick). The test shows good performance of Optical Head illumination capability and of the window cleanliness during the drilling. Illumination spot is focused at the nominal distance of 0.2 mm from the sapphire window. During the ExoMars Pasteur Rover mission, the Ma_Miss experiment will allow collecting valuable data of the drilled stratigraphic column, will document "in-situ" the nature of the samples that will be delivered to the Pasteur Laboratory and will be able to identify hydrated minerals, sedimentary materials and different kind of diagnostic materials of Martian subsurface.

Combined hormonal contraceptive trials: variable data collection and bleeding assessment methodologies influence study outcomes and physician perception.

PubMed

Mishell, Daniel R; Guillebaud, John; Westhoff, Carolyn; Nelson, Anita L; Kaunitz, Andrew M; Trussell, James; Davis, Ann Jeanette

2007-01-01

Initially approved for use in the United States nearly 50 years ago, oral hormonal contraceptives containing both estrogen and progestin have undergone steady improvements in safety and convenience. Concurrent with improvements in safety associated with decreasing doses of both steroids, there has been an increased incidence of unscheduled bleeding and spotting. There exist no standards regarding data collection techniques and methods, and reporting and analysis of bleeding and spotting events during combined hormonal contraceptive (CHC) trials. For the regulatory review of hormonal contraceptives, data regarding the incidence of bleeding and spotting events are not included in either of the traditional categories of efficacy and safety. Standardization of methods for collecting and analyzing data about cycle control in all clinical trials of CHCs is long overdue. Until such standards are developed and implemented, clinicians need to familiarize themselves with the techniques used in each study in order to provide correct information to their patients about the frequency of bleeding and spotting associated with different formulations and delivery systems.

Proteomic Profiling Comparing the Effects of Different Heat Treatments on Camel (Camelus dromedarius) Milk Whey Proteins

PubMed Central

Benabdelkamel, Hicham; Masood, Afshan; Alanazi, Ibrahim O.; Alzahrani, Dunia A.; Alrabiah, Deema K.; AlYahya, Sami A.; Alfadda, Assim A.

2017-01-01

Camel milk is consumed in the Middle East because of its high nutritional value. Traditional heating methods and the duration of heating affect the protein content and nutritional quality of the milk. We examined the denaturation of whey proteins in camel milk by assessing the effects of temperature on the whey protein profile at room temperature (RT), moderate heating at 63 °C, and at 98 °C, for 1 h. The qualitative and quantitative variations in the whey proteins before and after heat treatments were determined using quantitative 2D-difference in gel electrophoresis (DIGE)-mass spectrometry. Qualitative gel image analysis revealed a similar spot distribution between samples at RT and those heated at 63 °C, while the spot distribution between RT and samples heated at 98 °C differed. One hundred sixteen protein spots were determined to be significantly different (p < 0.05 and a fold change of ≥1.2) between the non-heated and heated milk samples. Eighty protein spots were decreased in common in both the heat-treated samples and an additional 25 spots were further decreased in the 98 °C sample. The proteins with decreased abundance included serum albumin, lactadherin, fibrinogen β and γ chain, lactotransferrin, active receptor type-2A, arginase-1, glutathione peroxidase-1 and, thiopurine S, etc. Eight protein spots were increased in common to both the samples when compared to RT and included α-lactalbumin, a glycosylation-dependent cell adhesion molecule. Whey proteins present in camel milk were less affected by heating at 63 °C than at 98 °C. This experimental study showed that denaturation increased significantly as the temperature increased from 63 to 98 °C. PMID:28350354

Identification of serum biomarkers in dogs naturally infected with Babesia canis canis using a proteomic approach

PubMed Central

2014-01-01

Background Canine babesiosis is a tick-borne disease that is caused by the haemoprotozoan parasites of the genus Babesia. There are limited data on serum proteomics in dogs, and none of the effect of babesiosis on the serum proteome. The aim of this study was to identify the potential serum biomarkers of babesiosis using proteomic techniques in order to increase our understanding about disease pathogenesis. Results Serum samples were collected from 25 dogs of various breeds and sex with naturally occurring babesiosis caused by B. canis canis. Blood was collected on the day of admission (day 0), and subsequently on the 1st and 6th day of treatment. Two-dimensional electrophoresis (2DE) of pooled serum samples of dogs with naturally occurring babesiosis (day 0, day 1 and day 6) and healthy dogs were run in triplicate. 2DE image analysis showed 64 differentially expressed spots with p ≤ 0.05 and 49 spots with fold change ≥2. Six selected spots were excised manually and subjected to trypsin digest prior to identification by electrospray ionisation mass spectrometry on an Amazon ion trap tandem mass spectrometry (MS/MS). Mass spectrometry data was processed using Data Analysis software and the automated Matrix Science Mascot Daemon server. Protein identifications were assigned using the Mascot search engine to interrogate protein sequences in the NCBI Genbank database. A number of differentially expressed serum proteins involved in inflammation mediated acute phase response, complement and coagulation cascades, apolipoproteins and vitamin D metabolism pathway were identified in dogs with babesiosis. Conclusions Our findings confirmed two dominant pathogenic mechanisms of babesiosis, haemolysis and acute phase response. These results may provide possible serum biomarker candidates for clinical monitoring of babesiosis and this study could serve as the basis for further proteomic investigations in canine babesiosis. PMID:24885808

Application Values of T-SPOT.TB in Clinical Rapid Diagnosis of Tuberculosis.

PubMed

Zhu, Feng; Ou, Qinfang; Zheng, Jian

2018-01-01

This paper aims to explore the application value of tuberculosis-specific enzyme-linked immunospot assay (T-SPOT.TB) in the diagnosis of tuberculosis. Fifty one patients with tuberculosis (TB) admitted to Wuxi No.5 People's Hospital, Wuxi, China from June 2015 to June 2017 were selected as the TB group, and 40 patients without tuberculosis admitted in the same period were randomly selected as the non-TB group. Patients in the two groups received T-SPOT.TB, TB antibody (TB-Ab) test and mycobacterium TB deoxyribonucleic acid (TB-DNA) test, and the results were compared. Comparisons of the sensitivity of the three methods showed that the sensitivity of T-SPOT.TB was the highest, followed by TB-DNA from sputum samples, and that of TB-Ab was the lowest. The specificity of TB-Ab was the highest, followed by T-SPOT.TB, and that of TB-DNA from sputum samples was the lowest. In the receiver operating characteristic (ROC) curve analysis, the area under curve (AUC) of T-SPOT.TB (0.896) was the highest, followed by TB-DNA from sputum samples (0.772), and that of sputum smears (0.698) was the lowest. T-SPOT.TB can quickly and accurately determine the presence of tuberculosis infection, and it is a non-invasive examination, which can further assist in the diagnosis and guide the treatment.

Dried blood spot on-card derivatization: an alternative form of sample handling to overcome the instability of thiorphan in biological matrix.

PubMed

Mess, Jean-Nicholas; Taillon, Marie-Pierre; Côté, Cynthia; Garofolo, Fabio

2012-12-01

Thiorphan, the active metabolite of racecadotril, can undergo oxidation in biological matrices such as blood and plasma. In bioanalysis, a general approach for the stabilization of such a molecule is to derivatize the thiol group to a more stable thioether, often requiring complex handling procedures at the clinical site. In this research, the concept of dried blood spot (DBS) on-card derivatization was evaluated to stabilize thiorphan. DBS cards were in-house pre-treated with 2-bromo-3'-methoxyacetophenone and left to dry prior to blood spotting. Thiorphan was shown to be effectively derivatized to thiorphan-methoxyacetophenone once applied on the in-house pre-treated cards. Thiorphan-methoxyacetophenone was extracted by soaking a 6 mm DBS punch in methanol containing the internal standard (thiorphan-methoxyacetophenone-D₅). Chromatographic separation was achieved on a Waters XBridge C₁₈ column with a gradient elution of 5 mM NH₄HCO₃ and methanol in 2.5 min and detection by ESI(+)/MS/MS. A linear (weighted 1/x²) relationship was obtained over a concentration range of 5.00-600.00 ng/mL. The assay met regulatory guidelines acceptance criteria for sensitivity, selectivity, precision and accuracy, matrix effect, recovery, dilution integrity and multiple stability evaluations. The DBS on-card derivatization has shown to be an easy and reliable alternative form of sample collection for the quantification of thiorphan. Copyright © 2012 John Wiley & Sons, Ltd.

Dried blood spot assay for the quantification of phenytoin using Liquid Chromatography-Mass Spectrometry.

PubMed

Villanelli, Fabio; Giocaliere, Elisa; Malvagia, Sabrina; Rosati, Anna; Forni, Giulia; Funghini, Silvia; Shokry, Engy; Ombrone, Daniela; Della Bona, Maria Luisa; Guerrini, Renzo; la Marca, Giancarlo

2015-02-02

Phenytoin (PHT) is one of the most commonly used anticonvulsant drugs for the treatment of epilepsy and bipolar disorders. The large amount of plasma required by conventional methods for drug quantification makes mass spectrometry combined with dried blood spot (DBS) sampling crucial for pediatric patients where therapeutic drug monitoring or pharmacokinetic studies may be difficult to realize. DBS represents a new convenient sampling support requiring minimally invasive blood drawing and providing long-term stability of samples and less expensive shipment and storage. The aim of this study was to develop a LC-MS/MS method for the quantification of PHT on DBS. This analytical method was validated and gave good linearity (r(2)=0.999) in the range of 0-100mg/l. LOQ and LOD were 1.0mg/l and 0.3mg/l, respectively. The drug extraction from paper was performed in a few minutes using a mixture composed of organic solvent for 80%. The recovery ranged from 85 to 90%; PHT in DBS showed to be stable at different storage temperatures for one month. A good correlation was also obtained between PHT plasma and DBS concentrations. This method is both precise and accurate and appears to be particularly suitable to monitor treatment with a simple and convenient sample collection procedure. Copyright © 2014 Elsevier B.V. All rights reserved.

In Situ X-ray Diffraction of Forsterite Under Shock Compression to 52 GPa: Time Resolved Observation of Changes in Crystal Structure and Phase

NASA Astrophysics Data System (ADS)

Akin, M. C.; Maddox, B.; Teruya, A.; Asimow, P. D.

2015-12-01

The Earth's mantle is composed primarily of ferromagnesian silicates, of which Forsterite (Fo) is the magnesium-rich end member of the dominant upper mantle phase, olivine. Fo is thought to undergo a chemical decomposition associated with a structural phase transition when dynamically loaded to 40-71 GPa, but previous inferences about such decomposition have been based only on pressure-density data with no direct phase identification. To obtain direct data on the phase evolution of shocked Fo, synthetic single crystal samples of Mg2SiO4 Fo were loaded to pressures of 52 GPa using a two stage light gas gun. X-ray diffraction (XRD) patterns were collected on the static and the loaded samples in situ using a single pulse Mo Kα anode to provide a 17 keV X-ray source. X-ray polycapillary optics were used to couple the source to the sample. Clear Laue spots were observed in the static images, while the dynamic images show the appearance of new spots at early times and powder-like rings at late times. The angles of the dynamically driven spots and rings overlap with each other and indicate the change in phase of forsterite under pressure through a process that begins with the formation of single crystals and ends with polycrystalline material. Efforts are underway to identify the high-pressure phases from among the library of dense magnesium silicates, and further experiments covering a larger pressure range will be completed shortly. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344.

Development and validation of UHPLC-MS/MS methods for the quantification of colistin in plasma and dried plasma spots.

PubMed

Cangemi, Giuliana; Barco, Sebastiano; Castagnola, Elio; Tripodi, Gino; Favata, Fabio; D'Avolio, Antonio

2016-09-10

Quantification of colistin in plasma samples may be very useful in optimizing therapy especially in special patients' population. Nevertheless, therapeutic drug monitoring of colistin is still limited probably for the low number of laboratories which perform this analysis and for high shipment costs. We developed and validated new UHPLC-MS/MS methods to quantify colistin in plasma and in dried plasma spots (DPS) collected on dried sample spots devices (DSSD). Colistin A, Colistin B and polimixin B, used as internal standard, were detected using multiple reaction monitoring (MRM) of the following specific transitions: 585.5→534.9; 576, 578.5→527.9; 568.9 and 602.5→100.9, 551.9, 592.8, respectively. Colistin A and B were extracted from plasma using protein precipitation and from DSSD using an extraction basic solution. Both methods were validated, and the mean intra and inter-day accuracies and precisions were in accordance with FDA and EMA guidelines. Colistin in DPS was found to be stable for at least one week at room temperature (20-25°C). A statistically significant linear correlation was found between colistin extracted from plasma and from DPS [r(2) 0.9864 (P<0.0001, 95% CI 0.9699-0.9939) for colistin A and 0.9695 (P<0.0001, 95% CI 0.9310-0.9866) for colistin B, respectively]. DPS on DSSD represents a safe and cheap strategy to store and ship at room temperature plasma samples. Thus, it is suited for pharmacokinetic studies and therapeutic drug monitoring of colistin. Copyright © 2016 Elsevier B.V. All rights reserved.

Delta spots and great flares

NASA Technical Reports Server (NTRS)

Zirin, Harold; Liggett, Margaret A.

1987-01-01

The development of delta spots and the great flares they produce are reviewed based on 18 years of observations. Delta groups are found to develop in three ways: (1) by the eruption of a single complex active region formed below the surface; (2) by the eruption of large satellite spots near a large older spot; and (3) by the collision of spots of opposite polarity from different dipoles. It is shown that the present sample of 21 delta spots never separate once they lock together, and that the driving force for the shear is spot motion. Indicators for the prediction of the occurrence of great flares are identified.

Multiple Reaction Monitoring Enables Precise Quantification of 97 Proteins in Dried Blood Spots*

PubMed Central

Chambers, Andrew G.; Percy, Andrew J.; Yang, Juncong; Borchers, Christoph H.

2015-01-01

The dried blood spot (DBS) methodology provides a minimally invasive approach to sample collection and enables room-temperature storage for most analytes. DBS samples have successfully been analyzed by liquid chromatography multiple reaction monitoring mass spectrometry (LC/MRM-MS) to quantify a large range of small molecule biomarkers and drugs; however, this strategy has only recently been explored for MS-based proteomics applications. Here we report the development of a highly multiplexed MRM assay to quantify endogenous proteins in human DBS samples. This assay uses matching stable isotope-labeled standard peptides for precise, relative quantification, and standard curves to characterize the analytical performance. A total of 169 peptides, corresponding to 97 proteins, were quantified in the final assay with an average linear dynamic range of 207-fold and an average R2 value of 0.987. The total range of this assay spanned almost 5 orders of magnitude from serum albumin (P02768) at 18.0 mg/ml down to cholinesterase (P06276) at 190 ng/ml. The average intra-assay and inter-assay precision for 6 biological samples ranged from 6.1–7.5% CV and 9.5–11.0% CV, respectively. The majority of peptide targets were stable after 154 days at storage temperatures from −20 °C to 37 °C. Furthermore, protein concentration ratios between matching DBS and whole blood samples were largely constant (<20% CV) across six biological samples. This assay represents the highest multiplexing yet achieved for targeted protein quantification in DBS samples and is suitable for biomedical research applications. PMID:26342038

Dioxin concentrations in breast milk of Vietnamese nursing mothers: a survey four decades after the herbicide spraying.

PubMed

Tai, Pham The; Nishijo, Muneko; Kido, Teruhiko; Nakagawa, Hideaki; Maruzeni, Shoko; Naganuma, Rie; Anh, Nguyen Thi Nguyet; Morikawa, Yuko; Luong, Hoang Van; Anh, Tran Hai; Hung, Nguyen Ngoc; Son, Le Ke; Tawara, Kenji; Nishijo, Hisao

2011-08-01

In an operation by United States Armed Forces during 1961 to 1971, large quantities of herbicides were sprayed in South Vietnam. These herbicides contained 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-tetraCDD), the most toxic congener of dioxins. Several decades after the herbicide spraying ceased, dioxin concentrations in the environment and human remained elevated in the sprayed areas. Breast milk samples from 520 nursing mothers residing in areas including the hot spots as well as the sprayed and unsprayed areas were collected to quantify the levels of dioxins. The total toxic equivalents of 2,3,7,8-substitued PCDDs/PCDFs in breast milk of mothers living in the hot spots, and the sprayed and unsprayed areas were 14.10 pg/g lipid, 10.89 pg/g lipid, and 4.09 pg/g lipid for primiparae and 11.48 pg/g lipid, 7.56 pg/g lipid, and 2.84 pg/g lipid for multiparae, respectively, with significant differences in the values among the three areas. In the hot spots, dioxin levels were highly correlated with the residency of mothers after adjustment for their age and parity.

Identification of the hot-spot areas for sickle cell disease using cord blood screening at a district hospital: an Indian perspective.

PubMed

Dixit, Sujata; Sahu, Pushpansu; Kar, Shantanu Kumar; Negi, Sapna

2015-10-01

Sickle cell disease (SCD), a genetic disorder often reported late, can be identified early in life, and hot-spot areas may be identified to conduct genetic epidemiology studies. This study was undertaken to estimate prevalence and to identify hot spot area for SCD in Kalahandi district, by screening cord blood of neonates delivered at the district hospital as first-hand information. Kalahandi District Hospital selected for the study is predominated by tribal population with higher prevalence of SCD as compared to other parts of Odisha. Cord blood screening of SCD was carried out on 761 newborn samples of which 13 were screened to be homozygous for SCD. Information on area of parent's residence was also collected. Madanpur Rampur area was found to be with the highest prevalence of SCD (10.52 %) and the gene distribution did not follow Hardy-Weinberg Equation indicating un-natural selection. The approach of conducting neonatal screening in a district hospital for identification of SCD is feasible and appropriate for prioritizing area for the implementation of large-scale screening and planning control measures thereof.

Biodiversity hot spot on a hot spot: novel extremophile diversity in Hawaiian fumaroles.

PubMed

Wall, Kate; Cornell, Jennifer; Bizzoco, Richard W; Kelley, Scott T

2015-01-06

Fumaroles (steam vents) are the most common, yet least understood, microbial habitat in terrestrial geothermal settings. Long believed too extreme for life, recent advances in sample collection and DNA extraction methods have found that fumarole deposits and subsurface waters harbor a considerable diversity of viable microbes. In this study, we applied culture-independent molecular methods to explore fumarole deposit microbial assemblages in 15 different fumaroles in four geographic locations on the Big Island of Hawai'i. Just over half of the vents yielded sufficient high-quality DNA for the construction of 16S ribosomal RNA gene sequence clone libraries. The bacterial clone libraries contained sequences belonging to 11 recognized bacterial divisions and seven other division-level phylogenetic groups. Archaeal sequences were less numerous, but similarly diverse. The taxonomic composition among fumarole deposits was highly heterogeneous. Phylogenetic analysis found cloned fumarole sequences were related to microbes identified from a broad array of globally distributed ecotypes, including hot springs, terrestrial soils, and industrial waste sites. Our results suggest that fumarole deposits function as an "extremophile collector" and may be a hot spot of novel extremophile biodiversity. © 2015 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Biodiversity hot spot on a hot spot: novel extremophile diversity in Hawaiian fumaroles

PubMed Central

Wall, Kate; Cornell, Jennifer; Bizzoco, Richard W; Kelley, Scott T

2015-01-01

Fumaroles (steam vents) are the most common, yet least understood, microbial habitat in terrestrial geothermal settings. Long believed too extreme for life, recent advances in sample collection and DNA extraction methods have found that fumarole deposits and subsurface waters harbor a considerable diversity of viable microbes. In this study, we applied culture-independent molecular methods to explore fumarole deposit microbial assemblages in 15 different fumaroles in four geographic locations on the Big Island of Hawai'i. Just over half of the vents yielded sufficient high-quality DNA for the construction of 16S ribosomal RNA gene sequence clone libraries. The bacterial clone libraries contained sequences belonging to 11 recognized bacterial divisions and seven other division-level phylogenetic groups. Archaeal sequences were less numerous, but similarly diverse. The taxonomic composition among fumarole deposits was highly heterogeneous. Phylogenetic analysis found cloned fumarole sequences were related to microbes identified from a broad array of globally distributed ecotypes, including hot springs, terrestrial soils, and industrial waste sites. Our results suggest that fumarole deposits function as an “extremophile collector” and may be a hot spot of novel extremophile biodiversity. PMID:25565172

Urinary DAP metabolite levels in Thai farmers and their families and exposure to pesticides from agricultural pesticide spraying.

PubMed

Hanchenlaksh, Chalalai; Povey, Andrew; O'Brien, Sarah; de Vocht, Frank

2011-08-01

We conducted a biomarker study to characterise exposure to pesticides among farmers and their families in Thailand to assess the relative importance of the dermal exposure route and to identify important factors that determine exposure levels within farmers' families. Sixteen farmers' families (eight vegetable and eight fruit farmers) participated in the study. Three morning spot urine samples were collected during a pesticide spraying week. Spot samples were grouped by individual and analysed for dialkylphosphate (DAP) metabolites and creatinine. Additional information on exposure and lifestyle was collected by means of questionnaires. Dermal exposure was assessed using a semi-quantitative observational method (DREAM). Urinary DAP levels varied 20-fold between farmers, with average (geometric mean) levels of 51.1 μg/g for vegetable and 122.2 μg/g for fruit farmers. A moderate correlation (r(s)∼0.45) was found between log(e)-transformed DREAM scores and DAP levels. Farmers' urinary metabolite levels were not correlated with those of their spouses (r(s)∼-0.30) or children (r(s)∼-0.00) collected on the same days. Detectable spouses' DAP levels were on average (geometric mean) 13.0 μg/g and those of children 7.6 μg/g. Farmers in Thailand as well as their families are exposed to pesticides in the spraying season and dermal exposure is an important route. The main route of exposure for farmers' families seems to be through transfer from the farmer to family members or contamination of the home environment, rather than family members helping or playing on the farm. Showering or washing immediately after pesticide spraying greatly reduces the potential exposure of family members to pesticide residues.

Study on the paper substrate room temperature phosphorescence of theobromine, caffeine and theophylline and analytical application

NASA Astrophysics Data System (ADS)

Chuan, Dong; Yan-Li, Wei; Shao-Min, Shuang

2003-05-01

Paper substrate room temperature phosphorescence (RTP) of theobromine (TB), caffeine (CF) and theophylline (TP) were investigated. The method is based on fast speed quantitative filter paper as substrate and KI-NaAc as heavy atom perturber. Various factors affecting their RTP were discussed in detail. Under the optimum experimental conditions, the linear dynamic range, limit of detection (LOD), and relative standard deviation (R.S.D.) were 14.41˜576.54 ng per spot, 1.14 ng per spot, 4.8% for TB, 5.44˜699.08 ng per spot, 0.78 ng per spot, 1.56% for CF, 7.21˜360.34 ng per spot, 1.80 ng per spot, 3.80% for TP, respectively. The first analytical application for the determination of these compounds was developed. The recovery of standard samples added to commercial products chocolate, tea, coffee and aminophylline is in the range 92.80-106.08%. The proposed method was successfully applied to real sample analysis without separation.

Implementing DBS methodology for the determination of Compound A in monkey blood: GLP method validation and investigation of the impact of blood spreading on performance.

PubMed

Fan, Leimin; Lee, Jacob; Hall, Jeffrey; Tolentino, Edward J; Wu, Huaiqin; El-Shourbagy, Tawakol

2011-06-01

This article describes validation work for analysis of an Abbott investigational drug (Compound A) in monkey whole blood with dried blood spots (DBS). The impact of DBS spotting volume on analyte concentration was investigated. The quantitation range was between 30.5 and 10,200 ng/ml. Accuracy and precision of quality controls, linearity of calibration curves, matrix effect, selectivity, dilution, recovery and multiple stabilities were evaluated in the validation, and all demonstrated acceptable results. Incurred sample reanalysis was performed with 57 out of 58 samples having a percentage difference (versus the mean value) less than 20%. A linear relationship between the spotting volume and the spot area was drawn. The influence of spotting volume on concentration was discussed. All validation results met good laboratory practice acceptance requirements. Radial spreading of blood on DBS cards can be a factor in DBS concentrations at smaller spotting volumes.

Quantification of sulfatides in dried blood and urine spots from metachromatic leukodystrophy patients by liquid chromatography/electrospray tandem mass spectrometry.

PubMed

Barcenas, Mariana; Suhr, Teryn R; Scott, C Ronald; Turecek, Frantisek; Gelb, Michael H

2014-06-10

Treatments are being developed for metachromatic leukodystrophy (MLD), suggesting the need for eventual newborn screening. Previous studies have shown that sulfatide molecular species are increased in the urine of MLD patients compared to samples from non-MLD individuals, but there is no data using dried blood spots (DBS), the most common sample available for newborn screening laboratories. We used ultra-high performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) to quantify sulfatides in DBS and dried urine spots from 14 MLD patients and 50 non-MLD individuals. Several sulfatide molecular species were increased in dried urine samples from all MLD samples compared to non-MLD samples. Sulfatides, especially low molecular species, were increased in DBS from MLD patients, but the sulfatide levels were relatively low. There was good separation in sulfatide levels between MLD and non-MLD samples when dried urine spots were used, but not with DBS, because DBS from non-MLD individuals have measurable levels of sulfatides. Sulfatide accumulation studies in urine, but not in DBS, emerges as the method of choice if newborn screening is to be proposed for MLD. Copyright © 2013 Elsevier B.V. All rights reserved.

Detection of Streptococcus pneumoniae and Haemophilus influenzae Type B by Real-Time PCR from Dried Blood Spot Samples among Children with Pneumonia: A Useful Approach for Developing Countries

PubMed Central

Selva, Laura; Benmessaoud, Rachid; Lanaspa, Miguel; Jroundi, Imane; Moraleda, Cinta; Acacio, Sozinho; Iñigo, Melania; Bastiani, Alien; Monsonis, Manuel; Pallares, Roman; Bassat, Quique; Muñoz-Almagro, Carmen

2013-01-01

Background 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. Methods 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. Results 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). Conclusion 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. PMID:24116190

Simultaneous Quantification of Forskolin and Iso-Forskolin in Coleus forskohlii (Wild.) Briq. and Identification of Elite Chemotype, Collected from Eastern Ghats (India).

PubMed

Shukla, Pushpendra Kumar; Misra, Ankita; Kumar, Manish; Jaichand; Singh, Kuldeep; Akhtar, Juber; Srivastava, Sharad; Agrawal, Pawan K; Singh Rawat, Ajay K

2018-01-01

Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at R f of 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300-1200 ng/spot with the regression coefficient ( R 2 ) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop. 12 Samples are collected from different locations of the eastern ghat regionsQuantification of two major marker forskolin and iso forskolinThe maximum content of both the markers was found in NBC -31, from Thakurwada, MaharashtraIdentification of elite chemotype of collected samples may be useful for commercial prospection in industries.

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

PubMed Central

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

2014-01-01

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

Use of Dried Capillary Blood Sampling for Islet Autoantibody Screening in Relatives: A Feasibility Study

PubMed Central

Rafkin, Lisa E.; Matheson, Della; Steck, Andrea K.; Yu, Liping; Henderson, Courtney; Beam, Craig A.; Boulware, David C.

2015-01-01

Abstract Background: Islet autoantibody testing provides the basis for assessment of risk of progression to type 1 diabetes. We set out to determine the feasibility and acceptability of dried capillary blood spot–based screening to identify islet autoantibody–positive relatives potentially eligible for inclusion in prevention trials. Materials and Methods: Dried blood spot (DBS) and venous samples were collected from 229 relatives participating in the TrialNet Pathway to Prevention Study. Both samples were tested for glutamic acid decarboxylase, islet antigen 2, and zinc transporter 8 autoantibodies, and venous samples were additionally tested for insulin autoantibodies and islet cell antibodies. We defined multiple autoantibody positive as two or more autoantibodies in venous serum and DBS screen positive if one or more autoantibodies were detected. Participant questionnaires compared the sample collection methods. Results: Of 44 relatives who were multiple autoantibody positive in venous samples, 42 (95.5%) were DBS screen positive, and DBS accurately detected 145 of 147 autoantibody-negative relatives (98.6%). Capillary blood sampling was perceived as more painful than venous blood draw, but 60% of participants would prefer initial screening using home fingerstick with clinic visits only required if autoantibodies were found. Conclusions: Capillary blood sampling could facilitate screening for type 1 diabetes prevention studies. PMID:26375197

Isolated populations of Ixodes lividus ticks in the Czech Republic and Belgium host genetically homogeneous Rickettsia vini.

PubMed

Nováková, Markéta; Heneberg, Petr; Heylen, Dieter J A; Medvecký, Matej; Muñoz-Leal, Sebastián; Šmajs, David; Literák, Ivan

2018-03-01

In the last two decades, the advent of molecular methods has revealed a remarkable diversity of rickettsiae (Rickettsiales: Rickettsiaceae) in invertebrates. Several species of these obligate intracellular bacteria are known to cause human infections, hence more attention has been directed towards human-biting ectoparasites. A spotted fever group Rickettsia sp. was previously detected in Ixodes lividus ticks (Ixodidae) associated with sand martins (Hirundinidae: Riparia riparia). In order to identify whether this rickettsia varies among isolated tick populations, a total of 1758 I. lividus ticks and five Ixodes ricinus ticks (Ixodidae) were collected in the Czech Republic and 148 I. lividus ticks were collected in Belgium, from nests of sand martins, European bee-eaters (Meropidae: Merops apiaster), Eurasian tree sparrows (Passeridae: Passer montanus), and from captured sand martins. We screened 165 and 78 I. lividus ticks (from the Czech Republic and Belgium, respectively) and all five I. ricinus ticks for the presence of rickettsial DNA. Only I. lividus samples were positive for Rickettsia vini, a spotted fever group rickettsia that commonly infects the tree-hole tick Ixodes arboricola (Ixodidae). Maximum likelihood analysis of the rickettsial sequences showed that the most closely related organism to R. vini corresponds to an uncharacterized rickettsia detected in Argas lagenoplastis (Argasidae), a nidicolous soft tick of the fairy martin (Hirundinidae: Petrochelidon ariel) in Australia. The observed variability of R. vini sequences from isolated tick populations was low; all 85 sequenced samples were identical to each other in five out of six partial rickettsial genes, except for the sca4 sequence (99.9% identity, 808/809 nt) that differed in I. lividus ticks from two sampling sites in the Czech Republic. Copyright © 2018 Elsevier GmbH. All rights reserved.

Investigation of magnetic nanoparticles for the rapid extraction and assay of alpha-emitting radionuclides from urine: Demonstration of a novel radiobioassay method

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

O'Hara, Matthew J.; Carter, Jennifer C.; Maclellan, Jay A.

2011-08-01

In the event of an accidental or intentional release of radionuclides into a populated area, three things must occur in a timely manner: food and drinking water supplies must be determined to be safe to eat / drink, civilians and/or military personnel must be surveyed to ensure that they do not have external contamination, and they must be screened to ensure that significant ingestion or inhalation of radionuclides has not occurred (this paper is concerned with the latter). In the event of such a disaster, the volume of radiobioassays to be performed would be tremendous. If the event released significantmore » levels of β- or α-emitting radionuclides, in vivo assays would be ineffective. Therefore, highly efficient and rapid analytical methods for radionuclide detection from submitted spot urine samples (≤ 50 mL) would be required. At present, the quantitative determination of α-emitting radionuclides from urine samples is highly labor intensive, and requires significant sample preparation and analysis time. Sorbent materials that provide effective collection and enable rapid assay could significantly streamline the radioanalytical process. We have demonstrated the use of paramagnetic nanoparticles as a novel class of extracting media for four α-emitting radionuclides of concern (Po, Ra, Am, and U) from chemically unmodified and pH 2 human urine. Herein the initial experimental sorption results are presented along with a novel method that utilizes paramagnetic nanoparticles for the extraction of radionuclides from unmodified human urine followed by the magnetic field-induced collection of the particles for subsequent α-counting-source preparation. Additionally, we construct a versatile human dose model that determines the detector count times required to estimate internal human dose at specific protective action thresholds. The model provides a means to assess a method’s detection capabilities and use fundamental health physics parameters and actual experimental data as core variables. The modeling shows that with effective sorbent materials, rapid screening for internalized α-emitters is possible from a 50 mL spot urine sample volume collected within one week of exposure/intake.« less

Microscopic analysis of "iron spot" on blue-and-white porcelain from Jingdezhen imperial kiln in early Ming dynasty (14th-15th century).

PubMed

Wang, Wenxuan; Zhu, Jian; Jiang, Jianxin; Xu, Changqing; Wu, Shurong; Guan, Li; Zhang, Zhaoxia; Wu, Menglei; Du, Jingnan

2016-11-01

"Sumali," as an imported cobalt ore from overseas, was a sort of precious and valuable pigment used for imperial kilns only, which produces characteristic "iron spot" to blue-and-white porcelain in early Ming Dynasty (A.D. 14th-15th century). Although there were some old studies on it, the morphology and formation of iron spot has not been fully investigated and understood. Therefore, five selected samples with typical spot from Jingdezhen imperial kiln in Ming Yongle periods (A.D. 1403-1424) were analyzed by various microscopic analysis including 3D digital microscope, SEM-EDS and EPMA. According to SEM images, samples can be divided into three groups: un-reflected "iron spot" without crystals, un-reflected "iron spot" with crystals and reflected "iron spot" with crystals. Furthermore, 3D micro-images revealed that "iron spots" separate out dendritic or snow-shaped crystals of iron only on and parallel to the surface of glaze for which "iron spot" show strong metallic luster. Combining with microscopic observation and microanalysis on crystallization and non-crystallization areas, it indicates that firing oxygen concentration is the ultimate causation of forming reflective iron spot which has a shallower distribution below the surface and limits crystals growing down. More details about characters of "iron spot" used "Sumali" were found and provided new clues to coloration, formation mechanism and porcelain producing technology of imperial kiln from 14th to 15th centuries of China. © 2016 Wiley Periodicals, Inc.

Design and implementation of an external quality assessment program for HIV viral load measurements using dried blood spots.

PubMed

Prach, Lisa M; Puren, Adrian; Lippman, Sheri A; Carmona, Sergio; Stephenson, Sophie; Cutler, Ewalde; Barnhart, Scott; Liegler, Teri

2015-03-01

An external quality assurance program was developed for HIV-1 RNA viral load measurements taken from dried blood spots using a reference panel and field-collected specimens. The program demonstrated that accurate and reproducible quantitation can be obtained from field-collected specimens. Residual proviral DNA may confound interpretation in virologically suppressed subjects. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Rocky Mountain spotted fever, Colombia.

PubMed

Hidalgo, Marylin; Orejuela, Leonora; Fuya, Patricia; Carrillo, Pilar; Hernandez, Jorge; Parra, Edgar; Keng, Colette; Small, Melissa; Olano, Juan P; Bouyer, Donald; Castaneda, Elizabeth; Walker, David; Valbuena, Gustavo

2007-07-01

We investigated 2 fatal cases of Rocky Mountain spotted fever that occurred in 2003 and 2004 near the same locality in Colombia where the disease was first reported in the 1930s. A retrospective serosurvey of febrile patients showed that > 21% of the serum samples had antibodies aaainst spotted fever group rickettsiae.

Rocky Mountain Spotted Fever, Colombia

PubMed Central

Hidalgo, Marylin; Orejuela, Leonora; Fuya, Patricia; Carrillo, Pilar; Hernandez, Jorge; Parra, Edgar; Keng, Colette; Small, Melissa; Olano, Juan P.; Bouyer, Donald; Castaneda, Elizabeth; Walker, David

2007-01-01

We investigated 2 fatal cases of Rocky Mountain spotted fever that occurred in 2003 and 2004 near the same locality in Colombia where the disease was first reported in the 1930s. A retrospective serosurvey of febrile patients showed that >21% of the serum samples had antibodies against spotted fever group rickettsiae. PMID:18214179

7 CFR 28.425 - Low Middling Spotted Color.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 2 2013-01-01 2013-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples in...

7 CFR 28.422 - Strict Middling Spotted Color.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 2 2014-01-01 2014-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of samples...

7 CFR 28.425 - Low Middling Spotted Color.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 7 Agriculture 2 2014-01-01 2014-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples in...

7 CFR 28.422 - Strict Middling Spotted Color.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 2 2012-01-01 2012-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of samples...

7 CFR 28.422 - Strict Middling Spotted Color.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of samples...

7 CFR 28.425 - Low Middling Spotted Color.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 7 Agriculture 2 2012-01-01 2012-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples in...

7 CFR 28.425 - Low Middling Spotted Color.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 7 Agriculture 2 2011-01-01 2011-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples in...

7 CFR 28.422 - Strict Middling Spotted Color.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 7 Agriculture 2 2013-01-01 2013-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of samples...

7 CFR 28.425 - Low Middling Spotted Color.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Low Middling Spotted Color. 28.425 Section 28.425 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Low Middling Spotted Color is color which is within the range represented by a set of samples in...

7 CFR 28.422 - Strict Middling Spotted Color.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 7 Agriculture 2 2010-01-01 2010-01-01 false Strict Middling Spotted Color. 28.422 Section 28.422 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards... Color. Strict Middling Spotted Color is color which is within the range represented by a set of samples...

Validation of spot-testing kits to determine iodine content in salt.

PubMed Central

Pandav, C. S.; Arora, N. K.; Krishnan, A.; Sankar, R.; Pandav, S.; Karmarkar, M. G.

2000-01-01

Iodine deficiency disorders are a major public health problem, and salt iodization is the most widely practised intervention for their elimination. For the intervention to be successful and sustainable, it is vital to monitor the iodine content of salt regularly. Iodometric titration, the traditional method for measuring iodine content, has problems related to accessibility and cost. The newer spot-testing kits are inexpensive, require minimal training, and provide immediate results. Using data from surveys to assess the availability of iodized salt in two states in India, Madhya Pradesh and the National Capital Territory of Delhi, we tested the suitability of such a kit in field situations. Salt samples from Delhi were collected from 30 schools, chosen using the Expanded Programme on Immunization (EPI) cluster sampling technique. A single observer made the measurement for iodine content using the kit. Salt samples from Madhya Pradesh were from 30 rural and 30 urban clusters, identified by using census data and the EPI cluster sampling technique. In each cluster, salt samples were collected from 10 randomly selected households and all retailers. The 15 investigators performing the survey estimated the iodine content of salt samples in the field using the kit. All the samples were brought to the central laboratory in Delhi, where iodine content was estimated using iodometric titration as a reference method. The agreement between the kit and titration values decreased as the number of observers increased. Although sensitivity was not much affected by the increase in the number of observers (93.3% for a single observer and 93.9% for multiple observers), specificity decreased sharply (90.4% for a single observer and 40.4% for multiple observers). Due to the low specificity and resulting high numbers of false-positives for the kit when used by multiple observers ("real-life situations"), kits were likely to consistently overestimate the availability of iodized salt. This overestimation could result in complacency. Therefore, we conclude that until a valid alternative is available, the titration method should be used for monitoring the iodine content of salt at all levels, from producer to consumer, to ensure effectiveness of the programme. PMID:10994281

Spot test analysis of microbial contents during composting of kitchen- and garden biowaste: sampling procedures, bacterial reductions, time-temperature relationships, and their relevance for EU-regulations concerning animal by-products.

PubMed

Bijlsma, P B; de Wit, D H; Duindam, J W; Elsinga, G J; Elsinga, W

2013-01-30

This study was aimed to collect data and develop methodologies to determine if and how Dutch biowaste composting plants can meet the microbiological requirements set out in EU-Regulations (EC) 1774/2002 and (EC) 1069/2009, and to provide the European Food and Safety Authority (EFSA) with data and analysis for evaluation of these regulations. We examined twenty plant locations and four types of composting technologies, all with forced aeration and without an anaerobic digestion phase. Raw biowaste, material after sanitation and compost were sampled by spot test analysis according to a standard protocol, and according to an additional protocol with enhanced hygienic precautions. Samples were analyzed for Escherichia coli, Enterococcaceae and Salmonella content. The latter protocol resulted in improved bacterial reductions after sanitation, whereas in compost Enterococcus levels but not E. coli levels increased substantially with both protocols, due to more thermo-resistant regrowth. Salmonella presence in compost coincided with low temperatures and increased levels of E. coli and Enterococcus, absence of Salmonella was associated with absence of E. coli (74%), but not with absence of Enterococcus (17%). In compost, E. coli and Salmonella showed a comparable time-temperature inactivation pattern. A pilot study with co-composting of biowaste and poultry manure indicated a similar inactivation pattern for ESBL-containing bacteria. We conclude that the abundance of Enterococcus in compost is caused by regrowth and not by (re)contamination, and that E. coli is a more reliable indicator species for the absence/presence of Salmonella in compost. Compliance with current EU-regulations concerning biowaste composting can be shown by spot test analysis at all examined plants, provided that adequate hygienic precautions are taken during sampling. Copyright © 2012 Elsevier Ltd. All rights reserved.

Stability and Application of Reactive Nitrogen and Oxygen Species-Induced Hemoglobin Modifications in Dry Blood Spots As Analyzed by Liquid Chromatography Tandem Mass Spectrometry.

PubMed

Chen, Hauh-Jyun Candy; Fan, Chih-Huang; Yang, Ya-Fen

2016-12-19

Dried blood spot (DBS) is an emerging microsampling technique for the bioanalysis of small molecules, including fatty acids, metabolites, drugs, and toxicants. DBS offers many advantages as a sample format including easy sample collection and cheap sample shipment. Hemoglobin adducts have been recognized as a suitable biomarker for monitoring chemical exposure. We previously reported that certain modified peptides in hemoglobin derived from reactive chlorine, nitrogen, and oxygen species are associated with factors including smoking, diabetes mellitus, and aging. However, the stability of these oxidation-induced modifications of hemoglobin remains unknown and whether they can be formed artifactually during storage of DBS. To answer these questions, globin extracted from the DBS cards was analyzed, and the stability of the modifications was evaluated. After storage of the DBS cards at 4 °C or room temperature up to 7 weeks, we isolated globin from a quarter of the spot every week. The extents of 11 sites and types of post-translational modifications (PTMs), including nitration and nitrosylation of tyrosine and oxidation of cysteine and methionine residues, in human hemoglobin were measured in the trypsin digest by nanoflow liquid chromatography-nanospray ionization tandem mass spectrometry (nanoLC-NSI/MS/MS) using selected reaction monitoring. The extents of all these PTMs are stable within 14 days when stored on DBS at room temperature and at 4 °C, while those from direct extraction of fresh blood are stable for at least 8 weeks when stored as an aqueous solution at -20 °C. Extraction of globin from a DBS card is of particular importance for hemolytic blood samples. To our knowledge, this is the first report on the stability of oxidative modifications of hemoglobin on DBSs, which are stable for 14 days under ambient conditions (room temperature, in air). Therefore, it is feasible and convenient to analyze these hemoglobin modifications from DBSs in studies involving large populations.

Serologic evidence of the exposure of small mammals to spotted-fever Rickettsia and Rickettsia bellii in Minas Gerais, Brazil.

PubMed

Coelho, Marcella Gonçalves; Ramos, Vanessa do Nascimento; Limongi, Jean Ezequiel; de Lemos, Elba Regina Sampaio; Guterres, Alexandro; da Costa Neto, Sócrates Fraga; Rozental, Tatiana; Bonvicino, Cibele Rodrigues; D'Andrea, Paulo Sérgio; Moraes-Filho, Jonas; Labruna, Marcelo Bahia; Szabó, Matias Pablo Juan

2016-03-31

Sources of pathogenic Rickettsia in wildlife are largely unknown in Brazil. In this work, potential tick vectors and seroreactivity of small mammals against four spotted-fever group Rickettsia (R. rickettsii, R. parkeri, R. amblyommii and R. rhipicephali) and Rickettsia bellii from peri-urban areas of Uberlândia, a major town in Brazil, are described for the first time. Small mammals were captured and blood samples collected. Ticks were collected from the surface of the host and the environment and posteriorly identified. Reactivity of small mammal sera to Rickettsia was tested by indirect immunofluorescence assay (IFA) using crude antigens from five Brazilian Rickettsia isolates. Information was obtained from 416 small mammals (48 Marsupialia and 368 Rodentia). Forty-eight animals were parasitized and two tick species, Ixodes loricatus and Amblyomma dubitatum, were found on several host species, with a few tick-host relationships described for the first time. From the 416 tested sera, 70 reacted to at least one Rickettsia antigen (prevalence of 16.8%) and from these, 19 (27.1%) reacted to two or more antigens. Seroprevalence was higher for marsupials (39.6%) than for rodents (13.8%). Marsupial and Rhipidomys spp. sera reacted mainly (highest seroprevalence and titers) to R. bellii, and that of Necromys lasiurus mainly to R. rickettsii. Although the serologic assays poorly discriminate between closely related spotted-fever group Rickettsia, the observed small mammal seroreactivity suggests the circulation of Rickettsia in the peri-urban area of Uberlândia, albeit at low levels.

Development and validation of a new method to simultaneously quantify triazoles in plasma spotted on dry sample spot devices and analysed by HPLC-MS.

PubMed

Baietto, Lorena; D'Avolio, Antonio; Marra, Cristina; Simiele, Marco; Cusato, Jessica; Pace, Simone; Ariaudo, Alessandra; De Rosa, Francesco Giuseppe; Di Perri, Giovanni

2012-11-01

Therapeutic drug monitoring (TDM) of triazoles is widely used in clinical practice to optimize therapy. TDM is limited by technical problems and cost considerations, such as sample storage and dry-ice shipping. We aimed to develop and validate a new method to analyse itraconazole, posaconazole and voriconazole in plasma spotted on dry sample spot devices (DSSDs) and to quantify them by an HPLC system. Extraction from DSSDs was done using n-hexane/ethyl acetate and ammonia solution. Samples were analysed using HPLC with mass spectrometry (HPLC-MS). Accuracy and precision were assayed by inter- and intra-day validation. The stability of triazoles in plasma spotted on DSSDs was investigated at room temperature for 1 month. The method was compared with a validated standard HPLC method for quantification of triazoles in human plasma. Mean inter- and intra-day accuracy and precision were <15% for all compounds. Triazoles were stable for 2 weeks at room temperature. The method was linear (r(2) > 0.999) in the range 0.031-8 mg/L for itraconazole and posaconazole, and 0.058-15 mg/L for voriconazole. High sensitivity was observed; limits of detection were 0.008, 0.004 and 0.007 mg/L for itraconazole, posaconazole and voriconazole, respectively. A high degree of correlation (r(2) > 0.94) was obtained between the DSSD method and the standard method of analysis. The method that we developed and validated to quantify triazoles in human plasma spotted on DSSDs is accurate and precise. It overcomes problems related to plasma sample storage and shipment, allowing TDM to be performed in a cheaper and safer manner.

Antibodies for Rickettsia spp. in patients with negative serology for dengue virus, leptospirosis, and meningococcal disease in municipalities of São Paulo State, Brazil.

PubMed

Prata, Juliana Anacleto Cabral; Souza, Celso Eduardo de; Angerami, Rodrigo Nogueira; Barbosa, Taíse Marongio Cotrim de Moraes; Santos, Fabiana Cristina Pereira Dos; Colombo, Silvia; Guercio, Vânia Martins Fontes Del; Donalísio, Maria Rita

2016-01-01

Brazilian spotted fever is an infectious disease with a high mortality rate if not treated early. Differential diagnosis is difficult, as the first clinical signs are non-specific and can be confused with other diseases. The aim of the study was to investigate evidence of infection with Rickettsia rickettsii and Rickettsia parkeri in negative sera samples, collected in 2014, from patients with suspected leptospirosis, dengue fever, and meningococcal disease in Atibaia and Bragança Paulista municipalities of the State of São Paulo. The samples stored at the Institute Adolfo Lutz in Campinas were tested using an indirect immunofluorescence assay (IFA) with IgG and IgM against R. rickettsii and R. parkeri. Real-time polymerase chain reaction (PCR) testing was performed for the sera samples of patients who died (n = 3), those with initial suspicion of meningococcal disease (n = 6), and those with positive IFA results. Of 258 samples from Bragança Paulista, 4 (1.6%) were positive, with IgG titers of 1:64 and 1:128 against R. rickettsii and R. parkeri, respectively. Of 155 samples from Atibaia, 2 (1.3%) were positive, with IgG titers of 1:64 and 1:128 against R. rickettsii and R. parkeri, respectively. No sample showed positive PCR results. This serological investigation suggests there is evidence of exposure to Rickettsia spp. in residents of areas that have environmental conditions favorable to the spread of bacteria, in which Brazilian spotted fever incidence was not previously confirmed.

Quantitation of 25-hydroxyvitamin D in dried blood spots by 2D LC-MS/MS without derivatization and correlation with serum in adult and pediatric studies.

PubMed

Jensen, Berit P; Saraf, Rajneeta; Ma, Jing; Berry, Sarah; Grant, Cameron C; Camargo, Carlos A; Sies, Christiaan W

2018-06-01

Demand for measurement of 25-hydroxyvitamin D (25OHD) is growing and dried blood spot (DBS) sampling is attractive as samples are easier to collect, transport and store. A 2D LC-MS/MS assay without derivatization was developed. DBS punches (3.2 mm) were ultrasonicated with d 6 -25OHD 3 in 70% methanol followed by hexane extraction, dry-down and reconstitution. The assay was validated and applied to two studies comparing whole blood adult DBS with serum samples (n = 40) and neonatal whole blood DBS with cord serum samples (n = 80). The assay was validated in whole blood DBS over the range 13-106 nmol/L 25OHD 3 and 11-91 nmol/L 25OHD 2 with a limit of detection of 3 nmol/L. Intra- and inter-day imprecision was <13% CV and bias <12%. The assay had high recovery and minimal matrix effects. Triplicate DBS study samples had a mean CV of ≤13% for 25OHD 3. No 25OHD 2 was detected. DBS calculated serum 25OHD 3 concentrations correlated strongly with serum concentrations in the adult DBS/serum study (r = 0.94) and moderately in the neonatal DBS/cord serum study (r = 0.69). Direct quantitation of 25OHD in DBS by 2D LC-MS/MS without derivatization was found to be an alternative to serum quantitation applicable to clinical research studies on adult DBS samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Continuous motion scan ptychography: Characterization for increased speed in coherent x-ray imaging

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

Deng, Junjing; Nashed, Youssef S. G.; Chen, Si

Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object’s complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous “fly-scan” mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

Combining near infrared spectra of feces and geostatistics to generate forage nutritional quality maps across landscapes

NASA Astrophysics Data System (ADS)

Jean, Pierre-Olivier; Bradley, Robert; Tremblay, Jean-Pierre

2015-04-01

An important asset for the management of wild ungulates is the ability to recognize the spatial distribution of forage quality across heterogeneous landscapes. To do so typically requires knowledge of which plant species are eaten, in what abundance they are eaten, and what their nutritional quality might be. Acquiring such data may be, however, difficult and time consuming. Here, we are proposing a rapid and cost-effective forage quality monitoring tool that combines near infrared (NIR) spectra of fecal samples and easily obtained data on plant community composition. Our approach rests on the premise that NIR spectra of fecal samples collected within low population density exclosures reflect the optimal forage quality of a given landscape. Forage quality can thus be based on the Mahalanobis distance of fecal spectral scans across the landscape relative to fecal spectral scans inside exclosures (referred to as DISTEX). The Gi* spatial autocorrelation statistic can then be applied among neighbouring DISTEX values to detect and map 'hot-spots' and 'cold-spots' of nutritional quality over the landscape. We tested our approach in a heterogeneous boreal landscape on Anticosti Island (Qu

Continuous motion scan ptychography: Characterization for increased speed in coherent x-ray imaging

DOE PAGES

Deng, Junjing; Nashed, Youssef S. G.; Chen, Si; ...

2015-02-23

Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object’s complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous “fly-scan” mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

Continuous motion scan ptychography: characterization for increased speed in coherent x-ray imaging

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

Deng, Junjing; Nashed, Youssef S. G.; Chen, Si

2015-01-01

Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object's complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous "fly-scan" mode for ptychographic data collection in whichmore » the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.« less

Continuous motion scan ptychography: characterization for increased speed in coherent x-ray imaging.

PubMed

Deng, Junjing; Nashed, Youssef S G; Chen, Si; Phillips, Nicholas W; Peterka, Tom; Ross, Rob; Vogt, Stefan; Jacobsen, Chris; Vine, David J

2015-03-09

Ptychography is a coherent diffraction imaging (CDI) method for extended objects in which diffraction patterns are acquired sequentially from overlapping coherent illumination spots. The object's complex transmission function can be reconstructed from those diffraction patterns at a spatial resolution limited only by the scattering strength of the object and the detector geometry. Most experiments to date have positioned the illumination spots on the sample using a move-settle-measure sequence in which the move and settle steps can take longer to complete than the measure step. We describe here the use of a continuous "fly-scan" mode for ptychographic data collection in which the sample is moved continuously, so that the experiment resembles one of integrating the diffraction patterns from multiple probe positions. This allows one to use multiple probe mode reconstruction methods to obtain an image of the object and also of the illumination function. We show in simulations, and in x-ray imaging experiments, some of the characteristics of fly-scan ptychography, including a factor of 25 reduction in the data acquisition time. This approach will become increasingly important as brighter x-ray sources are developed, such as diffraction limited storage rings.

Fabrication of Biomimetic Fog-Collecting Superhydrophilic-Superhydrophobic Surface Micropatterns Using Femtosecond Lasers.

PubMed

Kostal, Elisabeth; Stroj, Sandra; Kasemann, Stephan; Matylitsky, Victor; Domke, Matthias

2018-03-06

The exciting functionalities of natural superhydrophilic and superhydrophobic surfaces served as inspiration for a variety of biomimetic designs. In particular, the combination of both extreme wetting states to micropatterns opens up interesting applications, as the example of the fog-collecting Namib Desert beetle shows. In this paper, the beetle's elytra were mimicked by a novel three-step fabrication method to increase the fog-collection efficiency of glasses. In the first step, a double-hierarchical surface structure was generated on Pyrex wafers using femtosecond laser structuring, which amplified the intrinsic wetting property of the surface and made it superhydrophilic (water contact angle < 10°). In the second step, a Teflon-like polymer (CF 2 ) n was deposited by a plasma process that turned the laser-structured surface superhydrophobic (water contact angle > 150°). In the last step, the Teflon-like coating was selectively removed by fs-laser ablation to uncover superhydrophilic spots below the superhydrophobic surface, following the example of the Namib Desert beetle's fog-collecting elytra. To investigate the influence on the fog-collection behavior, (super)hydrophilic, (super)hydrophobic, and low and high contrast wetting patterns were fabricated on glass wafers using selected combinations of these three processing steps and were exposed to fog in an artificial nebulizer setup. This experiment revealed that high-contrast wetting patterns collected the highest amount of fog and enhanced the fog-collection efficiency by nearly 60% compared to pristine Pyrex glass. The comparison of the fog-collection behavior of the six samples showed that the superior fog-collection efficiency of surface patterns with extreme wetting contrast is due to the combination of water attraction and water repellency: the superhydrophilic spots act as drop accumulation areas, whereas the surrounding superhydrophobic areas allow a fast water transportation caused by gravity. The presented method enables a fast and flexible surface functionalization of a broad range of materials including transparent substrates, which offers exciting possibilities for the design of biomedical and microfluidic devices.

Temporal variation of aqueous-extractable Ca, Mg and K in acidified forest mountainous soils under different vegetation cover

NASA Astrophysics Data System (ADS)

Tejnecky, V.; Bradová, M.; Boruvka, L.; Vasat, R.; Nemecek, K.; Ash, C.; Sebek, O.; Rejzek, J.; Drabek, O.

2012-12-01

Acidification of forest soils is a natural degradation process which can be significantly enhanced by anthropogenic activities. Inputs of basic cations (BC - Ca, Mg and K) via precipitation, litter and soil organic matter decomposition and also via inter-soil weathering may partially mitigate the consequences of this degradation process. The aim of this study is to assess the temporal variation of aqueous-extractable Ca, Mg and K in acidified forest mountainous soils under different vegetation cover. The Jizera Mountains region (Czech Republic, northern Bohemia) was chosen as a representative soil mountainous ecosystem strongly affected by acidification. Soil and precipitation samples were collected at monthly basis from April till October/ November during the years 2009-2011. Study spots were delimited under two contrasting vegetation covers - beech and spruce monoculture. Prevailing soil types were classified as Alumic Cambisols under beech and Entic Podzols under spruce stands (according to FAO classification). Soil samples were collected from surface fermentation (F) and humified (H) organic horizons and subsurface B horizons (cambic or spodic). The collected soil samples were analyzed immediately under laboratory condition in a "fresh" state. Unsieved fresh samples were extracted by deionised water. The content of main elements (Ca, Mg, K, Al and Fe) was determined by ICP-OES. The content of major anions (SO42-, NO3-, Cl- and F-) was determined by ion-exchange chromatography (IC). Content of major anions and main elements were determined in the precipitation samples (throughfall, stemflow and bulk) as well. Besides computing the basic statistical parameters (mean, median, variance, maximum, minimum, etc.) we also employed other statistical methods such as T-test and ANOVA to assess the differences between beech and spruce vegetation spots. To carry out the temporal variability in the data we used the time series analysis and short-term forecasting by Holt-Winters exponential smoothing and ARIMA models. Our results clearly exhibit differences in the horizontal and spatial distribution of BC. The influences of the study spot, i.e. the influence of stand factors e.g. vegetation covers on BC distribution are well pronounced. The highest amounts of aqueous extractable BC were identified in the F and H organic horizons. The contents of Ca and Mg were significantly higher under beech cover than under spruce cover. The influence of seasonality on BC content and distribution was the strongest in the upper organic horizons. The annual changes are less pronounced in inner mineral B horizons. We have observed a significant influence of the snow melting period - after this event the content of BC was the lowest. In contrast, the BC content rises during the summer period - the time of high biological activity and accelerated organic matter decomposition. This period is again followed by a BC content decrease during the fall period - the time of gradually decreasing biological activity and high precipitation. Generally, we can conclude that the seasonal variations are higher than annual and spatial for both sites.

Optimal sampling for radiotelemetry studies of spotted owl habitat and home range.

Treesearch

Andrew B. Carey; Scott P. Horton; Janice A. Reid

1989-01-01

Radiotelemetry studies of spotted owl (Strix occidentalis) ranges and habitat-use must be designed efficiently to estimate parameters needed for a sample of individuals sufficient to describe the population. Independent data are required by analytical methods and provide the greatest return of information per effort. We examined time series of...

RandomSpot: A web-based tool for systematic random sampling of virtual slides.

PubMed

Wright, Alexander I; Grabsch, Heike I; Treanor, Darren E

2015-01-01

This paper describes work presented at the Nordic Symposium on Digital Pathology 2014, Linköping, Sweden. Systematic random sampling (SRS) is a stereological tool, which provides a framework to quickly build an accurate estimation of the distribution of objects or classes within an image, whilst minimizing the number of observations required. RandomSpot is a web-based tool for SRS in stereology, which systematically places equidistant points within a given region of interest on a virtual slide. Each point can then be visually inspected by a pathologist in order to generate an unbiased sample of the distribution of classes within the tissue. Further measurements can then be derived from the distribution, such as the ratio of tumor to stroma. RandomSpot replicates the fundamental principle of traditional light microscope grid-shaped graticules, with the added benefits associated with virtual slides, such as facilitated collaboration and automated navigation between points. Once the sample points have been added to the region(s) of interest, users can download the annotations and view them locally using their virtual slide viewing software. Since its introduction, RandomSpot has been used extensively for international collaborative projects, clinical trials and independent research projects. So far, the system has been used to generate over 21,000 sample sets, and has been used to generate data for use in multiple publications, identifying significant new prognostic markers in colorectal, upper gastro-intestinal and breast cancer. Data generated using RandomSpot also has significant value for training image analysis algorithms using sample point coordinates and pathologist classifications.

Differentiating Medicated Patients Suffering from Major Depressive Disorder from Healthy Controls by Spot Urine Measurement of Monoamines and Steroid Hormones

PubMed Central

Wijaya, Chandra S.; Lee, Jovia J. Z.; Husain, Syeda F.; Ho, Cyrus S. H.; McIntyre, Roger S.; Tam, Wilson W.

2018-01-01

Introduction: Major Depressive Disorder (MDD) is a common psychiatric disorder. Currently, there is no objective, cost-effective and non-invasive method to measure biological markers related to the pathogenesis of MDD. Previous studies primarily focused on urinary metabolite markers which are not proximal to the pathogenesis of MDD. Herein, we compare urinary monoamines, steroid hormones and the derived ratios amongst MDD when compared to healthy controls. Methods: Morning urine samples of medicated patients suffering from MDD (n = 47) and healthy controls (n = 41) were collected. Enzyme-linked immunosorbent assay (ELISA) was performed to measure five biomarkers: cortisol, dopamine, noradrenaline, serotonin and sulphate derivative of dehydroepiandrosterone (DHEAS). The mean urinary levels and derived ratios of monoamines and steroid hormones were compared between patients and controls to identify potential biomarkers. The receiver operative characteristic curve (ROC) analysis was conducted to evaluate the diagnostic performance of potential biomarkers. Results: Medicated patients with MDD showed significantly higher spot urine ratio of DHEAS/serotonin (1.56 vs. 1.19, p = 0.004) and lower ratio of serotonin/dopamine (599.71 vs. 888.60, p = 0.008) than healthy controls. A spot urine serotonin/dopamine ratio cut-off of >667.38 had a sensitivity of 73.2% and specificity of 51.1%. Conclusions: Our results suggest that spot urine serotonin/dopamine ratio can be used as an objective diagnostic method for adults with MDD. PMID:29701669

Direct tandem mass spectrometry for the simultaneous assay of opioids, cocaine and metabolites in dried urine spots.

PubMed

Otero-Fernández, Mara; Cocho, José Ángel; Tabernero, María Jesús; Bermejo, Ana María; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

2013-06-19

A micro-analytical method based on spotting urine samples (20μL) onto blood/urine spot collection cards followed by air-drying and extraction (dried urine spot, DUS) was developed and validated for the screening/confirmation assay of morphine, 6-methylacetylmorphine (6-MAM), codeine, cocaine and benzoylecgonine (BZE). Acetonitrile (3 mL) was found to be a useful solvent for target extraction from DUSs under an orbital-horizontal stirring at 180 rpm for 10 min. Determinations were performed by direct electrospray ionization tandem mass spectrometry (ESI-MS/MS) under positive electrospray ionization conditions, and by using multiple reaction monitoring (MRM) with one precursor ion/product ion transition for the identification and quantification (deuterated analogs of each target as internal standards) of each analyte. The limits of detection of the method were 0.26, 0.94, 1.5, 1.1, and 2.0 ng mL(-1), for cocaine, BZE, codeine, morphine and 6-MAM, respectively; whereas, relative standard deviations of intra- and inter-day precision were lower than 8 and 11%, respectively, and intra- and inter-day analytical recoveries ranged from 94±4 to 105±3%. The small volume of urine required (20 μL), combined with the simplicity of the analytical technique makes it a useful procedure for screening/quantifying drugs of abuse. The method was successfully applied to the analysis of urine from polydrug abusers. Copyright © 2013 Elsevier B.V. All rights reserved.

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

USDA-ARS?s Scientific Manuscript database

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

Estimation of daily protein intake based on spot urine urea nitrogen concentration in chronic kidney disease patients.

PubMed

Kanno, Hiroko; Kanda, Eiichiro; Sato, Asako; Sakamoto, Kaori; Kanno, Yoshihiko

2016-04-01

Determination of daily protein intake in the management of chronic kidney disease (CKD) requires precision. Inaccuracies in recording dietary intake occur, and estimation from total urea excretion presents hurdles owing to the difficulty of collecting whole urine for 24 h. Spot urine has been used for measuring daily sodium intake and urinary protein excretion. In this cross-sectional study, we investigated whether urea nitrogen (UN) concentration in spot urine can be used to predict daily protein intake instead of the 24-h urine collection in 193 Japanese CKD patients (Stages G1-G5). After patient randomization into 2 datasets for the development and validation of models, bootstrapping was used to develop protein intake estimation models. The parameters for the candidate multivariate regression models were male gender, age, body mass index (BMI), diabetes mellitus, dyslipidemia, proteinuria, estimated glomerular filtration rate, serum albumin level, spot urinary UN and creatinine level, and spot urinary UN/creatinine levels. The final model contained BMI and spot urinary UN level. The final model was selected because of the higher correlation between the predicted and measured protein intakes r = 0.558 (95 % confidence interval 0.400, 0.683), and the smaller distribution of the difference between the measured and predicted protein intakes than those of the other models. The results suggest that UN concentration in spot urine may be used to estimate daily protein intake and that a prediction formula would be useful for nutritional control in CKD patients.

Understanding biological mechanisms underlying adverse birth outcomes in developing countries: protocol for a prospective cohort (AMANHI bio–banking) study

PubMed Central

Baqui, Abdullah H; Khanam, Rasheda; Rahman, Mohammad Sayedur; Ahmed, Aziz; Rahman, Hasna Hena; Moin, Mamun Ibne; Ahmed, Salahuddin; Jehan, Fyezah; Nisar, Imran; Hussain, Atiya; Ilyas, Muhammad; Hotwani, Aneeta; Sajid, Muhammad; Qureshi, Shahida; Zaidi, Anita; Sazawal, Sunil; Ali, Said M; Deb, Saikat; Juma, Mohammed Hamad; Dhingra, Usha; Dutta, Arup; Ame, Shaali Makame; Hayward, Caroline; Rudan, Igor; Zangenberg, Mike; Russell, Donna; Yoshida, Sachiyo; Polašek, Ozren; Manu, Alexander; Bahl, Rajiv

2017-01-01

Objectives The AMANHI study aims to seek for biomarkers as predictors of important pregnancy–related outcomes, and establish a biobank in developing countries for future research as new methods and technologies become available. Methods AMANHI is using harmonised protocols to enrol 3000 women in early pregnancies (8–19 weeks of gestation) for population–based follow–up in pregnancy up to 42 days postpartum in Bangladesh, Pakistan and Tanzania, with collection taking place between August 2014 and June 2016. Urine pregnancy tests will be used to confirm reported or suspected pregnancies for screening ultrasound by trained sonographers to accurately date the pregnancy. Trained study field workers will collect very detailed phenotypic and epidemiological data from the pregnant woman and her family at scheduled home visits during pregnancy (enrolment, 24–28 weeks, 32–36 weeks & 38+ weeks) and postpartum (days 0–6 or 42–60). Trained phlebotomists will collect maternal and umbilical blood samples, centrifuge and obtain aliquots of serum, plasma and the buffy coat for storage. They will also measure HbA1C and collect a dried spot sample of whole blood. Maternal urine samples will also be collected and stored, alongside placenta, umbilical cord tissue and membrane samples, which will both be frozen and prepared for histology examination. Maternal and newborn stool (for microbiota) as well as paternal and newborn saliva samples (for DNA extraction) will also be collected. All samples will be stored at –80°C in the biobank in each of the three sites. These samples will be linked to numerous epidemiological and phenotypic data with unique study identification numbers. Importance of the study AMANHI biobank proves that biobanking is feasible to implement in LMICs, but recognises that biobank creation is only the first step in addressing current global challenges. PMID:29163938

Evaluation of maternal serum alpha-foetoprotein assay using dry blood spot samples.

PubMed

González, C; Guerrero, J M; Elorza, F L; Molinero, P; Goberna, R

1988-02-01

The quantification of alpha-foetoprotein in dry blood spots from pregnant women was evaluated, using a conventional radioimmunoassay (RIA) with a monospecific antibody. The stability of alpha-foetoprotein in dry blood spots on filter paper was evaluated with respect to mailing, distances travelled, and the existence of high summer temperatures in our region. The results obtained show that the blood alpha-foetoprotein is stable on dry filter spots sent by mail and is stable for up to four weeks at 4, 25 and 37 degrees C. The analytical method used has a minimal detectable concentration of 10 +/- 1.9 international kilo-units/l. Both inter- and intra-assay variabilities are smaller than 10% and this method can provide results comparable with those of conventional serum assays. Results from dry blood spots and serum samples (the latter analysed by both RIA and two-site enzyme immunoassay) exhibited a good correlation (r = 0.98 and r = 0.97, p less than 0.001). The design of the assay and the nature of the samples make this method suitable for a screening programmes for the antenatal detection of open neural tube defects.

Energy dispersive X-ray fluorescence spectrometry for the direct multi-element analysis of dried blood spots

NASA Astrophysics Data System (ADS)

Marguí, E.; Queralt, I.; García-Ruiz, E.; García-González, E.; Rello, L.; Resano, M.

2018-01-01

Home-based collection protocols for clinical specimens are actively pursued as a means of improving life quality of patients. In this sense, dried blood spots (DBS) are proposed as a non-invasive and even self-administered alternative to sampling whole venous blood. This contribution explores the potential of energy dispersive X-ray fluorescence spectrometry for the simultaneous and direct determination of some major (S, Cl, K, Na), minor (P, Fe) and trace (Ca, Cu, Zn) elements in blood, after its deposition onto clinical filter papers, thus giving rise to DBS. For quantification purposes the best strategy was to use matrix-matched blood samples of known analyte concentrations. The accuracy and precision of the method were evaluated by analysis of a blood reference material (Seronorm™ trace elements whole blood L3). Quantitative results were obtained for the determination of P, S, Cl, K and Fe, and limits of detection for these elements were adequate, taking into account their typical concentrations in real blood samples. Determination of Na, Ca, Cu and Zn was hampered by the occurrence of high sample support (Na, Ca) and instrumental blanks (Cu, Zn). Therefore, the quantitative determination of these elements at the levels expected in blood samples was not feasible. The methodology developed was applied to the analysis of several blood samples and the results obtained were compared with those reported by standard techniques. Overall, the performance of the method developed is promising and it could be used to determine the aforementioned elements in blood samples in a simple, fast and economic way. Furthermore, its non-destructive nature enables further analyses by means of complementary techniques to be carried out.

The use of mass spectrometry to analyze dried blood spots.

PubMed

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

2016-01-01

Dried blood spots (DBS) typically consist in the deposition of small volumes of capillary blood onto dedicated paper cards. Comparatively to whole blood or plasma samples, their benefits rely in the fact that sample collection is easier and that logistic aspects related to sample storage and shipment can be relatively limited, respectively, without the need of a refrigerator or dry ice. Originally, this approach has been developed in the sixties to support the analysis of phenylalanine for the detection of phenylketonuria in newborns using bacterial inhibition test. In the nineties tandem mass spectrometry was established as the detection technique for phenylalanine and tyrosine. DBS became rapidly recognized for their clinical value: they were widely implemented in pediatric settings with mass spectrometric detection, and were closely associated to the debut of newborn screening (NBS) programs, as a part of public health policies. Since then, sample collection on paper cards has been explored with various analytical techniques in other areas more or less successfully regarding large-scale applications. Moreover, in the last 5 years a regain of interest for DBS was observed and originated from the bioanalytical community to support drug development (e.g., PK studies) or therapeutic drug monitoring mainly. Those recent applications were essentially driven by improved sensitivity of triple quadrupole mass spectrometers. This review presents an overall view of all instrumental and methodological developments for DBS analysis with mass spectrometric detection, with and without separation techniques. A general introduction to DBS will describe their advantages and historical aspects of their emergence. A second section will focus on blood collection, with a strong emphasis on specific parameters that can impact quantitative analysis, including chromatographic effects, hematocrit effects, blood effects, and analyte stability. A third part of the review is dedicated to sample preparation and will consider off-line and on-line extractions; in particular, instrumental designs that have been developed so far for DBS extraction will be detailed. Flow injection analysis and applications will be discussed in section IV. The application of surface analysis mass spectrometry (DESI, paper spray, DART, APTDCI, MALDI, LDTD-APCI, and ICP) to DBS is described in section V, while applications based on separation techniques (e.g., liquid or gas chromatography) are presented in section VI. To conclude this review, the current status of DBS analysis is summarized, and future perspectives are provided. © 2014 Wiley Periodicals, Inc.

A novel method for extracting nucleic acids from dried blood spots for ultrasensitive detection of low-density Plasmodium falciparum and Plasmodium vivax infections.

PubMed

Zainabadi, Kayvan; Adams, Matthew; Han, Zay Yar; Lwin, Hnin Wai; Han, Kay Thwe; Ouattara, Amed; Thura, Si; Plowe, Christopher V; Nyunt, Myaing M

2017-09-18

Greater Mekong Subregion countries are committed to eliminating Plasmodium falciparum malaria by 2025. Current elimination interventions target infections at parasite densities that can be detected by standard microscopy or rapid diagnostic tests (RDTs). More sensitive detection methods have been developed to detect lower density "asymptomatic" infections that may represent an important transmission reservoir. These ultrasensitive polymerase chain reaction (usPCR) tests have been used to identify target populations for mass drug administration (MDA). To date, malaria usPCR tests have used either venous or capillary blood sampling, which entails complex sample collection, processing and shipping requirements. An ultrasensitive method performed on standard dried blood spots (DBS) would greatly facilitate the molecular surveillance studies needed for targeting elimination interventions. A highly sensitive method for detecting Plasmodium falciparum and P. vivax 18S ribosomal RNA from DBS was developed by empirically optimizing nucleic acid extraction conditions. The limit of detection (LoD) was determined using spiked DBS samples that were dried and stored under simulated field conditions. Further, to assess its utility for routine molecular surveillance, two cross-sectional surveys were performed in Myanmar during the wet and dry seasons. The lower LoD of the DBS-based ultrasensitive assay was 20 parasites/mL for DBS collected on Whatman 3MM filter paper and 23 parasites/mL for Whatman 903 Protein Saver cards-equivalent to 1 parasite per 50 µL DBS. This is about 5000-fold more sensitive than standard RDTs and similar to the LoD of ≤16-22 parasites/mL reported for other ultrasensitive methods based on whole blood. In two cross-sectional surveys in Myanmar, nearly identical prevalence estimates were obtained from contemporaneous DBS samples and capillary blood samples collected during the wet and dry season. The DBS-based ultrasensitive method described in this study shows equal sensitivity as previously described methods based on whole blood, both in its limit of detection and prevalence estimates in two field surveys. The reduced cost and complexity of this method will allow for the scale-up of surveillance studies to target MDA and other malaria elimination interventions, and help lead to a better understanding of the epidemiology of low-density malaria infections.

Distinct Rayleigh scattering from hot spot mutant p53 proteins reveals cancer cells.

PubMed

Jun, Ho Joon; Nguyen, Anh H; Kim, Yeul Hong; Park, Kyong Hwa; Kim, Doyoun; Kim, Kyeong Kyu; Sim, Sang Jun

2014-07-23

The scattering of light redirects and resonances when an electromagnetic wave interacts with electrons orbits in the hot spot core protein and oscillated electron of the gold nanoparticles (AuNP). This report demonstrates convincingly that resonant Rayleigh scattering generated from hot spot mutant p53 proteins is correspondence to cancer cells. Hot spot mutants have unique local electron density changes that affect specificity of DNA binding affinity compared with wild types. Rayleigh scattering changes introduced by hot-spot mutations were monitored by localized surface plasmon resonance (LSPR) shift changes. The LSPR λmax shift for hot-spot mutants ranged from 1.7 to 4.2 nm for mouse samples and from 0.64 nm to 2.66 nm for human samples, compared to 9.6 nm and 15 nm for wild type and mouse and human proteins, respectively with a detection sensitivity of p53 concentration at 17.9 nM. It is interesting that hot-spot mutants, which affect only interaction with DNA, launches affinitive changes as considerable as wild types. These changes propose that hot-spot mutants p53 proteins can be easily detected by local electron density alterations that disturbs the specificity of DNA binding of p53 core domain on the surface of the DNA probed-nanoplasmonic sensor. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Urine osmolality in the US population: Implications for environmental biomonitoring

PubMed Central

Yeh, Hung-Chieh; Lin, Yu-Sheng; Kuo, Chin-Chi; Weidemann, Darcy; Weaver, Virginia; Fadrowski, Jeffrey; Neu, Alicia; Navas-Acien, Ana

2018-01-01

Background For many environmental chemicals, concentrations in spot urine samples are considered valid surrogates of exposure and internal dose. To correct for urine dilution, spot urine concentrations are commonly adjusted for urinary creatinine. There are, however, several concerns about the use of urine creatinine. While urine osmolality is an attractive alternative; its characteristics and determinants in the general population remain unknown. Our objective was to describe the determinants of urine osmolality and to contrast the difference between osmolality and creatinine in urine. Methods From the National Health and Nutrition Examination Survey (NHANES) 2009–2012, 10,769 participants aged 16 years or older with measured urine osmolality and creatinine were used in the analysis. Very dilute and very concentrated urine was defined as urine creatinine lower than 0.3 g/l and higher than 3 g/l, respectively. Linear and logistic regression analyses were performed to investigate the associations of interest. Results Urine osmolality and creatinine were highly correlated (Pearson correlation coefficient = 0.75) and their respective median values were 648 mOsm/kg and 1.07 g/l. The prevalence of very dilute and very concentrated urine samples was 8.1% and 3.1%, respectively. Factors associated in the same direction with both urine osmolality and urine creatinine included age, sex, race, body mass index (BMI), hypertension, water intake, and blood osmolality. The magnitude of associations expressed as percent change was significantly stronger with creatinine than osmolality. Compared to urine creatinine, urine osmolality did not vary by diabetes status but was affected by daily total protein intake. Participants with chronic kidney disease (CKD) had significantly higher urine creatinine concentrations but lower urine osmolality. Both very dilute and concentrated urine were associated with a diverse array of sociodemographic, medical conditions, and dietary factors. For instance, females were approximately 3.3 times more likely to have urine over-dilution than male [the adjusted odds ratios (95% CI) = 3.27 (2.10–5.10)]. Conclusion Although the determinants of urine osmolality were generally similar to those of urine creatinine, the relative influence of socio-demographic and medical conditions was less on urine osmolality than on urine creatinine. Protocols for spot urine sample collection could recommend avoiding excessive and insufficient water intake before urine sampling to improve urine adequacy. The feasibility of adopting urine osmolality adjustment and water intake recommendations before providing spot urine samples for environmental biomonitoring merits further investigation. PMID:25460670

Design and experimental research on a self-magnetic pinch diode under MV

NASA Astrophysics Data System (ADS)

Pengfei, ZHANG; Yang, HU; Jiang, SUN; Yan, SONG; Jianfeng, SUN; Zhiming, YAO; Peitian, CONG; Mengtong, QIU; Aici, QIU

2018-01-01

A self-magnetic pinch diode (SMPD) integrating an anode foil-reinforced electron beam pinch focus and a small high-dose x-ray spot output was designed and optimized. An x-ray focal spot measuring system was developed in accordance with the principle of pinhole imaging. The designed SMPD and the corresponding measuring system were tested under ∼MV, with 1.75 × 2 mm2 oval x-ray spots (AWE defined) and forward directed dose 1.6 rad at 1 m. Results confirmed that the anode foil can significantly strengthen the electron beam pinch focus, and the focal spot measuring system can collect clear focal spot images. This finding indicated that the principle and method are feasible.

Membrane materials for storing biological samples intended for comparative nanotoxicological testing

NASA Astrophysics Data System (ADS)

Metelkin, A.; Kuznetsov, D.; Kolesnikov, E.; Chuprunov, K.; Kondakov, S.; Osipov, A.; Samsonova, J.

2015-11-01

The study is aimed at identifying the samples of most promising membrane materials for storing dry specimens of biological fluids (Dried Blood Spots, DBS technology). Existing sampling systems using cellulose fiber filter paper have a number of drawbacks such as uneven distribution of the sample spot, dependence of the spot spreading area on the individual biosample properties, incomplete washing-off of the sample due to partially inconvertible sorption of blood components on cellulose fibers, etc. Samples of membrane materials based on cellulose, polymers and glass fiber with applied biosamples were studied using methods of scanning electron microscopy, FT-IR spectroscopy and surface-wetting measurement. It was discovered that cellulose-based membrane materials sorb components of biological fluids inside their structure, while membranes based on glass fiber display almost no interaction with the samples and biological fluid components dry to films in the membrane pores between the structural fibers. This characteristic, together with the fact that membrane materials based on glass fiber possess sufficient strength, high wetting properties and good storage capacity, attests them as promising material for dry samples of biological fluids storage systems.

n-Hexane metabolism in occupationally exposed workers.

PubMed Central

Mutti, A; Falzoi, M; Lucertini, S; Arfini, G; Zignani, M; Lombardi, S; Franchini, I

1984-01-01

Lung uptake and excretion of n-hexane were studied in ten workers in a shoe factory. Simultaneous samples of inhaled and alveolar air were collected with the aid of a Rhan-Otis valve, personal samplers, and charcoal tubes. Alveolar excretion was monitored during a six hour postexposure period. Uptake was calculated from lung ventilation, the retention coefficient, and environmental concentrations. The amount of exhaled n-hexane was calculated from the decay curve. According to the experimental data, alveolar retention was about 25% of the inhaled n-hexane, corresponding to a lung uptake of about 17%. The postexposure alveolar excretion was about 10% of the total uptake. The main metabolites of n-hexane were identified and measured by capillary GC/MS in spot urine samples collected before, at the end, and 15 hours after the same working shift. Urinary concentrations were low, though related to n-hexane in the air. 2,5-Hexanedione in the end of shift samples gave the best estimate of overall exposure. About 3 mg/g creatinine of 2,5-hexanedione would correspond to about 50 ppm of n-hexane in the air (mean daily exposure). PMID:6498115

The hot spot of vegetation canopies

NASA Technical Reports Server (NTRS)

Myneni, Ranga B.; Kanemasu, Edward T.

1988-01-01

A conventional radiometer is used to identify the hot spot (the peak in reflected radiation in the retrosolar direction) of vegetation. A multiwavelength-band radiometer collected radiances on fully grown dense wheat and maize canopies on several clear sunny days. It is noted that the hot spot is difficult to detect in the near IR wavelengths because the shadows are much darker. In general, the retrosolar brightness is found to be higher for smaller sun polar angles than for larger angles.

Dried blood spot measurement of pregnancy-associated plasma protein A (PAPP-A) and free β-subunit of human chorionic gonadotropin (β-hCG) from a low-resource setting.

PubMed

Browne, J L; Schielen, P C J I; Belmouden, I; Pennings, J L A; Klipstein-Grobusch, K

2015-06-01

The objectives of the article is to compare pregnancy-associated plasma protein A (PAPP-A) and free β-subunit of human chorionic gonadotropin (β-hCG) concentrations in dried blood spots (DBSs) with serum of samples obtained from a public hospital in a low-resource setting and to evaluate their stability. Serum and DBS samples were obtained by venipuncture and finger prick from 50 pregnant participants in a cohort study in a public hospital in Accra, Ghana. PAPP-A and β-hCG concentrations from serum and DBS were measured with an AutoDELFIA® (PerkinElmer, PerkinElmer, Turku, Finland) automatic immunoassay. Correlation and Passing-Bablok regression analyses were performed to compare marker levels. High correlation (>0.9) was observed for PAPP-A and β-hCG levels between various sampling techniques. The β-hCG concentration was stable between DBS and serum, PAPP-A concentration consistently lower in DBS. Our findings suggest that β-hCG can be reliably collected from DBS in low-resource tropical settings. The exact conditions of the clinical workflow necessary for reliable PAPP-A measurement in these settings need to be further developed in the future. These findings could have implications for prenatal screening programs feasibility in low-income and middle-income countries, as DBS provides an alternative minimally invasive sampling method, with advantages in sampling technique, stability, logistics, and potential application in low-resource settings. © 2015 John Wiley & Sons, Ltd.

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

PubMed Central

Batterman, Stuart; Chernyak, Sergei

2014-01-01

Dried blood spots (DBS) can provide accurate and valuable estimates of exposure to environmental toxicants, and the use of information derived from archived newborn DBS information has enormous potential to open up new research on the impacts of early chemical exposure on disease. Broad application of DBS for the purpose of quantitative exposure estimation requires robust and validated methods. This study investigates the suitability of DBS analyses for population studies of exposure to three chemical groups: polychlorinated biphenyls (PCBs), brominated flame retardants (BFRs), and chlorinated pesticides. It examines background (matrix) contamination, recovery and extraction variability, sensitivity, and storage stability. DBS samples prepared using 50 μL of adult blood were analyzed by GC/MS, and method performance was confirmed by using certified materials and paired DBS-blood samples from six volunteers. Several of the target compounds and their degradation products have not been previously measured in DBS. All target compounds were detected in DBS samples collected from the volunteers. Sample DBS cards showed background contamination of several compounds. When stored at room temperature, target compounds, excluding PBDEs, were stable for up to one month. When refrigerated or frozen, stability was acceptable for all compounds up to one year, and multiyear storage appears acceptable at colder (e.g., −80 °C) temperatures. Multicompartment models may be used to estimate or correct for storage losses. Considering concentrations of contaminants for adults and children reported in the literature, and experimental values of detection limits and background contamination, DBS samples are suitable for quantifying exposures to many PCBs, BFRs and persistent pesticides. PMID:25058892

Detection of antibodies against hepatitis A in blood spots dried on filter paper. Is this a reliable method for epidemiological studies?

PubMed Central

Gil, A.; González, A.; Dal-Ré, R.; Dominguez, V.; Astasio, P.; Aguilar, L.

1997-01-01

Diluted dried blood drops on filter paper were compared with serum samples as a specimen source for qualitative anti-HAV antibody determination by ELISA. A total of 298 serum samples and dried blood drops were collected from a population of healthy adolescents (15.3 +/- 1.2 years old). The prevalence of anti-HAV antibody obtained by testing serum samples was 7.7% (95% CI:4.8 10.1). Compared with serum sampling the sensitivity and specificity of diluted dried blood drops were 91.3 and 99.3%. The positive and negative predictive values were 91.3 and 99.3%, respectively, and the likelihood ratios of positive and negative results were 91 and 0.09. It is proposed that this test represents a reliable procedure for anti-HAV antibody testing. PMID:9129596

Diagnostic value of potassium level in a spot urine sample as an index of 24-hour urinary potassium excretion in unselected patients hospitalized in a hypertension unit

PubMed Central

Symonides, Bartosz; Wojciechowska, Ewa; Gryglas, Adam; Gaciong, Zbigniew

2017-01-01

Background Primary hyperaldosteronism may be associated with elevated 24-hour urinary potassium excretion. We evaluated the diagnostic value of spot urine (SU) potassium as an index of 24-hour urinary potassium excretion. Methods We measured SU and 24-hour urinary collection potassium and creatinine in 382 patients. Correlations between SU and 24-hour collections were assessed for potassium levels and potassium/creatinine ratios. We used the PAHO formula to estimate 24-hour urinary potassium excretion based on SU potassium level. The agreement between estimated and measured 24-hour urinary potassium excretion was evaluated using the Bland-Altman method. To evaluate diagnostic performance of SU potassium, we calculated areas under the curve (AUC) for SU potassium/creatinine ratio and 24-hour urinary potassium excretion estimated using the PAHO formula. Results Strongest correlation between SU and 24-hour collection was found for potassium/creatinine ratio (r = 0.69, P<0.001). The PAHO formula underestimated 24-hour urinary potassium excretion by mean 8.3±18 mmol/d (95% limits of agreement -28 to +44 mmol/d). Diagnostic performance of SU potassium/creatinine ratio was borderline good only if 24-hour urinary potassium excretion was largely elevated (AUC 0.802 for 120 mmol K+/24 h) but poor with lower values (AUC 0.696 for 100 mmol K+/24 h, 0.636 for 80 mmol K+/24 h, 0.675 for 40 mmol K+/24 h). Diagnostic performance of 24-hour urinary potassium excretion estimated by the PAHO formula was excellent with values above 120 mmol/d and good with lower values (AUC 0.941 for 120 mmol K+/24 h, 0.819 for 100 mmol K+/24 h, 0.823 for 80 mmol K+/24 h, 0.836 for 40 mmol K+/24 h). Conclusions Spot urine potassium/creatinine ratio might be a marker of increased 24-hour urinary potassium excretion and a potentially useful screening test when reliable 24-hour urine collection is not available. The PAHO formula allowed estimation of the 24-hour urinary potassium excretion based on SU measurements with reasonable clinical accuracy. PMID:28662194

Characterisation and detection of spoilage mould responsible for black spot in dry-cured fermented sausages.

PubMed

Lozano-Ojalvo, Daniel; Rodríguez, Alicia; Cordero, Mirian; Bernáldez, Victoria; Reyes-Prieto, Mariana; Córdoba, Juan J

2015-02-01

Moulds responsible for black spot spoilage of dry-cured fermented sausages were characterised. For this purpose, samples were taken from those dry-cured fermented sausages which showed black spot alteration. Most of the mould strains were first tentatively identified as Penicillium spp. due to their morphological characteristics in different culture conditions, with one strain as Cladosporium sp. The Cladosporium strain was the only one which provoked blackening in culture media. This strain was further characterised by sequencing of ITS1-5.8S-ITS2 rRNA and β-tubulin genes. This mould strain was able to reproduce black spot formation in dry-cured fermented sausage 'salchichón' throughout the ripening process. In addition, a specific and sensitive real-time PCR method was also developed to detect Cladosporium oxysporum responsible for the black spot formation in sausages. This method could be of great interest for the meat industry to detect samples contaminated with this mould before spoilage of product avoiding economic losses for this sector.

A comparison of mercury burdens between St. Marks National Wildlife Refuge and St. Andrew Bay, Florida: Evaluation of fish body burdens and physiological responses in largemouth bass, spotted seatrout, striped mullet, and sunfish

USGS Publications Warehouse

Huge, D.H.; Rauschenberger, R.H.; Wieser, F.M.; Hemming, J.M.

2011-01-01

Musculature from the dorsal region of 130 largemouth bass (Micropterus salmoides), 140 sunfish (Lepomis sp.), 41 spotted seatrout (Cynoscion nebulosus) and 67 striped mullet (Mugil cephalus) were collected from five estuarine and five freshwater sites within the St. Marks National Wildlife Refuge and two estuarine and two freshwater sites from St. Andrew Bay, Florida, United States of America. Musculature was analyzed for total mercury content, sagittal otoliths were removed for age determination and physiological responses were measured. Largemouth bass and sunfish from the refuge had higher mercury concentrations in musculature than those from the bay. Male spotted seatrout, male striped mullet, male and female sunfish and female largemouth bass had mercury burdens positively correlated with length. The majority of all four species of fish from both study areas contained mercury levels below 1.5 part per million, the limit for safe consumption recommended the Florida Department of Health. In comparison, a significant percentage of largemouth bass and sunfish from several sampled sites, most notably Otter Lake and Lake Renfroe within St. Marks National Wildlife Refuge, had mercury levels consistent with the health department's guidelines of 'limited consumption' or 'no consumption guidelines.'

Wound outcome in combat injuries is associated with a unique set of protein biomarkers

PubMed Central

2013-01-01

Background The ability to forecast whether a wound will heal after closure without further debridement(s), would provide substantial benefits to patients with severe extremity trauma. Methods Wound effluent is a readily available material which can be collected without disturbing healthy tissue. For analysis of potential host response biomarkers, forty four serial combat wound effluent samples from 19 patients with either healing or failing traumatic- and other combat-related wounds were examined by 2-D DIGE. Spot map patterns were correlated to eventual wound outcome (healed or wound failure) and analyzed using DeCyder 7.0 and differential proteins identified via LC-MS/MS. Results This approach identified 52 protein spots that were differentially expressed and thus represent candidate biomarkers for this clinical application. Many of these proteins are intimately involved in inflammatory and immune responses. Furthermore, discriminate analysis further refined the 52 differential protein spots to a smaller subset of which successfully differentiate between wounds that will heal and those that will fail and require further surgical intervention with greater than 83% accuracy. Conclusion These results suggest candidates for a panel of protein biomarkers that may aid traumatic wound care prognosis and treatment. We recommend that this strategy be refined, and then externally validated, in future studies of traumatic wounds. PMID:24192341

Global positioning system & Google Earth in the investigation of an outbreak of cholera in a village of Bengaluru Urban district, Karnataka.

PubMed

Masthi, N R Ramesh; Madhusudan, M; Puthussery, Yannick P

2015-11-01

The global positioning system (GPS) technology along with Google Earth is used to measure (spatial map) the accurate distribution of morbidity, mortality and planning of interventions in the community. We used this technology to find out its role in the investigation of a cholera outbreak, and also to identify the cause of the outbreak. This study was conducted in a village near Bengaluru, Karnataka in June 2013 during a cholera outbreak. House-to-house survey was done to identify acute watery diarrhoea cases. A hand held GPS receiver was used to record north and east coordinates of the households of cases and these values were subsequently plotted on Google Earth map. Water samples were collected from suspected sources for microbiological analysis. A total of 27 cases of acute watery diarrhoea were reported. Fifty per cent of cases were in the age group of 14-44 yr and one death was reported. GPS technology and Google Earth described the accurate location of household of cases and spot map generated showed clustering of cases around the suspected water sources. The attack rate was 6.92 per cent and case fatality rate was 3.7 per cent. Water samples collected from suspected sources showed the presence of Vibrio cholera O1 Ogawa. GPS technology and Google Earth were easy to use, helpful to accurately pinpoint the location of household of cases, construction of spot map and follow up of cases. Outbreak was found to be due to contamination of drinking water sources.

Inference about density and temporary emigration in unmarked populations

USGS Publications Warehouse

Chandler, Richard B.; Royle, J. Andrew; King, David I.

2011-01-01

Few species are distributed uniformly in space, and populations of mobile organisms are rarely closed with respect to movement, yet many models of density rely upon these assumptions. We present a hierarchical model allowing inference about the density of unmarked populations subject to temporary emigration and imperfect detection. The model can be fit to data collected using a variety of standard survey methods such as repeated point counts in which removal sampling, double-observer sampling, or distance sampling is used during each count. Simulation studies demonstrated that parameter estimators are unbiased when temporary emigration is either "completely random" or is determined by the size and location of home ranges relative to survey points. We also applied the model to repeated removal sampling data collected on Chestnut-sided Warblers (Dendroica pensylvancia) in the White Mountain National Forest, USA. The density estimate from our model, 1.09 birds/ha, was similar to an estimate of 1.11 birds/ha produced by an intensive spot-mapping effort. Our model is also applicable when processes other than temporary emigration affect the probability of being available for detection, such as in studies using cue counts. Functions to implement the model have been added to the R package unmarked.

Seroepidemiololgy of rickettsioses in Sri Lanka: a patient based study

PubMed Central

2011-01-01

Background Rickettsioses are emerging infections in Sri Lanka as shown by the increase in the number of clinically diagnosed rickettsial patients being reported to the Epidemiology Unit, Sri Lanka. However, mapping the disease for the whole island with laboratory confirmed cases has not been previously carried out. Methods 615 samples received from 23 hospital representing 8 provinces were tested using ELISA or IFA methods and clinical data was collected using a validated questionnaire. Results Rash was found among more spotted fever seropositive patients than scrub typhus seropositive patients while the opposite was true for the presence of eschar. Spotted fever and scrub typhus was found in a geographically restricted manner. Consistent temporal patterns were seen for the presentation of patients with rickettsioses in Kandy and Kurunegala districts for 2009 and 2010. Conclusions This study expanded knowledge on the distribution of rickettsioses in Sri Lanka and their clinical profiles which in turn helps in the clinical diagnosis of these infections. PMID:22118601

Rocky Mountain spotted fever in Argentina.

PubMed

Paddock, Christopher D; Fernandez, Susana; Echenique, Gustavo A; Sumner, John W; Reeves, Will K; Zaki, Sherif R; Remondegui, Carlos E

2008-04-01

We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or confirmed cases of spotted fever rickettsiosis in Jujuy Province and evaluated by polymerase chain reaction assays for spotted fever group rickettsiae. DNA of R. rickettsii was amplified from a pool of A. cajennense ticks and from tissues of one of four patients who died during 2003-2004 after illnesses characterized by high fever, severe headache, myalgias, and petechial rash. The diagnosis of spotted fever rickettsiosis was confirmed in the other patients by indirect immunofluorescence antibody and immunohistochemical staining techniques. These findings show the existence of RMSF in Argentina and emphasize the need for clinicians throughout the Americas to consider RMSF in patients with febrile rash illnesses.

Liquid Microjunction Surface Sampling Probe Electrospray Mass Spectrometry for Detection of Drugs and Metabolites in Thin Tissue Sections

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

Van Berkel, Gary J; Kertesz, Vilmos; Koeplinger, Kenneth A.

2008-01-01

A self-aspirating, liquid micro-junction surface sampling probe/electrospray emitter mass spectrometry system was demonstrated for use in the direct analysis of spotted and dosed drugs and their metabolites in thin tissue sections. Proof-of-principle sampling and analysis directly from tissue without the need for sample preparation was demonstrated first by raster scanning a region on a section of rat liver onto which reserpine was spotted. The mass spectral signal from selected reaction monitoring was used to develop a chemical image of the spotted drug on the tissue. The probe was also used to selectively spot sample areas of sagittal whole mouse bodymore » tissue sections that had been dosed orally (90 mg/kg) with R,S-sulforaphane 3 hrs prior to sacrifice. Sulforaphane and its glutathione and N-acetyl cysteine conjugates were monitored with selected reaction monitoring and detected in the stomach and various other tissues from the dosed mouse. No signal for these species was observed in the tissue from a control mouse. The same dosed tissue section was used to illustrate the possibility of obtaining a line scan across the whole body section. In total these results illustrate the potential for rapid screening of the distribution of drugs and metabolites in tissue sections with the micro-liquid junction surface sampling probe/electrospray mass spectrometry approach.« less

Evaluation of the human immunodeficiency virus type 1 and 2 antibodies detection in dried whole blood spots (DBS) samples.

PubMed

Castro, Andréa Cauduro de; Borges, Luiz Gustavo dos Anjos; Souza, Ricardo da Silva de; Grudzinski, Melina; D'Azevedo, Pedro Alves

2008-01-01

Human Immunodeficiency Virus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903). Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO) and immunofluorescence was the definitive confirmatory test. The samples were obtained from the Hospital Nossa Senhora da Conceição in Porto Alegre, RS - Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS-Brazil where the tests were performed. The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 degrees C, -20 degrees C and -70 degrees C, while the second was composed of two negative and three positive samples stored at 37 degrees C (humidity <50%). Each sample was screened every week for six weeks. These measurement results didn't show variation during the study period. The detected sensibility was 100%, specificity was 99.6%, the positive predictive value was 99.5% and negative predictive values were 100%. The results demonstrated high performance characteristics, opening a new perspective of dried whole blood spot utilization in HIV screening diagnosis.

Virological and molecular epidemiological investigations into the role of wild birds in the epidemiology of influenza A/H5N1 in central Thailand.

PubMed

Siengsanan-Lamont, J; Robertson, I; Blacksell, S D; Ellis, T; Fenwick, S; Saengchoowong, S; Suwanpukdee, S; Yongyuttawichai, P; Sariya, L; Prompiram, P; Chaichoun, K; Wiriyarat, W; Pothieng, D; Ratanakorn, P

2011-03-24

A serological and virological surveillance program to investigate the HPAI H5N1 virus in wild bird populations was undertaken from February 2007 to October 2008. The purpose of the survey was to investigate the infection status in free ranging wild birds in Banglane district, Nakhon Pathom province, central Thailand. Samples from wild birds were collected every two months. Choanal and cloacal swabs, serum and tissue samples were collected from 421 birds comprising 44 species. Sero-prevalence of the virus tested by H5N1 serum neutralization test (using a H5N1 virus clade 1; A/chicken/Thailand/vsmu-3-BKK/2004) was 2.1% (8 out of 385 samples; 95% CI 0.7, 3.5). Species that were antibody positive included rock pigeons (Columba livia), Asian pied starling (Gracupica contra), spotted dove (Streptopelia chinensis), oriental magpie robin (Copsychus saularis), blue-tailed bee-eater (Merops philippinus), myna (Acridotheres spp.), and pond heron (Ardeola spp.). Prevalence by H5N1 virus isolation was 0.5% (2 out of 421 samples; 95% CI 0.0, 1.1); the two H5N1 virus-positive samples were from Asian pied starling (Gracupica contra) and white vented myna (Acridotheres grandis). Positive virological samples were collected in June 2007 while all positive serology samples were collected between May and August except for one sample collected in December 2007. No positive samples were collected in 2008. Molecular studies revealed that the wild bird H5N1 viruses were closely related to poultry viruses isolated in other parts of Thailand. However, there was no poultry H5N1 prevalence study performed in the study site during the time of this wild bird survey. Interpretation of source of virus isolates would include spill-over of H5N1 viruses from contaminated sources due to movement of domestic poultry and/or fomites from other areas; or infection of wild birds within the outbreak locations and then translocation by wild bird movement and interaction with wild birds inhabiting distant locations. Copyright © 2010 Elsevier B.V. All rights reserved.

Day-to-day variability in spot urine albumin-creatinine ratio.

PubMed

Naresh, Chetana N; Hayen, Andrew; Weening, Alexander; Craig, Jonathan C; Chadban, Steven J

2013-12-01

Accurate quantification of albuminuria is important in the diagnosis and management of chronic kidney disease. The reference test, a timed urinary albumin excretion, is cumbersome and prone to collection errors. Spot urine albumin-creatinine ratio (ACR) is convenient and commonly used, but random day-to-day variability in ACR measurements has not been assessed. Prospective cohort study of day-to-day variability in spot urine ACR measurements. Clinically stable outpatients (N = 157) attending a university hospital clinic in Australia between July 2007 and April 2010. Spot urine ACR variability was assessed and repeatability limits were determined using fractional polynomials. ACRs were measured from spot urine samples collected at 9:00 am on consecutive days and 24-hour urine albuminuria was measured concurrently. Paired ACRs were obtained from 157 patients (median age, 56 years; 60% men; median daily albumin excretion, 226 [range, 2.5-14,000] mg/d). Day-to-day variability was substantial and increased in absolute terms, but decreased in relative terms, with increasing baseline ACR. For patients with normoalbuminuria (ACR < 3 mg/mmol [<27 mg/g]), a change greater than ±467% (0-17 mg/mmol [0-150 mg/g]) is required to indicate a significant change in albuminuria status with 95% certainty; for those with microalbuminuria (ACR of 3-30 mg/mmol [27-265 mg/g]), a change of ±170% (0-27 mg/mmol [0-239 mg/g]) is required; for those with macroalbuminuria (ACR > 30 mg/mmol [>265 mg/g]), a change of ±83% (5-55 mg/mmol [44-486 mg/g]) is required; and for those with nephrotic-range proteinuria (ACR > 300 mg/mmol [>2,652 mg/g]), a change of ±48% (158-443 mg/mmol [1,397-3,916 mg/g]) is needed to represent a significant change. These study results need to be replicated in other ethnic groups. Changes in chronic kidney disease status attributed to therapy or disease progression, when based solely on a change in ACR, may be incorrect unless the potential for day-to-day biological variation has been considered. Only relatively large changes are likely to indicate a change in disease status. Copyright © 2013 National Kidney Foundation, Inc. Published by Elsevier Inc. All rights reserved.

Ethnicity is important for creatinine excretion among Inuit and Caucasians in Greenland.

PubMed

Andersen, Stig; Dehnfeld, Marie; Laurberg, Peter

2015-01-01

Human nutrition, contamination and renal function are commonly assessed by the analysis of urine. A complete 24-hour urine sample is the ideal but it is inconvenient and unreliable. Thus, spot urine sampling with creatinine adjustment is widely used. Stratification for age and gender is recommended. Still, ethnicity may influence creatinine excretion. We collected 104 24-h urine samples among Inuit and non-Inuit living in Greenland. Completeness of sampling was checked by using para-amino benzoic acid (PABA) that also allowed for compensation of creatinine excretion when sampling was incomplete. We measured creatinine using the Jaffe method and PABA by the HPLC method. Participants were recruited from the capital city, a major town and a settlement (n = 36/48/20). They were aged 30-69 years with 78 Inuit and 26 non-Inuit. Inuit were smaller than non-Inuit (Caucasians): height, 163 vs. 177 cm, p < 0.001; weight, 71 vs. 84 kg, p = 0.001 with similar BMI. Creatinine excretion was lower in Inuit compared to non-Inuit (men, 1344/1807 mg/24 h; women 894/1259 mg/24 h; p = 0.002; 0.02). It was influenced by age (p < 0.001), gender (p < 0.001), weight (p = 0.001) and ethnicity (p = 0.030) while not by the intake of the protein-rich Inuit diet in the adjusted analysis. Creatinine excretion was described by: Inuit men, 1925 mg - (13.1 × age); Inuit women, 1701 mg - (17.0 × age). Inuit and Caucasians have different creatinine excretion. It is recommended to stratify by ethnicity in addition to adjustment for age and gender when using creatinine correction of spot urine samples.

Within- and between-child variation in repeated urinary pesticide metabolite measurements over a 1-year period.

PubMed

Attfield, Kathleen R; Hughes, Michael D; Spengler, John D; Lu, Chensheng

2014-02-01

Children are exposed to pesticides from many sources and routes, including dietary and incidental ingestion, dermal absorption, and inhalation. Linking health outcomes to these exposures using urinary metabolites requires understanding temporal variability within subjects to avoid exposure misclassification. We characterized the within- and between-child variability of urinary organophosphorus and pyrethroid metabolites in 23 participants of the Children's Pesticide Exposure Study-Washington over 1 year and examined the ability of one to four spot urine samples to categorize mean exposures. Each child provided urine samples twice daily over 7- to 16-day sessions in four seasons in 2003 and 2004. Samples were analyzed for five pyrethroid and five organophosphorus (OP) metabolites. After adjusting for specific gravity, we used a customized maximum likelihood estimation linear mixed-effects model that accounted for values below the limit of detection to calculate intraclass correlation coefficients (ICC) and conducted surrogate category analyses. Within-child variability was 2-11 times greater than between-child variability. When restricted to samples collected during a single season, ICCs were higher in the fall, winter, and spring than in summer for OPs, and higher in summer and winter for pyrethroids, indicating an increase in between-person variability relative to within-person variability during these seasons. Surrogate category analyses demonstrated that a single spot urine sample did not categorize metabolite concentrations well, and that four or more samples would be needed to categorize children into quartiles consistently. Urinary biomarkers of these short half-life pesticides exhibited substantial within-person variability in children observed over four seasons. Researchers investigating pesticides and health outcomes in children may need repeated biomarker measurements to derive accurate estimates of exposure and relative risks.

Quantitative determination of opioids in whole blood using fully automated dried blood spot desorption coupled to on-line SPE-LC-MS/MS.

PubMed

Verplaetse, Ruth; Henion, Jack

2016-01-01

Opioids are well known, widely used painkillers. Increased stability of opioids in the dried blood spot (DBS) matrix compared to blood/plasma has been described. Other benefits provided by DBS techniques include point-of-care collection, less invasive micro sampling, more economical shipment, and convenient storage. Current methodology for analysis of micro whole blood samples for opioids is limited to the classical DBS workflow, including tedious manual punching of the DBS cards followed by extraction and liquid chromatography-tandem mass spectrometry (LC-MS/MS) bioanalysis. The goal of this study was to develop and validate a fully automated on-line sample preparation procedure for the analysis of DBS micro samples relevant to the detection of opioids in finger prick blood. To this end, automated flow-through elution of DBS cards was followed by on-line solid-phase extraction (SPE) and analysis by LC-MS/MS. Selective, sensitive, accurate, and reproducible quantitation of five representative opioids in human blood at sub-therapeutic, therapeutic, and toxic levels was achieved. The range of reliable response (R(2)  ≥0.997) was 1 to 500 ng/mL whole blood for morphine, codeine, oxycodone, hydrocodone; and 0.1 to 50 ng/mL for fentanyl. Inter-day, intra-day, and matrix inter-lot accuracy and precision was less than 15% (even at lower limits of quantitation (LLOQ) level). The method was successfully used to measure hydrocodone and its major metabolite norhydrocodone in incurred human samples. Our data support the enormous potential of DBS sampling and automated analysis for monitoring opioids as well as other pharmaceuticals in both anti-doping and pain management regimens. Copyright © 2015 John Wiley & Sons, Ltd.

High seroprevalence of antibodies against spotted fever and scrub typhus bacteria in patients with febrile Illness, Kenya.

PubMed

Thiga, Jacqueline W; Mutai, Beth K; Eyako, Wurapa K; Ng'ang'a, Zipporah; Jiang, Ju; Richards, Allen L; Waitumbi, John N

2015-04-01

Serum samples from patients in Kenya with febrile illnesses were screened for antibodies against bacteria that cause spotted fever, typhus, and scrub typhus. Seroprevalence was 10% for spotted fever group, <1% for typhus group, and 5% for scrub typhus group. Results should help clinicians expand their list of differential diagnoses for undifferentiated fevers.

A Compact X-Ray System for Support of High Throughput Crystallography

NASA Technical Reports Server (NTRS)

Ciszak, Ewa; Gubarev, Mikhail; Gibson, Walter M.; Joy, Marshall K.; Whitaker, Ann F. (Technical Monitor)

2001-01-01

Standard x-ray systems for crystallography rely on massive generators coupled with optics that guide X-ray beams onto the crystal sample. Optics for single-crystal diffractometry include total reflection mirrors, polycapillary optics or graded multilayer monochromators. The benefit of using polycapillary optic is that it can collect x-rays over tile greatest solid angle, and thus most efficiently, utilize the greatest portion of X-rays emitted from the Source, The x-ray generator has to have a small anode spot, and thus its size and power requirements can be substantially reduced We present the design and results from the first high flux x-ray system for crystallography that combine's a microfocus X-ray generator (40microns FWHM Spot size at a power of 45 W) and a collimating, polycapillary optic. Diffraction data collected from small test crystals with cell dimensions up to 160A (lysozyme and thaumatin) are of high quality. For example, diffraction data collected from a lysozyme crystal at RT yielded R=5.0% for data extending to 1.70A. We compare these results with measurements taken from standard crystallographic systems. Our current microfocus X-ray diffraction system is attractive for supporting crystal growth research in the standard crystallography laboratory as well as in remote, automated crystal growth laboratory. Its small volume, light-weight, and low power requirements are sufficient to have it installed in unique environments, i.e.. on-board International Space Station.

An on-spot internal standard addition approach for accurately determining colistin A and colistin B in dried blood spots using ultra high-performance liquid chromatography-tandem mass spectrometry.

PubMed

Tsai, I-Lin; Kuo, Ching-Hua; Sun, Hsin-Yun; Chuang, Yu-Chung; Chepyala, Divyabharathi; Lin, Shu-Wen; Tsai, Yun-Jung

2017-10-25

Outbreaks of multidrug-resistant Gram-negative bacterial infections have been reported worldwide. Colistin, an antibiotic with known nephrotoxicity and neurotoxicity, is now being used to treat multidrug-resistant Gram-negative strains. In this study, we applied an on-spot internal standard addition approach coupled with an ultra high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify colistin A and B from dried blood spots (DBSs). Only 15μL of whole blood was required for each sample. An internal standard with the same yield of extraction recoveries as colistin was added to the spot before sample extraction for accurate quantification. Formic acid in water (0.15%) with an equal volume of acetonitrile (50:50v/v) was used as the extraction solution. With the optimized extraction process and LC-MS/MS conditions, colistin A and B could be quantified from a DBS with respective limits of quantification of 0.13 and 0.27μgmL -1 , and the retention times were < 2min. The relative standard deviations of within-run and between-run precisions for peak area ratios were all < 17.3%. Accuracies were 91.5-111.2% for lower limit of quantification, low, medium, and high QC samples. The stability of the easily hydrolyzed prodrug, colistin methanesulfonate, was investigated in DBSs. Less than 4% of the prodrug was found to be hydrolyzed in DBSs at room temperature after 48h. The developed method applied an on-spot internal standard addition approach which benefited the precision and accuracy. Results showed that DBS sampling coupled with the sensitive LC-MS/MS method has the potential to be an alternative approach for colistin quantification, where the bias of prodrug hydrolysis in liquid samples is decreased. Copyright © 2017 Elsevier B.V. All rights reserved.

Effect of post weld impact treatment (PWIT) on mechanical properties of spot-welded joint

NASA Astrophysics Data System (ADS)

Ghazali, F. A.; Salleh, Z.; Hyie, K. M.; Rozlin, N. M. Nik; Hamidi, S. H. Ahmad; Padzi, M. M.

2017-12-01

This paper focuses on the study of improvement for spot welding on the tensile shear and hardness by applying post weld impact treatment (PWIT) on the welded joint. The main objective of the research is to characterize and improve the mechanical properties of the joint. The method of PWIT used on the welded joint was Pneumatic Impact Treatment (PIT). The concept of PIT on spot welding is that it improves the mechanical properties of the welded zone. The working sample was undergoing a resistance spot welding of joining two similar in dimension and material of a steel plate before treated. The dimension of both plate are 110 mm × 45 mm × 1.2 mm and the material used were low carbon steel (LCS). All the welded samples were tested for its mechanical properties by performing the tensile-shear and hardness test. Tensile-shear test was conducted on the spot welded, both treated and as-welded samples using crosshead speed of 2 mm/min, while hardness test was performed using 1kgf load via Vickers hardness indenter. The effects of PIT on tensile-shear properties and hardness were evaluated and found that the implementation of PIT has increased tensile shear and hardness significantly.

The Application of Chinese High-Spatial Remote Sensing Satellite Image in Land Law Enforcement Information Extraction

NASA Astrophysics Data System (ADS)

Wang, N.; Yang, R.

2018-04-01

Chinese high -resolution (HR) remote sensing satellites have made huge leap in the past decade. Commercial satellite datasets, such as GF-1, GF-2 and ZY-3 images, the panchromatic images (PAN) resolution of them are 2 m, 1 m and 2.1 m and the multispectral images (MS) resolution are 8 m, 4 m, 5.8 m respectively have been emerged in recent years. Chinese HR satellite imagery has been free downloaded for public welfare purposes using. Local government began to employ more professional technician to improve traditional land management technology. This paper focused on analysing the actual requirements of the applications in government land law enforcement in Guangxi Autonomous Region. 66 counties in Guangxi Autonomous Region were selected for illegal land utilization spot extraction with fusion Chinese HR images. The procedure contains: A. Defines illegal land utilization spot type. B. Data collection, GF-1, GF-2, and ZY-3 datasets were acquired in the first half year of 2016 and other auxiliary data were collected in 2015. C. Batch process, HR images were collected for batch preprocessing through ENVI/IDL tool. D. Illegal land utilization spot extraction by visual interpretation. E. Obtaining attribute data with ArcGIS Geoprocessor (GP) model. F. Thematic mapping and surveying. Through analysing 42 counties results, law enforcement officials found 1092 illegal land using spots and 16 suspicious illegal mining spots. The results show that Chinese HR satellite images have great potential for feature information extraction and the processing procedure appears robust.

[Monitoring and SWOT analysis of Ascaris eggs pollution in soil of rural China].

PubMed

Zhu, Hui-hui; Zhou, Chang-hai; Zang, Wei; Zhang, Xue-qiang; Chen, Ying-dan

2014-06-01

To understand the status of Ascaris eggs pollution in soil at national monitoring spots of soil-transmitted nematodiasis, so as to provide the evidence for making countermeasures and evaluating the control effect. Ten households were selected from each of the 22 national monitoring spots annually according to the National Surveillance Program of Soil-Transmitted Nematodiasis (Trial), and the soil samples from vegetable gardens, toilet periphery, courtyards and kitchens were collected and examined by using the modified floatation test with saturated sodium nitrate. Fertilized or unfertilized eggs as well as live or dead fertilized eggs were discriminated and identified. In addition, a SWOT analysis of monitoring of Ascaris eggs pollution in the soil of rural China was carried out. A total of 1 090 households were monitored in 22 monitoring spots from 2006 to 2010. The total detection rate of Ascaris eggs in the soil was 30.73%, and the detection rates of fertilized, unfertilized and live fertilized eggs were 13.21%, 26.42% and 20.28%, respectively. The total detection rates of Ascaris eggs in the vegetable garden, toilet periphery, courtyard and kitchen were 16.51%, 13.49%, 14.22% and 10.73% respectively. The SWOT analysis demonstrated that the monitoring work had both advantages and disadvantages, and was faced with opportunities as well as threats. The pollution status of Ascaris eggs in the soil is still quite severe at some national monitoring spots, and the counter-measures such as implementing hazard-free treatment of stool, improving water supply and sanitation and reforming environment should be taken to protect people from being infected.

Size-segregated aerosol in a hot-spot pollution urban area: Chemical composition and three-way source apportionment.

PubMed

Bernardoni, V; Elser, M; Valli, G; Valentini, S; Bigi, A; Fermo, P; Piazzalunga, A; Vecchi, R

2017-12-01

In this work, a comprehensive characterisation and source apportionment of size-segregated aerosol collected using a multistage cascade impactor was performed. The samples were collected during wintertime in Milan (Italy), which is located in the Po Valley, one of the main pollution hot-spot areas in Europe. For every sampling, size-segregated mass concentration, elemental and ionic composition, and levoglucosan concentration were determined. Size-segregated data were inverted using the program MICRON to identify and quantify modal contributions of all the measured components. The detailed chemical characterisation allowed the application of a three-way (3-D) receptor model (implemented using Multilinear Engine) for size-segregated source apportionment and chemical profiles identification. It is noteworthy that - as far as we know - this is the first time that three-way source apportionment is attempted using data of aerosol collected by traditional cascade impactors. Seven factors were identified: wood burning, industry, resuspended dust, regional aerosol, construction works, traffic 1, and traffic 2. Further insights into size-segregated factor profiles suggested that the traffic 1 factor can be associated to diesel vehicles and traffic 2 to gasoline vehicles. The regional aerosol factor resulted to be the main contributor (nearly 50%) to the droplet mode (accumulation sub-mode with modal diameter in the range 0.5-1 μm), whereas the overall contribution from the two factors related to traffic was the most important one in the other size modes (34-41%). The results showed that applying a 3-D receptor model to size-segregated samples allows identifying factors of local and regional origin while receptor modelling on integrated PM fractions usually singles out factors characterised by primary (e.g. industry, traffic, soil dust) and secondary (e.g. ammonium sulphate and nitrate) origin. Furthermore, the results suggested that the information on size-segregated chemical composition in different size classes was exploited by the model to relate primary emissions to rapidly-formed secondary compounds. Copyright © 2017 Elsevier Ltd. All rights reserved.

Rickettsia parkeri in Amblyomma dubitatum ticks in a spotted fever focus from the Brazilian Pampa.

PubMed

Weck, Bárbara; Dall'Agnol, Bruno; Souza, Ugo; Webster, Anelise; Stenzel, Bárbara; Klafke, Guilherme; Martins, João Ricardo; Reck, José

2017-07-01

Spotted fever is an acute febrile illness, which is considered severely underreported and misdiagnosed in the Brazilian Pampa, caused by tick-borne Rickettsiae. Here, we report an eco-epidemiological investigation of Rickettsia spp. in ticks from a spotted fever focus in Toropi, southern Brazil. Ticks were collected from capybara carcasses and processed individually to obtain genomic DNA. Rickettsia was investigated using PCR that amplified the rickettsial fragments of the gltA, ompA and htrA genes. DNA from Rickettsia parkeri was found in four of 14 Amblyomma dubitatum ticks collected from capybara carcasses in Toropi and the nearby municipality of Quevedos. We also tested 210A. dubitatum ticks obtained from road-killed capybaras of other localities from the Pampa biome; none of them were positive for Rickettsiae. Thus, in Rio Grande do Sul, two Rickettsia species can be potentially associated to spotted fever: Rickettsia sp. strain Atlantic Rainforest, associated with Amblyomma ovale ticks in the Atlantic Rainforest biome, and R. parkeri, associated both with Amblyomma tigrinum and A. dubitatum ticks in the Pampa biome. Our results reinforce that R. parkeri may be the agent associated with spotted fever in the Brazilian Pampa. Copyright © 2017 Elsevier B.V. All rights reserved.

[Hypothermia as the cause of death in forensic pathology: autopsy study].

PubMed

Nikolić, Slobodan; Zivković, Magdalena; Zivković, Vladimir; Juković, Fehim

2010-01-01

The body cooling process goes through few clinical phases. These are followed by some morphological thanatological changes such as frost erythema and Wischnewsky's spots, which are used in diagnosis of death due to hypothermia. In such cases there is no any specific autopsy finding. To establish the frequency of hypothermia as the cause of death for a ten-year-period, and to analyze the sample according to gender and age, risk factors and autopsy findings of subjects. A retrospective autopsy study was performed for a ten-year-period (total of 12,765 forensic autopsies). The relevant data were collected from autopsy records, police reports and heteroanamnestic interviews. The sample was analyzed according to gender, age, scene of death, blood alcohol concentration, risk factors, and autopsy findings of all observed subjects. The sample included 67 subjects, 42 males and 25 females (chi2 = 4.31; p < 0.05), of average age 63.9 +/- 14.7 years (min=27, max=92; med=65, mod=55). Nineteen of subjects were found at in-door places. In 13 subjects blood alcohol concentration ranged from 0.50 to 3.32 promille (average 1.81 +/- 0.93). The younger the observed subject was, the higher the blood alcohol concentration (p = -0.251; p = 0.04). One third of the observed subjects were chronic alcohol abusers. Thirteen persons had psychiatric diseases. In 43 observed subjects the concomitant appearance of frost erythema and Wischniewsky's spots were established (chi2 = 49.59; df = 3; p < 0.001). In the analyzed ten-year period hypothermia was not often the cause of death; it was disclosed only in 0.5% of the total number of the studied autopsies. The most of the deceased were older males with cardiovascular problems found in unprotected open-air places. The most frequent thanatological findings in the analyzed subjects were frost erythema and Wischnewsky's spots.

An Investigation of the Measurement Properties of the Spot-the-Word Test In a Community Sample

ERIC Educational Resources Information Center

Mackinnon, Andrew; Christensen, Helen

2007-01-01

Intellectual ability is assessed with the Spot-the-Word (STW) test (A. Baddeley, H. Emslie, & I. Nimmo Smith, 1993) by asking respondents to identify a word in a word-nonword item pair. Results in moderate-sized samples suggest this ability is resistant to decline due to dementia. The authors used a 3-parameter item response theory model to…

Advanced High Brilliance X-Ray Source

NASA Technical Reports Server (NTRS)

Gibson, Walter M.

1998-01-01

The possibility to dramatically increase the efficiency of laboratory based protein structure measurements through the use of polycapillary X-ray optics was investigated. This project initiated April 1, 1993 and concluded December 31, 1996 (including a no cost extension from June 31, 1996). This is a final report of the project. The basis for the project is the ability to collect X-rays from divergent electron bombardment laboratory X-ray sources and redirect them into quasiparallel or convergent (focused) beams. For example, a 0.1 radian (approx. 6 deg) portion of a divergent beam collected by a polycapillary collimator and transformed into a quasiparallel beam of 3 millradian (0.2 deg) could give a gain of 6(exp 2)/0.2(exp 2) x T for the intensity of a diffracted beam from a crystal with a 0.2 deg diffraction width. T is the transmission efficiency of the polycapillary diffraction optic, and for T=0.5, the gain would be 36/0.04 x O.5=45. In practice, the effective collection angle will depend on the source spot size, the input focal length of the optic (usually limited by the source spot-to-window distance on the x-ray tube) and the size of the crystal relative to the output diameter of the optic. The transmission efficiency, T, depends on the characteristics (fractional open area, surface roughness, shape and channel diameter) of the polycapillary optic and is typically in the range 0.2-0.4. These effects could substantially reduce the expected efficiency gain. During the course of this study, the possibility to use a weakly focused beam (0.5 deg convergence) was suggested which could give an additional 10-20 X efficiency gain for small samples . Weakly focused beams from double focusing mirrors are frequently used for macromolecular crystallography studies. Furthermore the crystals are typically oscillated by as much as 2 deg during each X-ray exposure in order to increase the reciprocal space (number of crystal planes) sampled and use of a slightly convergent beam could, in principle, provide a similar sampling benefit without oscillation. Although more problematic, because of complications in analyzing the diffraction patterns, it was also suggested that even more extreme beam convergence might be used to give another order of magnitude intensity gain and even smaller focused spot size which could make it possible to study smaller protein crystals than can be studied using standard laboratory based X-ray diffraction systems. This project represents the first systematic investigation of these possibilities. As initially proposed, the contract included requirements for design, purchase, evaluation and delivery of three polycapillary lenses to the Laboratory for Structural Biology at MSFC and demonstration of such optics at MSFC for selected protein crystal diffraction applications.

Simultaneous Quantification of Forskolin and Iso-Forskolin in Coleus forskohlii (Wild.) Briq. and Identification of Elite Chemotype, Collected from Eastern Ghats (India)

PubMed Central

Shukla, Pushpendra Kumar; Misra, Ankita; Kumar, Manish; Jaichand; Singh, Kuldeep; Akhtar, Juber; Srivastava, Sharad; Agrawal, Pawan K; Singh Rawat, Ajay K

2017-01-01

Background: Coleus forskohlii is a well-known industrially important medicinal plant, for its high forskolin content. Objective: A simple, selective, and sensitive high-performance thin layer chromatography (HPTLC) method was developed and validated for simultaneous quantification of forskolin and iso-forskolin in C. forskohlii germplasm collected from the Eastern Ghats, India. Materials and Methods: Chromatographic separation of the targeted marker(s) was obtained on precoated silica plates using toluene: ethyl acetate: methanol (90:30:0.5, v/v/v) as the mobile phase. Results: Densitometric quantification of forskolin and iso-forskolin was carried out at 545 nm. Forskolin and iso-forskolin were identified by comparing the ultraviolet spectra of standard and sample track at Rf of 0.64 ± 0.02 and 0.36 ± 0.01, after derivatization with anisaldehyde sulfuric acid reagent. The linearity of both the analytes was obtained in the range of 300–1200 ng/spot with the regression coefficient (R2) of 0.991 and 0.986. Recovery of analyte (s) at three levels, namely, 100, 150, and 200 ng/spot was found to be 100.46% ± 0.29%, 99.64% ± 0.33%, 100.02% ± 0.76% and 99.76% ± 0.62%, 99.56% ± 0.35%, 100.02% ± 0.22%, respectively, for forskolin and iso-forskolin. The content of forskolin and iso-forskolin varies from 0.046% to 0.187% and 0.002% to 0.077%, respectively (dry weight basis), the maximum content of both the markers was found in NBC-31, from Thakurwada, Maharashtra. Conclusion: The developed HPTLC method was linear, accurate, and reliable as per the International Council for Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use guidelines. The study aids in the identification of elite chemotype for commercial prospection of industrially viable medicinal crop. SUMMARY 12 Samples are collected from different locations of the eastern ghat regionsQuantification of two major marker forskolin and iso forskolinThe maximum content of both the markers was found in NBC -31, from Thakurwada, MaharashtraIdentification of elite chemotype of collected samples may be useful for commercial prospection in industries. PMID:29491648

'FloraArray' for screening of specific DNA probes representing the characteristics of a certain microbial community.

PubMed

Yokoi, Takahide; Kaku, Yoshiko; Suzuki, Hiroyuki; Ohta, Masayuki; Ikuta, Hajime; Isaka, Kazuichi; Sumino, Tatsuo; Wagatsuma, Masako

2007-08-01

To investigate uncharacterized microbial communities, a custom DNA microarray named 'FloraArray' was developed for screening specific probes that would represent the characteristics of a microbial community. The array was prepared by spotting 2000 plasmid DNAs from a genomic shotgun library of a sludge sample on a DNA microarray. By comparative hybridization of the array with two different samples of genomic DNA, one from the activated sludge and the other from a nonactivated sludge sample of an anaerobic ammonium oxidation (anammox) bacterial community, specific spots were visualized as a definite fluctuating profile in an MA (differential intensity ratio vs. spot intensity) plot. About 300 spots of the array accounted for the candidate probes to represent anammox reaction of the activated sludge. After sequence analysis of the probes and examination of the results of blastn searches against the reported anammox reference sequence, complete matches were found for 161 probes (58.3%) and >90% matches were found for 242 probes (87.1%). These results demonstrate that 'FloraArray' could be a useful tool for screening specific DNA molecules of unknown microbial communities.

Roost sites of radio-marked Mexican spotted owls in Arizona and New Mexico: sources of variability and descriptive characteristics

Treesearch

Joseph L. Ganey; William M. Block; Rudy M. King

2000-01-01

To increase understanding of roosting habitat of Mexican Spotted Owls (Strix occidentalis lucida) and factors that influence use of roosting habitat, we sampled habitat characteristics at 1790 sites used for roosting by 28 radio-marked Mexican Spotted Owls in three study areas in Arizona and New Mexico. We explored potential patterns of variation in...

EPICS Controlled Collimator for Controlling Beam Sizes in HIPPO

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

Napolitano, Arthur Soriano; Vogel, Sven C.

2017-08-03

Controlling the beam spot size and shape in a diffraction experiment determines the probed sample volume. The HIPPO - High-Pressure-Preferred Orientation– neutron time-offlight diffractometer is located at the Lujan Neutron Scattering Center in Los Alamos National Laboratories. HIPPO characterizes microstructural parameters, such as phase composition, strains, grain size, or texture, of bulk (cm-sized) samples. In the current setup, the beam spot has a 10 mm diameter. Using a collimator, consisting of two pairs of neutron absorbing boron-nitride slabs, horizontal and vertical dimensions of a rectangular beam spot can be defined. Using the HIPPO robotic sample changer for sample motion, themore » collimator would enable scanning of e.g. cylindrical samples along the cylinder axis by probing slices of such samples. The project presented here describes implementation of such a collimator, in particular the motion control software. We utilized the EPICS (Experimental Physics Interface and Control System) software interface to integrate the collimator control into the HIPPO instrument control system. Using EPICS, commands are sent to commercial stepper motors that move the beam windows.« less

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed

Bullen, A; Patel, S S; Saggau, P

1997-07-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.

MALDI-typing of infectious algae of the genus Prototheca using SOM portraits.

PubMed

Wirth, Henry; von Bergen, Martin; Murugaiyan, Jayaseelan; Rösler, Uwe; Stokowy, Tomasz; Binder, Hans

2012-01-01

MALDI-typing has become a frequently used approach for the identification of microorganisms and recently also of invertebrates. Similarity-comparisons are usually based on single-spectral data. We apply self-organizing maps (SOM) to portray the MS-spectral data with individual resolution and to improve the typing of Prototheca algae by using meta-spectra representing prototypes of groups of similar-behaving single spectra. The MALDI-TOF peaklists of more than 300 algae extracts referring to five Prototheca species were transformed into colored mosaic images serving as molecular portraits of the individual samples. The portraits visualize the algae-specific distribution of high- and low-amplitude peaks in two dimensions. Species-specific pattern of MS intensities were readily discernable in terms of unique single spots of high amplitude MS-peaks which collect characteristic fingerprint spectra. The spot patterns allow the visual identification of groups of samples referring to different species, genotypes or isolates. The use of meta-peaks instead of single-peaks reduces the dimension of the data and leads to an increased discriminating power in downstream analysis. We expect that our SOM portray method improves MS-based classifications and feature selection in upcoming applications of MALDI-typing based species identifications especially of closely related species. Copyright © 2011. Published by Elsevier B.V.

Comparative Proteomics of Tumor and Paired Normal Breast Tissue Highlights Potential Biomarkers in Breast Cancer.

PubMed

Da Costa, Gustavo Góes; Gomig, Talita Helen Bombardelli; Kaviski, Rodrigo; Santos Sousa, Karla; Kukolj, Caroline; De Lima, Rubens Silveira; De Andrade Urban, Cicero; Cavalli, Iglenir J; Ribeiro, Enilze M S F

2015-01-01

Breast cancer is the most common type of cancer among women worldwide, and about 57,000 new cases are expected for the Brazilian population in 2015. Elucidation of protein expression and modification is essential for the biological understanding, early diagnosis and therapeutics of breast cancer. The main objectives of the study are comparison between the proteome of tumor and paired non-tumor breast cancer tissues, describing all identified proteins, highlighting the ones most differentially expressed and comparing the data with existing literature. The five paired samples from patients with invasive ductal carcinoma were analyzed by 2-DE and MS. We collected 161 identified spots corresponding to 110 distinct proteins. Forty-three differentially-expressed spots were common to at least two samples, and the ten proteins with the highest-fold changes were CASPE, ENOG, TPM1, CAPG, VIME, TPM3, TRFE, PDIA6, WDR61 and PDIA3. Metabolic enzymes and proteins with binding functions were the most representative functional classes of proteins with increased and decreased expression in tumor tissue respectively. Taking the fold change as a parameter, we point to future targets to be studied by functional methods in a search for biomarkers for initiation and progress of breast cancer. Copyright© 2015, International Institute of Anticancer Research (Dr. John G. Delinasios), All rights reserved.

2D-electrophoresis and the urine proteome map: where do we stand?

PubMed

Candiano, Giovanni; Santucci, Laura; Petretto, Andrea; Bruschi, Maurizio; Dimuccio, Veronica; Urbani, Andrea; Bagnasco, Serena; Ghiggeri, Gian Marco

2010-03-10

The discovery of urinary biomarkers is a main topic in clinical medicine. The development of proteomics has rapidly changed the knowledge on urine protein composition and probably will modify it again. Two-dimensional electrophoresis (2D-PAGE) coupled with mass spectrometry has represented for years the technique of choice for the analysis of urine proteins and it is time to draw some conclusions. This review will focus on major methodological aspects related to urine sample collection, storage and analysis by 2D-PAGE and attempt to define an advanced normal urine protein map. Overall, 1118 spots were reproducibly found in normal urine samples but only 275 were characterized as isoforms of 82 proteins. One-hundred height spots belonging to 30 proteins were also detected in plasma and corresponded to typical plasma components. The identity of most of the proteins found in normal urine by 2D-PAGE remains to be determined, the majority being low-molecular weight proteins (<30 kDa). Equalization procedures would also enhance sensitivity of the analysis and allow low abundance proteins to be characterized. Therefore, we are still on the way to define the normal urine composition. Technology advancements in concentrating procedure will improve sensitivity and give the possibility to purify proteins for mass spectrometry. Copyright (c) 2009 Elsevier B.V. All rights reserved.

Impact of tannery effluents on the aquatic environment of the Buriganga River in Dhaka, Bangladesh.

PubMed

Asaduzzaman, Mohammad; Hasan, Imtiaj; Rajia, Sultana; Khan, Nazneen; Kabir, Kazi Ahmed

2016-06-01

This study presents an overview of the existence and effects of six heavy metals, chromium (Cr), lead (Pb), cadmium (Cd), mercury (Hg), manganese (Mn), and aluminum (Al), in tannery effluents released to the Buriganga River in Dhaka, Bangladesh. The pollutants were found in three different sources, such as effluents from tanneries, contaminated river water and three species of fish-climbing perch (Anabas testudineus), spotted snakehead (Channa punctata), and Black tilapia (Oreochromis mossambicus) caught from the river. Tannery effluents, water, and fish samples were collected from three different factories, five sample stations, and three different harvesting points, respectively. Effluents from all three factories contained significant amounts of heavy metals, especially Cr (374.19 ppm in average), whereas lesser amounts were found in the tissues of the three fish species studied. The trends in tissue elemental concentrations of fish were Cr > Pb > Al > Hg > Mn > Cd. In most cases (Cr, Cd, Mn, and Al), heavy metal concentrations were found to be greater in climbing perch than in Black tilapia and spotted snakehead. Although the river water contained high concentrations of harmful heavy metals, the fish species under study had concentrations well below the permissible Food and Agriculture Organization/World Health Organization levels for those metals and seemed to be safe for human consumption. © The Author(s) 2014.

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

PubMed Central

Bullen, A; Patel, S S; Saggau, P

1997-01-01

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging. Images FIGURE 6 PMID:9199810

Sampling designs for contaminant temporal trend analyses using sedentary species exemplified by the snails Bellamya aeruginosa and Viviparus viviparus.

PubMed

Yin, Ge; Danielsson, Sara; Dahlberg, Anna-Karin; Zhou, Yihui; Qiu, Yanling; Nyberg, Elisabeth; Bignert, Anders

2017-10-01

Environmental monitoring typically assumes samples and sampling activities to be representative of the population being studied. Given a limited budget, an appropriate sampling strategy is essential to support detecting temporal trends of contaminants. In the present study, based on real chemical analysis data on polybrominated diphenyl ethers in snails collected from five subsites in Tianmu Lake, computer simulation is performed to evaluate three sampling strategies by the estimation of required sample size, to reach a detection of an annual change of 5% with a statistical power of 80% and 90% with a significant level of 5%. The results showed that sampling from an arbitrarily selected sampling spot is the worst strategy, requiring much more individual analyses to achieve the above mentioned criteria compared with the other two approaches. A fixed sampling site requires the lowest sample size but may not be representative for the intended study object e.g. a lake and is also sensitive to changes of that particular sampling site. In contrast, sampling at multiple sites along the shore each year, and using pooled samples when the cost to collect and prepare individual specimens are much lower than the cost for chemical analysis, would be the most robust and cost efficient strategy in the long run. Using statistical power as criterion, the results demonstrated quantitatively the consequences of various sampling strategies, and could guide users with respect of required sample sizes depending on sampling design for long term monitoring programs. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biomonitoring short- and long-term exposure to the herbicide terbuthylazine in agriculture workers and in the general population using urine and hair specimens.

PubMed

Mercadante, Rosa; Polledri, Elisa; Bertazzi, Pier Alberto; Fustinoni, Silvia

2013-10-01

The aim of this work was to evaluate short-term and long-term exposure to terbuthylazine (TBA) in agriculture workers (AW), rural residents (RR), and urban residents (UR) using urine and hair specimens. Twelve AW, 13 RR, and 17 UR were included in the study. Urine spot samples were collected with two different protocols. AW urine samples were collected before the application season (February, U0), at bedtime on the day of TBA application (March-May, U1), and prior to the next shift on the day after TBA application (U2). RR and UR urine samples were collected on any day during the application season (Ue). Hair samples were collected for all subjects before the application season (February, H0) and at the end of the season (June, H1). TBA and its metabolite desethylterbuthylazine (DET) were measured by liquid chromatography coupled with triple quadrupole mass spectrometry detection. DET was exclusively found in urine, while TBA was mostly found in the hair. In the AW, the urinary levels of DET were not detected in the U0 samples, and they increased to median levels of 1.81 and 2.94μg/L in the U1 and U2 samples, respectively (p<0.001). In the RR and UR, DET was not detected in the Ue samples. In the UR, TBA was not detected in the H0 samples, and the median levels of TBA were 0.01ng/mg hair in both the AW and RR. In the H1 samples, the median TBA levels were not detected, 0.01, and 0.08ng/mg hair in the UR, RR, and AW, respectively (p<0.001). Urinary DET and hair TBA are promising candidates for biomonitoring short- and long-term exposure to TBA. The use of this herbicide in agriculture leads to exposure in rural residents. © 2013.

Pine Bluff Arsenal Fish Survey.

DTIC Science & Technology

2000-03-01

6). Sites Collected: 39 Bowfins (Amiidae) Amia calva Linnaeus - Bowfin The bowfin is a primitive member of the monotypic family Amiidae, an...spotted gar (Lepisosteus oculatus), longnose gar (Lepisosteus osseus), bowfin ( Amia calva ), goldeye (Hiodon alosoides), goldfish (Carassius auratus...included the spotted gar (Lepisosteus oculatus),longnose gar (Lepisosteus osseus), bowfin ( Amia calva ), gizzard shad (Dorosoma cepedianum), threadfin

76 FR 29235 - Agency Information Collection Activities; Submission to OMB for Review and Approval; Comment...

Federal Register 2010, 2011, 2012, 2013, 2014

2011-05-20

.... Please note that EPA's policy is that public comments, whether submitted electronically or in paper, will... learning to perform quantitative hot-spot analyses; new burden associated with using the MOVES model for..., adjustment for increased burden associated with quantitative hot-spot analyses, an adjustment for the...

New record for the invasive Spotted Wing Drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae) in Anillaco, Argentina

USDA-ARS?s Scientific Manuscript database

The invasive Spotted Wing Drosophila (SWD), Drosophila suzukii Matsumura, is reported for the first time in La Rioja, Argentina. This represents a major range expansion for this species. The natural enemies of SWD, Leptopilina clavipes and Ganaspis hookeri were also collected with the SWD at the s...

The changes of serum proteome and tissular pathology in mouse induced by botulinum toxin E injection.

PubMed

Wang, J F; Mao, X Y; Zhao, C

2014-01-01

The experiment were performed to investigate the poisoning-related proteins and main pathological changes after mouse suffered from injection of botulinum toxin serotype E. Dose of 0.75 LD50 botulinum toxin serotype E per mice were administrated by intraperitoneal injection. Survival mouse were picked as experimental group. The blood were collected from orbital blood and serum sample was separated by centrifugation. The heart, liver, spleen, lung, kidney were fixed in 10 % neutral buffered formalin and then developed paraffin sections. Serum protein components were analyzed by SDS-PAGE gel electrophoresis coupled with 2-DE SDS-PAGE gel electrophoresis. Differentially expressed proteins were analyzed by PDQUest8.0 software and subjected to ion trap mass spectrometry equipped with a high performance liquid chromatography system. The observation of pathological section showed that heart, liver, spleen, lung, kidney exhibited pathological changes in different degree, especially in heart, liver and lung tissues. Heart muscle tissue display serious inflammatory response, heart muscle fiber compulsively expanded and filled with erythrocyte and inflammatory exudates, some heart muscle fiber ruptured, even necrosis; hepatic cell in edge of liver occur apoptosis and some hepatic cell have disintegrated, and even died; pulmonary alveoli broken and partial vein filled with blood. Serum proteins component present a significant changes between control serum and botulism in 24 h by SDS-PAGE gel electrophoresis and 2-DE-SDS-PAGE gel electrophoresis. Twenty differentially expressed protein spots were observed in 2-DE profiles, in which 14 protein spots were undetectable in serum proteome under botulism, 3 protein spots exclusively expressed in state of botulism, 3 protein spots were low-expressed in serum proteome under botulism. Fourteen proteins have been identified among 20 spots elected on two-dimensional electrophoresis gels. Crystal proteins family exclusively expressed in control group serum. Haptoglobin were low-expressed under botulism in serum protein components, however, serum amyloid A only expressed in serum sample under botulism in 24 h, which were verified by Western-blot. Identified proteins involved in energy metabolism, cellular stress response, transcription, body defense and cell proliferation. These findings represent the first report of BoNT-induced changes in serum proteome and histopathology, and reinforce the utility of applying proteomic tools to the study of system-wide biological processes in normal and botulism.

The Median Isn't the Message: Elucidating Nutrient Hot spots and Hot Moments in a Sierra Nevada Forest Soil

NASA Astrophysics Data System (ADS)

Barnes, M. E.; Hart, S. C.; Johnson, D. W.; Meadows, M. W.

2015-12-01

Most biogeochemical studies in forests have concentrated on nutrient pools and transformations occurring at relatively large spatial scales (i.e., stand or small catchment), over monthly or annual time scales. Many of these studies have also focused on the average or medial values observed across the spatial or temporal scale studied, discounting outliers. However, extremely high values found consistently (hot spot) or infrequently (hot moment) at a given soil microsite may be critical for nutrient acquisition by organisms and nutrient retention by terrestrial ecosystems. We have been evaluating soil nutrient hot-spot and hot-moment phenomena vertically (to a 60-cm depth) and horizontally (2-m sampling interval within a 6 m x 6 m grid) in two areas within a mixed-conifer, Sierran forest experiencing a Mediterranean-type climate. Nutrient fluxes in space and time were measured using ion exchange resin capsules placed at various depths and collected at two times (first significant precipitation in fall and post-snowmelt in spring) per year. Our previous work over a single year showed that fluxes of Ca2+ and Mg2+ in mineral soil were substantially greater in the spring than in the fall, suggesting that soil water was a major factor in controlling these nutrient fluxes. The opposite pattern was found for NH4+ and Na+, where greater fluxes occurred following the first precipitation event in fall. Here, we report new data over two additional years at these same sites. Over the entire 3-year study, nutrient fluxes were greater in the fall for all mineral soil nutrients except Ca2+ and Mg2+. Calcium fluxes were consistent with previous results; however, Mg2+ demonstrated no statistical significance between fall and spring sampling dates. Generally, the number of high statistical outliers persisted through time for Ca2+ and Mg2+, suggesting hot spots for these nutrients. In contrast, large seasonal and annual changes in the number of high statistical outliers occurred for NH4+, NO3-, and PO43-, nutrients whose availabilities are more mediated by microbial activity than base cations. Further elucidation of the mechanisms responsible for nutrient hot spot-hot moment phenomena within soil should be invaluable for improving the predictive capacity of biogeochemical models and for scaling these models across space and time.

Speciation of Soil Phosphorus Assessed by XANES Spectroscopy at Different Spatial Scales

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

Hesterberg, Dean; McNulty, Ian; Thieme, Juergen

Precise management of soil phosphorus (P) to meet competing demands of agriculture and environmental protection can benefit from more comprehensive characterization of P speciation in soils. Our objectives were to provide spatial context for spectroscopic analyses of soil P speciation in relation to molecular-scale species and landscape-scale management of P, and to compare soil P-species diversity from spectroscopic measurements at submicron and millimeter scales. The spatial range of ~26 orders of magnitude between atomic and field scales presents a challenge to upscaling and downscaling information from spectroscopic analyses of soils. Scanning fluorescence X-ray microscopy images of a 50-mm ´ 45-mmmore » area of an organic soil sample showed heterogeneous distributions of P, Al, and Si. Microscale X-ray absorption near edge structure (μ-XANES) spectra collected at the P K-edge from 12 spots on the soil sample exhibited diverse features that indicated variations in highly localized P speciation. Linear combination fitting analysis of the μ-XANES spectra included various proportions of three standards that appeared in fits for most spots and five standards that appeared in fits for one spot each. The fit to a bulk-soil spectrum was dominated by two of the common standards in the μ-XANES fits, and a fit to the sum of m-XANES spectra included four of the standards. Lastly, these results illustrate a gain in P species sensitivity from spatially resolved XANES analysis. Integrating spectroscopic analyses from multiple scales determines soil P species diversity and will ultimately help connect speciation to the chemical reactivity and mobility of P in soils.« less

Speciation of Soil Phosphorus Assessed by XANES Spectroscopy at Different Spatial Scales

DOE PAGES

Hesterberg, Dean; McNulty, Ian; Thieme, Juergen

2017-07-27

Precise management of soil phosphorus (P) to meet competing demands of agriculture and environmental protection can benefit from more comprehensive characterization of P speciation in soils. Our objectives were to provide spatial context for spectroscopic analyses of soil P speciation in relation to molecular-scale species and landscape-scale management of P, and to compare soil P-species diversity from spectroscopic measurements at submicron and millimeter scales. The spatial range of ~26 orders of magnitude between atomic and field scales presents a challenge to upscaling and downscaling information from spectroscopic analyses of soils. Scanning fluorescence X-ray microscopy images of a 50-mm ´ 45-mmmore » area of an organic soil sample showed heterogeneous distributions of P, Al, and Si. Microscale X-ray absorption near edge structure (μ-XANES) spectra collected at the P K-edge from 12 spots on the soil sample exhibited diverse features that indicated variations in highly localized P speciation. Linear combination fitting analysis of the μ-XANES spectra included various proportions of three standards that appeared in fits for most spots and five standards that appeared in fits for one spot each. The fit to a bulk-soil spectrum was dominated by two of the common standards in the μ-XANES fits, and a fit to the sum of m-XANES spectra included four of the standards. Lastly, these results illustrate a gain in P species sensitivity from spatially resolved XANES analysis. Integrating spectroscopic analyses from multiple scales determines soil P species diversity and will ultimately help connect speciation to the chemical reactivity and mobility of P in soils.« less

Analysis of biodiesel and biodiesel-petrodiesel blends by high performance thin layer chromatography combined with easy ambient sonic-spray ionization mass spectrometry.

PubMed

Eberlin, Livia S; Abdelnur, Patricia V; Passero, Alan; de Sa, Gilberto F; Daroda, Romeu J; de Souza, Vanderlea; Eberlin, Marcos N

2009-08-01

High performance thin layer chromatography (HPTLC) combined with on-spot detection and characterization via easy ambient sonic-spray ionization mass spectrometry (EASI-MS) is applied to the analysis of biodiesel (B100) and biodiesel-petrodiesel blends (BX). HPTLC provides chromatographic resolution of major components whereas EASI-MS allows on-spot characterization performed directly on the HPTLC surface at ambient conditions. Constituents (M) are detected by EASI-MS in a one component-one ion fashion as either [M + Na](+) or [M + H](+). For both B100 and BX samples, typical profiles of fatty acid methyl esters (FAME) detected as [FAME + Na](+) ions allow biodiesel typification. The spectrum of the petrodiesel spot displays a homologous series of protonated alkyl pyridines which are characteristic for petrofuels (natural markers). The spectrum for residual or admixture oil spots is characterized by sodiated triglycerides [TAG + Na](+). The application of HPTLC to analyze B100 and BX samples and its combination with EASI-MS for on-spot characterization and quality control is demonstrated.

Water-Soluble Dried Blood Spot in Protein Analysis: A Proof-of-Concept Study.

PubMed

Rosting, Cecilie; Gjelstad, Astrid; Halvorsen, Trine Grønhaug

2015-08-04

In the present work human chorionic gonadotropin (hCG) was used as a model protein in a proof-of-concept study combining water-soluble dried blood spot (DBS) material in liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based protein analysis. A water-soluble material consisting of commercially available carboxymethyl cellulose (CMC) was evaluated as sampling material for this purpose. The material dissolved readily at physiological pH. Different sample preparation methods were evaluated, and in the final method, 15 μL of whole blood was deposited and dried on CMC before the whole spot was dissolved prior to cleanup by immunoaffinity extraction, tryptic digest, and preconcentration by solid-phase extraction (SPE). The results indicated complete dissolution of hCG from the spots, acceptable limit of detection (LOD) (0.1 IU/mL), linearity (R(2) = 0.959), accuracy (16%), and precision (≤22%). Long-term stability (45 days) of hCG in dried spots at reduced temperatures (≤8 °C) was also demonstrated. The analyte recovery was comparable to the commercially available nonsolvable cellulose material (FTA DMPK-C card).

OSIRIS-REx Flight Dynamics and Navigation Design

NASA Astrophysics Data System (ADS)

Williams, B.; Antreasian, P.; Carranza, E.; Jackman, C.; Leonard, J.; Nelson, D.; Page, B.; Stanbridge, D.; Wibben, D.; Williams, K.; Moreau, M.; Berry, K.; Getzandanner, K.; Liounis, A.; Mashiku, A.; Highsmith, D.; Sutter, B.; Lauretta, D. S.

2018-06-01

OSIRIS-REx is the first NASA mission to return a sample of an asteroid to Earth. Navigation and flight dynamics for the mission to acquire and return a sample of asteroid 101955 Bennu establish many firsts for space exploration. These include relatively small orbital maneuvers that are precise to ˜1 mm/s, close-up operations in a captured orbit about an asteroid that is small in size and mass, and planning and orbit phasing to revisit the same spot on Bennu in similar lighting conditions. After preliminary surveys and close approach flyovers of Bennu, the sample site will be scientifically characterized and selected. A robotic shock-absorbing arm with an attached sample collection head mounted on the main spacecraft bus acquires the sample, requiring navigation to Bennu's surface. A touch-and-go sample acquisition maneuver will result in the retrieval of at least 60 grams of regolith, and up to several kilograms. The flight activity concludes with a return cruise to Earth and delivery of the sample return capsule (SRC) for landing and sample recovery at the Utah Test and Training Range (UTTR).

Association of drinking-water source and use characteristics with urinary antimony concentrations.

PubMed

Makris, Konstantinos C; Andra, Syam S; Herrick, Lisa; Christophi, Costas A; Snyder, Shane A; Hauser, Russ

2013-03-01

Environmental factors, such as storage time, frequency of bottle reuse and temperature, have been shown to facilitate antimony (Sb) leaching from water- and food-packaging materials. The globally escalating consumption of water packaged in Sb-containing bottles, such as that of polyethylene terephthalate (PET), could increase human daily Sb doses. This study set out to investigate the relationship between drinking-water source, use characteristics, and urinary Sb concentrations (U-Sb) accompanied with survey responses of a healthy (n=35) Cypriot participant pool. One spot urine sample was collected during administration of questionnaire, while a second spot urine sample was collected from the same individual about 7 days later. Urinary and water Sb concentrations were measured with an inductively coupled plasma mass spectrometer. Survey responses showed that bottled water summed over various volumes and plastic types, such as polycarbonate and PET contributed to an average 61% of daily water consumption. Water sources such as tap, mobile stations (explained in a following section), and well water contributed to 24%, 14%, and 2% of an individual's daily water consumption pattern, respectively. Average daily potable water use of both bottled and tap water by individuals consisted of 65% drinking-water, while the remaining 35% was water used for preparing cold and hot beverages, such as, tea, coffee, and juices. A significant (P=0.02) association between per capita water consumption from PET bottles and urinary creatinine-unadjusted concentrations was observed, but this relationship did not remain after inclusion of covariates in a multivariate regression model. In the creatinine-adjusted regression model, only gender (female) was a significant (P<0.01) predictor of U-Sb, after adjusting for several covariates. It is proposed that consumption data collection on various water uses and sources among individuals could perhaps decrease the uncertainty associated with derivations of acceptable daily Sb intakes.

Design of the sample cell in near-field surface-enhanced Raman scattering by finite difference time domain method

NASA Astrophysics Data System (ADS)

Li, Yaqin; Jian, Guoshu; Wu, Shifa

2006-11-01

The rational design of the sample cell may improve the sensitivity of surface-enhanced Raman scattering (SERS) detection in a high degree. Finite difference time domain (FDTD) simulations of the configuration of Ag film-Ag particles illuminated by plane wave and evanescent wave are performed to provide physical insight for design of the sample cell. Numerical solutions indicate that the sample cell can provide more "hot spots' and the massive field intensity enhancement occurs in these "hot spots'. More information on the nanometer character of the sample can be got because of gradient-field Raman (GFR) of evanescent wave.

Detection of hepatitis B virus infection markers in dried plasma spots among patients in Congo-Brazzaville.

PubMed

Alidjinou, Enagnon Kazali; Moukassa, Donatien; Sané, Famara; Twagirimana Nyenyeli, Séraphin; Akoko, Estina Chandrelle; Mountou, Michèle Valy; Bocket, Laurence; Ibara, Jean-Rosaire; Hober, Didier

2014-03-01

The detection of hepatitis B virus (HBV) infection markers by using dried plasma spots from 32 patients living in Congo has been assessed. Considering frozen plasma samples as gold standard, the sensitivity and specificity of HBV serologic markers detection in dried plasma eluted from filter paper were 100%. The sensitivity and the specificity of HBV DNA detection reached 96% and 100%, respectively, with plasma samples dried on filter paper compared to standard samples. Dried plasma samples can represent an alternative to conventional sampling for HBV detection and management of the infection in developing countries. Copyright © 2014 Elsevier Inc. All rights reserved.

Spectral reproducibility and quantification of peptides in MALDI of samples prepared by micro-spotting.

PubMed

Bae, Yong Jin; Park, Kyung Man; Ahn, Sung Hee; Moon, Jeong Hee; Kim, Myung Soo

2014-08-01

Previously, we reported that MALDI spectra of peptides became reproducible when temperature was kept constant. Linear calibration curves derived from such spectral data could be used for quantification. Homogeneity of samples was one of the requirements. Among the three popular matrices used in peptide MALDI [i.e., α-cyano-4-hydroxycinnamic acid (CHCA), 2,5-dihydroxybenzoic acid (DHB), and sinapinic acid (SA)], homogeneous samples could be prepared by conventional means only for CHCA. In this work, we showed that sample preparation by micro-spotting improved the homogeneity for all three cases.

Evaluation of HBsAg and anti-HBc assays in saliva and dried blood spot samples according HIV status.

PubMed

Flores, Geane Lopes; Cruz, Helena Medina; Potsch, Denise Vigo; May, Silvia Beatriz; Brandão-Mello, Carlos Eduardo; Pires, Marcia Maria Amendola; Pilotto, Jose Henrique; Lewis-Ximenez, Lia Laura; Lampe, Elisabeth; Villar, Livia Melo

2017-09-01

Influence of HIV status in HBV markers detection in saliva and dried blood spots (DBS) was not well established. This study aims to evaluate the performance of optimized commercial immunoassay for identifying HBsAg and anti-HBc in saliva and DBS according HIV status. A sum of 535 individuals grouped as HIV + , HBV + , HIV/HBV + and HIV/HBV- were recruited where 347 and 188 were included for HBsAg and anti-HBc evaluation, respectively. Serum, DBS collected in Whatman 903 paper and saliva obtained using salivette device were analyzed using EIA. Increased sample volume and ROC curve analysis for cut off determination were used for DBS and saliva testing. HBsAg detection in saliva and DBS exhibited sensitivities of 80.9% and 85.6% and specificities of 86.8% and 96.3%. Sensitivity of anti-HBc in saliva and DBS were 82.4% and 76.9% and specificities in saliva and DBS were 96.9% and 91.7%. Low sensitivities were observed for HBsAg (62%) and anti-HBc (47%) detection in saliva of HIV/HBV+ individuals. OD values were also lower for HBsAg detection in DBS and saliva of HIV/HBV+ individuals compared to their serum samples. Statistical significance was found for sensitivities in HBsAg detection between saliva and DBS demonstrating high sensitivity for DBS specimens. In conclusion, HIV status or antiretroviral treatment appears to interfere in the performance of HBsAg and anti-HBc detection in DBS and saliva samples using the adapted commercial EIA. Copyright © 2017 Elsevier B.V. All rights reserved.

Sports drug testing using complementary matrices: Advantages and limitations.

PubMed

Thevis, Mario; Geyer, Hans; Tretzel, Laura; Schänzer, Wilhelm

2016-10-25

Today, routine doping controls largely rely on testing whole blood, serum, and urine samples. These matrices allow comprehensively covering inorganic as well as low and high molecular mass organic analytes relevant to doping controls and are collecting and transferring from sampling sites to accredited anti-doping laboratories under standardized conditions. Various aspects including time and cost-effectiveness as well as intrusiveness and invasiveness of the sampling procedure but also analyte stability and breadth of the contained information have been motivation to consider and assess values potentially provided and added to modern sports drug testing programs by alternative matrices. Such alternatives could be dried blood spots (DBS), dried plasma spots (DPS), oral fluid (OF), exhaled breath (EB), and hair. In this review, recent developments and test methods concerning these alternative matrices and expected or proven contributions as well as limitations of these specimens in the context of the international anti-doping fight are presented and discussed, guided by current regulations for prohibited substances and methods of doping as established by the World Anti-Doping Agency (WADA). Focusing on literature published between 2011 and 2015, examples for doping control analytical assays concerning non-approved substances, anabolic agents, peptide hormones/growth factors/related substances and mimetics, β 2 -agonists, hormone and metabolic modulators, diuretics and masking agents, stimulants, narcotics, cannabinoids, glucocorticoids, and beta-blockers were selected to outline the advantages and limitations of the aforementioned alternative matrices as compared to conventional doping control samples (i.e. urine and blood/serum). Copyright © 2016 Elsevier B.V. All rights reserved.

Density-based kinetics for mesoscale simulations of detonation initiation in energetic materials

NASA Astrophysics Data System (ADS)

Jackson, Thomas Luther; Zhang, Ju

2017-07-01

In this work we present one- and two-dimensional mesoscale simulations of detonation initiation in energetic materials. We solve the reactive Euler equations, with the energy equation augmented by a power deposition term. The reaction rate at the mesoscale is modelled using a density-based kinetics scheme, adapted from standard 'Ignition and Growth' models. The deposition term is based on previous results of simulations of void collapse at the microscale, modelled at the mesoscale as hot spots. For an isolated hot spot in a homogeneous medium, it is found that a critical size of the hot spots exists. If the hot spots exceed the critical size, initiation of detonation can be achieved. For sub-critical hot-spot sizes, we show that it takes a collection of hot spots to achieve detonation. We also carry out two-dimensional mesoscale simulations of random packs of HMX crystals in a binder, and show that the transition between no detonation and detonation depends on the number density of the hot spots, the initial radius of the hot spot, the post-shock pressure of an imposed shock, and the amplitude of the power deposition term.

A short working distance multiple crystal x-ray spectrometer

USGS Publications Warehouse

Dickinson, B.; Seidler, G.T.; Webb, Z.W.; Bradley, J.A.; Nagle, K.P.; Heald, S.M.; Gordon, R.A.; Chou, I.-Ming

2008-01-01

For x-ray spot sizes of a few tens of microns or smaller, a millimeter-sized flat analyzer crystal placed ???1 cm from the sample will exhibit high energy resolution while subtending a collection solid angle comparable to that of a typical spherically bent crystal analyzer (SBCA) at much larger working distances. Based on this observation and a nonfocusing geometry for the analyzer optic, we have constructed and tested a short working distance (SWD) multicrystal x-ray spectrometer. This prototype instrument has a maximum effective collection solid angle of 0.14 sr, comparable to that of 17 SBCA at 1 m working distance. We find good agreement with prior work for measurements of the Mn K?? x-ray emission and resonant inelastic x-ray scattering for MnO, and also for measurements of the x-ray absorption near-edge structure for Dy metal using L??2 partial-fluorescence yield detection. We discuss future applications at third- and fourth-generation light sources. For concentrated samples, the extremely large collection angle of SWD spectrometers will permit collection of high-resolution x-ray emission spectra with a single pulse of the Linac Coherent Light Source. The range of applications of SWD spectrometers and traditional multi-SBCA instruments has some overlap, but also is significantly complementary. ?? 2008 American Institute of Physics.

Analysis of polyfluoroalkyl substances and bisphenol A in dried blood spots by liquid chromatography tandem mass spectrometry.

PubMed

Ma, Wanli; Kannan, Kurunthachalam; Wu, Qian; Bell, Erin M; Druschel, Charlotte M; Caggana, Michele; Aldous, Kenneth M

2013-05-01

Dried blood spots (DBS), collected as part of the newborn screening program (NSP) in the USA, is a valuable resource for studies on environmental chemical exposures and associated health outcomes in newborns. Nevertheless, determination of concentrations of environmental chemicals in DBS requires assays with great sensitivity, as the typical volume of blood available on a DBS with 16-mm diameter disc is approximately 50 μL. In this study, we developed a liquid-liquid extraction and high-performance liquid chromatography/tandem mass spectrometry method for the detection of perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and bisphenol A (BPA) in DBS. The method was validated for accuracy, precision, and sensitivity, by spiking of target chemicals at different levels on Whatman 903 filter cards, which is used in the collection of DBS by the NSP. Contamination arising from collection, storage, and handling of DBS is an important issue to be considered in the analysis of trace levels of environmental chemicals in DBS. For the evaluation of the magnitude of background contamination, field blanks were prepared from unspotted portions of DBS filter cards collected by the NSP. The method was applied for the measurement of PFOS, PFOA, and BPA in 192 DBS specimens provided by NSP of New York State. PFOS and PFOA were detected in 100 % of the specimens analyzed. The concentrations of PFOS and PFOA measured in DBS were similar to those reported earlier in the whole blood samples of newborns. BPA was also found in 86 % of the specimens at concentrations ranging from 0.2 to 36 ng/mL (excluding two outliers). Further studies are needed to evaluate the sources of BPA exposures and health outcomes in newborns.

Simultaneous determination of cocaine and opiates in dried blood spots by electrospray ionization tandem mass spectrometry.

PubMed

Antelo-Domínguez, Ángel; Cocho, José Ángel; Tabernero, María Jesús; Bermejo, Ana María; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

2013-12-15

A sample pre-treatment method based on blood spot collection filter cards was optimized as a means of using small volume samples for the screening and confirmation of cocaine and opiates abuse. Dried blood spots (DBSs) were prepared by dispersing 20 µL of whole blood specimens previously mixed with the internal standards (deuterated analogs of each target), and subjecting the whole DBS to extraction with 5 mL of methanol under orbital-horizontal shaking (180 rpm) for 10 min. Determinations were based on direct electrospray ionization tandem mass spectrometry (ESI-MS/MS) by injecting the re-dissolved methanol extract with the delivery solution (acetonitrile-water-formic acid, 80:19.875:0.125) at a flow rate of 60 µL min(-1), and using multiple reaction monitoring (MRM) mode with the m/z (precursor ion)→m/z (product ion) transitions for acquisition. Matrix effect has been found to be statistically significant (Multiple Range Test) when assessing cocaine, BZE, codeine and morphine, and the use of the standard addition method (dispersion of whole blood previously mixed with standards onto the filter papers) was needed for accurate determinations. The developed DBS-ESI-MS/MS procedure offered good intra-day and inter-day precisions (lower than 10% and 12%, respectively), as well as good intra-day and inter-day accuracies (inter-day absolute recoveries, expressed as the mean analytical recovery over three target concentration levels, of 103%, 100%, 101%, 98% and 100% for cocaine, BZE, codeine, morphine and 6-MAM, respectively). The high sensitivity inherent to MS/MS determinations combined with the minimal dilution of sample allowed low limits of quantification for all targets, and the developed method results therefore adequate for cocaine and opiates screening and confirmation purposes. The procedure was finally applied to DBSs prepared from whole blood from polydrug abusers, and results were compared with those obtained after a conventional sample pretreatment method based on solid phase extraction for plasma specimens and gas chromatography-mass spectrometry. © 2013 Elsevier B.V. All rights reserved.

Determining population and developmental pharmacokinetics of metronidazole using plasma and dried blood spot samples from premature infants.

PubMed

Cohen-Wolkowiez, Michael; Sampson, Mario; Bloom, Barry T; Arrieta, Antonio; Wynn, James L; Martz, Karen; Harper, Barrie; Kearns, Gregory L; Capparelli, Edmund V; Siegel, David; Benjamin, Daniel K; Smith, P Brian

2013-09-01

Limited pharmacokinetic (PK) data of metronidazole in premature infants have led to various dosing recommendations. Surrogate efficacy targets for metronidazole are ill-defined and therefore aimed to exceed minimum inhibitory concentration of organisms responsible for intra-abdominal infections. We evaluated the PK of metronidazole using plasma and dried blood spot samples from infants ≤32 weeks gestational age in an open-label, PK, multicenter (N = 3) study using population PK modeling (NONMEM). Monte Carlo simulations (N = 1000 virtual subjects) were used to evaluate the surrogate efficacy target. Metabolic ratios of parent and metabolite were calculated. Twenty-four premature infants (111 plasma and 51 dried blood spot samples) were enrolled: median (range) gestational age at birth 25 (23-31) weeks, postnatal age 27 (1-82) days, postmenstrual age 31 (24-39) weeks and weight 740 (431-1466) g. Population clearance (L/h/kg) was 0.038 × (postmenstrual age/30) and volume of distribution (L/kg) of 0.93. PK parameter estimates and precision were similar between plasma and dried blood spot samples. Metabolic ratios correlated with clearance. Simulations suggested the majority of infants in the neonatal intensive care unit (>80%) would meet the surrogate efficacy target using postmenstrual age-based dosing.

Dried blood spots for the enzymatic diagnosis of lysosomal storage diseases in dogs and cats.

PubMed

Sewell, Adrian C; Haskins, Mark E; Giger, Urs

2012-12-01

In people, lysosomal storage diseases (LSD) can be diagnosed by assaying enzyme activities in dried blood spots (DBS). The aim of this study was to evaluate the feasibility of using DBS samples from dogs and cats to measure lysosomal enzymatic activities and diagnose LSD. Drops of fresh whole blood collected in EDTA from dogs and cats with known or suspected LSD and from clinically healthy dogs and cats were placed on neonatal screening cards, dried, and mailed to the Metabolic Laboratory, University Children's Hospital, Frankfurt, Germany. Activities of selected lysosomal enzymes were measured using fluorescent substrates in a 2-mm diameter disk (~2.6 μL blood) punched from the DBS. Results were expressed as nmol substrate hydrolyzed per mL of blood per minute or hour. Reference values were established for several lysosomal enzyme activities in DBS from dogs and cats; for most enzymes, activities were higher than those published for human samples. Activities of β-glucuronidase, N-acetylglucosamine-4-sulfatase (arylsulfatase B), α-mannosidase, α-galactosidase, α-fucosidase, and hexosaminidase A were measureable in DBS from healthy cats and dogs; α-iduronidase activity was measureable only in cats. In samples from animals with LSD, markedly reduced activity of a specific enzyme was found. In contrast, in samples from cats affected with mucolipidosis II, activities of lysosomal enzymes were markedly increased. Measurement of lysosomal enzyme activities in DBS provides an inexpensive, simple, and convenient method to screen animals for suspected LSD and requires only a small sample volume. For diseases in which the relevant enzyme activity can be measured in DBS, a specific diagnosis can be made. © 2012 American Society for Veterinary Clinical Pathology.

Pre-analytical Factors Influence Accuracy of Urine Spot Iodine Assessment in Epidemiological Surveys.

PubMed

Doggui, Radhouene; El Ati-Hellal, Myriam; Traissac, Pierre; El Ati, Jalila

2018-03-26

Urinary iodine concentration (UIC) is commonly used to assess iodine status of subjects in epidemiological surveys. As pre-analytical factors are an important source of measurement error and studies about this phase are scarce, our objective was to assess the influence of urine sampling conditions on UIC, i.e., whether the child ate breakfast or not, urine void rank of the day, and time span between last meal and urine collection. A nationwide, two-stage, stratified, cross-sectional study including 1560 children (6-12 years) was performed in 2012. UIC was determined by the Sandell-Kolthoff method. Pre-analytical factors were assessed from children's mothers by using a questionnaire. Association between iodine status and pre-analytical factors were adjusted for one another and socio-economic characteristics by multivariate linear and multinomial regression models (RPR: relative prevalence ratios). Skipping breakfast prior to morning urine sampling decreased UIC by 40 to 50 μg/L and the proportion of UIC < 100 μg/L was higher among children having those skipped breakfast (RPR = 3.2[1.0-10.4]). In unadjusted analyses, UIC was less among children sampled more than 5 h from their last meal. UIC decreased with rank of urine void (e.g., first vs. second, P < 0.001); also, the proportion of UIC < 100 μg/L was greater among 4th rank samples (vs. second RPR = 2.1[1.1-4.0]). Subjects' breakfast status and urine void rank should be accounted for when assessing iodine status. Providing recommendations to standardize pre-analytical factors is a key step toward improving accuracy and comparability of survey results for assessing iodine status from spot urine samples. These recommendations have to be evaluated by future research.

Predictors and long-term reproducibility of urinary phthalate metabolites in middle-aged men and women living in urban Shanghai

PubMed Central

Starling, Anne P.; Engel, Lawrence S.; Calafat, Antonia M.; Koutros, Stella; Satagopan, Jaya M.; Yang, Gong; Matthews, Charles E.; Cai, Qiuyin; Buckley, Jessie P.; Ji, Bu-Tian; Cai, Hui; Chow, Wong-Ho; Zheng, Wei; Gao, Yu-Tang; Rothman, Nathaniel; Xiang, Yong-Bing; Shu, Xiao-Ou

2015-01-01

Phthalate esters are man-made chemicals commonly used as plasticizers and solvents, and humans may be exposed through ingestion, inhalation, and dermal absorption. Little is known about predictors of phthalate exposure, particularly in Asian countries. Because phthalates are rapidly metabolized and excreted from the body following exposure, it is important to evaluate whether phthalate metabolites measured at a single point in time can reliably rank exposures to phthalates over a period of time. We examined the concentrations and predictors of phthalate metabolite concentrations among 50 middle-aged women and 50 men from two Shanghai cohorts, enrolled in 1997-2000 and 2002-2006, respectively. We assessed the reproducibility of urinary concentrations of phthalate metabolites in three spot samples per participant taken several years apart (mean interval between first and third sample was 7.5 years [women] or 2.9 years [men]), using Spearman's rank correlation coefficients and intra-class correlation coefficients. We detected ten phthalate metabolites in at least 50% of individuals for two or more samples. Participant sex, age, menopausal status, education, income, body mass index, consumption of bottled water, recent intake of medication, and time of day of collection of the urine sample were associated with concentrations of certain phthalate metabolites. The reproducibility of an individual's urinary concentration of phthalate metabolites across several years was low, with all intra-class correlation coefficients and most Spearman rank correlation coefficients ≤ 0.3. Only mono(2-ethylhexyl) phthalate, a metabolite of di(2-ethylhexyl)phthalate, had a Spearman rank correlation coefficient ≥ 0.4 among men, suggesting moderate reproducibility. These findings suggest that a single spot urine sample is not sufficient to rank exposures to phthalates over several years in an adult urban Chinese population. PMID:26255822

The Use of Dried Blood Spots for Pharmacokinetic Monitoring of Vemurafenib Treatment in Melanoma Patients.

PubMed

Nijenhuis, Cynthia M; Huitema, Alwin D R; Marchetti, Serena; Blank, Christian; Haanen, John B A G; van Thienen, Johannes V; Rosing, Hilde; Schellens, Jan H M; Beijnen, Jos H

2016-10-01

Pharmacokinetic monitoring is increasingly becoming an important part of clinical care of tyrosine kinase inhibitor treatment. Vemurafenib is an oral tyrosine kinase inhibitor that inhibits mutated serine/threonine protein kinase B-Raf (BRAF) and is approved for the treatment of adult patients with BRAF V600 mutation-positive unresectable or metastatic melanoma. The aim of this study was to establish the relationship between dried blood spot (DBS) and plasma concentrations of vemurafenib to enable the use of DBS sampling, which is a minimally invasive form of sample collection. In total, 43 paired plasma and DBS samples (in duplicate) were obtained from 8 melanoma patients on vemurafenib therapy and were analyzed using high-performance liquid chromatography-tandem mass spectrometry. Plasma concentrations were predicted from the DBS concentrations using 2 methods: (1) individual hematocrit correction and blood cell-to-plasma partitioning and (2) the calculated slope explaining the relationship between DBS and plasma concentrations (without individual hematocrit correction). Vemurafenib DBS concentrations and plasma concentrations showed a strong correlation (r = 0.964), and the relationship could be described by ([vemurafenib]plasma = [vemurafenib]DBS /0.64). The predicted plasma concentrations were within ±20% of the analyzed plasma concentrations in 97% and 100% of the samples for the methods with and without hematocrit correction, respectively. In conclusion, DBS concentrations and plasma concentrations of vemurafenib are highly correlated. Plasma concentrations can be predicted from DBS concentration using the blood cell-to-plasma partition and the average hematocrit value of this cohort (0.40 L/L). DBS sampling for pharmacokinetic monitoring of vemurafenib treatment can be used in clinical practice. © 2016, The American College of Clinical Pharmacology.

PRACTICAL STRATEGIES FOR PROCESSING AND ANALYZING SPOTTED OLIGONUCLEOTIDE MICROARRAY DATA

EPA Science Inventory

Thoughtful data analysis is as important as experimental design, biological sample quality, and appropriate experimental procedures for making microarrays a useful supplement to traditional toxicology. In the present study, spotted oligonucleotide microarrays were used to profile...

3D plasmonic nanoarchitectures for extreme light concentration

NASA Astrophysics Data System (ADS)

Arnob, Md Masud Parvez; Zhao, Fusheng; Shih, Wei-Chuan

2017-08-01

Plasmonic nanomaterials are known to concentrate incident light to their surfaces by collective electron oscillation. Plasmonic hot-spot refers to locations where electromagnetic fields are particularly enhanced relative to the incident field. Traditional plasmonic nanomaterials are 1D (e.g., colloidal nanoparticles) or 2D (lithographically patterned nanostructure arrays) in nature, which typically result in sparse field concentration patterns. To improve efficiency and better utilization of hot-spots, we investigate 3D plasmonic nanoarchitecture where abundant hot-spots are formed in a 3D volumetric fashion, a feature drastically departing from traditional nanostructures.

Geographic Distribution of Leishmania Species in Ecuador Based on the Cytochrome B Gene Sequence Analysis.

PubMed

Kato, Hirotomo; Gomez, Eduardo A; Martini-Robles, Luiggi; Muzzio, Jenny; Velez, Lenin; Calvopiña, Manuel; Romero-Alvarez, Daniel; Mimori, Tatsuyuki; Uezato, Hiroshi; Hashiguchi, Yoshihisa

2016-07-01

A countrywide epidemiological study was performed to elucidate the current geographic distribution of causative species of cutaneous leishmaniasis (CL) in Ecuador by using FTA card-spotted samples and smear slides as DNA sources. Putative Leishmania in 165 samples collected from patients with CL in 16 provinces of Ecuador were examined at the species level based on the cytochrome b gene sequence analysis. Of these, 125 samples were successfully identified as Leishmania (Viannia) guyanensis, L. (V.) braziliensis, L. (V.) naiffi, L. (V.) lainsoni, and L. (Leishmania) mexicana. Two dominant species, L. (V.) guyanensis and L. (V.) braziliensis, were widely distributed in Pacific coast subtropical and Amazonian tropical areas, respectively. Recently reported L. (V.) naiffi and L. (V.) lainsoni were identified in Amazonian areas, and L. (L.) mexicana was identified in an Andean highland area. Importantly, the present study demonstrated that cases of L. (V.) braziliensis infection are increasing in Pacific coast areas.

Scanning optical microscope with long working distance objective

DOEpatents

Cloutier, Sylvain G.

2010-10-19

A scanning optical microscope, including: a light source to generate a beam of probe light; collimation optics to substantially collimate the probe beam; a probe-result beamsplitter; a long working-distance, infinity-corrected objective; scanning means to scan a beam spot of the focused probe beam on or within a sample; relay optics; and a detector. The collimation optics are disposed in the probe beam. The probe-result beamsplitter is arranged in the optical paths of the probe beam and the resultant light from the sample. The beamsplitter reflects the probe beam into the objective and transmits resultant light. The long working-distance, infinity-corrected objective is also arranged in the optical paths of the probe beam and the resultant light. It focuses the reflected probe beam onto the sample, and collects and substantially collimates the resultant light. The relay optics are arranged to relay the transmitted resultant light from the beamsplitter to the detector.

Validation of paper-based assay for rapid blood typing.

PubMed

Al-Tamimi, Mohammad; Shen, Wei; Zeineddine, Rania; Tran, Huy; Garnier, Gil

2012-02-07

We developed and validated a new paper-based assay for the detection of human blood type. Our method involves spotting a 3 μL blood sample on a paper surface where grouping antibodies have already been introduced. A thin film chromatograph tank was used to chromatographically elute the blood spot with 0.9% NaCl buffer for 10 min by capillary absorption. Agglutinated red blood cells (RBCs) were fixed on the paper substrate, resulting in a high optical density of the spot, with no visual trace in the buffer wicking path. Conversely, nonagglutinated RBCs could easily be eluted by the buffer and had low optical density of the spot and clearly visible trace of RBCs in the buffer wicking path. Different paper substrates had comparable ability to fix agglutinated blood, while a more porous substrate like Kleenex paper had enhanced ability to elute nonagglutinated blood. Using optimized conditions, a rapid assay for detection of blood groups was developed by spotting blood to antibodies absorbed to paper and eluted with 200 μL of 0.9% NaCl buffer directly by pipetting. RBCs fixation on paper accurately detected blood groups (ABO and RhD) using ascending buffer for 10 min or using a rapid elution step in 100/100 blood samples including 4 weak AB and 4 weak RhD samples. The assay has excellent reproducibility where the same blood group was obtained in 26 samples assessed in 2 different days. Agglutinated blood fixation on porous paper substrate provides a new, simple, and sensitive assay for rapid detection of blood group for point-of-care applications. © 2011 American Chemical Society

Normal Raman and surface enhanced Raman spectroscopic experiments with thin layer chromatography spots of essential amino acids using different laser excitation sources.

PubMed

István, Krisztina; Keresztury, Gábor; Szép, Andrea

2003-06-01

A comparative study of the feasibility and efficiency of Raman spectroscopic detection of thin layer chromatography (TLC) spots of some weak Raman scatterers (essential amino acids, namely, glycine and L-forms of alanine, serine, valine, proline, hydroxyproline, and phenylalanine) was carried out using four different visible and near-infrared (NIR) laser radiations with wavelengths of 532, 633, 785, and 1064 nm. Three types of commercial TLC plates were tested and the possibility of inducing surface enhanced Raman scattering (SERS) by means of Ag-sol was also investigated. The spectra obtained from spotted analytes adsorbed on TLC plates were of very different quality strongly depending on the excitation wavelength, the wetness of the samples, and the compounds examined. The best results were obtained with the simple silica TLC plate, and it has been established that the longest wavelength (lowest energy) NIR excitation of a Nd:YAG laser is definitely more suitable for generating normal Raman scattering of analyte spots than any of the visible radiations. Concerning SERS with application of Ag-sol to the TLC spots, 1-3 orders of magnitude enhancement was observed with wet samples, the greatest with the 532 nm radiation and gradually smaller with the longer wavelength excitations. It is shown, however, that due to severe adsorption-induced spectral distortions and increased sensitivity to microscopic inhomogeneity of the sample, none of the SERS spectra obtained with the dispersive Raman microscope operating in the visible region were superior to the best NIR normal FT-Raman spectra, as far as sample identification is concerned.

Normal Raman and surface enhanced Raman spectroscopic experiments with thin layer chromatography spots of essential amino acids using different laser excitation sources

NASA Astrophysics Data System (ADS)

István, Krisztina; Keresztury, Gábor; Szép, Andrea

2003-06-01

A comparative study of the feasibility and efficiency of Raman spectroscopic detection of thin layer chromatography (TLC) spots of some weak Raman scatterers (essential amino acids, namely, glycine and L-forms of alanine, serine, valine, proline, hydroxyproline, and phenylalanine) was carried out using four different visible and near-infrared (NIR) laser radiations with wavelengths of 532, 633, 785, and 1064 nm. Three types of commercial TLC plates were tested and the possibility of inducing surface enhanced Raman scattering (SERS) by means of Ag-sol was also investigated. The spectra obtained from spotted analytes adsorbed on TLC plates were of very different quality strongly depending on the excitation wavelength, the wetness of the samples, and the compounds examined. The best results were obtained with the simple silica TLC plate, and it has been established that the longest wavelength (lowest energy) NIR excitation of a Nd:YAG laser is definitely more suitable for generating normal Raman scattering of analyte spots than any of the visible radiations. Concerning SERS with application of Ag-sol to the TLC spots, 1-3 orders of magnitude enhancement was observed with wet samples, the greatest with the 532 nm radiation and gradually smaller with the longer wavelength excitations. It is shown, however, that due to severe adsorption-induced spectral distortions and increased sensitivity to microscopic inhomogeneity of the sample, none of the SERS spectra obtained with the dispersive Raman microscope operating in the visible region were superior to the best NIR normal FT-Raman spectra, as far as sample identification is concerned.

Vigabatrin in dried plasma spots: validation of a novel LC-MS/MS method and application to clinical practice.

PubMed

Kostić, Nađa; Dotsikas, Yannis; Jović, Nebojša; Stevanović, Galina; Malenović, Anđelija; Medenica, Mirjana

2014-07-01

This paper presents a LC-MS/MS method for the determination of antiepileptic drug vigabatrin in dried plasma spots (DPS). Due to its zwitterionic chemical structure, a pre-column derivatization procedure was performed, aiming to yield enhanced ionization efficiency and improved chromatographic behaviour. Propyl chloroformate, in the presence of propanol, was selected as the best derivatization reagent, providing a strong signal along with reasonable run time. A relatively novel sample collection technique, DPS, was utilized, offering easy sample handling and analysis, using a sample in micro amount (∼5μL). Derivatized vigabatrin and its internal standard, 4-aminocyclohexanecarboxylic acid, were extracted by liquid-liquid extraction (LLE) and determined in positive ion mode by applying two SRM transitions per analyte. A Zorbax Eclipse XDB-C8 column (150×4.6mm, 5μm particle size) maintained at 30°C, was utilized with running mobile phase composed of acetonitrile: 0.15% formic acid (85:15, v/v). Flow rate was 550μL/min and total run time 4.5min. The assay exhibited excellent linearity over the concentration range of 0.500-50.0μg/mL, which is suitable for the determination of vigabatrin level after per os administration in children and youths with epilepsy, who were on vigabatrin therapy, with or without co-medication. Specificity, accuracy, precision, recovery, matrix-effect and stability were also estimated and assessed within acceptance criteria. Copyright © 2014 Elsevier B.V. All rights reserved.

Detection of feral GT73 transgenic oilseed rape (Brassica napus) along railway lines on entry routes to oilseed factories in Switzerland.

PubMed

Hecht, Mirco; Oehen, Bernadette; Schulze, Jürg; Brodmann, Peter; Bagutti, Claudia

2014-01-01

To obtain a reference status prior to cultivation of genetically modified oilseed rape (OSR, Brassica napus L.) in Switzerland, the occurrence of feral OSR was monitored along transportation routes and at processing sites. The focus was set on the detection of (transgenic) OSR along railway lines from the Swiss borders with Italy and France to the respective oilseed processing factories in Southern and Northern Switzerland (Ticino and region of Basel). A monitoring concept was developed to identify sites of largest risk of escape of genetically modified plants into the environment in Switzerland. Transport spillage of OSR seeds from railway goods cars particularly at risk hot spots such as switch yards and (un)loading points but also incidental and continuous spillage were considered. All OSR plants, including their hybridization partners which were collected at the respective monitoring sites were analyzed for the presence of transgenes by real-time PCR. On sampling lengths each of 4.2 and 5.7 km, respectively, 461 and 1,574 plants were sampled in Ticino and the region of Basel. OSR plants were found most frequently along the routes to the oilseed facilities, and in larger amounts on risk hot spots compared to sites of random sampling. At three locations in both monitored regions, transgenic B. napus line GT73 carrying the glyphosate resistance transgenes gox and CP4 epsps were detected (Ticino, 22 plants; in the region of Basel, 159).

Prenatal Exposure to Phenols and Growth in Boys

PubMed Central

Philippat, Claire; Botton, Jérémie; Calafat, Antonia M.; Ye, Xiaoyun; Charles, Marie-Aline; Slama, Rémy

2016-01-01

Background Phenols interact with nuclear receptors implicated in growth and adipogenesis regulation. Only a few studies have explored their effects on growth in humans. Objectives We studied the associations of maternal exposure to phenols during pregnancy with prenatal and postnatal growth of male newborns. Methods Within a cohort of women recruited during pregnancy, we selected 520 mother–son pairs and quantified 9 phenols in spot urine samples collected during pregnancy. We used ultrasonography during pregnancy, together with birth measurements, to assess fetal growth. We modeled individual postnatal growth trajectories from repeated measures of weight and height in the first 3 years of life. Results Triclosan concentration was negatively associated with growth parameters measured at the third ultrasound examination but not earlier in pregnancy. At birth, this phenol tended to be negatively associated with head circumference (−1.2 mm for an interquartile range [IQR] increase in ln-transformed triclosan concentration [95% confidence interval = −2.6 to 0.3]) but not with weight or height. Parabens were positively associated with weight at birth. This positive association remained for 3 years for methylparaben (β = 193 g [−4 to 389]) for an IQR increase in ln-transformed concentrations. Conclusion We relied on only 1 spot urine sample to assess exposure; because of the high variability in phenol urinary concentrations reported during pregnancy, using only 1 sample may result in exposure misclassification, in particular for bisphenol A. Our study suggested associations between prenatal exposure to parabens and triclosan and prenatal or early postnatal growth. PMID:25061923

HIV drug resistance in infants increases with changing prevention of mother-to-child transmission regimens.

PubMed

Poppe, Lisa K; Chunda-Liyoka, Catherine; Kwon, Eun H; Gondwe, Clement; West, John T; Kankasa, Chipepo; Ndongmo, Clement B; Wood, Charles

2017-08-24

The objectives of this study were to determine HIV drug resistance (HIVDR) prevalence in Zambian infants upon diagnosis, and to determine how changing prevention of mother-to-child transmission (PMTCT) drug regimens affect drug resistance. Dried blood spot (DBS) samples from infants in the Lusaka District of Zambia, obtained during routine diagnostic screening, were collected during four different years representing three different PMTCT drug treatment regimens. DNA extracted from dried blood spot samples was used to sequence a 1493 bp region of the reverse transcriptase gene. Sequences were analyzed via the Stanford HIVDRdatabase (http://hivdb.standford.edu) to screen for resistance mutations. HIVDR in infants increased from 21.5 in 2007/2009 to 40.2% in 2014. Nonnucleoside reverse transcriptase inhibitor resistance increased steadily over the sampling period, whereas nucleoside reverse transcriptase inhibitor resistance and dual class resistance both increased more than threefold in 2014. Analysis of drug resistance scores in each group revealed increasing strength of resistance over time. In 2014, children with reported PMTCT exposure, defined as infant prophylaxis and/or maternal treatment, showed a higher prevalence and strength of resistance compared to those with no reported exposure. HIVDR is on the rise in Zambia and presents a serious problem for the successful lifelong treatment of HIV-infected children. PMTCT affects both the prevalence and strength of resistance and further research is needed to determine how to mitigate its role leading to resistance.

Hydrothermal venting on the flanks of Heard and McDonald islands, southern Indian Ocean

NASA Astrophysics Data System (ADS)

Lupton, J. E.; Arculus, R. J.; Coffin, M.; Bradney, A.; Baumberger, T.; Wilkinson, C.

2017-12-01

Heard Island and the nearby McDonald Islands are two sites of active volcanism associated with the so-called Kerguelen mantle plume or hot spot. In fact, it has been proposed that the Kerguelen hot spot is currently located beneath Heard Island. During its maiden maximum endurance voyage (IN2016_V01), the recently commissioned Australian R/V Investigator conducted a detailed bathymetric and water column survey of the waters around Heard Island and the McDonald Islands as well as other sites on the Kerguelen Plateau. Some 50 hydrographic profiles were completed using the CTD/rosette system equipped with trace metal sampling and a nephelometer for suspended particle concentrations. In addition to the hydrographic profiles, 244 bubble plumes were detected in the vicinity of the Heard and McDonald Islands using the ship's multibeam system. It is thought that the bubble plumes observed on sea knolls and other seafloor surrounding the McDonald Islands are likely hydrothermal in origin, while plumes northeast of Heard Island may be biogenic methane from cold seeps. At 29 of the hydrographic stations water samples for helium isotope measurements were drawn from the CTD rosette and hermetically sealed into copper tubing for subsequent shorebased mass spectrometer and gas chromatograph analysis. In this paper we report results for 3He/4He ratios and CO2 and CH4 concentrations in water samples collected with the CTD/rosette, and discuss how these results are correlated with suspended particle concentrations and temperature anomalies.

Status of tobacco viruses in Serbia and molecular characterization of tomato spotted wilt virus isolates.

PubMed

Stanković, I; Bulajić, A; Vučurović, A; Ristić, D; Milojević, K; Berenji, J; Krstić, B

2011-01-01

In a four-year survey to determine the presence and distribution of viruses in tobacco crops at 17 localities of the Vojvodina Province and Central Serbia, 380 samples were collected and analyzed by DAS-ELISA. Out of the seven viruses tested, tomato spotted wilt virus (TSWV), potato virus Y (PVY), tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), and alfalfa mosaic virus (AMV) were detected in 37.9, 33.4, 28.7, 23.9, and 15.5% of the total tested samples, respectively. TSWV was the most frequently found virus at the localities of Central Serbia, while PVY and CMV were the most frequent viruses in the Vojvodina Province. Single infections were prevalent in years 2005-2007 and the most frequent were those of PVY. A triple combination of those viruses was most frequent mixed infection type in 2008. The presence of all five detected viruses was confirmed in selected ELISA-positive samples by RT-PCR and sequencing. The comparisons of obtained virus isolate sequences with those available in NCBI, confirmed the authenticity of serologically detected viruses. Phylogenetic analysis based on partial nucleocapsid gene sequences revealed a joint clustering of Serbian, Bulgarian and Montenegrin TSWV isolates into one geographic subpopulation, which was distinct from the other subpopulation of TSWV isolates from the rest of the European countries. The high incidence of viruses in Serbian tobacco crops highlights the importance of enhancing farmers knowledge towards better implementation of control strategies for preventing serious losses.

A one-step multiplex RT-PCR assay for simultaneous detection of four viruses that infect peach.

PubMed

Yu, Y; Zhao, Z; Jiang, D; Wu, Z; Li, S

2013-10-01

A multiplex reverse transcription polymerase chain reaction (mRT-PCR) assay was developed to enable the simultaneous detection and differentiation of four viruses that infect peach, namely Apple chlorotic leaf spot virus (ACLSV), Cherry green ring mottle virus (CGRMV), Prunus necrotic ringspot virus (PNRSV) and Apricot pseudo-chlorotic leaf spot virus (APCLSV). In this study, four pairs of primers, one specific for each virus, were designed; the corresponding PCR products were 632, 439, 346 and 282 bp in length for ACLSV, CGRMV, PNRSV and APCLSV, respectively, and the fragments could be distinguished clearly by agarose gel electrophoresis. The sensitivity and specificity of the method were tested using individual RT-PCR and enzyme-linked immunosorbent assay (ELISA), and the identity of the RT-PCR amplification products was also confirmed by DNA sequencing. The results of RT-PCR and ELISA, along with batch detection using samples collected from peach orchards, revealed that this rapid and simple technique is an effective way to identify the four viruses simultaneously. The mRT-PCR assay described in this study was developed for the simultaneous detection of four peach viruses from infected peach samples is reliable and sensitive. In contrast to conventional uniplex RT-PCR, mRT-PCR is more efficient, reducing costs, time and handling when testing large numbers of samples. This rapid and simple method is useful for large-scale surveys of viruses that infect peach. © 2013 The Society for Applied Microbiology.

A data processing method based on tracking light spot for the laser differential confocal component parameters measurement system

NASA Astrophysics Data System (ADS)

Shao, Rongjun; Qiu, Lirong; Yang, Jiamiao; Zhao, Weiqian; Zhang, Xin

2013-12-01

We have proposed the component parameters measuring method based on the differential confocal focusing theory. In order to improve the positioning precision of the laser differential confocal component parameters measurement system (LDDCPMS), the paper provides a data processing method based on tracking light spot. To reduce the error caused by the light point moving in collecting the axial intensity signal, the image centroiding algorithm is used to find and track the center of Airy disk of the images collected by the laser differential confocal system. For weakening the influence of higher harmonic noises during the measurement, Gaussian filter is used to process the axial intensity signal. Ultimately the zero point corresponding to the focus of the objective in a differential confocal system is achieved by linear fitting for the differential confocal axial intensity data. Preliminary experiments indicate that the method based on tracking light spot can accurately collect the axial intensity response signal of the virtual pinhole, and improve the anti-interference ability of system. Thus it improves the system positioning accuracy.

Rickettsia (Rickettsiales: Rickettsiaceae) Vector Biodiversity in High Altitude Atlantic Forest Fragments Within a Semiarid Climate: A New Endemic Area of Spotted-Fever in Brazil.

PubMed

Moerbeck, Leonardo; Vizzoni, Vinícius F; Machado-Ferreira, Erik; Cavalcante, Robson C; Oliveira, Stefan V; Soares, Carlos A G; Amorim, Marinete; Gazêta, Gilberto S

2016-11-01

Rickettsioses are re-emerging vector-borne zoonoses with a global distribution. Recently, Rickettsia sp. strain Atlantic rainforest has been associated with new human spotted-fever (SF) cases in Brazil, featuring particular clinical signs: eschar formation and lymphadenopathy. These cases have been associated with the tick species, Amblyomma ovale From 2010 until 2015, the Brazilian Health Department confirmed 11 human SF cases in the Maciço de Baturité region, Ceará, Brazil. The present study reports the circulation of Rickettsia spp. in vectors from this entirely new endemic area for SF. A total of 1,727 ectoparasites were collected in this area from the environment, humans, and wild and domestic animals. Samples (n = 887) were screened by polymerase chain reaction (PCR), targeting the gltA and ompA rickettsial genes. Sequencing and phylogenetic analyses of gltA gene amplicons were carried out for 13 samples positive for both screening PCRs. Fragments of gltA and ompA from three samples were cloned, sequenced, and analyzed further. A. ovale and Rhipicephalus sanguineus specimens, collected from dogs, were found to be infected with Rickettsia sp. str. Atlantic rainforest, suggesting the importance of dogs in the epidemic cycle. Candidatus Rickettsia andeanae, Rickettsia felis, and Rickettsia bellii were also found infecting ticks and fleas in five municipalities, demonstrating the broad diversity of rickettsiae in circulation in the studied area. This study reports, for the first time, evidence of infection with Rickettsia sp. strain Atlantic rainforest in A. ovale and R. sanguineus in Ceará, and Ca. R. andeanae in an Atlantic rainforest environment of Brazil. © The Authors 2016. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Efficacy of a new spot-on formulation of selamectin plus sarolaner for cats against adult Ctenocephalides felis, flea egg production and adult flea emergence.

PubMed

Vatta, Adriano F; Everett, William R; Holzmer, Susan J; Cherni, Judith A; King, Vickie L; Rugg, Douglas; Geurden, Thomas

2017-04-01

A new spot-on formulation of selamectin plus sarolaner was evaluated against fleas for adulticidal efficacy, and for the effect on egg production and hatching when applied to flea-infested cats. Ten male and ten female adult domestic shorthair cats were randomly assigned to one of two treatment groups based on pre-treatment flea counts. Cats received topical treatment on Day 0 in a single spot to the dorsal scapular area with either a placebo formulation or with the combination formulation at the minimal dose of 6.0mg selamectin plus 1.0mg sarolaner per kg bodyweight. On Days -1, 5, 12, 19, 26 and 33, cats were infested with approximately 100 (±5) unfed Ctenocephalides felis fleas. At 24h after treatment or 48h after subsequent flea infestation, cats were housed for a 20-h period in a cage to allow collection of flea eggs. At the end of this period, flea eggs were collected from the cages and cats were combed to remove and count live fleas. Emerged viable larvae and emerged adult fleas were counted 3days and 35days, respectively, after egg collection. The new spot-on formulation of selamectin plus sarolaner provided 100% efficacy against adult fleas up to Day 36 following a single application. Fleas on placebo-treated cats produced large numbers of eggs throughout the study, with individual counts ranging from 110 to 1256 eggs. Following treatment, four flea eggs were collected from a single selamectin/sarolaner-treated cat on Day 29, but there were no eggs collected from any other selamectin/sarolaner-treated animal during the study. No larvae or adult fleas developed from these four eggs. From the eggs collected from the placebo-treated cats, the mean percentage of live larvae and adults that emerged ranged from 67.3% to 84.2% and from 50.7% to 81.8%, respectively. A single topical treatment with a new spot-on formulation of selamectin plus sarolaner at the minimum label dose thus controlled fleas on cats and was 100% effective in preventing flea reproduction for over one month after treatment. Copyright © 2017 Zoetis Services LLC. Published by Elsevier B.V. All rights reserved.

Diets of California spotted owls in the Sierra National Forest

Treesearch

Thomas E. Munton; Kenneth D. Johnson; George N. Steger; Gary P. Eberlein

2002-01-01

From May 1987 through October 1992 and from July through August 1998, we studied diets of California spotted owls (Strix occidentalis occidentalis). Regurgitated pellets were collected at roost and nest sites between 1,000 and 7,600 ft elevation in the Sierra National Forest and were examined for remnant bones, feathers, and insect exoskeletons....

Evaluation of a barley core collection for spot form net blotch reaction reveals distinct genotype specific pathogen virulence and host susceptibility

USDA-ARS?s Scientific Manuscript database

Spot form net blotch (SFNB) caused by Pyrenophora teres Drechs. f. maculata Smedeg., (anamorph Drechslera teres [Sacc.] Shoem.) is a major foliar disease of barley (Hordeum vulgare L.) worldwide. SFNB epidemics have recently been observed in major barley producing countries, suggesting that the loca...

Activity patterns of nesting Mexican Spotted Owls

Treesearch

David K. Delaney; Teryl G. Grubb; Paul Beier

1999-01-01

We collected 2,665 hr of behavioral information using video surveillance on 19 Mexican Spotted Owl (Strix occidentalis lucida) pairs between 25 April and 26 July 1996. Prey deliveries per day increased as the nesting season progressed, with an average of 2.68 prey deliveries during incubation, 4.10 items during brooding, and 4.51 items during the...

Cow-specific diet digestibility predictions based on near-infrared reflectance spectroscopy scans of faecal samples.

PubMed

Mehtiö, T; Rinne, M; Nyholm, L; Mäntysaari, P; Sairanen, A; Mäntysaari, E A; Pitkänen, T; Lidauer, M H

2016-04-01

This study was designed to obtain information on prediction of diet digestibility from near-infrared reflectance spectroscopy (NIRS) scans of faecal spot samples from dairy cows at different stages of lactation and to develop a faecal sampling protocol. NIRS was used to predict diet organic matter digestibility (OMD) and indigestible neutral detergent fibre content (iNDF) from faecal samples, and dry matter digestibility (DMD) using iNDF in feed and faecal samples as an internal marker. Acid-insoluble ash (AIA) as an internal digestibility marker was used as a reference method to evaluate the reliability of NIRS predictions. Feed and composite faecal samples were collected from 44 cows at approximately 50, 150 and 250 days in milk (DIM). The estimated standard deviation for cow-specific organic matter digestibility analysed by AIA was 12.3 g/kg, which is small considering that the average was 724 g/kg. The phenotypic correlation between direct faecal OMD prediction by NIRS and OMD by AIA over the lactation was 0.51. The low repeatability and small variability estimates for direct OMD predictions by NIRS were not accurate enough to quantify small differences in OMD between cows. In contrast to OMD, the repeatability estimates for DMD by iNDF and especially for direct faecal iNDF predictions were 0.32 and 0.46, respectively, indicating that developing of NIRS predictions for cow-specific digestibility is possible. A data subset of 20 cows with daily individual faecal samples was used to develop an on-farm sampling protocol. Based on the assessment of correlations between individual sample combinations and composite samples as well as repeatability estimates for individual sample combinations, we found that collecting up to three individual samples yields a representative composite sample. Collection of samples from all the cows of a herd every third month might be a good choice, because it would yield a better accuracy. © 2015 Blackwell Verlag GmbH.

Decadal changes in phenology of peak abundance patterns of woodland pond salamanders in northern Wisconsin

USGS Publications Warehouse

Donner, Deahn M.; Ribic, Christine; Beck, Albert J.; Higgins, Dale; Eklund, Dan; Reinecke, Susan

2015-01-01

Woodland ponds are important landscape features that help sustain populations of amphibians that require this aquatic habitat for successful reproduction. Species abundance patterns often reflect site-specific differences in hydrology, physical characteristics, and surrounding vegetation. Large-scale processes such as changing land cover and environmental conditions are other potential drivers influencing amphibian populations in the Upper Midwest, but little information exists on the combined effects of these factors. We used Blue-spotted (Ambystoma laterale Hallowell) and Spotted Salamander (A. maculatum Shaw) monitoring data collected at the same woodland ponds thirteen years apart to determine if changing environmental conditions and vegetation cover in surrounding landscapes influenced salamander movement phenology and abundance. Four woodland ponds in northern Wisconsin were sampled for salamanders in April 1992-1994 and 2005-2007. While Blue-spotted Salamanders were more abundant than Spotted Salamanders in all ponds, there was no change in the numbers of either species over the years. However, peak numbers of Blue-spotted Salamanders occurred 11.7 days earlier (range: 9-14 days) in the 2000s compared to the 1990s; Spotted Salamanders occurred 9.5 days earlier (range: 3 - 13 days). Air and water temperatures (April 13- 24) increased, on average, 4.8°C and 3.7°C, respectively, between the decades regardless of pond. There were no discernible changes in canopy openness in surrounding forests between decades that would have warmed the water sooner (i.e., more light penetration). Our finding that salamander breeding phenology can vary by roughly 10 days in Wisconsin contributes to growing evidence that amphibian populations have responded to changing climate conditions by shifting life-cycle events. Managers can use this information to adjust monitoring programs and forest management activities in the surrounding landscape to avoid vulnerable amphibian movement periods. Considering direct and indirect stressors such as changing habitat and environmental conditions simultaneously to better understand trends in space and time can help improve monitoring programs for this taxa, which is at major risk of continued declines.

Local diversity hot spots in the Middle Miocene of the Central Paratethys: influence of environment and sampling

NASA Astrophysics Data System (ADS)

Nawrot, Rafal; Zuschin, Martin; Harzhauser, Mathias; Kroh, Andreas; Mandic, Oleg

2015-04-01

Species richness captured by historical fossil inventories is a complex function of true local diversity, degree of outcrop-scale heterogeneity in species composition and sampling intensity. Disentangling these factors is hindered by the fact that the 'systematists follow the fossils' (Raup, 1977) and thus non-uniform research effort may both drive apparent diversity patterns and follow the actual presence of highly fossiliferous strata. The molluscan fauna of Lapugiu de Sus (Hunedoara District, Romania) constitutes one of the most diverse Early Badenian (Langhian) assemblages of the Paratethys Sea, with almost one thousand species reported during 170 years of extensive studies. We evaluate whether this exceptional richness reflects the actual diversity hot spot or just a long history of fossil-collecting by comparing the fauna of Lapugiu with other Paratethyan molluscan lagerstätten of similar age. The literature-derived species lists for each section were contrasted with independent abundance data based on a standardized sampling protocol (42 samples, 24,000 specimens, and 530 species from six localities). Although individual samples from other localities can exhibit comparable diversity levels, richness estimates for samples from Lapugiu are all consistently high, reflecting increasing evenness in more offshore depositional settings. This translates to the highest diversity at the outcrop scale when all samples are pooled. In contrast to other localities, however, for which data from historical inventories corresponds well to our quantitative estimates of total richness, the number of species described from Lapugiu is much higher than expected. This excessive richness likely reflects the 'Bonanza Effect' (sensu Dunhill et al., 2012), where uniformly species-rich deposits were attracting intensive taxonomic studies. The strong positive feedback between palaeontological sampling effort and fossil diversity may thus greatly overestimate the true differences in species richness between the most diverse faunas and more typical, background, fossil assemblages. References Raup, D.M., 1977. Paleobiology, 3: 328-329. Dunhill, A.M. et al., 2012. Palaeontology, 55: 1155-1175.

Mechanisms of Laser-Induced Dissection and Transport of Histologic Specimens

PubMed Central

Vogel, Alfred; Lorenz, Kathrin; Horneffer, Verena; Hüttmann, Gereon; von Smolinski, Dorthe; Gebert, Andreas

2007-01-01

Rapid contact- and contamination-free procurement of histologic material for proteomic and genomic analysis can be achieved by laser microdissection of the sample of interest followed by laser-induced transport (laser pressure catapulting). The dynamics of laser microdissection and laser pressure catapulting of histologic samples of 80 μm diameter was investigated by means of time-resolved photography. The working mechanism of microdissection was found to be plasma-mediated ablation initiated by linear absorption. Catapulting was driven by plasma formation when tightly focused pulses were used, and by photothermal ablation at the bottom of the sample when defocused pulses producing laser spot diameters larger than 35 μm were used. With focused pulses, driving pressures of several hundred MPa accelerated the specimen to initial velocities of 100–300 m/s before they were rapidly slowed down by air friction. When the laser spot was increased to a size comparable to or larger than the sample diameter, both driving pressure and flight velocity decreased considerably. Based on a characterization of the thermal and optical properties of the histologic specimens and supporting materials used, we calculated the evolution of the heat distribution in the sample. Selected catapulted samples were examined by scanning electron microscopy or analyzed by real-time reverse-transcriptase polymerase chain reaction. We found that catapulting of dissected samples results in little collateral damage when the laser pulses are either tightly focused or when the laser spot size is comparable to the specimen size. By contrast, moderate defocusing with spot sizes up to one-third of the specimen diameter may involve significant heat and ultraviolet exposure. Potential side effects are maximal when samples are catapulted directly from a glass slide without a supporting polymer foil. PMID:17766336

A screening method for biotinidase deficiency in newborns.

PubMed

Heard, G S; Secor McVoy, J R; Wolf, B

1984-01-01

We describe a method for neonatal screening for biotinidase (EC 3.5.1.12) deficiency. Biotinidase activity is assessed colorimetrically from dried samples of whole blood spotted on the same filter papers as used in the neonatal screening for phenylketonuria. After the reaction, samples from normal infants are characteristically purple, whereas those from affected individuals are straw-colored. To confirm the deficiency, the enzyme is quantitatively assayed in additional blood spots or serum. A pilot study has been initiated with samples obtained by the Commonwealth of Virginia for phenylketonuria testing.

Trehalose significantly enhances the recovery of serum and serum exosomal miRNA from a paper-based matrix.

PubMed

Neo, Shu Hui; Chung, Ka Yan; Quek, Jia Min; Too, Heng-Phon

2017-11-30

The preservation of nucleic acids from clinical samples is critical to facilitate accurate molecular diagnosis. The use of a paper matrix, Flinders Technology Associates (FTA) Elute cards, to archive DNA and viral RNA is well-documented. However, the feasibility of FTA Elute cards for archiving serum and serum exosomal microRNAs (miRNAs) remains unclear. Here, we performed a comprehensive evaluation of FTA Elute cards for miRNA storage and recovery in different pre-analytical conditions. The recovery of serum miRNA dry-spotted on FTA Elute cards by direct elution with water at high temperature was poor. However, serum miRNAs dry-spotted on the cards were isolated with about 40% yield when using QIAzol lysis reagent and recovery was improved remarkably (>80%) upon extraction from cards pre-treated with trehalose. miRNAs stored on the cards remained stable at room temperature and can be kept for prolonged periods. Furthermore, miRNAs could be similarly recovered from serum exosomes dry-spotted on the cards. Importantly, when using sera from gastric cancer (GC) patients, the miRNAs were efficiently recovered from trehalose pre-treated cards without affecting their representation. Collectively, we have demonstrated the potential of FTA Elute cards to archive serum and serum exosomal miRNAs, making it useful for biomarker discovery and diagnostics.

Diversity and genetic distance in populations of Steindachnerina in the upper Paraná river floodplain of Brazil.

PubMed

Oliveira, A V; Prioli, A J; Prioli, S M A P; Pavanelli, C S; Júlio, H F; Panarari, R S

2002-08-01

Whereas four species of the genus Steindachnerina occur in the Paraná river basin, S. insculpta was the only endemic species of the region under analysis, which is the third lower section of the upper Paraná river. Among other factors, this species has been characterised by the absence of spots in the basal region of the dorsal fin. However, various specimens with this characteristic appeared in the region after the construction of the Itaipu Hydroelectric Plant in 1982. An analysis of the genetic variability of Steindachnerina populations with or without spots is provided. Specimens were collected in different sites of the floodplain of the upper Paraná river and samples were compared by random amplified polymorphic DNA (RAPD) technique and morphological analyses. Ninety-eight amplified loci with nine random primers were analysed in 19 specimens of each phenotype. Data for genetic distance showed great divergences between the two phenotypes and indicate two different species. Spotted specimens may be identified as S. brevipinna, found in the region downstream Sete Quedas Falls. The species must have overcome the geographical barrier during the building of the Itaipu hydroelectric dam that submerged the waterfalls and which became an obstacle between the upper and middle Paraná river some 150 km downstream. Since phenotypes do not share dominant alleles, absence of gene flow has been suggested.

Drug 'hot-spots', alcohol availability and violence.

PubMed

Gorman, D M; Zhu, Li; Horel, Scott

2005-11-01

Ecological studies have shown a relationship between alcohol outlet densities and violence and between the location of crimes related to illicit drug use (so-called 'hot spots') and violence. To date, no study has compared the effects of alcohol outlets and drug hot spots on rates of violence. The present study examined this relationship in the City of Houston, Texas. An ecological study design was employed, using a sample of 439 census tracts from Houston, Texas. Neighborhood socio-structural, alcohol outlet density, drug crime density and violent crime density data were collected from archival sources and analyzed using multivariate and spatial statistics. Using ordinary least-squares analysis, the neighborhood socio-structural covariates explained about 40% of the variability in violent crime. Adding alcohol outlet density in the target census tracts explained an additional 6%, while the addition of drug crime density explained an additional 32%. In the final model, that controlled for the effects of autocorrelated error, both drug crime density in the target and adjacent census tracts remained significant predictors of violent crime, while only off-sale density in the target census tract remained significant in the model. The findings indicate that drug crime density explained a greater amount of variance in violent crime rates than the alcohol outlet density. The methodological and policy implications of these findings are discussed, along with the shortcomings of the analysis presented.

Community Structure of Tintinnid Ciliates of the Microzooplankton in the South West Pacific Ocean: Comparison of a High Primary Productivity with a Typical Oligotrophic Site.

PubMed

Dolan, John R; Gimenez, Audrey; Cornet-Barthaux, Veronique; de Verneil, Alain

2016-11-01

Transient 'hot spots' of phytoplankton productivity occur in the generally oligotrophic Southern Pacific Ocean and we hypothesized that the population structure of tintinnid ciliates, planktonic grazers, would differ from that of a typical oligotrophic sites. Samples were collected over a 1-wk period at each of two sites between Fiji and Tahiti: one of elevated chlorophyll a concentrations and primary productivity with an abundance of N-fixing cyanobacteria Trichodesmium, and a distant oligotrophic site. Tintinnid abundance differed between the sites by a factor of 2. A single species (Favella sp.), absent from the oligotrophic site, highly dominated the 'hot spot' site. However, total species richness was identical (71 spp.) as well as short-term temporal variability (2-4 d). At both sites, species abundance distributions most closely fit a log-series or log-normal distribution and the abundance distributions of ecological types, forms of distinct lorica oral diameter, were the typical geometric. Morphological diversity was only slightly lower at the high productivity site. We found that communities of these plankton grazers in 'hot spots' of phytoplankton productivity in oligotrophic systems, although harboring different species, differ little from surrounding oligotrophic areas in community structure. © 2016 The Author(s) Journal of Eukaryotic Microbiology © 2016 International Society of Protistologists.

[A comparative study of the glucose level in dry blood stains from donors and cadavers].

PubMed

Kachina, N N

1994-01-01

Glucose concentrations in dry spots of cadaveric and donor blood stored at room temperature for different periods were measured. Studies by glucose oxidase method revealed that glucose levels in dry spots of both cadaveric and donor blood gradually reduced until completely disappeared, but in comparison with glucose level lowering in liquid blood the period during which this carbohydrate completely disappeared from a dry blood spot was by several times longer. Effects of the velocity of blood spot drying and of microorganisms contaminating the sample may make the expert conclusions doubtful.

Improving strategies to assess competitive effects of barred owls on northern spotted owls in the Pacific Northwest

USGS Publications Warehouse

Wiens, J. David; Weekes, Anne

2011-01-01

A scientific study has determined that survey methods designed for spotted owls do not always detect barred owls that are actually present in spotted owl habitat. The researchers suggest that strategies to address potential interactions between spotted owls and barred owls will require carefully designed surveys that account for response behaviors and imperfect detection of both species. Species-specific sampling methods, which are proposed, can be used by forest managers to determine the occurrence and distribution of barred owls with high confidence. This fact sheet provides highlights of the research (Wiens and others, 2011).

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

PubMed

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

2016-09-23

Dried blood spot (DBS) sampling and analysis is increasingly being applied in bioanalysis. Although the use of DBS has many advantages, it is also associated with some challenges. E.g. given the limited amount of available material, highly sensitive detection techniques are often required to attain sufficient sensitivity. In gas chromatography coupled to mass spectrometry (GC-MS), derivatization can be helpful to achieve adequate sensitivity. Because this additional sample preparation step is considered as time-consuming, we introduce a new derivatization procedure, i.e. "microwave-assisted on-spot derivatization", to minimize sample preparation of DBS. In this approach the derivatization reagents are directly applied onto the DBS and derivatization takes place in a microwave instead of via conventional heating. In this manuscript we evaluated the applicability of this new concept of derivatization for the determination of two polar low molecular weight molecules, gamma-hydroxybutyric acid (GHB) and gabapentin, in DBS using a standard GC-MS configuration. The method was successfully validated for both compounds, with imprecision and bias values within acceptance criteria (<20% at LLOQ, <15% at 3 other QC levels). Calibration lines were linear over the 10-100μg/mL and 1-30μg/mL range for GHB and gabapentin, respectively. Stability studies revealed no significant decrease of gabapentin and GHB in DBS upon storage at room temperature for at least 84 days. Furthermore, DBS-specific parameters, including hematocrit and volume spotted, were evaluated. As demonstrated by the analysis of GHB and gabapentin positive samples, "microwave-assisted on-spot derivatization" proved to be reliable, fast and applicable in routine toxicology. Moreover, other polar low molecular weight compounds of interest in clinical and/or forensic toxicology, including vigabatrin, beta-hydroxybutyric acid, propylene glycol, diethylene glycol, 1,4-butanediol and 1,2-butanediol, can also be detected using this method. Copyright © 2016 Elsevier B.V. All rights reserved.

Sample exchange by beam scanning with applications to noncollinear pump-probe spectroscopy at kilohertz repetition rates.

PubMed

Spencer, Austin P; Hill, Robert J; Peters, William K; Baranov, Dmitry; Cho, Byungmoon; Huerta-Viga, Adriana; Carollo, Alexa R; Curtis, Anna C; Jonas, David M

2017-06-01

In laser spectroscopy, high photon flux can perturb the sample away from thermal equilibrium, altering its spectroscopic properties. Here, we describe an optical beam scanning apparatus that minimizes repetitive sample excitation while providing shot-to-shot sample exchange for samples such as cryostats, films, and air-tight cuvettes. In this apparatus, the beam crossing point is moved within the focal plane inside the sample by scanning both tilt angles of a flat mirror. A space-filling spiral scan pattern was designed that efficiently utilizes the sample area and mirror scanning bandwidth. Scanning beams along a spiral path is shown to increase the average number of laser shots that can be sampled before a spot on the sample cell is resampled by the laser to ∼1700 (out of the maximum possible 2500 for the sample area and laser spot size) while ensuring minimal shot-to-shot spatial overlap. Both an all-refractive version and an all-reflective version of the apparatus are demonstrated. The beam scanning apparatus does not measurably alter the time delay (less than the 0.4 fs measurement uncertainty), the laser focal spot size (less than the 2 μm measurement uncertainty), or the beam overlap (less than the 3.3% measurement uncertainty), leading to pump-probe and autocorrelation signal transients that accurately characterize the equilibrium sample.

The New Bedford Harbor Superfund Site Long Term ...

EPA Pesticide Factsheets

Background. New Bedford Harbor (NBH), located in southeastern Massachusetts, was designated as a marine Superfund site in 1983 due to sediment contamination by polychlorinated biphenyls (PCBs). Based on risks to human health and the environment, the first two phases of the site cleanup involved dredging PCB-contaminated sediments from the harbor. Therefore, a long-term monitoring program (LTM) was developed to measure spatial and temporal chemical and biological changes in sediment, water, and biota to assess the effects and effectiveness of the remedial activities. Approach. A systematic, probabilistic sampling design was used to select approximately 70 sediment sampling stations. Sediment was collected at each station and chemical (e.g., PCBs, metals), physical (e.g., grain size), and biological (e.g., benthic community) measurements were conducted on all samples. There have been six sample collections to date: 1993-baseline, 1995-post hot spot removal, 1999-prior to full scale dredging, and then at 5 year intervals: 2004, 2009, and 2014. Mussel (Mytilus edulis) bioaccumulation has also been measured twice yearly. Results. There is a decreasing spatial gradient in sediment PCB concentrations from the northern boundary (upper harbor) to the southern boundary (outer harbor) of the site. Along this same transect, there is an increase in biological condition (e.g., benthic community diversity). Temporally, the contaminant and biological gradients have been

Poro-elastic Properties of Whillan's Ice Stream Till: Implications for Basal Stick-Slip

NASA Astrophysics Data System (ADS)

Leeman, J.; Valdez, R. D.; Alley, R. B.; Anandakrishnan, S.; Saffer, D. M.

2016-12-01

Whillans ice stream, West Antarctica, flows rapidly from the West Antarctic ice sheet into the Ross Ice Shelf. Regions of highly compacted till, termed sticky-spots, pin the ice in place. Upstream ice flow increases driving stress, until minor changes in buttressing stresses from tides affecting the ice shelf cause the main sticky-spot to fail, triggering diurnal or semidiurnal stick-slip events. The mechanical and hydrological properties of the till partially control the basal conditions, generation and persistence of the sticky spots, and thus the dynamics of the rupture and healing processes. Here we present laboratory tests on core samples of the till beneath Whillan's Ice Stream collected in the 1989-1993 field seasons. Two types of tests were performed on till cores: stepped loading and cyclic loading. In the stepped loading test, the effective stress was increased from 0.1 to 10 MPa in a series of steps, and the permeability measured at each step. Cyclic loading tests consisted of a series of effective stress oscillations with 24 h period lasting 5-10 d each, increasing in amplitude from 20-150 kPa. The permeability was measured after each set of oscillations to investigate the role of cyclic loading in driving enhanced compaction. Compressional wave velocity (Vp) was also measured during both test sequences. We observe sample initial porosities of 30% and permeabilities of 3x10-17 m2. During stepped loading tests, porosity is reduced to 20% and permeability to 8x10-18 m2. Vp ranged from 2.2-2.95 km s-1 and was well fit by an effective medium model. Application of this model to Vp obtained by field seismic surveys is consistent with low ( 50 kPa) effective vertical stresses in the uppermost till. Cyclic loading sequences reduced porosity by 4% and permeability by an order of magnitude. A transient numerical model based on our data shows that over the tidal timescale, a layer of stiffened till 10 cm thick should develop. We suggest that this provides one mechanism to generate and maintain sticky spots and modify the stiffness of the system.

Contamination status by persistent organic pollutants of the Atlantic spotted dolphin (Stenella frontalis) at the metapopulation level.

PubMed

Méndez-Fernandez, Paula; Taniguchi, Satie; Santos, Marcos C O; Cascão, Irma; Quérouil, Sophie; Martín, Vidal; Tejedor, Marisa; Carrillo, Manuel; Rinaldi, Carolina; Rinaldi, Renato; Montone, Rosalinda C

2018-05-01

The Atlantic spotted dolphin (Stenella frontalis) is an endemic species of the tropical-temperate Atlantic Ocean with widespread distribution. Although this species has been the subject of a large number of studies throughout its range, it remains in the "data deficient" category of the International Union for Conservation of Nature (IUCN). Chemical pollution by persistent organic pollutants (POPs) has been listed as one of the major threats to this species, however, there is no information on a wide scale. Thus, the aim of the present study was to investigate the contamination status of spotted dolphins on the metapopulation level as well as determine spatial and temporal variations in POP concentrations and bio-accumulation. A total of 115 blubber samples collected from a large part of the Atlantic basin were analysed for PCBs, DDTs, PBDEs, chlordanes, HCB and mirex. Although PCBs and DDTs were the predominant compounds in all areas, inter-location differences in POP concentrations were observed. Dolphins found at São Paulo, southeastern coast of Brazil, had the highest PCB concentrations (median: 10.5 μg/g lw) and Canary Islands dolphins had the highest DDT concentrations (median: 5.13 μg/g lw). Differences in PCB patterns among locations were also observed. Dolphins from the Azores and São Paulo demonstrated a similar pattern, with relatively highly contributions of tetra- (6.8 and 5.2%, respectively) and penta-CBs (25.6 and 23.8%, respectively) and lower contributions of hepta-CBs (20.8 and 23.5%, respectively) in comparison to other areas. Moreover, the sex of the animals and the year in which sampling or capture occurred exerted an important influence on the majority of the POPs analysed. Comparisons with toxicity thresholds available in the literature reveal that the São Paulo and Canary Island dolphins are the most vulnerable populations and should be considered in future conservation and management programs for the Atlantic spotted dolphin. Copyright © 2018 Elsevier Ltd. All rights reserved.

Spectral and spatial resolution analysis of multi sensor satellite data for coral reef mapping: Tioman Island, Malaysia

NASA Astrophysics Data System (ADS)

Pradhan, Biswajeet; Kabiri, Keivan

2012-07-01

This paper describes an assessment of coral reef mapping using multi sensor satellite images such as Landsat ETM, SPOT and IKONOS images for Tioman Island, Malaysia. The study area is known to be one of the best Islands in South East Asia for its unique collection of diversified coral reefs and serves host to thousands of tourists every year. For the coral reef identification, classification and analysis, Landsat ETM, SPOT and IKONOS images were collected processed and classified using hierarchical classification schemes. At first, Decision tree classification method was implemented to separate three main land cover classes i.e. water, rural and vegetation and then maximum likelihood supervised classification method was used to classify these main classes. The accuracy of the classification result is evaluated by a separated test sample set, which is selected based on the fieldwork survey and view interpretation from IKONOS image. Few types of ancillary data in used are: (a) DGPS ground control points; (b) Water quality parameters measured by Hydrolab DS4a; (c) Sea-bed substrates spectrum measured by Unispec and; (d) Landcover observation photos along Tioman island coastal area. The overall accuracy of the final classification result obtained was 92.25% with the kappa coefficient is 0.8940. Key words: Coral reef, Multi-spectral Segmentation, Pixel-Based Classification, Decision Tree, Tioman Island

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

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

Thomson, S.; Wallace, A.M.; Cook, B.

1989-08-01

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

Decoding 2D-PAGE complex maps: relevance to proteomics.

PubMed

Pietrogrande, Maria Chiara; Marchetti, Nicola; Dondi, Francesco; Righetti, Pier Giorgio

2006-03-20

This review describes two mathematical approaches useful for decoding the complex signal of 2D-PAGE maps of protein mixtures. These methods are helpful for interpreting the large amount of data of each 2D-PAGE map by extracting all the analytical information hidden therein by spot overlapping. Here the basic theory and application to 2D-PAGE maps are reviewed: the means for extracting information from the experimental data and their relevance to proteomics are discussed. One method is based on the quantitative theory of statistical model of peak overlapping (SMO) using the spot experimental data (intensity and spatial coordinates). The second method is based on the study of the 2D-autocovariance function (2D-ACVF) computed on the experimental digitised map. They are two independent methods that are able to extract equal and complementary information from the 2D-PAGE map. Both methods permit to obtain fundamental information on the sample complexity and the separation performance and to single out ordered patterns present in spot positions: the availability of two independent procedures to compute the same separation parameters is a powerful tool to estimate the reliability of the obtained results. The SMO procedure is an unique tool to quantitatively estimate the degree of spot overlapping present in the map, while the 2D-ACVF method is particularly powerful in simply singling out the presence of order in the spot position from the complexity of the whole 2D map, i.e., spot trains. The procedures were validated by extensive numerical computation on computer-generated maps describing experimental 2D-PAGE gels of protein mixtures. Their applicability to real samples was tested on reference maps obtained from literature sources. The review describes the most relevant information for proteomics: sample complexity, separation performance, overlapping extent, identification of spot trains related to post-translational modifications (PTMs).

CdTe detector based PIXE mapping of geological samples

NASA Astrophysics Data System (ADS)

Chaves, P. C.; Taborda, A.; de Oliveira, D. P. S.; Reis, M. A.

2014-01-01

A sample collected from a borehole drilled approximately 10 km ESE of Bragança, Trás-os-Montes, was analysed by standard and high energy PIXE at both CTN (previous ITN) PIXE setups. The sample is a fine-grained metapyroxenite grading to coarse-grained in the base with disseminated sulphides and fine veinlets of pyrrhotite and pyrite. Matrix composition was obtained at the standard PIXE setup using a 1.25 MeV H+ beam at three different spots. Medium and high Z elemental concentrations were then determined using the DT2fit and DT2simul codes (Reis et al., 2008, 2013 [1,2]), on the spectra obtained in the High Resolution and High Energy (HRHE)-PIXE setup (Chaves et al., 2013 [3]) by irradiation of the sample with a 3.8 MeV proton beam provided by the CTN 3 MV Tandetron accelerator. In this paper we present results, discuss detection limits of the method and the added value of the use of the CdTe detector in this context.

Scanning Electron Microscopy with Samples in an Electric Field

PubMed Central

Frank, Ludĕk; Hovorka, Miloš; Mikmeková, Šárka; Mikmeková, Eliška; Müllerová, Ilona; Pokorná, Zuzana

2012-01-01

The high negative bias of a sample in a scanning electron microscope constitutes the “cathode lens” with a strong electric field just above the sample surface. This mode offers a convenient tool for controlling the landing energy of electrons down to units or even fractions of electronvolts with only slight readjustments of the column. Moreover, the field accelerates and collimates the signal electrons to earthed detectors above and below the sample, thereby assuring high collection efficiency and high amplification of the image signal. One important feature is the ability to acquire the complete emission of the backscattered electrons, including those emitted at high angles with respect to the surface normal. The cathode lens aberrations are proportional to the landing energy of electrons so the spot size becomes nearly constant throughout the full energy scale. At low energies and with their complete angular distribution acquired, the backscattered electron images offer enhanced information about crystalline and electronic structures thanks to contrast mechanisms that are otherwise unavailable. Examples from various areas of materials science are presented.

Topical treatment with fresh human milk versus emollient on atopic eczema spots in young children: a small, randomized, split body, controlled, blinded pilot study.

PubMed

Berents, Teresa Løvold; Rønnevig, Jørgen; Søyland, Elisabeth; Gaustad, Peter; Nylander, Gro; Løland, Beate Fossum

2015-05-04

Public health nurses report on effects of fresh human milk as treatment for conjunctivitis, rhinitis and atopic eczema (AE), the latter being highly prevalent in early childhood. Emollients and topical corticosteroids are first line treatment of AE. As many caregivers have steroid phobia, alternative treatment options for mild AE are of interest. The aim of this small pilot study was to assess the potential effects and risks of applying fresh human milk locally on eczema spots in children with AE. This was a split body, controlled, randomized and physician blinded pilot study, of children with AE with two similar contralateral eczema spots having a mother breastfeeding the child or a sibling. Fresh expressed milk and emollient was applied on the intervention spot and emollient alone on the control area, three times a day for four weeks. The severity and area of the eczema spots was evaluated weekly, and samples from milk and the spots were analysed weekly with respect to bacterial colonisation. Of nine patients included, six completed the study. Mean age at inclusion was 18.5 months. The spots examined were localized on the arms, legs or cheeks. The spots were similar in severity, but differed in area. In one patient the eczema ceased after inclusion. In four patients both control and intervention areas increased during the intervention. The relative change in eczema area compared to baseline showed less increase in the intervention spots in two patients, whereas the opposite was observed in three. In four children Staphylococcus aureus was found in their eczema once or more. In three of the 28 human milk samples, Staphylococcus aureus, alfa haemolytic streptococci or coagulase negative staphylococci were detected. Staphylococcus aureus was found once both in human milk and in the eczema spots, no clinical signs of infection were however observed. No secondary infection due to milk application was detected. In this small pilot study, no effect was found on eczema spots treated with topical application of fresh human milk. (ClinicalTrials.gov Identifier, NCT02381028 ).

Stability of benzodiazepines and cocaine in blood spots stored on filter paper.

PubMed

Alfazil, Abdulkareem A; Anderson, Robert A

2008-09-01

Previous studies have shown that drug concentrations in blood can change during storage, especially at room temperature, but even labile drugs such as cocaine may be stable in dried blood spots (DBS). A new method has been developed for the analysis of hydrolytically labile drugs in blood spots on filter paper in order to assess their degradation during a storage period of one month. The drugs selected included flunitrazepam, temazepam, oxazepam, lorazepam, nitrazepam, diazepam, and cocaine. A Guthrie card 903 was spotted with 100 microL of blood containing the drugs at concentrations of 1000 ng/mL and left overnight to dry at room temperature. The filter paper was suspended in extraction buffer for 1 h with ultrasonication. Drugs were then extracted from the buffer by solid-phase extraction using Clean Screen((R)) columns and analyzed by liquid chromatography-tandem mass spectrometry. Method validation showed that all calibration curves were linear over the concentration range 5-200 ng/spot with correlation coefficients of 0.994-0.999. Interday and intraday precisions at three concentrations (10, 50, and 100 ng/spot) were 1.6-18.3% and 2.8-14.7%, respectively. Limits of detection were 0.29-0.74 ng/spot, and lower limits of quantitation were 0.99-2.46 ng/spot. Recoveries of all analytes were in the range 81-106%. DBS were stored in duplicate at room temperature, 4 degrees C, and -20 degrees C for up to one month. Degradation of the drugs in DBS at all storage conditions was less than for the corresponding liquid blood samples stored under similar conditions and more than 80% of each analyte could be recovered from the samples.

Epimicrobiota Associated with the Decay and Recovery of Orbicella Corals Exhibiting Dark Spot Syndrome

PubMed Central

Meyer, Julie L.; Rodgers, John M.; Dillard, Brian A.; Paul, Valerie J.; Teplitski, Max

2016-01-01

Dark Spot Syndrome (DSS) is one of the most common diseases of boulder corals in the Caribbean. It presents as sunken brown lesions in coral tissue, which can spread quickly over coral colonies. With this study, we tested the hypothesis that similar to other coral diseases, DSS is a dysbiosis characterized by global shifts in the coral microbiome. Because Black Band Disease (BBD) was sometimes found following DSS lesions, we also tested the hypothesis that DSS is a precursor of BBD. To track disease initiation and progression 24 coral colonies were tagged. Of them five Orbicella annularis corals and three O. faveolata corals exhibited DSS lesions at tagging. Microbiota of lesions and apparently healthy tissues from DSS-affected corals over the course of 18 months were collected. Final visual assessment showed that five of eight corals incurred substantial tissue loss while two corals remained stable and one appeared to recover from DSS lesions. Illumina sequencing of the V6 region of bacterial 16S rRNA genes demonstrated no significant differences in bacterial community composition associated with healthy tissue or DSS lesions. The epimicrobiomes of both healthy tissue and DSS lesions contained high relative abundances of Operational Taxonomic Units assigned to Halomonas, an unclassified gammaproteobacterial genus, Moritella, an unclassified Rhodobacteraceae genus, Renibacterium, Pseudomonas, and Acinetobacter. The relative abundance of bacterial taxa was not significantly different between samples when grouped by tissue type (healthy tissue vs. DSS lesion), coral species, collection month, or the overall outcome of DSS-affected corals (substantial tissue loss vs. stable/recovered). Two of the tagged corals with substantial tissue loss also developed BBD during the 18-month sampling period. The bacterial community of the BBD layer was distinct from both healthy tissue and DSS lesions, with high relative abundances of the presumed BBD pathogen Roseofilum reptotaenium and an unclassified Bacteroidales genus, similar to previous results. Roseofilum was detected in all samples from this study, with the highest relative abundance in healthy tissue from DSS-affected corals sampled in August, suggesting that while DSS is not a precursor to BBD, DSS-affected corals are in a weakened state and therefore more susceptible to additional infections. PMID:27375605

ELIPGRID-PC: A PC program for calculating hot spot probabilities

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

Davidson, J.R.

1994-10-01

ELIPGRID-PC, a new personal computer program has been developed to provide easy access to Singer`s 1972 ELIPGRID algorithm for hot-spot detection probabilities. Three features of the program are the ability to determine: (1) the grid size required for specified conditions, (2) the smallest hot spot that can be sampled with a given probability, and (3) the approximate grid size resulting from specified conditions and sampling cost. ELIPGRID-PC also provides probability of hit versus cost data for graphing with spread-sheets or graphics software. The program has been successfully tested using Singer`s published ELIPGRID results. An apparent error in the original ELIPGRIDmore » code has been uncovered and an appropriate modification incorporated into the new program.« less

Hot Spots from Generated Defects in HMX Crystals

NASA Astrophysics Data System (ADS)

Sorensen, Christian; Cummock, Nicholas; O'Grady, Caitlin; Gunduz, I. Emre; Son, Steven

2017-06-01

There are several hot spot initiation mechanisms that have been proposed. However, direct observation of shock or impact compression of these mechanisms at macroscopic scale in explosives is difficult. Phase contrast imaging (PCI) may be applied to these systems. Here, high-speed video was used to record optical spectrum and for x-ray Phase Contrast Imaging (PCI) of shockwave interaction with low defect HMX crystals and crystals with engineered defects. Additionally, multiple crystals were arranged and observed under shock loading with PCI and optical high-speed video. Sample preparation techniques for generating voids and other engineered defects will be discussed. These methods include drilled holes and laser machined samples. Insight into hot spot mechanisms was obtained. Funding from ONR's PC@Xtreme MURI.

An old remedy for a new problem? Identification of Ooencyrtus kuvanae (Howard) (Hymenoptera: Encyrtidae) as egg parasitoids of spotted lanternfly, Lycorma delicatula (Hemiptera: Fulgoridae) in North America

USDA-ARS?s Scientific Manuscript database

Spotted Lanternfly (SLF), Lycorma delicatula (White) is a recently introduced pest of Tree-of-Heaven, Ailanthus altissima (Mill.) Swingle in North America. Natural enemy surveys for this pest in Pennsylvania in 2016 recovered an encyrtid egg parasitoid from both field collections and laboratory rear...

From SPOT 5 to Pleiades HR: evolution of the instrumental specifications

NASA Astrophysics Data System (ADS)

Rosak, A.; Latry, C.; Pascal, V.; Laubier, D.

2017-11-01

Image quality specifications should aimed to fulfil high resolution mission requirements of remote sensing satellites with a minimum cost. The most important trade-off to be taken into account is between Modulation Transfer Function, radiometric noise and sampling scheme. This compromise is the main driver during design optimisation and requirement definition in order to achieve good performances and to minimise the mission cost. For the SPOT 5 satellite, a new compromise had been chosen. The supermode principle of imagery (sampling at 2.5 meter with a pixel size of 5 meter) imp roves the resolution by a factor of four compared with the SPOT 4 satellite (10 meter resolution). This paper presents the image quality specifications of the HRG-SPOT 5 instrument. We introduce all the efforts made on the instrument to achieve good image quality and low radiometric noise, then we compare the results with the SPOT 4 instrument's performances to highlight the improvements achieved. Then, the in-orbit performance will be described. Finally, we will present the new goals of image quality specifications for the new Pleiades-HR satellite for earth observation (0.7 meter resolution) and the instrument concept.

Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis

PubMed Central

Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

2017-01-01

Background: As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Objective: Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Materials and Methods: Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). Results: The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Conclusions: Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. SUMMARY The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines PMID:28250651

Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis.

PubMed

Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

2017-01-01

As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used : SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis : CS, TCMs: Traditional Chinese medicines.

Verification of the anatomy and newly discovered histology of the G-spot complex.

PubMed

Ostrzenski, A; Krajewski, P; Ganjei-Azar, P; Wasiutynski, A J; Scheinberg, M N; Tarka, S; Fudalej, M

2014-10-01

To expand the anatomical investigations of the G-spot and to assess the G-spot's characteristic histological and immunohistochemical features. An observational study. International multicentre. Eight consecutive fresh human female cadavers. Anterior vaginal wall dissections were executed and G-spot microdissections were performed. All specimens were stained with haematoxylin and eosin (H&E). The tissues of two women were selected at random for immunohistochemical staining. The primary outcome measure was to document the anatomy of the G-spot. The secondary outcome measures were to identify the histology of the G-spot and to determine whether histological samples stained with H&E are sufficient to identify the G-spot. The anatomical existence of the G-spot was identified in all women and was in a diagonal plane. In seven (87.5%) and one (12.5%) of the women the G-spot complex was found on the left or right side, respectively. The G-spot was intimately fused with vessels, creating a complex. A large tangled vein-like vascular structure resembled an arteriovenous malformation and there were a few smaller feeding arteries. A band-like structure protruded from the tail of the G-spot. The size of the G-spot varied. Histologically, the G-spot was determined as a neurovascular complex structure. The neural component contained abundant peripheral nerve bundles and a nerve ganglion. The vascular component comprised large vein-like vessels and smaller feeding arteries. Circular and longitudinal muscles covered the G-complex. The anatomy of the G-spot complex was confirmed. The histology of the G-spot presents as neurovascular tissues with a nerve ganglion. H&E staining is sufficient for the identification of the G-spot complex. © 2014 Royal College of Obstetricians and Gynaecologists.

Validity of predictive equations for 24-h urinary sodium excretion in adults aged 18–39 y12345

PubMed Central

Wang, Chia-Yih; Chen, Te-Ching; Pfeiffer, Christine M; Elliott, Paul; Gillespie, Cathleen D; Carriquiry, Alicia L; Sempos, Christopher T; Liu, Kiang; Perrine, Cria G; Swanson, Christine A; Caldwell, Kathleen L; Loria, Catherine M

2013-01-01

Background: Collecting a 24-h urine sample is recommended for monitoring the mean population sodium intake, but implementation can be difficult. Objective: The objective was to assess the validity of published equations by using spot urinary sodium concentrations to predict 24-h sodium excretion. Design: This was a cross-sectional study, conducted from June to August 2011 in metropolitan Washington, DC, of 407 adults aged 18–39 y, 48% black, who collected each urine void in a separate container for 24 h. Four timed voids (morning, afternoon, evening, and overnight) were selected from each 24-h collection. Published equations were used to predict 24-h sodium excretion with spot urine by specimen timing and race-sex subgroups. We examined mean differences with measured 24-h sodium excretion (bias) and individual differences with the use of Bland-Altman plots. Results: Across equations and specimens, mean bias in predicting 24-h sodium excretion for all participants ranged from −267 to 1300 mg (Kawasaki equation). Bias was least with International Cooperative Study on Salt, Other Factors, and Blood Pressure (INTERSALT) equations with morning (−165 mg; 95% CI: −295, 36 mg), afternoon (−90 mg; −208, 28 mg), and evening (−120 mg; −230, −11 mg) specimens. With overnight specimens, mean bias was least when the Tanaka (−23 mg; 95% CI: −141, 95 mg) or Mage (−145 mg; −314, 25 mg) equations were used but was statistically significant when using the Tanaka equations among females (216 to 243 mg) and the Mage equations among races other than black (−554 to −372 mg). Significant over- and underprediction occurred across individual sodium excretion concentrations. Conclusions: Using a single spot urine, INTERSALT equations may provide the least biased information about population mean sodium intakes among young US adults. None of the equations evaluated provided unbiased estimates of individual 24-h sodium excretion. This trial was registered at clinicaltrials.gov as NCT01631240. PMID:24047921

Proteomic Analysis of Albumins and Globulins from Wheat Variety Chinese Spring and Its Fine Deletion Line 3BS-8

PubMed Central

Ma, Chao-Ying; Gao, Li-Yan; Li, Ning; Li, Xiao-Hui; Ma, Wu-Jun; Appels, Rudi; Yan, Yue-Ming

2012-01-01

The relationship between chromosome deletion in wheat and protein expression were investigated using Chinese Spring and fine deletion line 3BS-8. Through 2-DE (2-D electrophoresis) analysis, no differentially expressed proteins (DEPs) were found in leaf samples; however, 47 DEPs showed at least two-fold abundance variation (p < 0.05) in matured wheat grains and 21 spots were identified by tandem MALDI-TOF/TOF-MS. Among the identified spots, four were cultivar-specific, including three (spots B15, B16, and B21) in Chinese Spring and one in 3BS-8 (spot B10). Among variety-different DEPs between Chinese Spring and 3BS-8, most spots showed a higher express profile in CS; only four spots showed up-regulated expression tendency in 3BS-8. An interesting observation was that more than half of the identified protein spots were involved in storage proteins, of which 11 spots were identified as globulins. According to these results, we can presume that the encoded genes of protein spots B15, B16, and B21 were located on the chromosome segment deleted in 3BS-8. PMID:23202959

Increasing a Community's Knowledge about Drought, Watershed Ecosystems, and Water Quality Through Educational Activities Added to Coastal Cleanup Day Events

NASA Astrophysics Data System (ADS)

Brinker, R.; Allen, L.; Cole, P.; Rho, C.

2016-12-01

International Coastal Cleanup Day, held each September, is an effective campaign to bring volunteers together to clean trash from beaches and waterways and document results. Over 500,000 participants cleared over 9 million pounds of trash in 2015. To build on the enthusiasm for this event, the city of Livermore, California's Water Resource Department, the Livermore Valley Joint Unified School District, Livermore Area Recreation and Parks Department created a water education program to embed within the city's Coastal Cleanup Day events. Goals of the education program are to increase awareness of the local watershed and its geographic reach, impacts of climate change and drought on local water supplies, pollution sources and impacts of local pollution on the ocean, positive impacts of a recent plastic bag ban, water quality assessment, and action steps citizens can take to support a healthy watershed. Volunteers collect and test water samples (when water is in the creek) using modified GLOBE and World Water Monitoring Day protocols. Test results are uploaded to the World Water Monitoring Day site and documented on the program web site. Volunteers report that they did not know about watersheds, impacts of local pollution, and water quality components before the education program. Volunteers are encouraged to adopt a creek spot for one year, and continue to collect and document trash. High school and middle school science classes added the water quality testing into curriculum, and regularly visit creek sites to clean the spots and monitor habitats. Each year for the past five years, about 300 volunteers have worked on creek clean-up events, 20 have adopted creek sites, and collected over 4,000 gallons of trash annually. As a result of these efforts, sites have been downgraded from a trash hot spot of concern. Strategies will be shared to expand an established (or start a new) Coastal Cleanup Day event into a successful watershed and climate awareness citizen science program. Include are: Data collection and review; Watershed posters and maps created for the event; Water quality test kits, equipment and protocols; Promotional items; Uses of social media and apps; Connecting with teachers; Recruiting interns; Program costs and supporting agencies. Attendees should feel that the program could be duplicated at their location.

Integrating spot short-term measurements of carbon emissions and backward dietary energy partition calculations to estimate intake in lactating dairy cows fed ad libitum or restricted.

PubMed

Pereira, A B D; Utsumi, S A; Dorich, C D; Brito, A F

2015-12-01

The objective of this study was to use spot short-term measurements of CH4 (QCH4) and CO2 (QCO2) integrated with backward dietary energy partition calculations to estimate dry matter intake (DMI) in lactating dairy cows. Twelve multiparous cows averaging 173±37d in milk and 4 primiparous cows averaging 179±27d in milk were blocked by days in milk, parity, and DMI (as a percentage of body weight) and, within each block, randomly assigned to 1 of 2 treatments: ad libitum intake (AL) or restricted intake (RI=90% DMI) according to a crossover design. Each experimental period lasted 22d with 14d for treatments adaptation and 8d for data and sample collection. Diets contained (dry matter basis): 40% corn silage, 12% grass-legume haylage, and 48% concentrate. Spot short-term gas measurements were taken in 5-min sampling periods from 15 cows (1 cow refused sampling) using a portable, automated, open-circuit gas quantification system (GreenFeed, C-Lock Inc., Rapid City, SD) with intervals of 12h between the 2daily samples. Sampling points were advanced 2h from a day to the next to yield 16 gas samples per cow over 8d to account for diurnal variation in QCH4 and QCO2. The following equations were used sequentially to estimate DMI: (1) heat production (MJ/d)=(4.96 + 16.07 ÷ respiratory quotient) × QCO2; respiratory quotient=0.95; (2) metabolizable energy intake (MJ/d)=(heat production + milk energy) ± tissue energy balance; (3) digestible energy (DE) intake (MJ/d)=metabolizable energy + CH4 energy + urinary energy; (4) gross energy (GE) intake (MJ/d)=DE + [(DE ÷ in vitro true dry matter digestibility) - DE]; and (5) DMI (kg/d)=GE intake estimated ÷ diet GE concentration. Data were analyzed using the MIXED procedure of SAS (SAS Institute Inc., Cary, NC) and Fit Model procedure in JMP (α=0.05; SAS Institute Inc.). Cows significantly differed in DMI measured (23.8 vs. 22.4kg/d for AL and RI, respectively). Dry matter intake estimated using QCH4 and QCO2 coupled with dietary backward energy partition calculations (Equations 1 to 5 above) was highest in cows fed for AL (22.5 vs. 20.2kg/d). The resulting R(2) were 0.28 between DMI measured and DMI estimated by gaseous measurements, and 0.36 between DMI measured and DMI predicted by the National Research Council model (2001). Results showed that spot short-term measurements of QCH4 and QCO2 coupled with dietary backward estimations of energy partition underestimated DMI by 7.8%. However, the approach proposed herein was able to significantly discriminate differences in DMI between cows fed for AL or RI. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

An enhanced droplet-based liquid microjunction surface sampling system coupled with HPLC-ESI-MS/MS for spatially resolved analysis

DOE PAGES

Van Berkel, Gary J.; Weiskittel, Taylor M.; Kertesz, Vilmos

2014-11-07

Droplet-based liquid microjunction surface sampling coupled with high-performance liquid chromatography (HPLC)-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) for spatially resolved analysis provides the possibility of effective analysis of complex matrix samples and can provide a greater degree of chemical information from a single spot sample than is typically possible with a direct analysis of an extract. Described here is the setup and enhanced capabilities of a discrete droplet liquid microjunction surface sampling system employing a commercially available CTC PAL autosampler. The system enhancements include incorporation of a laser distance sensor enabling unattended analysis of samples and sample locations of dramatically disparatemore » height as well as reliably dispensing just 0.5 μL of extraction solvent to make the liquid junction to the surface, wherein the extraction spot size was confined to an area about 0.7 mm in diameter; software modifications improving the spatial resolution of sampling spot selection from 1.0 to 0.1 mm; use of an open bed tray system to accommodate samples as large as whole-body rat thin tissue sections; and custom sample/solvent holders that shorten sampling time to approximately 1 min per sample. Lastly, the merit of these new features was demonstrated by spatially resolved sampling, HPLC separation, and mass spectral detection of pharmaceuticals and metabolites from whole-body rat thin tissue sections and razor blade (“crude”) cut mouse tissue.« less

An enhanced droplet-based liquid microjunction surface sampling system coupled with HPLC-ESI-MS/MS for spatially resolved analysis

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

Van Berkel, Gary J.; Weiskittel, Taylor M.; Kertesz, Vilmos

Droplet-based liquid microjunction surface sampling coupled with high-performance liquid chromatography (HPLC)-electrospray ionization (ESI)-tandem mass spectrometry (MS/MS) for spatially resolved analysis provides the possibility of effective analysis of complex matrix samples and can provide a greater degree of chemical information from a single spot sample than is typically possible with a direct analysis of an extract. Described here is the setup and enhanced capabilities of a discrete droplet liquid microjunction surface sampling system employing a commercially available CTC PAL autosampler. The system enhancements include incorporation of a laser distance sensor enabling unattended analysis of samples and sample locations of dramatically disparatemore » height as well as reliably dispensing just 0.5 μL of extraction solvent to make the liquid junction to the surface, wherein the extraction spot size was confined to an area about 0.7 mm in diameter; software modifications improving the spatial resolution of sampling spot selection from 1.0 to 0.1 mm; use of an open bed tray system to accommodate samples as large as whole-body rat thin tissue sections; and custom sample/solvent holders that shorten sampling time to approximately 1 min per sample. Lastly, the merit of these new features was demonstrated by spatially resolved sampling, HPLC separation, and mass spectral detection of pharmaceuticals and metabolites from whole-body rat thin tissue sections and razor blade (“crude”) cut mouse tissue.« less

Bacterial Leaf Spot of Lettuce: Request for Samples

USDA-ARS?s Scientific Manuscript database

Bacterial leaf spot of lettuce caused by by Xanthomonas campestris pv. vitians has been affecting coastal California crops for many years and has become a chronic problem. Differences in pathogen genotypes have been demonstrated and correlated to disease responses on resistant and susceptible cultiv...

Spot Sampling and Exposure Surrogate Selection as Sources of Bias in Environmental Epidemiology Studies

EPA Science Inventory

Spot measurements of chemical biomarkers are often used as quantitative exposure surrogates in environmental epidemiology studies. These measures can be expressed a number of different ways – for example, urinary biomarkers can be expressed in units of concentration (&micr...

High resolution multiple excitation spot optical microscopy

NASA Astrophysics Data System (ADS)

Dilipkumar, Shilpa; Mondal, Partha Pratim

2011-06-01

We propose fundamental improvements in three-dimensional (3D) resolution of multiple excitation spot optical microscopy. The excitation point spread function (PSF) is generated by two interfering counter-propagating depth-of-focus beams along the optical axis. Detection PSF is obtained by coherently interfering the emitted fluorescent light (collected by both the objectives) at the detector. System PSF shows upto 14-fold reduction in focal volume as compared to confocal, and almost 2-fold improvement in lateral resolution. Proposed PSF has the ability to simultaneously excite multiple 3D-spots of sub-femtoliter volume. Potential applications are in fluorescence microscopy and nanobioimaging.

The opto-thermal effect on encapsulated cholesteric liquid crystals

NASA Astrophysics Data System (ADS)

Liu, Yu-Sung; Lin, Hui-Chi; Yang, Kin-Min

2017-12-01

In this study, we implemented a micro-encapsulated CLC electronic paper that is optically addressed and electrically erasable. The mechanism that forms spot diameters on the CLC films is discussed and verified through various experimental parameters, including the thickness of CLCs and Poly(2,3-dihydrothieno-1,4-dioxin)-poly(styrenesulfonate) (PEDOT:PSS), pump intensity, and pumping time. The opto-thermal effect, brought on by the PEDOT:PSS absorbing layer, causes the spot diameters on the cholesteric liquid crystal thin films to vary. According to our results, the spot diameter is larger for a sample with a thinner cholesteric liquid crystal layer with the same excitation conditions and same thickness of the PEDOT layer. The spot diameter is also larger for a sample with a thicker PEDOT under the same excitation conditions and same thickness of the cholesteric liquid crystal layer. We proposed a simple heat-conducting model to explain the experimental results, which qualitatively agree with this theoretical model.

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

PubMed

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

2016-02-01

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

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

PubMed Central

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

2016-01-01

Molecular approaches offer a means of testing archived samples stored as dried blood spots in settings where standard blood cultures are not possible. Peripheral blood films are one suggested source of material, although the sensitivity of this approach has not been well defined. Thin blood smears and dried blood spots from a severe pediatric malaria study were assessed using specific polymerase chain reaction (PCR) primers to detect non-typhoidal Salmonella (NTS; MisL gene), Streptococcus pneumoniae (lytA), and Plasmodium falciparum (18S rRNA). Of 16 cases of NTS and S. pneumoniae confirmed on blood culture, none were positive by PCR using DNA extracts from blood films or dried blood spots. In contrast, four of 36 dried blood spots and two of 178 plasma samples were PCR positive for S. pneumoniae, despite negative bacterial blood cultures, suggesting false positives. Quantitative assessment revealed that the effective concentration of P. falciparum DNA in blood films was three log orders of magnitude lower than for dried blood spots. The P. falciparum kelch13 gene could not be amplified from blood films. These findings question the value of blood PCR-based approaches for detection of NTS and S. pneumoniae, and show that stored blood films are an inefficient method of studying P. falciparum. PMID:26711525

Leaf spot disease adversely affects human health-promoting constituents and withanolide biosynthesis in Withania somnifera (L.) Dunal.

PubMed

Singh, V; Singh, B; Sharma, A; Kaur, K; Gupta, A P; Salar, R K; Hallan, V; Pati, P K

2017-01-01

The present work investigates the implication of leaf spot disease on the antioxidant potential and commercial value of pharmaceutically important constituents of Withania somnifera, a high-valued medicinal plant. Leaf spot disease was induced in W. somnifera by inoculating Alternaria alternata (Fr.) Keiss. pathogen. Total polyphenolic content and antioxidant potential showed a significant decrease during leaf spot disease. Evaluation of pharmaceutically active constituents withaferin A, withanone and withanolide A utilizing high-performance liquid chromatography showed a significant decrease in diseased samples as compared to healthy ones. Quantitative expression of major genes involved in withanolide biosynthesis also showed down-regulation in diseased samples. Alterations in the ultra-structure of chloroplasts were also analysed under transmission electron microscopy to get a better insight into the changes of withanolide biosynthesis in leaf during disease infestation. The present work suggests that when the pathogenic fungus invades the host plants, it evokes multiple responses, which could be studied at various levels. The knowledge gained from this work will provide appropriate rationale for controlling the bio-deterioration of the pharmaceutically active metabolites in W. somnifera and development of suitable strategies against leaf spot disease. This is the first study to investigate the effect of leaf spot disease on the human health-promoting constituents and withanolide biosynthesis in this high-valued medicinal plant. © 2016 The Society for Applied Microbiology.

Iodine Intakes of Victorian Schoolchildren Measured Using 24-h Urinary Iodine Excretion

PubMed Central

Beckford, Kelsey; Margerison, Claire; Skeaff, Sheila A.

2017-01-01

Mandatory fortification of bread with iodized salt was introduced in Australia in 2009, and studies using spot urine collections conducted post fortification indicate that Australian schoolchildren are now replete. However an accurate estimate of daily iodine intake utilizing 24-h urinary iodine excretion (UIE μg/day) has not been reported and compared to the estimated average requirement (EAR). This study aimed to assess daily total iodine intake and status of a sample of primary schoolchildren using 24-h urine samples. Victorian primary school children provided 24-h urine samples between 2011 and 2013, from which urinary iodine concentration (UIC, μg/L) and total iodine excretion (UIE, μg/day) as an estimate of intake was determined. Valid 24-h urine samples were provided by 650 children, mean (SD) age 9.3 (1.8) years (n = 359 boys). The mean UIE of 4–8 and 9–13 year olds was 94 (48) and 111 (57) μg/24-h, respectively, with 29% and 26% having a UIE below the age-specific EAR. The median (IQR) UIC was 124 (83,172) μg/L, with 36% of participants having a UIC < 100 μg/L. This convenience sample of Victorian schoolchildren were found to be iodine replete, based on UIC and estimated iodine intakes derived from 24-h urine collections, confirming the findings of the Australian Health Survey. PMID:28867787

CT Angiography Spot Sign, Hematoma Expansion, and Outcome in Primary Pontine Intracerebral Hemorrhage.

PubMed

Morotti, Andrea; Jessel, Michael J; Brouwers, H Bart; Falcone, Guido J; Schwab, Kristin; Ayres, Alison M; Vashkevich, Anastasia; Anderson, Christopher D; Viswanathan, Anand; Greenberg, Steven M; Gurol, M Edip; Romero, Javier M; Rosand, Jonathan; Goldstein, Joshua N

2016-08-01

The computed tomography angiography (CTA) spot sign is a validated predictor of hematoma expansion and poor outcome in supratentorial intracerebral hemorrhage (ICH), but patients with brainstem ICH have typically been excluded from the analyses. We investigated the frequency of spot sign and its relationship with hematoma expansion and outcome in patients with primary pontine hemorrhage (PPH). We performed a retrospective analysis of PPH cases obtained from a prospectively collected cohort of consecutive ICH patients who underwent CTA. CTA first-pass readings for spot sign presence were analyzed by two trained readers. Baseline and follow-up hematoma volumes on non-contrast CT scans were assessed by semi-automated computer-assisted volumetric analysis. Sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), positive and negative likelihood ratio, and accuracy of spot sign for prediction of in-hospital mortality were calculated. 49 subjects met the inclusion criteria of whom 11 (22.4 %) showed a spot sign. In-hospital mortality was higher in spot sign-positive versus spot sign-negative subjects (90.9 vs 47.4 %, p = 0.020). Spot sign showed excellent specificity (95 %) and PPV (91 %) in predicting in-hospital mortality. Absolute hematoma growth, defined as parenchymal and intraventricular hematoma expansion of any amount, was significantly higher in spot sign-positive versus spot sign-negative subjects (13.72 ± 20.93 vs 3.76 ± 8.55 mL, p = 0.045). As with supratentorial ICH, the CTA spot sign is a common finding and is associated with higher risk of hematoma expansion and mortality in PPH. This marker may assist clinicians in prognostic stratification.

Field application of passive SBSE for the monitoring of pesticides in surface waters.

PubMed

Assoumani, A; Coquery, M; Liger, L; Mazzella, N; Margoum, C

2015-03-01

Spot sampling lacks representativeness for monitoring organic contaminants in most surface waters. Passive sampling has emerged as a cost-effective complementary sampling technique. We recently developed passive stir bar sorptive extraction (passive SBSE), with Twister from Gerstel, for monitoring moderately hydrophilic to hydrophobic pesticides (2.18 < log K ow < 5.11) in surface water. The aims of the present study were to assess this new passive sampler for the determination of representative average concentrations and to evaluate the contamination levels of two French rivers. Passive SBSE was evaluated for the monitoring of 16 pesticides in two rivers located in a small vineyard watershed during two 1-month field campaigns in spring 2010 and spring 2011. Passive SBSE was applied for periods of 1 or 2 weeks during the field campaigns and compared with spot sampling and weekly average automated sampling. The results showed that passive SBSE could achieve better time-representativeness than spot sampling and lower limits of quantification than automated sampling coupled with analytical SBSE for the pesticides studied. Finally, passive SBSE proved useful for revealing spatial and temporal variations in pesticide contamination of both rivers and the impact of rainfall and runoff on the river water quality.

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

PubMed

Hirtz, Christophe; Lehmann, Sylvain

2015-01-01

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

Mass Spectrometric Imaging Using Laser Ablation and Solvent Capture by Aspiration (LASCA)

NASA Astrophysics Data System (ADS)

Brauer, Jonathan I.; Beech, Iwona B.; Sunner, Jan

2015-09-01

A novel interface for ambient, laser ablation-based mass spectrometric imaging (MSI) referred to as laser ablation and solvent capture by aspiration (LASCA) is presented and its performance demonstrated using selected, unaltered biological materials. LASCA employs a pulsed 2.94 μm laser beam for specimen ablation. Ablated materials in the laser plumes are collected on a hanging solvent droplet with electric field-enhanced trapping, followed by aspiration of droplets and remaining plume material in the form of a coarse aerosol into a collection capillary. The gas and liquid phases are subsequently separated in a 10 μL-volume separatory funnel, and the solution is analyzed with electrospray ionization in a high mass resolution Q-ToF mass spectrometer. The LASCA system separates the sampling and ionization steps in MSI and combines high efficiencies of laser plume sampling and of electrospray ionization (ESI) with high mass resolution MS. Up to 2000 different compounds are detected from a single ablation spot (pixel). Using the LASCA platform, rapid (6 s per pixel), high sensitivity, high mass-resolution ambient imaging of "as-received" biological material is achieved routinely and reproducibly.

Plankton Biomass Models Based on GIS and Remote Sensing Technique for Predicting Marine Megafauna Hotspots in the Solor Waters

NASA Astrophysics Data System (ADS)

Putra, MIH; Lewis, SA; Kurniasih, EM; Prabuning, D.; Faiqoh, E.

2016-11-01

Geographic information system and remote sensing techniques can be used to assist with distribution modelling; a useful tool that helps with strategic design and management plans for MPAs. This study built a pilot model of plankton biomass and distribution in the waters off Solor and Lembata, and is the first study to identify marine megafauna foraging areas in the region. Forty-three samples of zooplankton were collected every 4 km according to the range time and station of aqua MODIS. Generalized additive model (GAM) we used to modelling zooplankton biomass response from environmental properties.Thirty one samples were used to build a model of inverse distance weighting (IDW) (cell size 0.01°) and 12 samples were used as a control to verify the models accuracy. Furthermore, Getis-Ord Gi was used to identify the significance of the hotspot and cold-spot for foraging area. The GAM models was explain 88.1% response of zooplankton biomass and percent to full moon, phytopankton biomassbeing strong predictors. The sampling design was essential in order to build highly accurate models. Our models 96% accurate for phytoplankton and 88% accurate for zooplankton. The foraging behaviour was significantly related to plankton biomass hotspots, which were two times higher compared to plankton cold-spots. In addition, extremely steep slopes of the Lamakera strait support strong upwelling with highly productive waters that affect the presence of marine megafauna. This study detects that the Lamakera strait provides the planktonic requirements for marine megafauna foraging, helping to explain why this region supports such high diversity and abundance of marine megafauna.

Identification of Fungal Communities Associated with the Biodeterioration of Waterlogged Archeological Wood in a Han Dynasty Tomb in China

PubMed Central

Liu, Zijun; Wang, Yu; Pan, Xiaoxuan; Ge, Qinya; Ma, Qinglin; Li, Qiang; Fu, Tongtong; Hu, Cuiting; Zhu, Xudong; Pan, Jiao

2017-01-01

The Mausoleum of the Dingtao King (termed ‘M2’) is a large-scale huangchang ticou tomb that dates to the Western Han Dynasty (206 B.C.–25 A.D.). It is the highest-ranking Han Dynasty tomb discovered to date. However, biodeterioration on the surface of the tomb M2 is causing severe damage to its wooden materials. The aim of the present study was to give insight into the fungal communities colonized the wooden tomb. For this purpose, seven samples were collected from different sections of the tomb M2 which exhibited obvious biodeterioration in the form of white spots. Microbial structures associated with the white spots were observed with scanning electron microscopy. Fungal community structures were assessed for seven samples via a combination of high-throughput sequencing and culture-dependent techniques. Sequencing analyses identified 114 total genera that belonged to five fungal phyla. Hypochnicium was the most abundant genus across all samples and accounted for 98.61–99.45% of the total community composition. Further, Hypochnicium sp. and Mortierella sp. cultures were successfully isolated from the tomb samples, and were distinguished as Hypochnicium sp. WY-DT1 and Mortierella sp. NK-DT1, respectively. Cultivation-dependent experiments indicated that the dominant member, Hypochnicium sp. WY- DT1, could grow at low temperatures and significantly degraded cellulose and lignin. Thus, our results taken together suggest that this fungal strain must be regarded as a serious threat to the preservation of the wooden tomb M2. The results reported here are useful for informing future contamination mitigation efforts for the tomb M2 as well as other similar cultural artifacts. PMID:28890715

Atmospheric Electron-induced X-Ray Spectrometer (AEXS) Instrument Development

NASA Technical Reports Server (NTRS)

Urgiles, E.; Wilcox, J. Z.; Toda, R.; Crisp, J.; George, T.

2005-01-01

Introduction: This paper describes the progress in data acquisition and establishing the observational capability of the AEXS instrument. The AEXS is a miniature instrument[1-4] based on the excitation of characteristic X-Ray Fluorescence (XRF) and luminescence spectra using a focused electron beam which enables nondestructive evaluation of sample surfaces in planetary ambient atmospheres. In situ operation is obtained through the use of a thin electron transmissive membrane to isolate the vacuum of the AEXS source from the outside ambient atmosphere. Thus eliminating the need for a vacuum pumped sample chamber as is common in all laboratory SEM s. The transmitted electrons impinge on the sample exciting XRF spectra from the irradiated spot on in-situ or collected samples with sub-mm to cm-scale spatial resolution at Mars atmospheric pressure. The AEXS system (Fig 1) consists of a high-energy (>10keV) electron gun encapsulated by the isolation membrane, an XRF detection and analyzer system, and a high voltage power supply. The XRF data are analyzed to determine the elemental abundance for the irradiated spots. The approach to demonstrating a proof of concept of the AEXS has been through 1) demonstrating the viability of microfabricated membranes, 2) assembling AEXS setups with increasingly integrated functional components, and 3) simulating the AEXS observational capabilities. The development of the instrument is described in detail in the poster paper[4] at this conference. This paper focuses on describing the progress of the AEXS instrument to acquire XRF data and using commercially available software to analyze the data streams and determine the accuracy, precision and resolution of the analysis compared to the certified elemental abundance.

Field evaluation of Abbott Real Time HIV-1 Qualitative test for early infant diagnosis using dried blood spots samples in comparison to Roche COBAS Ampliprep/COBAS TaqMan HIV-1 Qual Test in Kenya

PubMed Central

Chang, Joy; Omuomo, Kenneth; Anyango, Emily; Kingwara, Leonard; Basiye, Frank; Morwabe, Alex; Shanmugam, Vedapuri; Nguyen, Shon; Sabatier, Jennifer; Zeh, Clement; Ellenberger, Dennis

2016-01-01

Timely diagnosis and treatment of infants infected with HIV are critical for reducing infant mortality. High-throughput automated diagnostic tests like Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Qual Test (Roche CAPCTM Qual) and the Abbott Real Time HIV-1 Qualitative (Abbott Qualitative) can be used to rapidly expand early infant diagnosis testing services. In this study, the performance characteristics of the Abbott Qualitative were evaluated using two hundred dried blood spots (DBS) samples (100 HIV-1 positive and 100 HIV-1 negative) collected from infants attending the antenatal facilities in Kisumu, Kenya. The Abbott Qualitative results were compared to the diagnostic testing completed using the Roche CAPCTM Qual in Kenya. The sensitivity and specificity of the Abbott Qualitative were 99.0% (95% CI: 95.0–100.0) and 100.0% (95% CI: 96.0–100.0), respectively, and the overall reproducibility was 98.0% (95% CI: 86.0–100.0). The limits of detection for the Abbott Qualitative and Roche CAPCTM Qual were 56.5 and 6.9 copies/mL at 95% CIs (p = 0.005), respectively. The study findings demonstrate that the Abbott Qualitative test is a practical option for timely diagnosis of HIV in infants. PMID:24726703

Field evaluation of Abbott Real Time HIV-1 Qualitative test for early infant diagnosis using dried blood spots samples in comparison to Roche COBAS Ampliprep/COBAS TaqMan HIV-1 Qual test in Kenya.

PubMed

Chang, Joy; Omuomo, Kenneth; Anyango, Emily; Kingwara, Leonard; Basiye, Frank; Morwabe, Alex; Shanmugam, Vedapuri; Nguyen, Shon; Sabatier, Jennifer; Zeh, Clement; Ellenberger, Dennis

2014-08-01

Timely diagnosis and treatment of infants infected with HIV are critical for reducing infant mortality. High-throughput automated diagnostic tests like Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Qual Test (Roche CAPCTM Qual) and the Abbott Real Time HIV-1 Qualitative (Abbott Qualitative) can be used to rapidly expand early infant diagnosis testing services. In this study, the performance characteristics of the Abbott Qualitative were evaluated using two hundred dried blood spots (DBS) samples (100 HIV-1 positive and 100 HIV-1 negative) collected from infants attending the antenatal facilities in Kisumu, Kenya. The Abbott Qualitative results were compared to the diagnostic testing completed using the Roche CAPCTM Qual in Kenya. The sensitivity and specificity of the Abbott Qualitative were 99.0% (95% CI: 95.0-100.0) and 100.0% (95% CI: 96.0-100.0), respectively, and the overall reproducibility was 98.0% (95% CI: 86.0-100.0). The limits of detection for the Abbott Qualitative and Roche CAPCTM Qual were 56.5 and 6.9copies/mL at 95% CIs (p=0.005), respectively. The study findings demonstrate that the Abbott Qualitative test is a practical option for timely diagnosis of HIV in infants. Published by Elsevier B.V.

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

PubMed

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

2016-02-05

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. Copyright © 2015 Elsevier B.V. All rights reserved.

Multivariate and geo-spatial approach for seawater quality of Chidiyatappu Bay, south Andaman Islands, India.

PubMed

Jha, Dilip Kumar; Vinithkumar, Nambali Valsalan; Sahu, Biraja Kumar; Dheenan, Palaiya Sukumaran; Das, Apurba Kumar; Begum, Mehmuna; Devi, Marimuthu Prashanthi; Kirubagaran, Ramalingam

2015-07-15

Chidiyatappu Bay is one of the least disturbed marine environments of Andaman & Nicobar Islands, the union territory of India. Oceanic flushing from southeast and northwest direction is prevalent in this bay. Further, anthropogenic activity is minimal in the adjoining environment. Considering the pristine nature of this bay, seawater samples collected from 12 sampling stations covering three seasons were analyzed. Principal Component Analysis (PCA) revealed 69.9% of total variance and exhibited strong factor loading for nitrite, chlorophyll a and phaeophytin. In addition, analysis of variance (ANOVA-one way), regression analysis, box-whisker plots and Geographical Information System based hot spot analysis further simplified and supported multivariate results. The results obtained are important to establish reference conditions for comparative study with other similar ecosystems in the region. Copyright © 2015 Elsevier Ltd. All rights reserved.

Statistical Analysis And Treatment Of Accident Black Spots: A Case Study Of Nandyal Mandal

NASA Astrophysics Data System (ADS)

Sudharshan Reddy, B.; Vishnu Vardhan Reddy, L.; Sreenivasa Reddy, G., Dr

2017-08-01

Background: Increased, economic activity raised the consumption levels of the people across the country. This created scope for increase in travel and transportation. The increase in the vehicles since last 10 years has put lot of pressure on the existing roads and ultimately resulting in road accidents. Nandyal Mandal is located in the Kurnool district of Andhra Pradesh and well developed in both agricultural and industrial sectors after Kurnool. 567 accidents occurred in the last seven years at 143 locations shows the severity of the accidents in the Nandyal Mandal. There is a need to carry out some work in the Nandyal Mandal to improve the accidents black spots for reducing the accidents. Methods: Last seven years (2010-2016) of accident data collected from Police Stations. Weighted Severity Index (WSI), a scientific method is used for identifying the accident black spots. Statistical analysis has carried out for the collected data using Chi-Square Test to determine the independence of accidents with other attributes. Chi-Square Goodness of fit test conducted for test whether the accidents are occurring by chance or following any pattern. Results: WSI values are determined for the 143 locations. The Locations with high WSI are treated as accident black spots. Five black spots are taken for field study. After field observations and interaction with the public, some improvements are suggested for improving the accident black spots. There is no relationship between the severity of accidents and the other attributes like month, season, day, hours in day and the age group except type of vehicle. Road accidents are distributed throughout the Year, Month and Season. Road accidents are not distributed throughout the day.

The effects of habitat, climate, and Barred Owls on long-term demography of Northern Spotted Owls

Treesearch

Katie M. Dugger; Eric D. Forsman; Alan B. Franklin; Raymond J. Davis; Gary C. White; Carl J. Schwarz; Kenneth P. Burnham; James D. Nichols; James E. Hines; Charles B. Yackulic; Paul F. Doherty; Larissa Bailey; Darren A. Clark; Steven H. Ackers; Lawrence S. Andrews; Benjamin Augustine; Brian L. Biswell; Jennifer Blakesley; Peter C. Carlson; Matthew J. Clement; Lowell V. Diller; Elizabeth M. Glenn; Adam Green; Scott A. Gremel; Dale R. Herter; J. Mark Higley; Jeremy Hobson; Rob B. Horn; Kathryn P. Huyvaert; Christopher McCafferty; Trent McDonald; Kevin McDonnell; Gail S. Olson; Janice A. Reid; Jeremy Rockweit; Viviana Ruiz; Jessica Saenz; Stan G. Sovern

2016-01-01

Estimates of species’ vital rates and an understanding of the factors affecting those parameters over time and space can provide crucial information for management and conservation. We used mark–recapture, reproductive output, and territory occupancy data collected during 1985–2013 to evaluate population processes of Northern Spotted Owls (Strix occidentalis...

Array biosensor for detection of toxins

NASA Technical Reports Server (NTRS)

Ligler, Frances S.; Taitt, Chris Rowe; Shriver-Lake, Lisa C.; Sapsford, Kim E.; Shubin, Yura; Golden, Joel P.

2003-01-01

The array biosensor is capable of detecting multiple targets rapidly and simultaneously on the surface of a single waveguide. Sandwich and competitive fluoroimmunoassays have been developed to detect high and low molecular weight toxins, respectively, in complex samples. Recognition molecules (usually antibodies) were first immobilized in specific locations on the waveguide and the resultant patterned array was used to interrogate up to 12 different samples for the presence of multiple different analytes. Upon binding of a fluorescent analyte or fluorescent immunocomplex, the pattern of fluorescent spots was detected using a CCD camera. Automated image analysis was used to determine a mean fluorescence value for each assay spot and to subtract the local background signal. The location of the spot and its mean fluorescence value were used to determine the toxin identity and concentration. Toxins were measured in clinical fluids, environmental samples and foods, with minimal sample preparation. Results are shown for rapid analyses of staphylococcal enterotoxin B, ricin, cholera toxin, botulinum toxoids, trinitrotoluene, and the mycotoxin fumonisin. Toxins were detected at levels as low as 0.5 ng mL(-1).

Spot test for 1,3,5-triamino-2,4,6-trinitrobenzene, TATB

DOEpatents

Harris, Betty W.

1986-01-01

A simple, sensitive and specific spot test for 1,3,5-triamino-2,4,6-trinitrobenzene, TATB, is described. Upon the application of the composition of matter of the present invention to samples containing in excess of 0.1 mg of this explosive, a bright orange color results. Interfering species such as TNT and Tetryl can be removed by first treating the sample with a solvent which does not dissolve much of the TATB, but readily dissolves these explosives.

Spot test for 1,3,5-triamino-2,4,6-trinitrobenzene, TATB

DOEpatents

Harris, B.W.

1984-11-29

A simple, sensitive and specific spot test for 1,3,5-triamino-2,4,6-trinitrobenzene, TATB, is described. Upon the application of the composition of matter of the subject invention to samples containing in excess of 0.1 mg of this explosive, a bright orange color results. Interfering species such as TNT and Tetryl can be removed by first treating the sample with a solvent which does not dissolve the TATB, but readily dissolves these interfering explosives.

Serum Dried Samples to Detect Dengue Antibodies: A Field Study.

PubMed

Maldonado-Rodríguez, Angelica; Rojas-Montes, Othon; Vazquez-Rosales, Guillermo; Chavez-Negrete, Adolfo; Rojas-Uribe, Magdalena; Posadas-Mondragon, Araceli; Aguilar-Faisal, Leopoldo; Cevallos, Ana Maria; Xoconostle-Cazares, Beatriz; Lira, Rosalia

2017-01-01

Dried blood and serum samples are useful resources for detecting antiviral antibodies. The conditions for elution of the sample need to be optimized for each disease. Dengue is a widespread disease in Mexico which requires continuous surveillance. In this study, we standardized and validated a protocol for the specific detection of dengue antibodies from dried serum spots (DSSs). Paired serum and DSS samples from 66 suspected cases of dengue were collected in a clinic in Veracruz, Mexico. Samples were sent to our laboratory, where the conditions for optimal elution of DSSs were established. The presence of anti-dengue antibodies was determined in the paired samples. DSS elution conditions were standardized as follows: 1 h at 4°C in 200  µ l of DNase-, RNase-, and protease-free PBS (1x). The optimal volume of DSS eluate to be used in the IgG assay was 40  µ l. Sensitivity of 94%, specificity of 93.3%, and kappa concordance of 0.87 were obtained when comparing the antidengue reactivity between DSSs and serum samples. DSS samples are useful for detecting anti-dengue IgG antibodies in the field.

Serum Dried Samples to Detect Dengue Antibodies: A Field Study

PubMed Central

Maldonado-Rodríguez, Angelica; Rojas-Montes, Othon; Chavez-Negrete, Adolfo; Rojas-Uribe, Magdalena; Posadas-Mondragon, Araceli; Aguilar-Faisal, Leopoldo; Xoconostle-Cazares, Beatriz

2017-01-01

Background Dried blood and serum samples are useful resources for detecting antiviral antibodies. The conditions for elution of the sample need to be optimized for each disease. Dengue is a widespread disease in Mexico which requires continuous surveillance. In this study, we standardized and validated a protocol for the specific detection of dengue antibodies from dried serum spots (DSSs). Methods Paired serum and DSS samples from 66 suspected cases of dengue were collected in a clinic in Veracruz, Mexico. Samples were sent to our laboratory, where the conditions for optimal elution of DSSs were established. The presence of anti-dengue antibodies was determined in the paired samples. Results DSS elution conditions were standardized as follows: 1 h at 4°C in 200 µl of DNase-, RNase-, and protease-free PBS (1x). The optimal volume of DSS eluate to be used in the IgG assay was 40 µl. Sensitivity of 94%, specificity of 93.3%, and kappa concordance of 0.87 were obtained when comparing the antidengue reactivity between DSSs and serum samples. Conclusion DSS samples are useful for detecting anti-dengue IgG antibodies in the field. PMID:28630868

Abundance and population characteristics of Northern Spotted Owls (Strix occidentalis caurina) in Olympic National Park, Washington

Treesearch

D. Erran Seaman

1997-01-01

We monitored the threatened Northern Spotted Owl (Strix occidentalis caurina) in Olympic National Park from 1992 through 1996. We used a stratified random sampling scheme to survey 35 plots totaling 236 km?, approximately 10 percent of the forested area of the park.

Simultaneous determination of trimethoprim and sulfamethoxazole in dried plasma and urine spots.

PubMed

Gonzalez, Daniel; Melloni, Chiara; Poindexter, Brenda B; Yogev, Ram; Atz, Andrew M; Sullivan, Janice E; Mendley, Susan R; Delmore, Paula; Delinsky, Amy; Zimmerman, Kanecia; Lewandowski, Andrew; Harper, Barrie; Lewis, Kenneth C; Benjamin, Daniel K; Cohen-Wolkowiez, Michael

2015-01-01

Trimethoprim-sulfamethoxazole (TMP-SMX) is an antimicrobial drug combination commonly prescribed in children and adults. The study objectives were to validate and apply an HPLC-MS/MS method to quantify TMP-SMX in dried plasma spots (DPS) and dried urine spots (DUS), and perform a comparability analysis with liquid matrices. For TMP the validated range was 100-50,000 ng/ml for DPS and 500-250,000 ng/ml for DUS; for SMX, the validated range was 1000-500,000 ng/ml for both DPS and DUS. Good agreement was noted between DPS/DUS and liquid plasma and urine samples for TMP, while only modest agreement was observed for SMX in both matrices. A precise, accurate and reproducible method was developed to quantify TMP-SMX in DPS and DUS samples.

Risk estimation and annual fluxes of emerging contaminants from a Scottish priority catchment to the estuary and North Sea.

PubMed

Zhang, Zulin; Lebleu, Melanie; Osprey, Mark; Kerr, Christine; Courtot, Estelle

2017-06-28

Emerging contaminants (ECs) such as endocrine-disrupting chemicals (EDCs) and pharmaceuticals and personal care products (PPCPs) attracted global concern during the last decades due to their potential adverse effects on humans and ecosystems. This work is the first study to assess the spatiotemporal changes, annual fluxes and ecological risk of ECs (4 EDCs and 6 PPCPs) by different monitoring strategies (spot and passive sampling) over 12 months in a Scottish priority catchment (River Ugie, Scotland, 335 km 2 ). Overall, the total concentration in water ranged from 1. This suggests that mitigation measures might need to be taken to reduce the input of emerging contaminants into the river and its adjacent estuary and sea.

White noise effects of U.S. crude oil spot prices on stock prices of a publicly traded company: A case study cross-correlation analysis based on green energy management theory

NASA Astrophysics Data System (ADS)

Roberts, Peter M.

The purpose of this study was to examine white noise effects of U.S. crude oil spot prices on the stock prices of a green energy company. Epistemological, Phenomenological, Axiological and Ontological assumptions of Green Energy Management (GEM) Theory were utilized for selecting Air Products and Chemicals Inc. (APD) as the case study. Exxon Mobil (XOM) was used as a control for triangulation purposes. The period of time examined was between January of 1999 and December of 2008. Monthly stock prices for APD and XOM for the ten year period of time were collected from the New York Stock Exchange. Monthly U.S. crude oil spot prices for the ten year period of time were collected from the US Energy Information Administration. The data was entered into SPSS 17.0 software in order to conduct cross-correlation analysis. The six cross-correlation assumptions were satisfied in order to conduct a Cross-correlation Mirror Test (CCMT). The CCMT established the lag time direction and verified that U.S. crude oil spot prices serve as white noise for stock prices of APD and XOM. The Theory of Relative Weakness was employed in order to analyze the results. A 2 year period of time between December, 2006 and December, 2008 was examined. The correlation coefficient r = - .155 indicates that U.S. crude oil spot prices lead APD stock prices by 4 months. During the same 2 year period of time, U.S. crude oil spot prices lead XOM stock prices by 4 months at r = -.283. XOM stock prices and APD stock prices were positively correlated with 0 lag in time with a positive r = .566. The 4 month cycle was an exact match between APD stock prices, XOM stock prices and U.S. crude oil spot prices. The 4 month cycle was due to the random price fluctuation of U.S. crude oil spot prices that obscured the true stock prices of APD and XOM for the 2 year period of time.

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

PubMed

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

2015-01-01

The 1.2 mm Electric Coring Tool (e-Core™) was developed to increase the throughput of FTA(™) sample collection cards used during forensic workflows and is similar to a 1.2 mm Harris manual micro-punch for sampling dried blood spots. Direct short tandem repeat (STR) DNA profiling was used to compare samples taken by the e-Core tool with those taken by the manual micro-punch. The performance of the e-Core device was evaluated using a commercially available PowerPlex™ 18D STR System. In addition, an analysis was performed that investigated the potential carryover of DNA via the e-Core punch from one FTA disc to another. This contamination study was carried out using Applied Biosystems AmpflSTR™ Identifiler™ Direct PCR Amplification kits. The e-Core instrument does not contaminate FTA discs when a cleaning punch is used following excision of discs containing samples and generates STR profiles that are comparable to those generated by the manual micro-punch. © 2014 American Academy of Forensic Sciences.

Ladybirds as Teaching Aids: 1 Collecting and Culturing.

ERIC Educational Resources Information Center

Majerus, M. E. N.; And Others

1989-01-01

Described are methods for locating, collecting and culturing ladybird beetles. Included are necessary equipment for feeding and breeding the insects and modifications for various species found in Great Britain. Overwintering spots of various species are also listed. (CW)

Isolation and characterization of bioactive components from Mirabilis jalapa L. radix

PubMed Central

Gogoi, Jyotchna; Nakhuru, Khonamai Sewa; Policegoudra, Rudragoud S.; Chattopadhyay, Pronobesh; Rai, Ashok Kumar; Veer, Vijay

2015-01-01

The present investigation was carried out to isolate and characterize bioactive components from Mirabilis jalapa L. radix (紫茉莉根 zǐ mò lì gēn). Thin-layer chromatography was used for the separation of spots from fractions of the crude extract. Separated spots were collected for identification of their activities. Free-radical scavenging activity was evaluated by spraying thin-layer chromatography plates (spotted with fractions) with 0.2% of 2,2-diphenyl-1-picrylhydrazyl solution. Activity against human pathogens such as Staphylococcus aureus and Candida albicans were determined using the agar diffusion method. Potential spots were subjected to infrared (IR) analysis and gas chromatography for characterization. Two spots (5F1 and 1F3) showed free-radical scavenging activity. The 1F3 spot was active against both S. aureus and C. albicans, whereas the 5F1 spot was active against S. aureus only. IR spectral analysis indicated that 5F1 spot to be a triterpenoid. Using IR spectral analysis and an IR library search, the 1F3 spot was identified to be a flavone, which may have a hydroxyl group in ring “A” of the flavone nucleus. Our results indicated that the 1F3 and 5F1 spots are potential free-radical scavengers. Both 1F3 and 5F1 exhibited antimicrobial activity. IR spectral analysis coupled with an IR library search indicated 1F3 and 5F1 to be a flavone and a triterpenoid, respectively. PMID:26870679

Nasal protein profiles in work-related asthma caused by different exposures.

PubMed

Suojalehto, H; Lindström, I; Wolff, H; Puustinen, A

2018-03-01

The mechanisms of work-related asthma (WRA) are incompletely delineated. Nasal cell samples may be informative about processes in the lower airways. Our aim was to determine the nasal protein expression profiles of WRA caused by different kind of exposures. We collected nasal brush samples from 82 nonsmoking participants, including healthy controls and WRA patients exposed to (i) protein allergens, (ii) isocyanates and (iii) welding fumes the day after relevant exposure. The proteome changes in samples were analysed by two-dimensional difference gel electrophoresis, and the differentially regulated proteins found were identified by mass spectrometry. Immunological comparison was carried out using Western blot. We detected an average of 2500 spots per protein gel. Altogether, 228 protein spots were chosen for identification, yielding 77 different proteins. Compared to the controls, exposure to protein allergens had the largest effects on the proteome. Hierarchical clustering revealed that protein allergen- and isocyanate-related asthma had similar profiles, whereas asthma related to welding fumes differed. The highly overrepresented functional categories in the asthma groups were defence response, protease inhibitor activity, inflammatory and calcium signalling, complement activation and cellular response to oxidative stress. Immunological analysis confirmed the found abundance differences in galectin 10 and protein S100-A9 between the groups. Work-related asthma patients exposed to protein allergens and isocyanates elicit similar nasal proteome responses and the profiles of welders and healthy controls were alike. Revealed biological activities of the protein expression changes are associated with allergic inflammation and asthma. © 2017 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

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

PubMed

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

2015-09-01

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

Hemoglobin Adducts of Benzene Oxide in Neonatal and Adult Dried Blood Spots

PubMed Central

Funk, William E.; Waidyanatha, Suramya; Chaing, Shu H.; Rappaport, Stephen M.

2010-01-01

Adducts of reactive chemicals with hemoglobin (Hb) or human serum albumin can be used as biomarkers of internal doses of carcinogens. Since dried blood spots (DBS) are easier to collect and store than conventional venous blood samples, they encourage applications of biomarkers of exposure in large epidemiology studies. Also, neonatal DBS can be used to investigate chemical exposures in utero. Here, we report a simple method to isolate Hb from DBS with high recovery and purity using the addition of ethanol to aqueous DBS extracts. To prove the concept that DBS-derived proteins can be used to assay for adducts, we measured Hb adducts of benzene oxide, a reactive metabolite of the ubiquitous air pollutant, benzene, in 9 neonatal and 9 adult DBS (from volunteer subjects), using a gas chromatography-mass spectrometry method that we had previously developed. For comparison, benzene oxide-Hb adducts (BO-Hb) were measured in the same 9 adult subjects, using Hb that had been isolated and purified using our conventional method for venous blood. The geometric mean BO-Hb levels in all DBS samples ranged from 27.7 to 33.1 pmol/g globin. Neither of the comparisons of mean (logged) BO-Hb levels between sources (adult conventional vs. adult DBS and adult DBS vs. newborn DBS) showed a significant difference. Based upon the estimated variance of the BO-Hb levels, we had 80% power to detect a 1.7-fold difference in geometric mean levels of BO-Hb in our samples of 9 subjects. PMID:18708378

A combinatory approach for analysis of protein sets in barley sieve-tube samples using EDTA-facilitated exudation and aphid stylectomy.

PubMed

Gaupels, Frank; Knauer, Torsten; van Bel, Aart J E

2008-01-01

This study investigated advantages and drawbacks of two sieve-tube sap sampling methods for comparison of phloem proteins in powdery mildew-infested vs. non-infested Hordeum vulgare plants. In one approach, sieve tube sap was collected by stylectomy. Aphid stylets were cut and immediately covered with silicon oil to prevent any contamination or modification of exudates. In this way, a maximum of 1muL pure phloem sap could be obtained per hour. Interestingly, after pathogen infection exudation from microcauterized stylets was reduced to less than 40% of control plants, suggesting that powdery mildew induced sieve tube-occlusion mechanisms. In contrast to the laborious stylectomy, facilitated exudation using EDTA to prevent calcium-mediated callose formation is quick and easy with a large volume yield. After two-dimensional (2D) electrophoresis, a digital overlay of the protein sets extracted from EDTA solutions and stylet exudates showed that some major spots were the same with both sampling techniques. However, EDTA exudates also contained large amounts of contaminative proteins of unknown origin. A combinatory approach may be most favourable for studies in which the protein composition of phloem sap is compared between control and pathogen-infected plants. Facilitated exudation may be applied for subtractive identification of differentially expressed proteins by 2D/mass spectrometry, which requires large amounts of protein. A reference gel loaded with pure phloem sap from stylectomy may be useful for confirmation of phloem origin of candidate spots by digital overlay. The method provides a novel opportunity to study differential expression of phloem proteins in monocotyledonous plant species.

Study on the Distribution of Disease-Resistant Shrimp Identified by DNA Markers in Respect to WSSV Infection in Different Seasons Along the Entire East Coast of India Aiming to Prevent White Spot Disease in Penaeus monodon.

PubMed

Mallik, A; Chakrabarty, U; Dutta, S; Mondal, D; Mandal, N

2016-02-01

White spot disease caused by white spot syndrome virus (WSSV) is responsible for harming shrimp aquaculture industry and results in a pandemic throughout the world. Undeniably, the knowledge on geographic distribution, transmission, virulence, and seasonal prevalence of this disease alongside information on the distribution of disease-resistant shrimps may be helpful to understand important aspects of disease biology. This study was intended to estimate WSSV prevalence by qualitative and quantitative PCR method among the Penaeus monodon samples collected from four different places namely Digha, West Bengal; Chilika, Orissa; Visakhapatnam, Andhra Pradesh; and Chennai, Tamil Nadu at three different seasons in the period of 2011-2013 from east coast of India. Along with this, the disease-resistant prevalence was also investigated using earlier developed 71 bp microsatellite and 457 bp RAPD-SCAR DNA marker among the collected shrimps. Qualitative PCR depicted that the cumulative WSSV prevalence at four places was the lowest (0%) at pre-monsoon, whereas, it was the highest (21.2%) during post-monsoon season. Quantitative real-time PCR showed the average copy number of WSSV to be the highest (~10(3) copy μg(-1) shrimp genomic DNA) at post-monsoon season. Additionally, estimated disease-resistant prevalence was the highest in Visakhapatnam (79%) and lowest in Digha (21%). It is well known to all that a trait cannot be identified using a single genetic pattern. This study will significantly contribute insight to develop specific pathogen-resistant (SPR) seeds of P. monodon simultaneously using two DNA markers that would be a cost-effective and safer approach towards disease prevention instead of conventional trends of seed generation from unselected wild broodstock. © 2014 Blackwell Verlag GmbH.

Feasibility study of the application of radially polarized illumination to solid immersion lens-based near-field optics.

PubMed

Yoon, Yong-Joong; Kim, Wan-Chin; Park, No-Cheol; Park, Kyoung-Su; Park, Young-Pil

2009-07-01

We analyzed the behavior of the electric field in a focal plane consisting of a solid immersion lens (SIL), an air gap, and a measurement sample for radially polarized illumination in SIL-based near-field optics with an annular aperture. The analysis was based on the Debye diffraction integral and multiple beam interference. For SIL-based near-field optics whose NA is higher than unity, radially polarized light generates a smaller beam spot on the bottom surface of a SIL than circularly polarized light; however, the beam spot on the measurement sample is broadened with a more dominant transverse electric field. By introducing an annular aperture technique, it is possible to decrease the effects of the transverse electric field, and therefore the size of the beam spot on the measurement sample can be small. This analysis could have various applications in near-field optical storage, near-field microscopy, lithography at ultrahigh resolution, and other applications that use SILs for high resolution.

Great Red Spot Rotation (animation)

NASA Image and Video Library

2017-12-11

Winds around Jupiter's Great Red Spot are simulated in this JunoCam view that has been animated using a model of the winds there. The wind model, called a velocity field, was derived from data collected by NASA's Voyager spacecraft and Earth-based telescopes. NASA's Juno spacecraft acquired the original, static view during passage over the spot on July 10, 2017. Citizen scientists Gerald Eichstädt and Justin Cowart turned the JunoCam data into a color image mosaic. Juno scientists Shawn Ewald and Andrew Ingersoll applied the velocity data to the image to produce a looping animation. An animation is available at https://photojournal.jpl.nasa.gov/catalog/PIA22178

A Simple Method to Quantitate IP-10 in Dried Blood and Plasma Spots

PubMed Central

Aabye, Martine G.; Eugen-Olsen, Jesper; Werlinrud, Anne Marie; Holm, Line Lindebo; Tuuminen, Tamara; Ravn, Pernille; Ruhwald, Morten

2012-01-01

Background Antigen specific release of IP-10 is an established marker for infection with M.tuberculosis. Compared to IFN-γ, IP-10 is released in 100-fold higher concentrations enabling the development of novel assays for detection. Dried blood spots are a convenient sample for high throughput newborn screening. Aim To develop a robust and sensitive ELISA-based assay for IP-10 detection in plasma, dried blood spots (DBS) and dried plasma spots (DPS); to validate the ELISA in clinically relevant samples; and to assess the performance of the assay for detection of Cytomegalovirus (CMV) and M.tuberculosis specific immune responses. Method We raised mice and rat monoclonal antibodies against human IP-10 and developed an ELISA. The assay was validated and applied to the detection of CMV and M.tuberculosis specific responses in 18 patients with immune reactivity towards M.tuberculosis and 32 healthy controls of which 22 had immune reactivity towards CMV and none towards M.tuberculosis. We compared the performance of this new assay to IFN-γ. Results The ELISA was reliable for IP-10 detection in both plasma and filter paper samples. The linear range of the ELISA was 2.5–600 pg/ml. IFN-γ was not readily detectable in DPS samples. IP-10 was stabile in filter paper samples for at least 4 weeks at 37°C. The correlation between IP-10 detected in plasma, DPS and DBS samples was excellent (r2>0.97). Conclusions This newly developed assay is reliable for IP-10 quantification in plasma, DBS and DPS samples from antigen stimulated and non-stimulated whole blood. The filter paper assays enable easy sample acquisition and transport at ambient temperature e.g. via the postal system. The system can potentially simplify diagnostic assays for M.tuberculosis and CMV infection. PMID:22761744

Spots and activity of solar-type stars from Kepler observations

NASA Astrophysics Data System (ADS)

Savanov, I. S.; Dmitrienko, E. S.

2017-05-01

The spot coverages S for 2846 solar-type stars with effective temperatures from 5700 K to 5800 K and gravities from 4.4 to 4.5 have been measured. An analysis based on the MAST catalog, which presents photometric measurements obtained with the Kepler Space Telescope during Q9 is presented. The existence of two groups of solar-type stars, with S values between 0.001 and 0.007 and with S > 0.007, is inferred. The second group (active stars) contains 279 stars (about 10% of the total number of stars analyzed). The mean S parameter for the entire sample is 0.004, comparable to the mean spot coverage of the Sun. In general, the dependence of S on the rotation period for solar-type stars has characteristics similar to those found earlier for stars with exoplanets. For the vast majority of the stars in the sample, the activity is constant, and independent of age. The activity of the small number of active stars with S > 0.007 decreases with age. The age variations of the chromospheric activity index R'HK are compared to variations of the spot coverage S. The relations analyzed have common characteristic features. It is likely that both the spot activity level and the chromospheric activity level abruptly decrease for stars older than 4 billion yrs.

Diets and foraging behavior of northern Spotted Owls in Oregon

USGS Publications Warehouse

Forsman, E.D.; Anthony, R.G.; Meslow, E.C.; Zabel, C.J.

2004-01-01

We describe local, regional, and annual variation in diets of northern Spotted Owls (Strix occidentalis caurina) in Oregon based on 24 497 prey collected at 1118 owl territories in 1970-2003. The sample included 91.5% mammals, 4.3% birds, 4.1% insects, and 0.1% other prey. The diet included ???131 species, including 49 mammals, 41 birds, 3 reptiles, 1 frog, 1 crayfish, 1 scorpion, 2 snails, and 33 species of insects. On average, 91.9 ?? 0.3% (SE) of prey in the diet were nocturnal animals, 3.3 ?? 0.2% were diurnal, and 4.8 ?? 0.2% were active both day and night. Of the prey captured, 50.5 ?? 0.8% were arboreal, 18.7 ?? 0.7% were scansorial, 4.8 ?? 0.2% were aerial, and 26.0 = 0.7% were terrestrial. Mean mass of prey was 116.6 ?? 6.5 g. Diets varied among owl territories, geographic regions, and years; but were generally dominated by four to six species of nocturnal mammals, including northern flying squirrels (Glaucomys sabrinus), woodrats (Neotoma fuscipes and N. cinerea), red tree voles (Arborimus longicaudus), western red-backed voles (Clethrionomys californicus), deer mice (Peromyscus maniculatus), or gophers (Thomomys spp.). Estimates of dietary evenness were low, indicating diets dominated by a few species of mammals. Forest management practices that produce healthy populations of arboreal and scansorial mammals such as flying squirrels, woodrats, and red tree voles should benefit northern Spotted Owls in Oregon and Washington. ?? 2004 The Raptor Research Foundation, Inc.

Gas chromatography-electron ionization-mass spectrometry quantitation of valproic acid and gabapentin, using dried plasma spots, for therapeutic drug monitoring in in-home medical care.

PubMed

Ikeda, Kayo; Ikawa, Kazuro; Yokoshige, Satoko; Yoshikawa, Satoshi; Morikawa, Norifumi

2014-12-01

A simple and sensitive gas chromatography-electron ionization-mass spectrometry (GC-EI-MS) method using dried plasma spot testing cards was developed for determination of valproic acid and gabapentin concentrations in human plasma from patients receiving in-home medical care. We have proposed that a simple, easy and dry sampling method is suitable for in-home medical patients for therapeutic drug monitoring. Therefore, in the present study, we used recently developed commercially available easy handling cards: Whatman FTA DMPK-A and Bond Elut DMS. In-home medical care patients can collect plasma using these simple kits. The spots of plasma on the cards were extracted into methanol and then evaporated to dryness. The residues were trimethylsilylated using N-methyl-N-trimethylsilyltrifluoroacetamide. For GC-EI-MS analysis, the calibration curves on both cards were linear from 10 to 200 µg/mL for valproic acid, and from 0.5 to 10 µg/mL for gabapentin. Intra- and interday precisions in plasma were both ≤13.0% (coefficient of variation), and the accuracy was between 87.9 and 112% for both cards within the calibration curves. The limits of quantification were 10 µg/mL for valproic acid and 0.5 µg/mL for gabapentin on both cards. We believe that the present method will be useful for in-home medical care. Copyright © 2014 John Wiley & Sons, Ltd.

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

Sun, Cheng-Jun, E-mail: cjsun@aps.anl.gov; Brewe, Dale L.; Heald, Steve M.

X-ray diffraction (XRD) and X-ray absorption spectroscopy (XAS) are two main x-ray techniques in synchrotron radiation facilities. In this Note, we present an experimental setup capable of performing simultaneous XRD and XAS measurements by the application of a pixel-array area detector. For XRD, the momentum transfer in specular diffraction was measured by scanning the X-ray energy with fixed incoming and outgoing x-ray angles. By selecting a small fixed region of the detector to collect the XRD signal, the rest of the area was available for collecting the x-ray fluorescence for XAS measurements. The simultaneous measurement of XRD and X-ray absorptionmore » near edge structure for Pr{sub 0.67}Sr{sub 0.33}MnO{sub 3} film was demonstrated as a proof of principle for future time-resolved pump-probe measurements. A static sample makes it easy to maintain an accurate overlap of the X-ray spot and laser pump beam.« less

HIV-1 viral load measurement in venous blood and fingerprick blood using Abbott RealTime HIV-1 DBS assay.

PubMed

Tang, Ning; Pahalawatta, Vihanga; Frank, Andrea; Bagley, Zowie; Viana, Raquel; Lampinen, John; Leckie, Gregor; Huang, Shihai; Abravaya, Klara; Wallis, Carole L

2017-07-01

HIV RNA suppression is a key indicator for monitoring success of antiretroviral therapy. From a logistical perspective, viral load (VL) testing using Dried Blood Spots (DBS) is a promising alternative to plasma based VL testing in resource-limited settings. To evaluate the analytical and clinical performance of the Abbott RealTime HIV-1 assay using a fully automated one-spot DBS sample protocol. Limit of detection (LOD), linearity, lower limit of quantitation (LLQ), upper limit of quantitation (ULQ), and precision were determined using serial dilutions of HIV-1 Virology Quality Assurance stock (VQA Rush University), or HIV-1-containing armored RNA, made in venous blood. To evaluate correlation, bias, and agreement, 497 HIV-1 positive adult clinical samples were collected from Ivory Coast, Uganda and South Africa. For each HIV-1 participant, DBS-fingerprick, DBS-venous and plasma sample results were compared. Correlation and bias values were obtained. The sensitivity and specificity were analyzed at a threshold of 1000 HIV-1 copies/mL generated using the standard plasma protocol. The Abbott HIV-1 DBS protocol had an LOD of 839 copies/mL, a linear range from 500 to 1×10 7 copies/mL, an LLQ of 839 copies/mL, a ULQ of 1×10 7 copies/mL, and an inter-assay SD of ≤0.30 log copies/mL for all tested levels within this range. With clinical samples, the correlation coefficient (r value) was 0.896 between DBS-fingerprick and plasma and 0.901 between DBS-venous and plasma, and the bias was -0.07 log copies/mL between DBS-fingerprick and plasma and -0.02 log copies/mL between DBS-venous and plasma. The sensitivity of DBS-fingerprick and DBS-venous was 93%, while the specificity of both DBS methods was 95%. The results demonstrated that the Abbott RealTime HIV-1 assay with DBS sample protocol is highly sensitive, specific and precise across a wide dynamic range and correlates well with plasma values. The Abbott RealTime HIV-1 assay with DBS sample protocol provides an alternative sample collection and transfer option in resource-limited settings and expands the utility of a viral load test to monitor HIV-1 ART treatment for infected patients. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA.

PubMed

Ferraz, Aline S; Belo, Elza F T; Coutinho, Ligia M C C; Oliveira, Ana P; Carmo, Andréia M S; Franco, Daniele L; Ferreira, Tatiane; Yto, André Y; Machado, Marta S F; Scola, Monica C G; De Gaspari, Elizabeth

2008-03-06

A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20 degrees C after spotting on filter paper, for subsequent serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA. Monoclonal antibodies (Mabs) were spotted within a 0.5-1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20 degrees C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20 degrees C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin. Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20 degrees C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper. The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked.

Successful prediction of genetic richness at wild potato collection sites in southeastern Arizona

USDA-ARS?s Scientific Manuscript database

Much time, money, and effort is needed to collect even a fraction of the potential geographic range of wild potato species, so there is efficiency to gain if one could predict and prioritize spots particularly rich in unique alleles for collecting. A previous experiment that used AFLP markers to com...

Ionization Suppression and Recovery in Direct Biofluid Analysis Using Paper Spray Mass Spectrometry

NASA Astrophysics Data System (ADS)

Vega, Carolina; Spence, Corina; Zhang, Chengsen; Bills, Brandon J.; Manicke, Nicholas E.

2016-04-01

Paper spray mass spectrometry is a method for the direct analysis of biofluid samples in which extraction of analytes from dried biofluid spots and electrospray ionization occur from the paper on which the dried sample is stored. We examined matrix effects in the analysis of small molecule drugs from urine, plasma, and whole blood. The general method was to spike stable isotope labeled analogs of each analyte into the spray solvent, while the analyte itself was in the dried biofluid. Intensity of the labeled analog is proportional to ionization efficiency, whereas the ratio of the analyte intensity to the labeled analog in the spray solvent is proportional to recovery. Ion suppression and recovery were found to be compound- and matrix-dependent. Highest levels of ion suppression were obtained for poor ionizers (e.g., analytes lacking basic aliphatic amine groups) in urine and approached -90%. Ion suppression was much lower or even absent for good ionizers (analytes with aliphatic amines) in dried blood spots. Recovery was generally highest in urine and lowest in blood. We also examined the effect of two experimental parameters on ion suppression and recovery: the spray solvent and the sample position (how far away from the paper tip the dried sample was spotted). Finally, the change in ion suppression and analyte elution as a function of time was examined by carrying out a paper spray analysis of dried plasma spots for 5 min by continually replenishing the spray solvent.